INTRODUCTION

The identification and estimation of different types of crime exhibits such as drugs, cement, petroleum product and their residues, liquor, egetable oil and fats, dyes, gold, chemicals etc in forensic science laboratories required high degree of s!ill and expertise" The forensic scientists are follo#ing arious methods for the chemical analysis of these substances in the laboratories" The $or!ing %rocedure &anual of Chemistry had been made at C'(), *yderabad #ith the consultation of a team of arious 'orensic (cientists of the country in the year +,,,"

SECTION - 1 CEMENT, MORTAR AND CONCRETE 1.1. Title- Cement, mortar and concrete 1.2. Scope: Detection and estimation of the constituents of cement, mortar and concrete in crime cases" 1.3. Purpose: To !no# the precise chemical composition of cement, mortar and concrete and to assist in estigating agencies and la# enforcement bodies" 1.4 Respo si!ilit": Reporting officers and supporting scientific staff" 1.#. I tro$uctio : 'orensic (cience )aboratories recei e cement cases under different crime heads li!e .ssential Commodities /ct", Indian %enal Code 0I%C1 and occasionally under Cement Orders /ct" Cement is one of the most important materials in building construction" In chemical examination, testing of cement is done to chec! its purity and characteri2ation" *ence only a fe# parameters li!e soluble calcium oxide, silica etc" are determined as per Indian (tandards specifications" Concrete and &ortar are referred to 'orensic (cience )aboratories in cases of building collapse" 1.% Ce&e t %ortland cement may be defined 3 1' as a product obtained by intimately mixing together calcareous and argillaceous, or other silica, alumina, and iron oxide4bearing materials, burning them at a clin!ering temperature and grinding the resulting clin!er" In +567 8oseph /spdin ga e the name portland

cement because this product resembled the colour of the stones from %ortland, .ngland" Cements are mainly mono silicates of calcium, soluble in dilute acids and al!ali" 1.%.1. COMPOSITION O( PORT)AND CEMENT: CaO (iO6 /l6O> 'e6O> &gO N*2O + -2O (O> CaO 9:49;< +;46=< :>4:5< :"=4:9< :"+4:7< ..2-.1/ :+46";=< ::4:+<

)ime (ilica /lumina Iron Oxide &agnesia So$* + Pot*s, (ulfur Trioxide 'ree )ime

The &ain Constituents of Cement are, Dicalcium Tricalcium Tricalcium (ilicate (ilicate /luminate 6 CaO(iO6 > CaO(iO6 > CaO/l6O> 7 CaO, /l6O>, 'e6O> ++< ++< >:< 7:<

Tetracalcium /lumino 'errite 1.%.2. T"pes o0 Ce&e ts 12': -

There are many types of cements a ailable, but fe# of them are discussed here" 1.%.2.1.R*pi$ 2*r$e i 3 Portl* $ Ce&e t- is similar to that of ordinary %ortland cement, but is grounded finer and slightly altered in composition" Its setting time is similar, but it de elops its strength more rapidly" 1.%.2.2.4uic5 setti 3 Portl* $ Ce&e t - it differs from normal %ortland cement in its setting time, #hich is less , compared to portland cement" Its rate of hardening may be similar to that of ordinary or rapid hardening %ortland Cement"

1.%.2.3.6,ite Portl* $ Ce&e t - is an ordinary %ortland cement containing a lo# proportion of iron oxide, so that its colour is #hite instead of grey" 1.%.2.4.6*ter proo0 Portl* $ Ce&e t- is ordinary %ortland Cement in #hich at grinding stage small proportion of calcium stearate or non4 saponifiable oil is added" 1.%.2.#.2"$rop,o!ic Ce&e t - is a material obtained by grinding %ortland Cement clin!er #ith a #ater repellent film forming substance such as fatty acid in order to reduce the rate of deterioration under fa ourable storage or transport conditions" 1.%.2.%.)o7-,e*t Portl* $ Ce&e t - is a material in #hich chemical composition has been so ad?usted as to reduce the heat of hydration" 1.%.2.8. Portl* $ Po99olo * Ce&e t : in ordinary %ortland cement, a po22olonic material li!e bric! po#der, flyash etc" are added in the range of 6:47:<" This cement is called %ortland %o22olona Cement and is generally used in the preparation of plaster materials" 1.%.2.; OT2ER <=I)DIN> MATERIA)S 1.%.2.;.1. Sto e po7$er, is poly silicates of calcium, magnesium and iron etc" and is practically insoluble in dilute acids" 1.%.2.;.2.Co cret, is the hard mass obtained by solidification of the inert material li!e sand, coarse stone, #ater and cement" 1.%.2.;.3.Mort*r, is the mixture of sand and cement for plastering the bric!#or!" 1.%.2.;.4. S* $ , is mostly silica in defined form and insoluble in dilute mineral acids, should be clean, strong, durable uncoated #ell4graded particles" The particles should be free from al!ali, organic matter, loam or other deterious substances" The diameter of the sand particles should not be abo e 9 mm" 1.%.2.;.#.A33re3*te, The aggregate should consist of crushed roc!, gra el or other inert material" The particles should be clean, though, hard and durable" It should not contain soft, flat and elongated material" The maximum length of aggregate should not be more than ++6 mm" 1.%.2.;.%.<ric5

@ric! is an important building material" Aarieties of bric!s are a ailable in the mar!et" Their qualities are determined on the basis of physical parameters" There is no much chemical examination in ol ed in deciding their fitness etc" 1.%.3. SAMP)IN> 1.%.3.1.Ce&e t: 1.%.3.1*..$hen the sample is dra#n from a cement bag, the details printed on the bag and another mar!ing thereon should be carefully noted and incorporated in the for#arding letter" + !g sample of cement should be sent in an airtight plastic ?ar if a ailable or it should be securely pac!ed in polythene bag and then in bro#n paper to a oid exposure to moisture" (ampling is done as per the procedure as laid do#n in the Indian (tandard %rocedures of random sampling" 1.%.3.1!.S*&pli 3 o0 S&*ll 4u* tities 0)ess than +6 bags or pac!ages1$hen number of bags or other pac!ages containing the cement bears the same label on all the pac!ages and are appearing to be similar, in such cases about +!g sample of cement 0in an air tight plastic ?ar1 shall be dra#n from each bag and sent for analysis" 1.%.3.1c.S*&pli 3 o0 l*r3e 4u* tities 0&ore than +6 bags or pac!ages1$hen number of bags containing the cement bear the same labels on the pac!ages and are appearing to be similar, in such cases the grouping must be done" .ach group should contain about equal no" of bags and 6: percent of sample #eighing + !g 0in an air tight plastic ?ar1 from each group shall be dra#n into air tight plastic ?ar and sent for analysis" 1.%.3.2.MORTAR: +46 Bg of mortar sample accompanied by + !g each of cement and C or lime and sand if a ailable from the field shall be sent for analysis" . ery article should be independently pac!ed" 1.%.3.3.CONCRETE: Concrete lumps, about >4=!g accompanied by + !g each of cement, sand and aggregate if a ailable from the field shall be sent for analysis" . ery article should be independently pac!ed" 1.%.4. MET2ODS O( ANA)?SIS 1E@PERIMENTA) re0.3':

1.%.4.1.Ce&e t:

1.%.4.1.1.T,"&olp,t,*lei Test: 0Thymolphthalein Indicator :"+< in ethyl alcohol1 Ta!e +::4+=: mg of cement sample in a test tube, add +46 ml #ater and 6 drops of indicator, de elopment of blue colour indicates the presence of cement, No colour indicates that the sample is stone po#der" 1.%.4.1.2.2e*ti 3 test: Ta!e :"= gm of sample, heat it for about 6: min" on a steel plate" a1" adulterated cement" b1" unadulterated cement" Change in colour No change in colour

1.%.4.1.3.Per0or&* ce Test&a!e thic! slurry of cement #ith about + part of cement #ith one part of #ater and put in an empty matchbox" The cement gets hardened" The performance is tested after 67 hrs" ?ust by remo ing matchbox and chec!ing approx" strength of the cement by fingers, if the bloc! brea!s easily, the setting property is said to be poor" If the bloc! does not brea! by fingers" The performance is said to be good" 1.%.4.1.4.Aci$ i solu!leTa!e :"= 4+": gm cement in a +:: ml bea!er add 6: ml #ater to ma!e a paste follo#ed by = ml conc" hydrochloric acid, add 6: ml #ater, stir, digest on #ater bath for fi e minutes, no lumps should be formed" Digest further for another +: minutes, filter through ashless filter paper till chloride free" Residue dried in o en and further incinerated in furnace at 5::DC4,::DC for + hr" #eigh the residue, till constant #eight" Calculate percent acid insoluble" 1.%.4.1.#.Silic*Concentrate filtrate from 6"9"7"+"7 on hot plate to dryness, further dry completely #ithout charring, then add 6: ml +-+ *ydrochloric acid and digest on #ater bath for +: minutes stir #ell, and filter on ashless filter paper till chloride free" Dry the residue 0%recipitated silica1 in o en for +hr and then incinerate in furnace at 5::DC4,::DC for +hr"#eigh the residue, till constant #eight" $eight of silica obtained is noted" 06:< (ilica E +::< cement1" 1.%.4.1.%.Co&!i e$ (erric OAi$e * $ Alu&i *: Concentrate the filtrate from 6"9"7"+"= to about 6::4ml by boiling then add 64 > drops of nitric acid to oxidise any ferrous iron to ferric condition" /dd +46

grams of ammonium chloride, stir, and then treat the filtrate #ith conc" ammonia solution till smell of ammonia persists then boil the solution containing the ppts" of 'e and /l hydroxides for fe# minute" 'ilter and #ash the ppt" #ith hot #ater" Dry the ppt" in o en and ignite in platinum crucible till constant #eight at +:=:DC to ++::DC" $eigh as alumina and ferric oxide" 1.%.4.1.8.Deter&i *tio o0 BC*lciu&B!" EDTA Titr*tio - 4

0@y %atton ReederFs indicator1 Ta!e filtrate from ferric oxide and alumina determination in 6=: ml Aol" flas! ad?ust Aol" to 6=: ml" Ta!e out 6=ml soln" in titration flas! add =: ml #ater, =ml 0+-+1 glycerol #ith constant stirring then add =ml diethylamine, further add =49 pellets of NaO* 0p* should be more than +61 sha!e #ell, further add =:mg of %atton ReederFs indicator, sha!e #ell and titrate against :":+& .DT/ soln" colour change iolet to blue" +ml :":+ & .DT/ E :"=9:5 mg of CaO" 09: < CaO E +::< Cement1 and CaO< E > (ilica < P*tto Ree$er`s I $ic*tor: Grind +:mg of the indicator #ith +:gm of sodium sulphate 0/"R"1 and store in an airtight bottle" 1.%.4.1.; Direct Ce&e t / !" *ci$ titr*tio Ta!e :"= gm cement in a conical flas! add =:ml of :"= N *Cl, digest on #ater bath for >: minutes, add =: ml #ater and titrate against :"= N NaO* using phenolphthalein as an indicator" /lso perform a blan! titration" Colour change is colourless to pin!" Cement < E 65 H N H Diff" in Reading 0NE Normality of NaO*1 1.8. Mort*r: &ortar is the blend of cement and sand in arious proportions used for arious purposes" The mortar used for bric!#or! in house #alls is generally +-7 in proportions" 'or testing of mortar and bric! good piece of mortar adhering to bric! from debris should be collected" 1.8.1 Testi 3 o0 Mort*r:

*eat good piece of mortar approx" 6:: gms is heated in o en at ++:DC for += min" cool and then #eigh" (eparate the cement portion from sand by slo#ly grinding the lump in iron mortar" 0to separate sand from cement lumps"1 (ie e the material and ma!e three fractions" %o#der, fine sand and coarse sand" $eigh indi idually and record" Ta!e about =4+: gm of each fraction in bea!er, add =4+: ml >"> N *Cl till all the material is #et #ith *Cl, if required,

add further =4+: ml *Cl, to dissol e the material" The cement portion gets dissol ed and sand portion gets separated from cement, digest on #ater bath for +: minutes I filter the liquid through filter paper, #ash #ith #ater till chloride free" The filtrate is e aporated and silica determined as in earlier part, from silica cement portion in each fraction is determined, from total #eight, #eight of sand obtained by subtracting #t" Of cement and hence the ratio of cement to sand is calculated" /lso < of cement in the sample is calculated" 1.8.2 EDTA Titr*tio s: 'or filtrate same as cement normally the ratio of cement- sand used in plastering #or! is +-7 0The ratio used for compound #alls and such other #or! is +-9 to +-51" 1.;. Co crete:

1.;.1. Co crete is a blend of cement, sand and aggregate in different proportions used for different purposes" Normally samples from debris selected are pieces of beams and slabs ta!en for analysis" /bout +C6!g sample is required for analysis dry the piece from slabCbeam in o en at ++:DC for += minutes", cool, and #eigh" Then grind the sample so that cement particle gets separated from sand and aggregate" (ie e the bul! #ith different mesh si2e sie es, to separate po#der, fine sand, coarse sand and aggregate" $eigh the fractions so separated indi idually" Ta!e about =4 9gm from po#der fraction and fine sand fraction, about =:49: gm from coarse sand and about +::4+=: gms of aggregate fraction for actual silica and calcium oxide determination" Ta!e all the four fractions as abo e in 6=: ml bea!ers, add sufficient quantity of >"> N *Cl to dissol e the adhering cement particles" Then digest on #ater bath for +:4+= minutes and filter" The filtrate so obtained is used for silica determination" 1.;.2. Silic*

