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Water for Pharmaceutical Uses: Quality control


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Chapter 22

Water for Pharmaceutical Uses: Quality Control and


Validation

Ashish Baldi', Sumit Kumar'


'Department of
Quality Assurance,I.S.F. College of pharmacy,
Moga, Punjab

Introduction
Water is the one of the major commodities used by the
pharmaceutical industry. It is widely used as a raw material, ingredient
[], and solvent in the processing, formulation, and manufacture of
phanpaceutical products, active pharmaceutical ingredients (APIs) and
internrediates, and analytical reagent [2].lt may also present as an
excipient, or used for reconstitution of products, during synthesis, during
production of finished product, or as a cleaning agent for rinsing vessels,
equipment and primary packing materials etc. There are many different
grades of water used for pharmaceutical purposes [3]. Several are
described in USP monographs that speciff uses, acceptable methods of
preparation, and quality attributes [4]. These classes of water can be
divided into trvo general types: bulk waters, which are typically produced
on site where they are used; and packaged waters, which are produced,
il,
packaged, and sterilized to preserve microbial quality throughout their
tl packaged shelf life. There are several specialized types of packaged
it:
iF waters, differing in their designated applications, packaging limitations,
i,
and other quality attributes.
lt
li Water is the most widely used substance, raw material or starting
ii material in the production, processing and formulation of pharmaceutical
products. It has unique chemical properties due to its polarity and
hydrogen bonds. This means it is able to dissolve, absorb, adsorb or
suspend many different compounds [5]. These include contaminants that
may represent hazards in themselves or that may be able to react with
intended product substances, resulting in hazards to health. Different
grades of water quality are required depending on the uses and route of
administration of the pharmaceutical products.
Control of the quality of rvater throughout the production,
storage and distribution processes, including microbiological and
chemical quality, is a major concern. Unlike other product and process
236
ingredients, water is usually drawn from a system on demand. and is not
subject to testing and batch or lot release before use. Assurance of
quality to meet the on-demand expectation is, therefore, essential [6].
Additionally, certain microbiological tests may require periods of
incubation and, therefore, the results are likely to lag behind the water
use. Some types of microorganism may proliferate in water treatment
components and in the storage and distribution systems. It is very
important to minimize microbial contamination by routine sanitization
and taking appropriate measures to prevent microbial proliferation.
Pharmaceutical water production, storage and distribution
systems should be designed, installed, commissioned, validated and
maintained to ensure the reliable production of water of an appropriate
quality. These systems should not be operated beyond their designed
Water should be produced, stored and distributed in a manner
"upu"ity.
that prevents unacceptable microbial, chemical or physical contamination
(e.g. with dust and dirt) [7].
water sources and treated water should be monitored regularly
for quality and for chemical, microbiological and, as appropriate,
endoioxin contamination. The performance of water purfication, storage
and distribution systems should also be monitored. Records of the
monitoring results and any actions taken should be maintained for an'
appropriate length of time.
where chemical sanitization of the water systems is part of the
biocontamination control programme, a validated procedure should be
followed to ensure that the sanitizing agent has been effectively
removed.
Types of Water Used
Water is the most common aqueous vehicle used in
pharmaceuticals [8]. Figure I shows various types of water used in
pharmaceutical companies. There are several types of water are used in
the preparation oldrug product such as:
. Non pctable water
r Potable water
Water for special pharmaceutical purposes
o Purified water
r Water for injection
. Sterile water for injection
o Bacteriostatic water for injection
r Sterile water for inhalation
. Sterile water for irrigation
237

I
r Water for hemodialysis

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Figure 1: Types of water for pharmaceutical applications