'iltrate e aporate the filtrate to dryness on hot plate, dry silica remains in the bea!er along#ith calcium and aluminium salts" Then add >"> N *Cl and digest on #ater bath for = mins" 'ilter the silica through ashless filter paper, #ash #ith #ater until chloride free" Dry the silica in o en and further in furnace at 5::DC4+:::DC for 6 hrs, $ash the silica so obtained" 'rom silica calculate the #eight of cement obtained in different fractions 06:< silica E +::< cement1" .ach fraction contains some cement portion and rest being fine sand, sand and aggregate respecti ely" Ta!e sand and fine sand fractions together" Thus calculate the total cement, total sand and total aggregates present in the sample, and hence calculate the ratio of cementsand- aggregate also calculate the cement percentage" 'rom filtrate of silica, calculate CaO < as detailed in cement, from CaO< also calculate the < cement in each fraction, and hence get the ratio of cement- sand- aggregate,

and also < of cement in the sample" Compare the results obtained by silica and CaO" 1.C. A)TERNATIDE MET2OD 1re0. 4+#': 1.C.1. Beep the sample in the o en for += minutes and then !eep them in desiccator for cooling" $eigh +: gms of sample" /dd distilled #ater and sha!e #ell" /dd 6:< *Cl and boil the solution" Distilled #ater and dilute *Cl 0i"e, 6:<1 should be added upto +=ml, if required" /dd a fe# drops of conc" *Cl, #arm4#aterCdistilled #ater o er the dissol ed residue so that complete CaO dissol es in the filtrate" &a!e the solution upto =:: ml after adding distilled #ater in the flas!" Transfer the solution after sha!ing #ell in the bea!er" %ipette out +:ml of the solution each in > bea!ers in separately" /dd +:4+= ml dilute nitric acid 06:<1 in each bea!er" /dd 6:ml distilled #ater in each and boil, add ammonium chloride 0nearly + J 4 6 tea spoon1 and boil, cool the solution and then add ammonium hydroxide and boil the solution for the precipitation to be formed in the III rd group" Remo e the bea!er and allo# precipitates to settle do#n" 'ilter the solution, ma!e saturated solution of ammonium oxalate in a bea!er and distilled #ater nearly >::ml of solution is formed" Nearly +::ml each of ammonium oxalate solution are added in each bea!er after filtering the precipitates" %recipitates as of interfering radical are being remo ed in the IIIrd group" /fter adding ammonium oxalate solution, boil the solution for the precipitates to be formed of CaO" 'ilter CaO precipitate" Note: - Three separate solutions in bea!er are ta!en as mean of the titration reading is ta!en as single reading gi es error. No#, #ash the precipitates #ith #arm #ater till it is fee from oxalate" Ta!e fe# drops of filtrate in a test tube, and CaCl 6 solution fe# drops, if precipitate forms then solution is not free from Oxalate" $ash till the residue is free from Oxalate" Beeping the filter paper carrying the precipitate in the bea!er, #ash the filter paper #ith distilled #ater so that, complete precipitates are transferred in a bea!er" /dd dilute sulphuric acid as CaO is soluble in dilute sulphuric acid" /dd fe# drops of concentrated sulphuric acid if precipitates are not dissol ed in dilute sulphuric acid completely, #arm the solution" %ut NC+: B&nO7 in a burette and titrate it against #arm solution at nearly =:4 9:DC i"e", till pin! colour appears" No the initial and final reading and mean amount of B&nO7 used" < Calcium Oxide 0CaO1 < Cement E &ean H +"7 E CaO < H 0+::C961

/ssume the pure Cement CaO < E 96"

1.C.2.I solu!le Resi$ues: Transfer the residue of the pre ious test in a bea!er" /dd >: ml hot #ater and >: ml 6N Na6CO> solution" *eat the solution belo# boiling point for +: minutes" 'ilter it, #ash the residue on the filter paper #ith dil"*Cl 0+-,1 and finally #ith hot #ater till the residue is free from chlorides" Ignite the residue in a tarred crucible at ,:: 4 +:::DC, cool in a descicator and #eigh" 1.C.3. C,ec5 0or C,lori$es:

Ta!e fe# drops of filtrate and add 64> drops of /gNO > solution, #hite precipitates formed indicates the presence of Chlorides, #ash the residue #ith hot #ater till it is free from chlorides" 1.1.. CONC)=SION:

Cement is complex mixture and testing of cement is a difficult tas!" In 'orensic Context one has to certify #hether the sample is cement or not, and if so, the percentage of cement in the sample" The initial tests li!e Thymolphthalein test, *eat test, %erformance test along #ith percentage of acid insoluble, calcium oxide and silica data can allo# one to frame a report regarding the cement percentage #ith acid insoluble and cement #ith non cementitious material" If stone po#der is used for adulteration the acid insoluble amount for adulteration percentage but if lime or other material is used for adulteration then acid insoluble #ill be less and calcium oxide #ill be more, the correct cement percentage can be assayed from silica content" The relati e standard de iations of :"77 < and reco ery of ,, < #as obtained for .DT/ titration" The relati e standard de iations of :"55 and reco ery of ,7 < #as obtained for silica determination" *ence the results of the analysis of cement by the abo e methods are quite accurate and reproducible" In case of &ortar and Concrete the ratio of Cement- (and and Cement- (and- /ggregate is ery important, hence selection of the sample plays an important role, finally the cement content determined by abo e method assures accuracy and reproducibility"

Ot,er i stru&e t*l tec, iEues 0or t,e ce&e t * *l"sis +" 6" IC% KRD

These technqiues can be used for the analysis of contentsCelements of the cement, as per the literature a ailable in the laboratory R.'.R.NC.(-

0+1" (tandard &ethods of Chemical /nalysis, by $ilfred $" (cott" 'ifth .dition, +,>:" 061" The Chemistry of Cement and Concrete, by 'rederic! &" )ea", Third .dition, +,=:" 0>1" Indian (tandard (pecification for Ordinary, Rapid, *ardening and )o# heat %ortland Cement 0re ised1" I( 69,4+,=5" 071" Technical &ethods of /nalysis, .dited by Ra?as Besle Griffren &c" Gra# *ill, NL" 0=1" *andboo! on Concrete mixtures based on Indian (tandards" I(I %ublishers"

SECTION - 2 (IRE + ARSON 2.1. Title- .xamination of fireCarson cases" 2.2. Scope- Inflammable materials li!e petrol, !erosene, diesel, alcohols, thinners, sol ents etc" 2.3. Purpose- Detection and identification of inflammable materials or their residues in the exhibits of fireCarson cases" 2.4. Respo si!ilit"- Reporting officers and supporting scientific staff" 2.#. I tro$uctio /rson means the crime of putting property on fire deliberately" The fire #ith mollified intention may be for getting economicCfinancial gain, do#ry death or communal purposes" This chapter also co ers the examination of burnt materials collected in connection #ith accidental fire cases" The cause of fire may be either #ith the use of explosi es material or accelerants" The collection of rele antCright physical e idence from scene of crime is a difficult tas!" The in estigating officerCteam collect arious types of exhibits in ol ed in fireCarson incidents and send them to the laboratory for examination" 'lammable petroleum products such as petrol, !erosene, diesel etc" are commonly used as accelerants in fireCarson cases" (ometimes, alcohol paintsCthinner, industrial sol ent and other inflammable material are also encountered in the incidents of fireCarson" The type of exhibits sent to the 'orensic (cience )aboratories comprise unburnt, partially burnt and badly burnt materials" 2.%. Met,o$s- Depending upon the nature 0shape, si2e and state1 the appropriate portion of the exhibits such as paper, hair, different types of clothes, #ood etc" are made into small pieces and are steam distilled 1" /pproximately =: ml of distillate is collected" .xtract the distillate #ith approximately >: ml of diethyl ether in 64> extractions" Combine the extracts and concentrate by e aporating the extract at room temperature to a small olume 0approx" :"=4+ ml1" The standard samples of petrol, !erosene and diesel 0by adding about :"= ml of each in distillation flas!1 and blan! sample are also prepared simultaneously in the same #ay" /lternati ely, the direct extraction of the exhibits, standard and blan! can also be carried out #ith diethyl ether" The concentrated extract is used for Gas Chromatographic /nalysis in both the cases" 2.6.1 Gas Chromatography2

/ppropriate olume of the concentrated extracts of the exhibit, standards and blan! samples are in?ected into a gas chromatograph #ith the follo#ing conditions- 4 Column- %ac! (. M >:, /pie2on ) or its equi alent column, #hich can used for the separation of petroleum products etc" Detector- 'lame Ioni2ation Detector Carrier Gas- Nitrogen 'lo# Rate- >: mlCminute O en Temperature %rogrammeiF: (or petrol M (tarts at ;::C 0isothermal for = minutes1N increases #ith : = Cminute upto +6: C" 0;:4+6::C1 iiF (or 5erose e M (tarts from ,::C and continues at +::CCminute upto : : : 66: C 0,:466: C1N hold for ="= minutes at 66: C iiiF (or : $iesel 4 (tarts from ,::C and continues at +::CCminute upto 67::C : 0,:467: C1N hold for +:"= minutes at 67: C Detector TemperatureiF (or petrol M +;::C iiF (or 5erose e M 6>::C iiiF (or $iesel 4 67::C In?ector TemperatureiF (or petrol M +;::C iiF (or 5erose e M 6>::C iiiF (or $iesel 4 67::C 2.%.2 >*s C,ro&*to3r*p,"3 Oper*ti 3 Co $itio s Column- 9O x :"+6=P (( %ac!ing- OA +, (. >: or /pie2on ) or its equi alent column, #hich can be used for the separation of petroleum products etc" Detector- 'lame Ioni2ation Detector 0'ID1 Carrier Gas- Nitrogen 'lo# Rate- >: mlCminute be

O en Temperature- %rogrammed from 9:oC to 6=:oC at 7oC per min" Detector Temperature- 6;=oC In?ector Temp- 6=:oC 2.%.3 C*pill*r" >*s C,ro&*to3r*p," Oper*ti 3 Co $itio s Column- =: M +::m Glass Capillary x :":+:P ID %ac!ing- OA +:+, or its equi alent column Detector- 'lame Ioni2ation Detector 0'ID1 Carrier Gas- Nitrogen 'lo# Rate- + mlCminute O en Temperature- %rogrammed from 9:oC to 6=:oC at 6oC per min" Detector Temperature- 6;=oC In?ector Temp- 6=:oC Re0ere ces: +" @aggi TR, &urthy *RB, %rasad Aeena /, Detection and characteri2ation of petroleum residues by Thin )ayer Chromatography, 8" Indian /cad" 'orens" (ci",+,5;, ol"69 No"6, pp >94>5 6" /rora BB, Golani *C, Narayans#ami B, Detection of adulteration in petroleum products by gas liquid chromatography, 8" Indian /cad" 'orens" (ci" +,;=N+7- pp 74+>" >" / unified scheme for the analysis of light petroleum products used as fire accelerants by Da id $illson, 'orensic (cience, +,;;, ol"+:, pp 67>46=6" +" /B, @hat @(, %andya A(, Detection of !erosene residue on T)C, 8" Indian /cad" 'orens" (ci",+,;;, ol"+9 No"+, pp =74=9" 6" Chhabra B(, Ra? anshi /C, Narain (ri, Thin )ayer Chromatography method for identification of petrol as fire accelerants, Indian 8ournal of 'orensic (ciences, +,,+, ol"=, pp+;>4+;=" >" Narayans#ami B, @ami *), Gas4chromatography in detection of !erosene residues, International Criminal %olice Re ie#, +,;+, No"6=+, pp 6+94 66+" 7" &attoo @N, %ai %%, $ani /B, (pectroscopic detection of !erosene residues in forensic #or!, 8" Indian /cad" 'orens" (ci",+,;:, ol",, No"+, pp 745"
3

=" /rora @@, Transmission .lectron &icroscopic (tudies of &orphology and Crystallography of %articulate .missions in (mo!es, 'orensic (cience International, >6, +,59, pp +5=4+,6

SECTION - 3 OI)( I '/T( 1.1 Title - Oils and 'ats" 3.2 Scope- /nalysis and characteri2ation of arious oils and fats" 3.3 Purpose- To determine the quality and adulteration of oils and fats" 3.4. Respo si!ilit"- Reporting officer and supporting scientific staff" 3.#. I tro$uctio Oils and fats analysis co ers a ma?or area in forensic chemistry as these are often adulterated in arious #ays i2 adulteration of lo#er grade, different origin 0 egetable oil by animal fat or ice4 ersa1 or misbranding or contrabanding of products etc" the cases as abo e may occur frequently as oils and fats are extremely used as coo!ing media and also in industries li!e paints, arnishes, pharmaceutical industries" There may be theft cases or illegal possession related to oils and fats" In the abo e case the samples are frequently sent for their examinations for different purposes under .ssential Commodities /ct 0."C /ct1" Oils and fats may be di ided into t#o classes based on their origin, egetable oil and animal fat" The #ords oil and fat are synonymous" They differ in their physical state i"e oil is liquid and fat is solid" The egetable oils include groundnut, sunflo#er, linseed, coconut, mustard, palm, sesame, cottonseed, rapeseed, soyabean, castor, pea4nut, rice bran oils etc" there may be hardened fat from egetable oil by the hydrogenation of oils 0product4 Aanaspati1" The animal fat is a ailable in the form of butter, ghee, tallo# and lard" (ome times tallo# or lard is added to egetableC desi4ghee or anaspati" In case of analysis of samples for#arded to forensic laboratories under ."C"/ct, the samples are ery often required to be examined for arious parameters based on physical and chemical properties of indi idual oil and fat to ascertain the nature, quality, adulteration etc" (ometimes chemical tests are performed to identify a specific oil, adulteration, or rancidity 0fouling of oils1" The examination of oils and fats is based on their composition" Oils and fats are tri4glycerides of fatty acids of saturated and unsaturated acids 0oleic, linoleic, linolenic1 in arying proportions" These may contain fatty acids of

definite carbon number in higher proportion in some oils i2 arachidic acid in arachis oil, groundnut, nut and peanut oil, crucic acid 0in mustard, rapeseed and rape oil1, normal and iso propionic and butyric acid in butter and ghee 0both animal fat1 or ary in the percentage of free acids 0 due to hydrolytic splitting of glycerides of en2ymes1 or ary in the nature and percentage of sterols in the unsaponifiable matter in animal fats and egetable oils 0cholesterol in animal fat and phytosterols in egetable oils1 or ary in drying properties 0 the tendency to form solid film due to unsaturation"1 or ary in their tendency to undergo addition #ith iodine or iodine monochloride 0due to ariation in unsaturation present1" The abo e ariation in fatty acid profile and other not fatty acid or non glyceride constituent 0unsaponifiable matter i"e" sterols- higher alcohols1 are responsible to impart effect on their physical and their chemical properties i2" colour, refracti e index, melting point, saponification point, al!aline hydrolysis beha iour, additi e reactions etc" the differences in physical beha iour and parameters based on chemical characteristics 0saponification alue, acid alue, acetyl alue, Reichert4 &eissel alue, polens!e alue etc"1 pro ide analytical guideline or clues to the classification or characteri2ation of different oils and fats" Chemical tests are often performed to detect adulteration or rancidity" *o#e er, quality assurance or adulteration chec!ing become ery often difficult #hen alues on some parameters o erlap" Then the help of other physical characteristics or special parameter i2" Reichert4&eissel alue, polens!y alue in case of adulteration of ghee or anaspati by hard and tallo# or by instrumental technique specially GC for fatty acid profiling" Aarious options are to be applied profitably to get the analysis done in a short time" The interpretation of the alues on parameters requires special !no#ledge and expertise" 3.%. Met,o$s- 3.%.1. SAMP)IN>: T,e s*&pli 3 o0 t,e eA,i!its to !e collecte$ 0or * *l"sis &*" !e $o e *s per t,e proce$ure l*i$ $o7 i t,e I $i* St* $*r$ Proce$ures o0 S*&pli 3 o0 Oils + (*ts. >"; /N/)L(I(>";"+" (ample %reparation>";"+"+" )iquid oils'ilter the oil sample through $hatmann filter paper no"+" >";"+"6" (olid and (emi solid oils&elt the solid or semi solid samples of oils and fats heated in an o en to +: :C abo e the melting point of the fat" 'ilter the melted sample through $hatmann filter paper no"+"

3.8.2. C2EMICA) TESTS: 3.8.2.1 >RO=ND N=T OI):

1

Ta!e one ml of the sample of oil in a conical flas!, add = ml of +"= N alcoholic potash and saponify completely on a boiling #ater bath using an air condenser to a oid loss of alcohol" It ta!es about +: minute" During the saponification s#irl the flas! many times, cool, add :"+ ml of phenolphthalein indicator" Neutrali2e the solution exactly by adding dil" /cetic acid 0One olume of glacial acetic acid Q t#o olumes of distilled #ater1 follo#ed by an extra amount of :"7 ml of acetic acid" &ix =: ml of ;:< alcohol fit a thermometer into the flas! #ith the help of a cor! in such a #ay that the bulb of thermometer is dipped in the liquid but does not touch the bottom of the flas!" *eat the flas! on a #ater bath until the temperature reaches =: :C to become solution clear" Beep the flas! for cooling in air #ith frequent sha!ing until the temperature of the solution comes to 7: :C" /ppearance of turbidity in bet#een >, to 7::C indicates the presence pure groundnut oil" Cool the flas! #ith constant sha!ing in a cooling bath at += :Q +: C in such a #ay that the temperature falls roughly at the rate of 6 :C per minute" Note the turbidity temperature, #hich is the temperature at #hich the first distinct turbidity appears" This turbidity temperature is once again confirmed by a little further cooling #hich results in the deposition of precipitate" Dissol e the precipitate by heating the contents at =::C on a #ater bath" /gain cool as abo e and ma!e an experiment for duplicate determination of turbidity temperature" Duplicate shall agree #ith in Q :"=:C" Re*3e ts1. Alco,olic Pot*s, G1.# NF: Dissol e 5"= g of potassium hydroxide in +:: ml purified rectified spirit" The solution may be !ept in dar! colour bottle preferably" 6" %urifiedC Rectified (piritReflux +"6 litre of rectified spirit for half an hour in a distillation flas! containing +:g of caustic potash and 9 g of granulated aluminium or aluminium foil pieces" Distil and collect one litre after discarding the first =:mm, this is purified, rectified spirit used for reagents" 3. P,e olp,t,*lei I $ic*tor:

Ta!e :"= g of phenolphthalein and dissol e in =:ml of purified rectified spirit and mix the solution #ith =:ml of distilled #ater" 3.8.2.2. SESAME OI):
1

Ta!e = ml of the oil sample or melted fat in a cylinder or test tube #ith a glass stopper add = ml of hydrochloric acid and :"=ml of furfural solution 06< freshly distilled furfural in ethanol1" 'it the glass stopper and sha!e igorously for t#o minutes" Beep the mixture separate" The appearance of pin! or red colour in the lo#er acid layer indicates the positi e tests for the presence of sesame oil" Confirm it by adding = ml of #ater and sha!e again" If the colour remains the sesame oil is present if colour disappears the sesame oil is absent"

3.8.2.3. COTTON SEED OI): Ta!e about =ml of the oil sample or melted fat in a test tube and add equal olume of sulphur solution 0+< #C solution of sulphur in carbon disulphide and add an equal olume of amyl alcohol1" (ha!e thoroughly and heat it on a #ater bath 0;:: to 5::C 1 for fe# minutes #ith occasional sha!ing" To boiled off carbon disulphide and stop foaming" Beep the test tube in an oil bath at ++:4++=:C for about t#o and a half hours" /ppearance of red colour at the end of the period indicates the positi e test for the presence of cottonseed oil" 3.8.2.4. PA)MO)EIN IN >RO=ND N=T OI): Ta!e 9:4;= gms of samples and heat it to +>: :C before its testing" /dd ca 7=ml of the heated fat into an oil sample bottle" Beep the bottle in #ater bath" Cool the bottle in #ater bath #ith stirring using the thermometer to !eep the temperature uniform" $hen the sample reaches a temperature ca +::C abo e the cloud point, being stirring steadily and rapidly in a circular motion so as to pre ent super4cooling and solidification of fat crystals on the sides or bottom of the bottle" Do not remo e the thermometer from the sample at this stage" The test bottle is maintained in such a position that the upper le els of the sample in the bottle and the #ater in the bath are about the same" Remo e the bottle from the bath and read the temperature" The cloud point is that temperature at #hich that portion of the thermometer immersed in the oil is no longer isible #hen ie#ed hori2ontally through the bottle" This test is useful for the detection of palmolein in groundnut oil" %resence of palmolein o er +:< in groundnut oil readily gi es cloud at a higher temperature than that of groundnut oil due to the presence of palmitic gly4 cerides in higher amounts in palmoleinCpalm oil" 3.8.2.#. )INSEED OI):

Ta!e one ml of the oil sample in a boiling tube of about +:: ml capacity" /dd =ml chloroform and about one ml of bromine drop4#ise till the mixture becomes deep red" Cool the test4tube in an ice #ater4bath" /dd to it +"=ml of rectified spirit drop #ise #ith sha!ing the mixture until the precipitate #hich is first formed ?ust dissol es and then add +:ml of diethyl ether" &ix the contents and !eep the tube in the ice #ater4bath for half an hour" /ppearance of precipitate indicates the positi e test for the presence of linseed oil" This test is not applicable for the detection of linseed oil in presence of mohua oil" 3.8.2.%. RICE-<RAN OI) IN OT2ER EDI<)E DE>ETA<)E OI)S: Ta!e 6:ml of oil sample in a +::ml separating funnel" /dd to it equal olume of aqueous potassium hydroxide solution0>:<1" (ha!e the content gently and constantly for +: minutes" /llo# the separating funnel to !eep for about 7= minutes for the separation of al!ali layer" Ta!e out the al!ali layer and neutrali2e it #ith dilute hydrochloric acid solution" Neutrali2ation is confirmed #ith blue litmus paper" .xtract this solution #ith diethyl ether 06:ml x > times1" $ash the extract #ith distilled #ater and dry on anhydrous sodium sulfate" . aporate the sol ent on hot #ater bath and dissol e the residue in chloroform and use it for Thin )ayer Chromatography #ith follo#ing conditions" St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness" DeHelopi 3 solHe t: - ben2ene and acetic acid 0+::-+1" DeHelop&e t: : +=cm Disu*li9*tio : M Iodine fuming O!serH*tio : - Rf alue of Rice4bran oil M :";: to :";= 0Use of control sample of Rice4bran oil is recommended1" This spot is absent in all the commonly a ailable eatable oils in the mar!et" The Rice4bran oil in other edible egetable oils can be detected by this T)C method up to a minimum of =< le el" 3.8.2.8. CASTOR OI) IN EDI<)E OI)S: A. T,i )*"er C,ro&*to3r*p,"

St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness"

DeHelopi 3 solHe t: - %etroleum ether 07:49::C1, diethyl ether and acetic acid 09:-7:-6, C 1" DeHelop&e t: : +:cm Disu*li9*tio : : Iodine apours O!serH*tio s: - Rf alue of castor oil M /bout :"6= S*&ple prep*r*tio : : Chloroform / prepared sample of an oil containing +< added castor oil may be used as a control sample for identifying the spot" The spot shall be noted in tan! since it fades on remo ing the plate" This method has a sensiti ity of +<" This method is specific for castor oil but ransid or oxidi2ed groundnut oil gi es a strea!" <. CASTOR OI) AND ITS DI((ERENTIATION (ROM RANCID OI)S: Ta!e about =ml of suspected ransid oil sample in a round bottom flas! add to it about 6 gms acti ated charcoal" &ix the content thoroughly and heat it on a boiling #ater bath for about half an hour #ith constant sha!ing" The bleached oil is filtered on a filter paper" The filtered oil may no# be passed through a mini column made up of neutral alumina M+: gms using =:ml hexane as eluent" This bleached and neutrali2ed oil is used for T)C as gi en in the detection of castor oil besides as reference standard" 3.8.2.;. AR>EMONE OI) IN EDI<)E OI)S: Met,o$ 1 : (erric c,lori$e test Ta!e about =ml of filtered oil and dissol e in =ml of toluene in a stoppered glass test tube" /dd to it = ml of concentrated hydrochloric acid" (ha!e igorously and allo# the acid layer to separate" (eparate the acid layer #ith the help of separating funnel to a test tube" /dd + ml of 'erric chloride solution0 +: gm 'erric chloride in +:ml concentrated hydrochloric acid and add ,:ml of #ater" If required filter it1 through the side #all of the test tube" &ix the reagent and acid layer #ell" This can be achie ed by gently rotating the test tube bet#een the palms of the hand" *eat the test tube boiling #ater bath for about +: minutes" 'ormation of needle shaped 0straightCcur ed1 reddish bro#n crystals in clusters on cooling indicates the positi e test for the presence of /rgemone oil" The method is not sensiti e enough to detect argemone oil in less than :"+< le el"

Met,o$ 2: T,i )*"er C,ro&*to3r*p,"

S*&ple Prep*r*tio Ta!e about =ml of suspected oil sample in about = ml of diethyl ether in stoppered flas!" /dd = ml of conc" *cl and sha!e the mixture igorously for +: minutes" *eat the flas! on a hot #ater bath ha ing temperature 7:o for about +: minutes" (eoarate the acid layer using separating funnel in a +: ml bea!er" Beep the bea!er on a boiling #ater bath and e aporate to dryness" Dissol e the residue in + ml of chloroform and acetic acid mixture 0,-+1 and used for T)C #ith follo#ing conditions" St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness" DeHelopi 3 solHe ts: : +" @utanol, acetic acid, and #ater 0;-6-+1" 6" *exane and acetone 09-71 >" *eptane DeHelop&e t: : +:cm Disu*li9*tio : : U"A" )ight O!serH*tio : - @right yello# spot at about Rf alue :"5 in system +, :"7 to :"= in system 6" The plate sho#s spot of blue fluorescence if it seen under U A )ight after sprayed #ith 6:< aqueous (olution of sodium hydroxide" The abo e obser ations indicate the positi e test for the presence of argemone oil" The detection limit is up to =: ppm" Met,o$ 3:2i3, Per0or&* ce )iEui$ C,ro&*to3r*p,"2 3.8.2.C. -ARANIA OI) G PON>AMIA >)A<RAF S*&ple Prep*r*tio : Ta!e about 6: ml of suspected oil sample in a +:: ml separating funnel and +: ml of conc" *cl" (ha!e the mixture gently and constantly for += minutes" Beep the funnel aside for about J hour to separate the acid layer" Ta!e out the acid layer in a bea!er and !eep it on a boiling #ater bath for the e aporation to dryness" Dissol e the residue in

about :"= ml of chloroform and used it to carry out the T)C #ith follo#ing conditions" St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness" DeHelopi 3 solHe t: : %etroleum ether 07:49::C1 , diethyl ether and acetic acid0 9-7-+1" DeHelop&e t: : 6: minutes Disu*li9*tio : : Dry the plate at room temp" and #atch under U"A" )ight O!serH*tio : +" /ppearance of > bluish green spots at about Rf alues:">7, :"66, and :"+; on the plate indicates the positi e test for the presence of Baran?a oil" The detection limit is up to :":+ <" 6" De elopment of a yello# color in the acid layer after extraction also indicates the positi e test for the presence of Baran?a oil" >" %alm argemone oil do not interfere" the oil

7" The T)C may also be carried out directly on chloroform solution of instead of extracted solution but the test may not be sensiti e" >";"6"+:" &IN.R/) OI)( IN .DI@). OI)(-

Met,o$ 1- Ta!e 66 ml of the :"= N alcoholic BO* solution in a conical flas!, add +ml of the suspected oil sample" @oil it on a #ater bath using an air or #ater cooled condenser till the mixture becomes clear and free from any oil drops on the side #all of the flas! also" Ta!e out the flas!, transfer the contents to a #ide mouthed #arm test tube, add 6= ml of boiling #ater along the side of the #all #ith care" (ha!e the tube lightly from side to side #hile adding #ater" The appearance of turbidity indicates the positi e test for the presence of mineral oil" Met,o$ 2 : T,i )*"er C,ro&*to3r*p,"

St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness" DeHelopi 3 solHe t: : %etroleum ether 07:49::C or 9:45:oC1"

DeHelop&e t: : /bout 7 minutes, C 9 cm Disu*li9*tio : : Dry the plate in air at room temperature, spray #ith fluorescin solution 0 6< soln" Of 6O,;O4dichloro4fluoresein in ,9 < ethanol and #atch under U"A" )ight O!serH*tio : - /ppearance of yello# fluorescent spots on the sol ent front indicates the positi e test for the presence of mineral oil" The egetable oil gi es a yello# strea! about 64> cm long from the base" 3.8.2.11. DETECTION O( RANCIDIT? IN EDI<)E OI)S &ethod +4 Bries TestTa!e about =ml of suspected oil sample and sha!e igorously #ith =ml of :"+< %hlorogucinol solution in diethyl ether and = ml of conc" *Cl for 6: seconds" /ppearance of pin! color indicates the presence of incipient rancidity" Met,o$ 2 : =D Spectrop,oto&etric &et,o$: Ta!e an accurately #eighed amount of oil sample in a 6= ml olumetric flas! so that the absorbance of its solution in iso4octane lies bet#een :"6 to :"5 using a + cm quart2 cell" (can the sample by UA (pectrophotometer using iso4octane as blan!Creference bet#een 66: to >6: nm" (ample gi es three #a elength l max" at 6>:,695, and 6;5 nm" 'or quantitation select the #a elength l max" of maximum absorption near orC at 6>:,695, and 6;5 nm, and the absorbance 0/1 at these points" Speci0ic A!sor!* ce E1 1/ c& Gl &*A.F E /bsorbance 0/1Cc K d $here c E concentration of the sample solution in gmC+::ml d E length of the cell in cm 3.8.3 (T-IR SPECTROSCOPIC MET2OD4 &a?or pea!s G c&-1F of some of the oils are gi en belo# Must*r$ oil: >::;, 6,67, 65=7, +;7;,+79=, +7+5, +>;5, +6>5, ++9>, +++,, ,95, ;66" De3et*!le (*t: >::=, 6,66, 65=7, +;7;, +79=, +7+5, +>;5, +6>9, ++9>, +++,, +:,,, ,9;, ;66

>rou $ ut oil: >::;, 6,=7, 6,69, 65=7, +;7;, +79=, +7+5, +>;5, +6>;, ++9>, +++,,+:,,, ,99, ;66 Coco ut oil: 6,==, 6,67, 65=7, +;7;, +799, +7+5, +>;5, +665, ++9+, ,96, ;66" 3.8.4. DETECTION O( ACID AND CO)ORS IN EDI<)E OI)S: 3.8.4.1. DETECTION O( 2?DROC?ANIC ACID IN EDI<)E OI)S: It is present sometimes as an impurity in mustard oil" Ta!e += M>: ml of suspected oil sample in a 6=: ml conical flas!, add about =: ml of #ater and mix #ell" /dd to it += ml of +:< tartaric acid solution and mix" %ut the stopper to the flas!, #ith a el et cor! from #hich hangs a picric acid paper 0(oa! the filter paper in a saturated aq" solution of picric acid, draining the excess liquid and dry the paper in air1 of about ;= mm long #etted #ith a drop of =< sodium carbonate solution" The flas! is !ept on a hot #ater bath by the side of the steam ent but not directly on the steam for >:to 7= min" The red colour on picric paper indicates the presence of hydrocyanic acid" Ignore the pin! or reddish hue, #hich may at times, appears at the periphery of the picric acid paper" >";"7"6" D.T.CTION O' CO/) T/R OI) (O)U@). CO)OR( IN .DI@). OI)(Ta!e about = ml of suspected sample of oil in test tubes, add about += ml of petroleum ether to each tube follo#ed by =ml of *Cl of different concentrations 07-+, >-+, 6-+, and +-+ *cl and #ater1 to different tubes" Change in color indicate the presence of coal tar oil soluble colors in the sample" 3.8.4.3. ISO)ATATION AND DETECTION O( OI) SO)=<)E CO)ORS: Ta!e about =ml of suspected sample of oil stoppered conical flas!, add to it 6= ml of hexane follo#ed by +: gms of silica gel of column chromatography grade and 6 gms of anhydrous sodium sulphate" (tir the mixture and !eep for = minutes" Decant off the sol ent" /dd another 6= ml of hexane and stir #ell and decant the sol ent" Repeat the process >47 times #ith 6= ml of hexane and draining out the sol ent each time to remo e almost all the oil lea ing behind the silica gel in the flas!" .lute the coloring matter absorbed by the silica gel in the flas! by sha!ing #ith 6: ml of diethyl ether 64> times" Collect the diethyl ether extract in a bea!er" . aporate the sol ent on a hot #ater bath to concentrate, #hich can used to carry out the T)C #ith follo#ing conditions" St*tio *r" p,*se: - (ilica gel G, :"6=mm thic!ness"

DeHelopi 3 solHe t: : @en2ene, hexane and acetic acid 09-7-+1" DeHelop&e t: : +64+= cm Disu*li9*tio : : Dry the plate at room temp" and heat the plate at +::: oc in an o en for + hour" O!serH*tio s: - Natural colors li!e carotenes fades a#ay lea ing oil soluble coal tar colors" >";"=" D.T.R&IN/TION O' %*L(IC/) %RO%.RTI.( O' OI)(3.8.#.1. Deter&i *tio o0 &oisture * $ Hol*tile su!st* ces:

Carry out the determination of moisture and olatiles in duplicate" Met,o$-1 $eigh =4+: gms of suspected sample of oil or fat in a tared metal dish of about ;cm dia and >47 cm deep #ith lid" &ixed the sample thoroughly bi stirring" )oose the lid and heat in an o en at +:>Q 6oC for > hours" Ta!e out the dish from the o en and close its lid" Cool it in desicator ha ing phosphorous pentoxide and #eigh" *eat once again for + hour and #eigh" Repeat the process to get concurrent readings #ith a difference of 6 mg /*3e o0 &oisture J $eight )ossC$eight of oil ta!en K +::

Met,o$-2 $eigh about +: gms but accurately of suspected sample of oil or fat in a tared glass bea!er #ith glass rod, cool it in a desiccator ha ing phosphorous pentoxide and #eigh" *eat the bea!er #ith sample on an electric hot plate #ith continuous stirring by glass rod" The complete remo al of moisture is indicated by the cessation of the rising bubbles and by absence of foam, #hich can also be tested by !eeping a clean and dry #atch glass on the top of the bea!er and obser e the #ater apours, if any" Cool the bea!er to room temperature in the desiccator and #eigh" / *3e !" 6t. o0 &oisture * $ Hol*tiles co te t E $t" )ossC$t"of sample ta!en K +:: 3.8.#.2 Deter&i *tio o0 Speci0ic >r*Hit" - 4 Determine the specific gra ity of the oil by specific gra ity C density bottle or a py!nometer method at ambient temperature by noting do#n the #eight of specific gra ity bottle #ith oil, #ith #ater and the empty bottle" $eight of bottle #ith oil 4 $eight empty bottle Speci0ic >r*Hit" E 44444444444444444444444444444444444444444444444444444444

$eight bottle #ith #ater M $t of empty bottle 3.8.%. C2EMICA) C2ARACTERISTCS 3.8.%.1 Aci$ D*lue: It is the number of milligrams of potassium hydroxide required to neutralise the free fatty acid present in one gm of the oil or fat" The acid alue is determined by directly titrating the material in an alcoholic medium #ith aqueous sodium or potassium hydroxide solution" Re*3e ts*F Et,"l *lco,ol 0,=< C 1 or rectified spirit neutral to phenolphthalein indicator" !F P,e olp,t,*lei I $ic*tor - Dissol e one gm of phenolphthalein po#der in +:: ml of ethyl alcohol" cF St* $*r$ *Eueous *l5*li Solutio : /queous potassium hydroxide or sodium hydroxide solution of :"+or :"= N" Proce$ure- 4 &ix the oil or melted fat thoroughly before #eighing" $eigh accurately a suitable quantity 0 6": to 6"= gms1 of the sample of oil or fat in a 6=:4ml conical flas! and add to it =: to +:: ml of freshly neutralised hot ethyl alcohol, add about + ml of phenolphthalein indicator solution" @oil the mixture for about = minutes and titrate #hile hot #ith standard aqueous al!ali solution" (ha!e igorously during titration" The #eight of the oil or fat ta!en and the strength of al!ali used for titration shall be such that the olume of al!ali required for titration does not exceed +: ml" C*lcul*tio - 4 Aci$ D*lue J =9"+ A N C $ $here, A E Aolume of standard al!ali hydroxide solution in ml, N E Normality of standard al!ali used, and $ E $eight of the sample of oil or fat ta!en in gm" 3.8.%.2 S*po i0ic*tio H*lue- 4 It is the number of milligrams of potassium hydroxide required to saponify completely one gram of oil or fat" The material is saponified by refluxing #ith a !no#n excess of alcoholic potassium hydroxide solution" The al!ali solution consumed for saponification is determined by iterating excess al!ali #ith standard hydrochloric acid"

Re*3e ts: *F Alco,olic pot*ssiu& ,"$roAi$e solutio :- Dissol e >= to 7: gm of potassium hydroxide in 6: ml of distilled #ater, and add sufficient aldehyde free rectified spirit to ma!e up +::: ml" /llo# to stand o ernight, decant the clear liquid and !eep in a bottle closed tight #ith cor! or rubber stopper" !F P,e olp,t,*lei I $ic*tor solutio : phenolphthalein in +:: ml of rectified spirit" Dissol e +": gm of

cF St* $*r$ ,"$roc,loric *ci$: -approximately :"= N" Proce$ure&elt the sample, if it is not already liquid, and filter through a filter paper to remo e any impurities and the last traces of moisture" &a!e sure the sample is completely dry, mix the sample thoroughly, and #eigh by difference about +"= to 6": gm of the sample in a conical flas!" /dd 6= ml of the alcoholic potassium hydroxide solution #ith pipette and connect the reflux air condenser to the flas!" *eat the flas! on #ater bath or an electric hot plate for not more than one hour" @oil gently but steadily until the sample is completely saponified as indicated by absence of any oily matter and appearance of clear solution" /fter the flas! and condenser ha e cooled some#hat, #ash do#n the inside of the condenser #ith about +: ml of hot ethyl alcohol neutral to phenolphthalein" /dd about + ml of phenolphthalein indicator solution and titrate #ith standard hydrochloric acid" Conduct a parallel blan! determination at the same time" C*lcul*tio s: S*po i0ic*tio D*lue E =9"+ 0@4(1 N $ $here, @ E Aolume of standard hydrochloric acid required for the blan! in ml, ( E Aolume of standard hydrochloric acid required for sample in ml, N E Normality of the standard hydrochloric acid,and $ E $eight of the sample of oilCfat ta!en in gms"

3.8.%.3 Io$i e H*lue- 4 Iodine alue is the number of gram of iodine absorbed by +:: gm of oil or fat" &any methods for its determination are in use" Out of these $i?s is #idely accepted and recommended method by the @I(" It gi es the information about the amount of unsaturation or number of double bonds in a fat sample" 3.8.%.3. *F Deter&i *tio o0 Io$i e D*lue G6iKsF

>e er*l: -The material is treated in carbon tetrachloride medium #ith a !no#n excess of iodine mono chloride solution in glacial acetic acid 0$i?s solution1" The excess of iodine mono chloride is treated #ith potassium iodide and liberated iodine estimated by titration #ith sodium thiosulphate solution" ReagentsiF Pot*ssiu& Dic,ro&*te GARF GiiF. 2"$roc,loric *ci$ - Concentrated GiiiF. Pot*ssiu& io$i$e solutio 4 %repare a fresh solution by dissol ing +: gm potassium iodide free from potassium iodate, in ,: ml of #ater" GiHF. St*rc, solutio 4 Triturate = gm of starch and :":+gm of mercuric iodide #ith >: ml cold #ater and slo#ly pour it #ith stirring into + litre of boiling #ater" @oil it for three minutes, allo# it to cool and decant off the supernatant clear liquid" GHF. St* $*r$ So$iu& T,iosulp,*te solutio G..1NF M Dissol e approximately 67"5 gm of sodium thiosulphate 0Na6 (6O>"=*6O1 in distilled #ater and ma!e up to +::: ml" /nd standardise it as belo# $eigh accurately about =": gm of pre iously dried to a constant #eight at +:= Q 6DC potassium dichromate into a clean one litre olumetric flas!" Dissol e in #ater, ma!e up to mar!, and sha!e thoroughly" %ipette out 6= ml of this solution into a clean 6=:4ml conical flas!" /dd = ml of conc" *Cl and += ml of +: < potassium iodide solution" /llo# it to stand in the dar! for = minutes and titrate the mixture #ith solution of sodium thiosulphate solution using starch solution as an internal indicator to#ards the end" The end point is ta!en #hen the blue colour changes to green" Calculate the normality 0N1 of the sodium thiosulphate solution as bello# N E 6= $ C 7,":>A $here,

$ E $eight of the potassium dichromate in gm, A E Aolume of sodium thiosulphate solution required for the titration in ml i1 Glacial acetic acid 4 'ree from alcohol" ii1 Carbon tetrachloride iii1 Iodine &onochloride 0 ICl1 ix1 $i?Os Iodine &onochloride (oution Dissol e +: ml of iodine monochloride in about +5:: ml of glacial acetic acid and sha!e igorously" %ipette = ml of this solution, add +: ml of potassium iodide solution and titrate #ith :"+ N standard sodium thiosulphate solution using starch solution as an indicator" /d?ust the olume of the solution till it is approx" :"6 N" %rocedure &elt the sample if it is not already completely liquid, and filter through a filter paper to remo e any impurities and the last traces of moisture" Use absolutely clean and dry glass#are" $eigh accurately by difference an approx" quantity of the oil or fat bet#een the limits indicated in col" 6 and > of Table4+ into a clean dry 6=:C=:: ml iodine flas! or #ell ground glass stoppered bottle to #hich 6= ml of carbon tetra chloride ha e been added and agitate to dissol e the contents" The #eight of the sample shall be such that there is an excess of =: to 9: percent of $i?Os solution o er that actually needed" /dd 6= ml of the $i?Os solution and replace the glass stopper after #etting #ith potassium iodide solution, s#irl for intimate mixing and allo# the flas! to stand in the dar! for >: minutes in the case of non4drying and semidrying oils and one hour in the case of drying oils" Carry out a blan! test simultaneously under similar experimental conditions" /fter standing, add += ml of potassium iodide solution and +:: ml of #ater, rinsing in the stopper also, and titrate the liberated iodine #ith standard thiosulphate solution, s#irling the contents of the bottle continuously to a oid any local excess until the colour of the solution is stra# yello#" /dd one millilitre of the starch solution and continue the titration until the blue colour formed disappears after thorough sha!ing #ith the stopper on" T*!le 1 - $eight of oil or fat for determination of iodine alue"

Io$i e H*lue

Mi i&u& 6ei3,tM*Ai&u& o0 s*&ple 3& 7ei3,t s*&ple 3& G1F G2F G3F )ess than > +: +: = =":;; 9">79 +: 6"=>57 >"+;> =: :"=655 :"99+6 +:: :"6=>5 :">+;> +=: :"+;:: :"6+6= 6:: :"+69, :"+=59 C*lcul*tio :Io$i e D*lue J 12.%C G< - SF N 6 $here,

o0

@ E Aolume of standard sodium thiosulphate solution required blan! in ml, ( E Aolume of standard sodium thiosulphate solution required oil C fat sample in ml, and N E Normality of the standard sodium thiosulphate solution, $ E $eight of the oil C fat sample ta!en for the test in gm"

for the for the

T*!le 2- %roperties of some commonly encountered Aegetable Oils"#,% De3. Oil Sp. >r*Hit" 3.L 3.LC Groundnu t oil Coconut oil (esame oil &ustard oil :",:,4 :",+> :",+=4 :",6: :",+=4 :",+, 4444444444444 Re0r*ctiHe S*p. I $eA *t D*lue 4.LC M Io$i e D*lue G6iKNsF M5; 4 ,5

+"796:4 +"797: +"775:4 +77,: +"797=4 +"799= +"79794 +"7999

+55 +,= 67=469: ;"=4+:

+5=4+,> +:=4++= +9,4+;; ,54++:

Cottonsee d oil (oyabean oil (unflo#er oil &ahua oil

:",+:4 +"79>:4 :",6: +"799: 4444444444444 +"79=:4 +7;+: 4444444444444 +"797:4 +75:: :"5964 +"7=,:4 :"5;= +"79+: Ricebran :",+:4 +"79::4 oil :",6: +"7;:: %alm oil 4444444444444 +"77,+4 +"7==6 %almolein 4444444444444 +"7==:4 +"79+: Castor oil :",=74 +"79=,4 :",9: +"7;>: )inseed :",6>4 +"7;6:4 oil :",65 +"7;=: Re0ere ces:

+,:4+,5 ,54++6 +5,4+,= +6=4+7: +554+,7 +::4+7: +5;4+,9 =54;: +5:4+,= ,:4+:= +,=46:= 7=4=9 +,=46:= =7496 +;;4+5> 5645, +,:4+,9 +;:46:6

1. &anual of &ethods of Tests and /nalysis for food 0Oils and 'ats1 , Directorate General of *ealth (er ices, &inistry of *ealth and 'amily $elfare, Go ernment of India, Ne# Delhi" 2. (a?id *usain, R" Narsimha, and R" Nages#ara Rao, (eparation, Identification and determination of (anguinarine in argemone and other adulterated edible oils by re ersed4phase high perfomance liquid chromatography, 8ournal of Chromatography /, 59> 0+,,,1, pp"+6> to+69" 3. &ethods of sampling and test for oil and fats, I(- =75 0%art I1, +,97" 4. R( Aerma, /B Dalela and .( @abu, 'T4IR (pectrophotometric analysis of organic explosi e formulations, %roceedings of KIII /ll India 'orensic (cience Conference, /hmedabad, Gu?rat, from , to ++, No ember, 6::+ %% ++;4+79" #F" Indian (tandard InstitutionRs (pecifications for respecti e egetable oils" %F" &erc! Index" .le enth edition" OT2ER RE(ERENCES: +1" @ulletins of Indian Industrial Research, No" +: , Indian Aegetable OilsN by N" @rodie , +,>5" 61" I(- =75 0%art I1 4 +,97, &ethods of (ampling and test for oils and fats"

>1" I(- =75 0%art II1 4 %urity tests" 71" Indian (tandard InstitutionRs (pecifications for respecti e egetable oils" =1" &erc! Index" .le enth edition"

SECTION - 4 /N/)L(I( O' GO)D I OT*.R &.T/)( IN C*./TING C/(.( 4.1. Title - .xamination of Gold and other metals" 4.2. Scope - Gold I other metals in crime exhibits" 4.3. Purpose - Sualitati e and quantitati e estimation of Gold" 4.4. Responsibility - Reporting officers and supporting scientif" staff" 4.#. I tro$uctio Gold is a precious and aluable metal" The human beings are fascinated for their ornaments" These ornaments are required their maintenance #hich includes repairing and polishing etc" The criminals often polish the ornaments #ith polishing po#der and by using aqua4regia solution" The gold is not soluble in most of the acids, it is soluble in aqua4regia #hich is a mixture of one part of nitric acid and three parts of hydrochloric acid" Generally, criminals dupe the house#i es in the name of cleaning and polishing their ornament and dissol e the gold from their ornament in aqua4 regia during the process of cleaningC polishing" This aqua4regia solution and gold etc" sei2ed from the scene of crime by the police are referred to forensic laboratory under I%C (ection 76:" These exhibits are to be examined in the laboratory for the analysis of gold" Colour test and modern sophisticated analytical techniquesCmethods, depending upon the nature of samples and examination, are used for the analysis of samples in the laboratory" 7"9 (ample preparation4.%.1. The aqua4regia solution, about 6=: ml from the crime scene should be sent in glass bottle #ith plastic lid or plastic bottle" 4.%.2. &aterial such as polishing po#ders from the crime scene should be pac!ed in polythene bag and sent to the laboratory" 4.8. Met,o$s o0 A *l"sis7";"+ Test for aqua4regia Test the aqua4regia solution for acidic nature, nitrate ions and chloride ions"

4.8.1.1 Test 0or Aci$ic N*ture : *F p2 P*per Test: &oist the p* paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the p*" p* less than ; indicates the presence of acid and more than ; indicates the presence of al!ali" /lternate &ethod *F )it&us P*per Test : &oist the blue litmus paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the colour change of the paper" Colour changing from blue to red indicates the presence of acid" If red litmus paper changes to blue, than it indicates the presence of al!ali" 4.8.1.2 Test for Chlori e1.2 : *F SilHer Nitr*te Test 1,2: Ta!e the appropriate portion of the exhibit in a test tube and add + drop of nitric acid follo#ed by fe# drops of :"6 & sil er nitrate solution" / #hite curdy precipitate is obtained #hich is soluble in excess of ammonium hydroxide solution but insoluble in #ater and dilute nitric acid" !F Test 6it, Sulp,uric Aci$3- Ta!e the appropriate portion of the exhibit in a test tube and add fe# drops of conc" (ulphuric acid if required #arm it" *ydrogen chloride gas is e ol ed, #hich can be tested by blue litmus paper turns to red or by formation of #hite clouds of ammonium chloride #hen a glass rod moistened #ith ammonia solution is brought near the mouth of test tube" 4.8.3 Test for !itric "ci 4.8.3.1 Test 0or Aci$ic N*ture: *F p2 P*per Test: &oist the p* paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the p*" %* less than ; indicates the presence of acid" /lternate &ethod !F )it&us P*per Test: &oist the blue litmus paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the colour change of the paper" Colour changing from blue to red indicates the presence of acid"

4.8.3.2 Test 0or Nitr*te: Depending upon the nature of exhibits and a ailability of resources, any one of the follo#ing methods can be used for the detection of nitrate" Ta!e the appropriate portion of the exhibit in a bea!er add distilled #ater, sha!e #ell and filter it" The filtrate may be used for performing the tests" *F Ri 3 Test3: 0i1 /dd about > ml of a freshly prepared saturated solution of ferrous sulphate to about 6 ml of the nitrate solution 0filtrate of the exhibit1 in a test tube and pour >4= ml conc" sulphuric acid slo#ly do#n the side of the test tube so that acid forms a layer beneath the mixture" / bro#n ring #ill form #here the liquids meet #hich indicates the positi e test for the presence of nitrate" 0ii1 /dd about 7 ml of conc" sulphuric acid slo#ly to about 6 ml of the nitrate solution 0filtrate of the exhibit1 in a test tube and mix the liquids thoroughly and cool the mixture in tap #ater" No# a saturate solution of ferrous sulphate in added slo#ly do#n the inner side#all of the test tube to form a layer on the top of the liquid" 'ormation of a bro#n ring at the ?unction of t#o liquids indicates the presence of nitrate"3 !F Dip,e "l*&i e Re*3e t Test3: To a small amount of diphenylamine reagent 0dissol e :"= gm" Of diphenylamine in 5= ml conc" sulphuric acid and dilute to +:: ml #ith #ater1 in a test tube add a small portion of the filtrate carefully to the side of the test tube, a blue ring at the ?unction indicates the presence of nitrate" cF (errous Sulp,*te Test4 : To a drop of filtrate on a spot plate add a crystal of ferrous sulphate 0pin head1, a drop of conc" sulphuric acid is allo#ed to run in at the side" In the presence of nitrate, a bro#n ring is formed around the ferrous sulphate crystal" $F <ruci e Test4 : To a fe# drop of filtrate, a solution of brucine 0:":6< in sulphuric acid, prepare immediately before use1 is added on a spot plate, in the presence of nitrate a red colour is de eloped, on standing it changes to yello#ish red" 4.8.2 A *l"sis o0 >ol$ Polis,i 3 Po7$er Generally the gold polishing po#der is reddish in color and sho#s the presence of inorganic radicals such as copper, sodium, potassium, ammonium, iron, chloride, sulphate, nitrate etc" the analysis of gold polishing po#der is carried out by the methods used for the analysis of inorganic qualitati e analysis" The gold polishing po#der may also be tested for the presence of gold" .xtract the polishing po#der #ith #ater and the #ater extract is tested for cations and anions"

4.8.2.1 Tests 0or *&&o iu& io s: a1 Nessler Reagent Test 7Ta!e one drop of test sample or extract, add one drop of conc" NaO* solution 0=gms in = ml #ater13 on a #atch glass or in a small test tube" Ta!e out a micro drop from this and !ept on a filter paper and add to it one drop of Nessler reagent" /ppearance of a yello# or orange red stain or ring indicates the positi e test for the presence of ammonium ions" Prep*r*tio o0 Nessler re*3e t 3-

(olution +- Dissol e +: gms of BI in +: ml of #ater" (olution 6- Dissol e 9 gms of mercury 0II1 chloride in +:: ml of #ater" (olution >- Dissol e 7= gms of NaO* in #ater and dilute to 5: ml" No# add solution 6 to solution + drop #ise until a slight permanent ppt" is formed then add solution >, mix and dilute #ith #ater to 6:: ml" Beep it for o ernight and decant the clear solution" The solution may be used for one month" !F ta!e an appropriate amount of the suspected sample , add to it fe# drop of NaO* solution in a test tube and heat it" (mell of ammonia is obser ed" This can be confirmed by bringing a glass rod dipped in *Cl acid on the mouth of the test tube" $hite fumes are produced"% 4.8.2.2 Test 0or C,lori$e1.2 : *F SilHer Nitr*te Test: Ta!e the appropriate portion of the exhibit in a bea!er add distilled #ater, sha!e #ell and filter it" Ta!e fe# ml" of the filtrate in a test tube and add + drop of nitric acid follo#ed by fe# drops of :"6 & sil er nitrate solution" / #hite curdy precipitate is obtained #hich is soluble in excess of ammonium but insoluble in #ater and dilute nitric acid" !F Test 6it, Sulp,uric Aci$: /s mentioned under section 6"="+"6 b" 4.8.2.3 Test 0or Sulp,*te#.$ : *F <*riu& c,lori$e Test: Ta!e the appropriate portion of the exhibit in a bea!er add distilled #ater, sha!e #ell and filter it" Ta!e fe# ml" of the filtrate in a test tube and add fe# drops of dilute hydrochloric acid follo#ed by :"6= & barium chloride solution" $hite precipitate, #hich is insoluble in #ater, indicates the presence of sulphate"

*F R,o$i9o *te Test3: ta!e a drop of barium chloride solution 0:"6=& M 9+"+ g barium chloride dihydrate diluted in + litre of #ater1 on a filter paper and add a drop of fresh solution of sodium rhodi2onate 0=<1" Reddish bro#n colour spot appears" No# add a drop of acid or al!aline test solution" Disappearance of colour spot indicates the positi e test for the presence of (ulphate" 4.8.2.4 Test 0or So$iu& io s: Color Test: =r* "l Oi c Acet*te Test 8: Ta!e a portion of exhibit solution and ma!e it neutrali2ed #ith acetic acid" /dd fe# drops of uranyl 2inc acetate reagent, sha!eC stir #ith glass rod" 'ormation of yello# precipitate or cloudiness indicates positi e test for the presence of sodium" Prep*r*tio o0 ur* "l 9i c *cet*te ;:

Ta!e +: gms of uranyl acetate in ==ml of #ater, >: gms of 2inc acetate, and , ml of acetic acid" *eat to dissol e and dilute #ith #ater to ma!e up to +:: ml" /llo# to stand for 67 hours, and filter" Alter *te &et,o$ 0or t,e prep*r*tio C: Solutio A: necessary o0 ur* "l 9i c *cet*te re*3e t

Ta!e +: grams uranyl acetate in 9 gms of >:< acetic acid" If #arm it, dilute #ith distilled #ater to =:ml "

Solutio <: >: grams 2inc acetate is stirred #ith >gms >:< acetic acid and dilute it #ith distilled #ater to =: ml" &ix the abo e t#o solutions / and @" $arm if required" /dd a trace of sodium chloride, !eep it for 67 hours and filtered" 'iltrate is used as abo e reagent" Alter *te Met,o$s 0or So$iu& (l*&e test 1.: Ta!e appropriate portion of the exhibit as such or its #ater 0distilled1 extract e aporate to dryness, moisten #ith a fe# drops of conc" *ydrochloric acid to ma!e past" Ta!e a small portion of paste #ith the platinum #ire and introduce into the non4luminous flame of a semi4micro burner" / persistence golden yello# flame indicates the presence of sodium"

Alter *te Met,o$ o0 (l*&e Test11 Ta!e a platinum or nichrom #ire and #et it #ith conc" *ydrochloric acid and heat it in the non4luminous flame of the burner until the yello# colour of the flame disappears" Dip the #ire into test exhibit solution as such or its distilled #ater extract 0or po#der if exhibit is solid1 and heat it in the non4 luminous flame of the burner" Obser e the color of the flame as abo e" / persistence golden yello# flame indicates the presence of sodium" 4.8.2.# Test 0or Pot*ssiu& io s: iF Dipicr"l*&i e Re*3e t Test 4:

%repare a drop reaction paper by soa!ing the filter paper in sodium dipicrylaminate reagent 0:"6 g dipicrylamine in 6 ml of 6N sodium carbonate and += ml #ater1 and dried in a blast of heated air" %lace a drop of neutral test solution on the drop reaction paper, dry it in a current of hot air" Beep the paper in :"+N nitric acid" 'ormation of red flec! or ring at site of the spot indicates the positi e test for the presence of %otassium" iiF (l*&e Test: %otassium gi es iolet color #hen it is tested by flame test as described in the testing of sodium" 4.8.2.% Test 0or Copper: iF Test 7it, *&&o iu& ,"$roAi$e Ta!e the test sample solution 0dissol e the sample in #ater or dil *cl or conc" *Cl or dilCconc" *NO> or aqua4regia to form original solution1 and add to it ammonium hydroxide" /ppearance of blue color indicates the positi e test for the presence of Copper" iiF Test 7it, Ali9eri !lue 4:

Ta!e a drop of reagent 0saturated solution of ali2erin blue in pyridine1 in a depression spot plate and add to it a drop of test solution" Run a parallel blan! test #ith #ater also" @lue color appears in both the cases" No# add +46 drops of acetic anhydride, the color in the blan! test con erts into yello# but remaining of blue4 iolet precipitate in test sample indicates the positi e test for the presence of Copper" Alter *te Met,o$:

Ta!e +46 drops of test solution in depression spot plate and e aporate it to dryness" /dd a drop of the reagent and follo#ed by a drop of glacial acetic acid" /ppearance of blue4 iolet color indicates the positi e test for the presence of Copper" iiiF (l*&e Test: Copper gi es bluish green color #hen it is tested by flame test as described in the testing of sodium" 4.8.2.8 Test 0or Iro iF Test 7it, Pot*ssiu& 0erroc"* i$e: Ta!e fe# drops of test solution or original solution, add to it a drop of conc" Nitric acid and boil for fe# minutes" /dd %otassium ferrocyanide solution" /ppearance of blue color indicates the positi e test for the presence of Iron" iiF Test 7it, t,ioc"* *te %: Ta!e fe# drops of test solution or original solution, add to it a drop of conc" Nitric acid and boil for fe# minutes" /dd ammonium or potassium thiocyanate solution" /ppearance of red color indicates the positi e test for the presence of Iron" The abo e tests can be performed on filter paper or spot tile as belo# 4 %ut a drop of test solution on a filter paper or spot plate, add to it a drop of %otassium ferrocyanide solution" /ppearance of blue color on paper or plate indicates the positi e test for the presence of Iron" %ut a drop of test solution on a spot plate, add to it a drop of potassium thiocyanate solution 0+<1" /ppearance of red color indicates the positi e test for the presence of Iron" Ot,er *lter *te &et,o$s: I1 IC%-/.( &ethod ii1 Ion Chromatogaphy iii1 /tomic /bsorption (pectrophotometry 0//(1 i 1 /tomic .mission (pectrography 1 'luroscence4KRD etc" These techniques C methods can also be used for the detection of abo e cations and anions depending upon the nature of the exhibits 4.8.3. TEST (OR >O)D:

4.8.3.1. Test %ith &en'i ine( : 'e# drops of the exhibit solution is placed on a filter paper and add +46 drops of ben2idine reagent solution on it" / blue colour appears on the filter paper, #hich indicates the presence of gold" Re*3e t: :":=< solution of ben2idine in +:< acetic acid 4.8.3.2. Test %ith Rho amine &( : 'e# drops of the exhibit solution is placed is ta!en in a micro test tube and add one4t#o drops of *Cl acid and +4 6 drops of rhodamine @ reagent solution and mix properly" The test tube is sha!en #ith about 54+: drops of ben2ene" /ppearance of red4 iolet to pin! colour in ben2ene layer indicates the presence gold" /fter about +46 minutes it displays an orange fluorescence if it is obser ed under quart2 lamp" Re*3e t: +1 :":+ g" rhodamine @ dye stuff in +:: ml #ater 61 :"6< aq" rhodamine @ solution 0gi en under detection of gold in alloys, coating etc"1 the sensiti e and specific test for gold #ith rhodamine @" 4.8.3.3. Test %ith )*alic "ci $ : /n appropriate portion of the exhibit generally heated till NO6 fumes remo ed" This solution is made al!aline by adding sodium hydroxide solution" (olid oxalic acid #as added and generally heated" @ro#nish blac! precipitate indicates the presence of gold" 4.8.3.4 T,i l*"er c,ro&*to3r*p," 0or 3ol$ 13:

The gold can be detected by T)C technique #hich in ol es the gold in 0*/uCl7 >*6O1 form free from nitrate ions is spotted on T)C plate along #ith standard gold soln and then the T)C plate is run in sol ent system of @en2ene- @utanol- /cetic acid 0>"=- 7"=- +1" The plate is then sprayed #ith =< $CA of (odium carbonate soln" @luish blac! coloured spots of gold oxide 0/u6O>1 similar to gold standard sample are de eloped for gold ions" This semi quantitati e technique is sensiti e and the detection limit is +mgCml of gold in the solution" 4.8.3.#.4u* tit*tiHe esti&*tio o0 3ol$ i *Eu*-re3i* 14:

Treat 6: ml of aqua4regia soln #ith dil *Cl and heat the resulting soln" on #ire gau2e" &a!e the soln" nitrate free and test for nitrate ion by adding a drop on spot tile, follo#ed by conc"* 6 (O7 soln" and then a pinch of brucine po#der" Nitrate ion gi es orange colour" Continue the addition of dil *Cl and heating #ith bunsen burner on #ire gau2e till the soln is free from nitrate ions" Reduce the nitrate free ion soln #ith =< hydroquinone soln in #ater and #arm on #ire gau2e" 'ilter the resulting soln" and the gold in precipitated form is #ashed #ith #arm #ater till it is chloride free, and then ignites in silica crucible in furnace at 5=:DC for about 6 hrs" and then #eigh the gold" This method estimates about :"+ gm< 0#C 1 of gold in aqua4regia" *o#e er for the determination of traces of gold in aqua4regia soln" is done

by modern analytical techniques such as IC% 0Inducti ely Coupled %lasma technique1" Ot,er *lter *te &et,o$s: 4.8.1. +CP,"-S .etho 1$ : This methodCtechnique is used for the qualitati e and quantitati e analysis of gold in arious types of crime exhibits" It in ol es the sample preparation follo#ed by IC%4/.( analysis" /ppropriate amount of solidCpo#der exhibit #as dissol ed in aqua4regia 0> parts *ydrochloric acid and +part of Nitric acid1" Dilute it in appropriate olume #ith distilled #ater and analysed by IC%4/.(" Run a parallel blan! sample along #ith the crime exhibit" ii1 Ion Chromatogaphy iii1 /tomic /bsorption (pectrophotometry 0//(1 i 1 /tomic .mission (pectrography 1 'luroscence4KRD etc" Re0ere ces : +" /"I" Aogel, T/ text boo! of macro and semi4micro quantitati e inorganic analysisP, +,9,, p">9>" 6" G"*" 8effery, TAogelOs text boo! of quantitati e chemical analysisP = th edition, +,5,, p"75:" >" G" ( ehla, TAogelOs text boo! of macro and semi4micro qualitati e inorganic analysisP, =th edition, +,;,, p">>74>>=, >>9, >67, >>9, =5=, =;,, >794>7;, >75 7" 'rit2 feigl, T(pot tests in inorganic analysisP, = th edition, +,=,, p">69, >654 >6,, +65, +6,, 6>5, 6>6, ,>, +9+, +97," =" Gurudeep I *arish, %ractical Inorganic Chemistry, 9 th edition, +,5545,, pp"+=>" 9" (ubhash4(atish, / hand boo! of analytical chemistry, +> th edition, +,;54 ;,, pp ,6 ;" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"9>497 5" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"=>;

," (pot test in inorganic analysis by 'rit2 'eigl, fifth edition, .lse ier %ublishing Company, Netherlands, +,=5, pp" 6>:" +:" AogelOs text boo! of macro and semi4micro qualitati e inorganic analysis by G" ( ehla, =th edition, )ongman Group )imited, Great @ritain, +,;,, pp" 79=499" ++" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"9=" +6" &"Thompson and 8"N"$alsh, / hand boo! of Inducti ely Coupled %lasma (pectrometry, +,5> +>" R"@" %udale et" al" T)C method for the detection and (emiquantitati e determination of gold in forensic #or!, %roceeding of K th /ll India 'orensic (cience conference, @hubanes#ar, +,,;" +7" / textboo! of Suantitati e Inorganic /nalysis by Aogel" 6nd edition , +,=+" +=" &"Thompson and 8"N"$alsh, / hand boo! of Inducti ely Coupled %lasma (pectrometry, +,5>

SECTION - # ACIDS + A)-A)IS #.1. Title - .xamination of corrosi e chemicals li!e hydrochloric acid, sulphuric acid, and nitric and al!alies in crime exhibits of acidCal!ali thro#ing cases" #.2. Scope - Crime exhibits containing corrosi e chemicals" #.3. Purpose - To detect the presence of hydrochloric acid, sulphuric acid, nitric acid and al!alies in crime exhibits" #.4. Responsibility - Reporting officers and supporting scientific staff" #.#. .etho s - The follo#ing methods are to be used for the detection of the constituents of the hydrochloric acid, sulphuric acid and nitric acid in the crime exhibits"

#.#.1 Test for /y rochloric "ci #.#.1.1 Test for "ci ic !ature : *F p/ Paper Test : &oist the p* paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the p*" p* less than ; indicates the presence of acid and more than ; indicates the presence of al!ali" /lternate &ethod !F 0itmus Paper Test : &oist the blue litmus paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the colour change of the paper" Colour changing from blue to red indicates the presence of acid" If red litmus paper changes to blue, than it is indicates the presence of al!ali"

#.#.1.2 Test for Chlori e1 : Ta!e the appropriate portion of the exhibit in a bea!er add distilled #ater, sha!e #ell and filter it" Ta!e fe# ml" of the filtrate in a test tube and add + drop of nitric acid follo#ed by fe# drops of :"6 & sil er nitrate solution" / #hite curdy precipitate is obtained #hich is soluble in excess of ammonium hydroxide solution" #.#.2 Test for Sulphuric "ci #.#.2.1 Test for "ci ic !ature: *F p/ Paper Test : &oist the p* paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the p*" %* less than ; indicates the presence of acid" /lternate &ethod !F 0itmus Paper Test : &oist the blue litmus paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the colour change of the paper" Colour changing from blue to red indicates the presence of acid" #.#.2.2 Test for Sulphate1 : Ta!e the appropriate portion of the exhibit in a bea!er add distilled #ater, sha!e #ell and filter it" Ta!e fe# ml" of the filtrate in a test tube and add fe# drops of concentrated hydrochloric acid follo#ed by:"6= & barium chloride solution" $hite precipitate indicates the presence of sulphate" #.#.3 Test for !itric "ci

#.#.3.1 Test for "ci ic !ature : !F p/ Paper Test : &oist the p* paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the p*" %* less than ; indicates the presence of acid" /lternate &ethod !F 0itmus Paper Test : &oist the blue litmus paper #ith distilled #ater and impregnate it #ith exhibit as such or its distilled #ater extract and obser e the colour change of the paper" Colour changing from blue to red indicates the presence of acid" #.#.3.2 Test for !itrate : Depending upon the nature of exhibits and a ailability of resources, any one of the follo#ing methods can be used for the detection of nitrate" Ta!e the appropriate portion of the exhibit in a bea!er, add distilled #ater, sha!e #ell and filter it" The filtrate may be used for performing the tests" a1 Ring Test2: 0i1 /dd about > ml of a freshly prepared saturated solution of ferrous sulphate to about 6 ml of the nitrate solution 0filtrate of the exhibit1 in a test tube and pour >4= ml conc" sulphuric acid slo#ly do#n the side of the test tube so that acid forms a layer beneath the mixture" / bro#n ring #ill form #here the liquids meet #hich indicates the positi e test for the presence of nitrate" 0ii1 /dd about 7 ml of conc" sulphuric acid slo#ly to about 6 ml of the nitrate solution 0filtrate of the exhibit1 in a test tube and mix the liquids thoroughly and cool the mixture in tap #ater" No# a saturate solution of ferrous sulphate in added slo#ly do#n the inner side#all of the test tube to form a layer on the top of the liquid" 'ormation of a bro#n ring at the ?unction of t#o liquids indicates the presence of nitrate" cF 2iphenylamine Reagent Test2 : To a small amount of diphenylamine reagent 0dissol e :"= gm" Of diphenylamine in 5= ml conc" sulphuric acid and dilute to +:: ml #ith #ater1 in a test tube add a small portion of the filtrate carefully to the side of the test tube, a blue ring at the ?unction indicates the presence of nitrate" c1 3errous Sulphate Test# : To a drop of filtrate on a spot plate add a crystal of ferrous sulphate 0pin head1, a drop of conc" sulphuric acid is allo#ed to run in at the side" In the presence of nitrate, a bro#n ring is formed around the ferrous sulphate crystal"

$F

&rucine Test# : To a fe# drop of filtrate, a solution of brucine 0:":6< in sulphuric acid, prepare immediately before use1 is added on a spot plate, in the presence of nitrate a red colour is de eloped, on standing it changes to yello#ish red"

References : +" G"*" 8effery et al, TAogelOs text boo! of quantitati e inorganic analysisP 7th edition, +,59, p"7>>, =:7" 2. G" ( ehla, TAogelOs text boo! of macro and semi4micro qualitati e inorganic analysisP, =th edition, +,;,, p">>74>>=, >794 >7;, >>9" 3. 'rit2 feigl, T(pot tests in inorganic analysisP, = th edition, +,=,, p">69, >654>6,"

SECTION - % E@AMINATION O( C2EMICA)S =SED IN TRAP CASES %.1. Title - .xamination of chemicals used in Trap Cases" %.2. Scope: %henolphthalein, anthracene, sodium carbonate and calcium hydroxide in crime exhibits"

%.3. Purpose- Detection and identification of phenolphthalein, sodium ions, carbonate ions, calcium ions, anthracene etc" %.4. Respo si!ilit"- Reporting officers and supporting scientific staff" %.#. I tro$uctio )a# .nforcement /gencies arrange a trap for illegal transaction 0bribe1 #ith the help of the complainant #here in the accused is gi en currency notes on #hich chemical li!e phenolphthalein or anthracene po#der has been applied" No# a days phenolphthalein is being used in most of the trap cases" If the accused touches the notes then the part of the chemical, #hich may be in traces, is transferred on his hands or fingers" If he !eeps these currency notes in his poc!et, bag, briefcase or file etc, this chemical is also transferred to these ob?ects 0)ocardOs %rinciple of exchange1" In case of phenolphthalein the ob?ect 0hand , bag, poc!et etc1 are #ashed #ith a colorless solution of sodium carbonate 0or sometimes #ith lime #ater1, #hich becomes immediately pin! confirming the touching of currency notesC transferred of phenolphthalein to the ob?ect" These #ashings are being collected and sent to the forensic laboratories along #ith other rele ant articles to establish the presence of phenolphthalein #hich can be considered as ital e idence in the court" The pin! color of this solution persists for some days or months depending on the quantity of the phenolphthalein and strength of the al!ali solution" It gradually fades and sometimes becomes colorless at the time of trial in the court" This creates unnecessary doubt and in estigating officer is put in an a#!#ard position" *o#e er this phenomenon can be explained by the scientist on scientific basis that the color of the phenolphthalein fades due to its brea!ing do#n into 607 Mhydroxy ben2oyl1 ben2oic acid and phenol in al!ali medium" /s mentioned earlier, anthracene po#der is also rarely used for this purpose in trap cases as it does not pose such problem of color fading and has an ad antage because of its fluorescence property" The hands, clothes etc of the suspect can be immediately examined under u" " light, ioletCblue fluorescence1+2 can be clearly seen" This pro es direct contact of the suspect #ith currency notes" %ure anthracene exhibits blue fluorescence but impure anthracene due to presence of tetracene, naphthacene etc exhibits yello# #ith green fluorescence"

%.%. Met,o$s 9"9"+ " %henolphthalein-

The hand #ashings, bag #ashings, cloth #ashings etc of the suspect collected in dilute sodium carbonate4#ater solution or lime #ater along #ith other rele ant articles from the scene of crime such as currency notes, clothes, bags etc shall be sent to the laboratory for the chemical examination" %.%.1.1. In case of untreated ob?ects, ethyl alcoholic #ashC extract of the appropriate portion of the exhibits can be ta!en for the examination for the detection of the phenolphthalein" /lternati ely, dilute solution of al!ali 0sodium carbonate1 in #ater can also be used for #ashingCextracting the exhibits" These #ashing shall be used only for the detection of the phenolphthalein and not for the detection of the sodium and carbonate ions" %.%.1.2. In case of al!ali treated ob?ects, #ash the appropriate portion of the exhibits #ith #ater and used for the detection of the phenolphthalein, sodium and carbonate ions etc" %.%.2. A t,r*ce e: /rticles from the scene of crime such as currency notes, clothes, bags etc along #ith traces of po#der collected by carefully brushing the suspected area of contact of accused shall be sent to the laboratory for the examination" %.%.2.1. In case of the ob?ect of anthracene, the appropriate portion of the ob?ectC exhibit 0after examination under u" " light1 can be #ash #ith ethyl alcohol for the examination" %.8. Det*ils o0 t,e Met,o$s o0 * *l"sis : The follo#ing methods can be used for the examination of the trap case for the detection of required constituents depending upon the case historyC nature of the examination of the case" %.8.1. Test 0or P,e olp,t,*lei %.8.1.1. Color Tests : %.8.1.1.1. p2 Test2,3 : Obser e the p* of the solution exhibit #ith the p* paper" &ore than p* , 0p* range 5">4+:1 #ith pin! Cred color indicates the positi e test for the presence of phenolphthalein" %.8.1.1.2. Aci$ :Al5*li Test3, 4: :

Ta!e an appropriate portion of the exhibit solution" /dd fe# drops dilute hydrochloric acid" The pin! color of the exhibit disappears" No# add fe# drops of dilute solution of sodium hydroxide in #ater, the pin! color reappears" If required, this test can also be performed on residue obtained after e aporation of ethanol extract of the exhibit, but in this case first add al!ali solution and then acid" /ppearing and disappearing of pin! color indicates the positi e test for the presence of phenolphthalein" %.8.1.2. EAtr*ctio
#46

Ta!e appropriate amount of the exhibit solution" &a!e the solution acidic #ith the addition of dilute hydrochloric acid drop #ise #ith stirring till p* about 7 to = 0alternati ely till acidic to congo red1" .xtract #ith 6:46= ml" of sol ent ether t#o times and e aporate to concentrate and used for other tests" The ether extract can also be e aporated to dryness and the residue can be dissol ed in ethanol, #hich can be used for other tests" %.8.1.3. Thin 0ayer Chromatography: (tationary phase - T)C plate coated #ith silica gel G or silica gel G '6=7" &obile phase /ny one of the follo#ings-4

+" @en2ene, Dioxan and acetic acid 0;=-+=-:"=1% 6" Chloroform and /cetone 0 5:-6:18 >" .thyl acetate, &ethanol and (trong /mmonia 06; to >:<#C#18 05"=-+":-:"=1 Aisuali2ation +" (pray #ith dilute sodium hydroxide solution 6" /cidified potassium permanganate solution 8 0+< soln" of potassium permanganate in :"6= & sulphuric acid1 >" U"A" light" %.8.1 I stru&e t*l tec, iEues %.8.1.4. Spectrop,oto&etric eA*&i *tio
#

Ta!e a portion of the exhibit solution, filtered and scan to note its l max absorbance alue by spectrophotometer in appropriate dilution using a standard solution of phenolphthalein in aqueous al!ali 0sodium carbonate1 for comparison" The pin! color of phenolphthalein in aq" (odium carbonate solution gi es the lambda max in bet#een around ==:4=== nm. /queous solution of sodium carbonate is used as blan! solution for the experiment" Other instrumental techniques li!e *%)C, 'TIR, GC4&( are also being used for the detection of %henolphthalein" %.8.1.#. Alter *te Test 0or P,e olp,t,*lei : 3olin,Cioc*lteuPs re*3e t test#: Ta!e about + ml of alcoholic extract of the exhibit obtained as per required extraction procedure" /dd I ml of folin Mciocalteu reagent follo#ed by 6 ml of 6:< sodium carbonate solution" @lue color indicates the positi e test for the presence of phenolphthalein" %.8.2. Test 0or A t,r*ce e : %.8.2.1.Color Test : Obser ation under u" " light M AioletCblueCgreen fluorescence" %.8.2.2.T,i )*"er C,ro&*to3r*p," ;:

(ample preparation - In ethyl alcohol (tationary phase &obile phase - (ilica gel G - 0/ny t#o1

+" *eptane 6" *exane >" Carbon tetrachloride Aisuali2ation - +" U"A" light 6" 'ormaldehyde4(ulphuric acid reagent 0:"6 ml of >;< formaldehyde solution in +: ml of conc" (ulphuric acid1 A)TERNATE MET2ODS %.8.2. Sep*r*tio &*teri*lC: * $ puri0ic*tio o0 A t,r*ce e 0ro& sei9e$

Currency notes, shirts, pant, hand!erchiefs, diaries, boo!s etc in anti4 corruption casesCbribe trap cases collected o er a year #ere examined a long #ith commercial anthracene by T)C technique using Chloroform de eloping sol ent" The suspected portion after locating under UA lamp of e ery exhibit #as initially extracted in ethanolCether, then sub?ected to T)CCG)C examination" %.8.2.1. T/+! 0"5-R C/R)."T)GR"P/56: (ample preparation - In ethyl alcohol 3,=< C U (tationary phase &obile phase Aisuali2ation - (ilica gel G 3/cti ated at ++:: C for + hourU - Chloroform - +" U"A" light

%.8.2.2 = D Spectrophotometry6: The separated spot of the analyte corresponding to reference spot of anthracene #as scrapped off from the preparati e T)C" To the scrapped silica gel appropriate amount of ethanol #as added" 'iltered or decanted it" Concentrate the filtrate to appropriate olume" @lan! #as similarly prepared from the silica gel scrapped off from the same plate" The filtrate #as sub?ected to U A spectroscopic study" /lternati ely the exhibit cab also be extracted #ith ,=< of ethanol and sub?ected to U A (pectroscopy study" / control sample of anthracene in ethanol cab be used as a standard sample for comparison of U A (pectrum" %.8.2.3 >*s )iEui$ C,ro&*to3r*p," C: Detector Column %ac!ing Carrier Gas 'lo# rate In?ector temperature Column temperature Detector temperature - 'lame ioni2ation - ( ( column +C5 inch dia" 6 meter length - +:< ( . >: 5:C+:: chromosorb $4*% - Nitrogen - 6= mlCmin - 6=::C - 66::C Isothermal - 6=::C

(ample preparation - In ethanol %.8.3. Test for Carbonate ions : %.8.3.1. Color Tests : %.8.3.1.1. Test %ith aci
17

To a potion of the exhibit solution add fe# drops of dilute hydrochloric acid, effer escence are obser ed" If needed, the resulting gas can be passed through baryata #aterC lime #ater" TurbidityC curdy #hite precipitate appears" If the gas is passed for long time, the precipitate or turbidity slo#ly disappears" The exhibit may be gently heated #ith dilute hydrochloric acid to produce sufficient effer escenceC gas" %.8.3.1.1. &arium Chlori e Test
1.

To a portion of the exhibit solution add fe# drops of barium chloride solution 0about =49 < barium chloride in #ater" 'ormation of #hite precipitate, #hich is soluble in mineral acids, indicates the positi e test for the presence of carbonate ions" @icarbonate ions do not form #hite precipitate, as they do not react #ith barium chloride solution" "0T-R!"T- .-T/)2S: %.8.3.1.1 Sil8er !itrate Test17: To a portion of the exhibit solution add fe# drops of sil er nitrate solution 0about 6 < in #ater1" 'ormation of #hite precipitate, #hich is soluble in ammonia solution, indicates the positi e test for the presence of carbonate ions" %.8.3.1.2 .agnesium Sulphate Test17: Ta!e appropriate portion of the exhibit as such or its #ater 0distilled1 extract" /dd magnesium sulphate in the cold condition" If no precipitate is obtained that it indicates the presence of bicarbonate" If a #hite precipitate is formed then it is a positi e test for the presence of carbonate" %.8.4 Test for So ium:

%.8.4.1 Color Test: %.8.4.1.1 9ranyl :inc "cetate Test11:

Ta!e a portion of exhibit solution and ma!e it neutrali2ed #ith acetic acid" /dd fe# drops of uranyl 2inc acetate reagent, sha!eC stir #ith glass rod" 'ormation of yello# precipitate or cloudiness indicates positi e test for the presence of sodium" Preparation of uranyl 'inc acetate12: Ta!e +: gms of uranyl acetate in ==ml of #ater, >: gms of 2inc acetate, and , ml of acetic acid" *eat to dissol e and dilute #ith #ater to ma!e up to +:: ml" /llo# to stand for 67 hours, and filter" "0T-R!"T- .-T/)2 3)R T/- PR-P"R"T+)! )3 9R"!50 :+!C "C-T"T- R-"G-!T 13: Solutio A:

Ta!e +: grams uranyl acetate in 9 gms of >:< acetic acid" If necessary #arm it, dilute #ith distilled #ater to =:ml " Solutio <: >: grams 2inc acetate is stirred #ith >gms >:< acetic acid and dilute it #ith distilled #ater to =: ml" &ix the abo e t#o solutions / and @" $arm if required" /dd a trace of sodium chloride, !eep it for 67 hours and filtered" 'iltrate is used as abo e reagent" %.8.#. Test for Calcium Test 7it, So$iu& R,o$i9o *te14: Ta!e one drop of neutral or #ea!ly acid test solution add a drop of freshly prepared :"6< a sodium rhodi2onate solution add one drop of :"= N sodium hydroxide solution, a iolet colour indicates the presence of calcium" Alter *te Met,o$s 0or So$iu& + C*lciu&: 3lame test1$: Ta!e appropriate portion of the exhibit as such or its #ater 0distilled1 extract e aporate to dryness, moisten #ith a fe# drops of conc" *ydrochloric acid to ma!e past" Ta!e a small portion of paste #ith the platinum #ire and introduce into the non4luminous flame of a semi4micro burner" / persistence golden yello# flame indicates the presence of sodium and a bric! red 0yello#ish red1 flame indicates the presence of calcium" "lternate .etho of 3lame Test16

Ta!e a platinum or nichrom #ire and #et it #ith conc" *ydrochloric acid and heat it in the non4luminous flame of the burner until the yello# colour of the flame disappears" Dip the #ire into test exhibit solution as such or its distilled #ater extract 0or po#der if exhibit is solid1 and heat it in the non4 luminous flame of the burner" Obser e the colour of the flame as abo e" R-3-R-!C-S: +" Organic Chemistry Aolume + The 'undamental %rinciples by I")" 'innar, 9th edition, The .nglish )anguage @oo! (ociety and )ongman Group )imited, )ondon, +,;>, pp" 5+7" 6" The &erc! Index ++th edition by (usan @uda ari, %ublished by &erc! I Co" Inc", Rah#ay, U(/, +,5,, pp"+:5 0/cc" No">7:71" >" AogelOs Text @oo! of Suantitati e Inorganic /nalysis, 7th edition, by 8" @assett, R"C" Denney, G"*" 8effery, 8" &endham, .nglish )anguage @oo! (ociety C )ongman, Great @ritain, +,59, pp" 67+ 7" The &erc! Index ++th edition by (usan @uda ari, %ublished by &erc! I Co" Inc", Rah#ay, U(/, +,5,, pp"++=: 0/cc" No">7:71 =" Isolation and identification of drugs by ."G"C" Clar!e, ol"+, The pharmaceutical %ress, )ondon, +,;7, pp"755 0/cc" No"+>6=1" 9" @rea! do#n products of phenolphthalein in al!ali media by B" Narayans#amy, B"(" Chha ra, @" &andal I *")" @ami, 'orensic (cience 08ournal1, +,;5, Aol"+; No"6, pp",64,7" 7" Clar!eOs Isolation and Identification of Drugs by /"C" &offat, second edition, The pharmaceutical %ress, )ondon, +,59, pp" 55= I +95 0/cc" No">,5+1" 5" Thin )ayer Chromatography by .gon (tahl, second edition, George /llen I Un#in )td", )ondon, +,9,, pp"995499, 0/cc" No"=,51" ," 'orensic .xamination of /nthracene Traces on (ei2ures in /nticorruption Offence by T)C, UA and G)C Techniques by / G @hoi and B / /mbade, 8ournal of Indian /cademy of 'orensic (ciences 0+,,+1 %age no" +, to 69" +:" AogelOs text boo! of macro and semi4micro qualitati e inorganic analysis by G" ( ehla, =th edition, )ongman Group )imited, Great @ritain, +,;,, pp" 6,54>:+ 0/cc"No6>=;1" ++" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"9>497

+6" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"=>; +>" (pot test in inorganic analysis by 'rit2 'eigl, fifth edition, .lse ier %ublishing Company, Netherlands, +,=5, pp" 6>: 0/cc"No">>1" +7" (pot test in inorganic analysis by 'rit2 'eigl, fifth edition, .lse ier %ublishing Company, Netherlands, +,=5, pp" 666466> 0/cc" No" >>1" +=" AogelOs text boo! of macro and semi4micro qualitati e inorganic analysis by G" ( ehla, =th edition, )ongman Group )imited, Great @ritain, +,;,, pp" 79=499" +9" /nalytical Chemistry by (" (hapiro I La" Gur ich, &IR %ublishers, &osco#, +,;6, pp"9="

Sectio

-8

/N/)L(I( O' /)CO*O) IN )ISUOR(CDRINB( 8.1. Title /nalysis of alcohol in liquorsCdrin!s" arious types of alcoholic drin!sCliquor in crime

8.2. Scope - /nalysis of exhibits"

8.3. Purpose - Sualitati e and quantitati e estimation of ethyl alcohol in arious alcoholic preparations" 8.4. Responsibility - Reporting officers and supporting scientific staff" 8.#. I tro$uctio )iquor is normally !no#n as a mixture of #ater and alcohol" The term alcohol is often used for ethyl alcohol" The liquor is manufactured by the fermentation process in #hich carbohydrates are fermented in presence of en2ymes as per their specifications gi en in @ureau of Indian (tandards 0@I(1" Country made liquor is alcoholic product usually prepared from fermentation of carbohydrates present in cereals, ?aggery, fruits, mahua, palm, molasses etc" The liquors are sold in the mar!et in arious brands and co ered under .xcise /ct" The possession, sale, transportation of liquor is allo#ed only as per the Rules and Regulations of .xcise and %rohibition" &any times these liquors are being smuggled from one (tate to another (tate, illegal possession, transported #ithout proper alid documents" These samples are sei2ed by the %olice and submitted to the 'orensic )aboratory for their examination" The liquor is examined in the laboratory for t#o purposesN firstly, for .xcise purpose #here, mainly the presence of alcohol plays an important role and accordingly the examination of liquor samples for the qualitati e and quantitati e analysis is the main purpose of the in estigation" (econdly, the liquor is examined for quality controlCduplicate samples, #hich are being sold in the mar!et in #hich the examination is carried out for other parameters also apart from alcohol contents" The alcohol contents are also reported in percentage of proof spirit, percentage of alcohol 0#eight by olume1 and percentage of alcohol 0 olume by olume1" 8.%. .etho s : 8.%.1 ;ualitati8e "nalysis of 0i<uor : 8.%.1.1 Test for -thyl "lcohol : The follo#ing tests are to be carried out for the detection of ethyl alcohol in the exhibits"

G*F +o oform Test1 - Ta!e about + ml or appropriate of sample 0distilled or as such depending upon the nature of sample and concentration of ethanol1 and add about + ml of =< sodium hydroxide solution and then add iodine solution 06: gm %otassium iodide Q +: gm Iodine in +:: ml #ater1 drop4#ise #ith sha!ing until the liquid becomes persistent dar! bro#n in colour" Beep it for 64> minutes" If the iodine colour disappears add more drops of iodine solution until persistent bro#n colour of iodine" /dd fe# drops of dilute sodium hydroxide solution to remo e extra iodine" /dd equal olume of #ater, !eep it for ten minutes" Lello# crystalline precipitate indicates the positi e test for the presence of ethanol" G!F 2ichromate Test2 : To about + ml or appropriate amount of sample 0distilled or as such depending upon the nature of samples and concentration of ethanol1 is added about :"6 ml of 6< potassium dichromate solution follo#ed by about + ml of concentration sulphuric acid" The yello# colour of the dichromate changes to green or blue indicates the presence of ethanol" 8.%.1.2 Test for .ethanol# : *F C,ro&otropic Aci$ Test : Ta!e about + ml or appropriate amount of sample 0distilled or as such depending upon the nature of sample and concentration of methanol1 in a test tube add about 6 ml of potassium permanganate solution 0> gm potassium permanganate and += ml of phosphoricCortho phosphoric acid in +:: ml distilled #ater1 and sha!e #ell" No# add fe# crystals of sodium bisulphate #ith sha!ing till disappearance of colour 0potassium permanganate colour1 of the solution" /dd about + ml of chromotropic acid 0=< of aqueous solution of sodium salt of chromotropic acid1 and add concentrate sulphuric acid slo#ly #ith inner side#all of the test tube to the extent of += ml" /ppearance of iolet colour indicates the presence of methanol" !F Sc,i00Ps Re*3e t Test12' : Ta!e about 7"= ml of sample 0distilled or as such depending upon the nature of sample1 in a test tube and add :"= ml of ethanol 0if the concentration of ethanol is high in the sample, the sample is fortified accordingly so that = ml olume should contain only :"= ml ethanol" /dd 6 ml of >< %otassium %ermanganate solution and "6ml of phosphoric acid" Beep it for +: minutes" /dd + ml of +:< oxalic acid follo#ed by +ml of concentrated sulphuric acid" The contents are cold at room temperature" No# add = ml of schiffOs reagent, !eep it for half an hour and obser e the colour" /ppearance of purple colour indicates positi e test for the presence of methanol" The parallel experiments may also been carried out #ith control sample containing :"= ml solution 0:"=< methanol in rectified spiritCethanol1 mixed #ith 7"= ml of #ater and a blan! sample ha ing = ml #ater" The colour appeared in the test sample may be matched #ith the colour of the controlCstandard sample of methanol #hich is eEuiH*le t to 2 &3 o0 &et,* ol " Thus the semi4quantitati e examination of methanol may be carried out"

8.%.1.3 Test for Copper = +ron( : Ta!e about = ml or appropriate amount of sample add + drop of Nitric acid and + ml of :":6= & potassium 'errocyanide solution" %russian blue colour indicates presence of iron and chocolate colour indicates the presence of copper" 8.%.1.4 Test for 3urfural# : Ta!e about = ml or appropriate amount of sample 0distilled or as such depending upon the nature of sample and concentration of furfural1 in a test tube, add about + ml aniline and about :"= ml hydrochloric acid and !eep it for += minutes" /ppearance of red colour indicates the presence of furfural" "lternati8e metho for testing furfural :

*F Ta!e about 6 ml or appropriate amount of the sample distilled or as such depending upon the nature of sample in a test tube, add about :"6 ml of aniline and about :"7 ml of glacial acetic acid" If the furfural is present in the sample, red colour de elops in a fe# seconds I reaches its maximum intensity in =4+: minutes#" !F Ta!e about 6 ml or appropriate amount of the sample distilled or as such depending upon the nature of sample in a test tube, add about + ml of aniline acetate solution 0+:< ACA solution of aniline in glacial acetic acid1" /ppearance of red colour indicates the presence of furfural" The colour de elops at room temperature of 6=4>: :C and reaches its maximum intensity in +4= minutes%" 8.%.2. ;uantitati8e "nalysis of 0i<uor : 8.%.2.1.2etermination of -thanol : Depending upon the nature of sample and examination required and resources a ailable any one of the follo#ing methods can be used for the quantitati e analysis of ethyl alcohol" Determine the specific gra ity of the distilled sample by specific gra ity bottle and apply the bottle correction and temperature correction for 9: :' as per the table sho#ing #eight in grains of the spirit of temperatures 9+ : to +::: gi en under the instructions for ascertaining the real alcoholic strength of drugs chemicals, medicines dietetics and toilet preparations entered for test prepared by R")" 8en!s, '"I"C", .xaminer for Customs and .xcise, Calcutta 0Central @oard of Re enue1 +,+74+,65 Re ised4+,>9 and find out the quantity of alcohol 0< of proof spirit and < of alcohol ACA1 from the table sho#ing the relation bet#een the specific gra ity of spirits at 9: :C9:: ' and the percentage of ethyl alcohol by #eight and by olume #ith the corresponding percentage of proof spirit issued under the authority of the Commissioner of *er &a?estyOs Customs and .xcise, )ondon, *er &a?estyOs stationery office +,==, Reprinted +,;+"

.xample of Calculation'ECK+"5Q>6E8%.1o( +" 6" >" 7" =" 9" ;" 5" ," +:" ++" gm +6" +>" +7" +=" +9"

/t

temperature

of

;9 :'

067"=oC

$eight of =: ml specific gra ity bottle - 69"95;: gm $eight of bottle plus distilled #ater - ;9"9;6: gm $eight of #ater - ;9"9;6:469"95;: E 7,",5=: gm $eight of +::: ml #ater - 7,",5=: x 6: E ,,,"; gm 041 Temperature correction - +"97 0to be added1 at ;9o' $eight of + liter #ater - ,,,"; Q +"97 E +::+">7 @ottle correction - +::+">7 M +::: E +">7Q $eight of exhibit and bottle - ;>"56=: gm $eight of the exhibit - ;>"56=: M 69"95;: E 7;"+>5 gm $eight of the +::: ml exhibit - 7;"+>5 x 6: E ,76";9 gm $eight of exhibits #ith bottle correction - ,76";9 M +">7 E ,7+"76 Temperature correction - 9":: 0at ;9:'1 Corrected #eight of the exhibit - ,7:"79 Q 9"::E ,79"79 gm (pecific gra ity of the exhibit - ,79"79C+::: E :",7979 %ercentage of proof spirit - ;="9, %ercentage of alcohol ACA - 7>"6=

"0T-R"!"T+>- .-T/)2S 3)R -T/50 "0C)/)0: G*F Gas Chromatography1 : )perating con itions :, Column - %orapac! polymer bead 5:4+:: mesh or its equi alent, #hich can separateCresol e the ethanol Column Temperature - +9::C Carrier Gas - Nitrogen Gas Rate of gas flo# - =: mlCmin Detector - 'ID 0'lame Ioni2ation Detector1 "lternati8e )perating Con itions? : Column - :">< Carbo#ax 6: & on 5:4+:: mesh Carbopa! C, 6m x 6mm ID or its equi alent Column Temperature - >=:C for 6 minutes and then programmed at = :C per minute to +;=:C and hold for at least 5 minutes

Carrier Gas - Nitrogen at >: mlCmin G!F The percentage 0contents1 of the alcohol can also be found from the table gi en in Official &ethods of /nalysis of the /ssociation of Official /gricultural Chemists 0/"O"/"C"1; after calculating the specific gra ity of the liquor samples" 8.%.2.2. 2etermination of .ethyl "lcohol : Gas Chromatography1 : )perating con itions :, Column - %orapac! polymer bead 5:4+:: mesh or its equi alent, #hich can separateCresol e the ethanol Column Temperature - +9::C Carrier Gas - Nitrogen Gas Rate of gas flo# - =: mlCmin Detector - 'ID 0'lame Ioni2ation Detector1 "lternati8e )perating Con itions? : Column - :">< Carbo#ax 6: & on 5:4+:: mesh Carbopa! C or its equi alent Column Temperature - >=: for 6 minutes and then programmed at = : per minute to +;=: and hold for at least 5 minutes Carrier Gas - Nitrogen at >: mlCmin 8.%.2.3 )ther 2eterminations: The follo#ing determinations 0some or all depending upon the nature of examination and infrastructure a ailable1, if required, may be carried out as per I(->;=6-+,55" iF Determination of ash iiF Determination of total acidity iiiF Determination of olatile acidity iHF Determination of fixed acidity HF Determination of residues on e aporation HiF Determination of esters HiiFDetermination of higher alcohols HiiiF Determination of aldehydes iAF Determination of copper

AF

Determination of furfural

References :+" /rthor I Aogel, T/ text boo! of practical organic chemistryP, >rd edition, +,=9, page +:954+:9, 6" (#arup Narain Ti#ari, T/nalytical Toxicology, @ureau of %olice Research I de elopmentP +,;9, %age 6:I6+ >" I( >;=6 N +,55 Indian (tandard /lcoholic Drin!s M &ethods of Test, 'irst Re ision 7" G" ( ehla, TAogelOs text boo! of &acro and (emi4micro qualitati e inorganic analysisP, =th edition, +,;,, %age 6+5 I 679 =" I( - >6> M +,=,, Indian (tandard specification for rectified spirit, Re ised, ,th Reprint December, +,5,, page 6546, 9" %"(" Ra?u and ("&" Chatter?ee, T/ (pectrophotometric &ethod for determining furfuraldehyde in illicit liquorsP, 8ournal of Indian /cademy of 'orensic (cience, Aol"7, No"+, 8an", +,9= ;" /"C" &offat et"al, TClar!eOs Isolation and Identification of DrugsP" The %harmaceutical %ress, )ondon, 6nd .dition, +,59"