Non potable water


' Non-potable water is the water that is not for drinking water
quality but which may still be used for many other purposes, depending
on its quality [9]. Non-potable water is generally all raw water that is
unheated, such as that form lakes, rivers, ground water, springs, and
ground wells !01.
Potable water
Potable rvater is not suitable for general pharmaceutical used
because of the ccnsiderable amount of dissolved solids present [ll].
These dissolved solid consists chiefly of the chlorides, sulfates and
bicarbonates of Na, K, Mg, and Ca [2]. A 100 ml of portion of official
water contains not rnore than 100 mg of residue (0J%) after evaporation
to dryness on a stem bath I l3].
Purified water (PW)
238
PLu'ifiec'l r.vlllei is Lt*\L-(l in illr- t.:'r. l',:1l'atiorr itf all nre ilication
containing water except anrpotrlcs, rn.icetion, attd sotlrc ofllcial extcrnal
preparation such as liniurents ll -11 Ptrrified $'ater ntust rneet tl')e
requirement for ionic anC organic cltemiual purity :tnci must be protected
from microbial contamination [ 5]. T'he minimal quality of source or
feed u'ater for the production of purified rvater is drinking water.
PW should be prepared from a potable water sourcc as a minimum-
quality feed-water, should meet the pharmacopoeial specfications for
chemical and microbiological purity, and should be protected from
recontamination and microbial proliferation [6]. PW is prepared by
distillation, by ion exchange or by any other suitable method, from water
that complies with the regulations on water intended for human
consumption [17].
Highly purified water (HPW)
Highly purfied water (HPW) should be prepared from potable
water as a minimum-quality feed-water. HPW is a unique spedcation
for *'ater found only in the European Pharmacopoeia. This grade of
water must meet the same quality standard as water for injections (WFI)
including the limit for endotoxins, but the water-treatment methods are
not considered to be as reliable as distillation [l8].
Water for injection (WFI)
WFI is a solvent used in used in the production of parenteral and
other preparations where product endotoxin content must be controlled
and in other pharmaceutical application. WFI is sterile, non-pyrogenic,
distilled water for the preparation of products for parenteral used. It
contains no added substance and meets all the requirements of the
pyrogen test. The finished water must meet all the chemical requirements
for PW as well as bacterial endotoxin specification [19]. Since
endotoxins are produced by the kinds of microorganisms that are prone
to inhabit water, the equipment and procedures use by the system to
puriry, store and distribute. Water for injection must be designed to
minimize and prevent microbial contamination as well as remove
incoming endotoxins for the starting water [20]. WFI is water for the
preparation of medicines for parenteral adrninistration when water is
used as a vehicle (water for injections in bulk) and for dissolving or
diluting substances or preparations for parenteral administration before
use (sterilized WFI). Water for injection system must be validated to
reliably and consistently produce and distributes this quality of water.
Control of the chemical purity of WFI presents few major
problems. The critical issue is that of ensuring consistent microbiological

239
quality with respect to removal of bacteria and bacterial endotoxinl,
Distillation has a long history of reliable performance and can bo
validated as a unit operation, hence it currently remains the only officiul
method for WFI.
WFI in bulk is obtained from water that complies with tho
regulation on water intended for human consumption laid down by lhc
competent authority, or from purified water, by distillation in un
apparatus of which the parts in contact with the water are of neutral
glass, quartz or suitable metal and which is fitted with an effective devicc
to prevent the entrainment of droplets. The correct maintenance of thc
apparatus is essential. During production and storage, appropriatc
measures are taken to ensure that the total viable aerobic count is
adequately controlled and monitored [21].
Storage conditions
r It can be stored for a period up to a month in special tanks containing
ultraviolet lamps [22]. when this freshly prepared water is stored and
sterilized in hermitically sealed containers, it will remain in good
condition indefinitely [23].
e If autoclave is not available, freshly distilled water may be sterilized
by boiling the water for at least 60 minutes in a flask stoppered with a
plug of purified non absorbent cotton covered with gauze [24], tin-
foil or stout non absorbent paper, or the neck of the flask may be
covered with cellophane and tightly fastened with cord [25].
Sterile WFI
Its specification are provided in USP monograph for water for
injection, sterilized and packaged in suitable single dose containers
preferably of type I glass of not larger than 1000 ml size. Its meets the
requirements of the sterility test and pyrogen test and other tests under
purified water [26]. SWFI is sterile, non pyrogenic, distilled water used
in pharmaceutical industry. A container of a sterile preparation for
parenteral use that contains many single doses. No antimicrobial or other
substance has been added. pH 5.5 (5.0 to 7.0)
Bacteriostatic WFI
Bacteriostatic WFI, USP is a steriie, nonpyrogenic preparation of
water for injection containing 0.9 % (9 mg/ml) of benzyl alcohol added
as a bacteriostatic preservative. It is supplied in single dose containers of
not larger than 5 ml size and in multiple-dose containers of not larger
than 30 mL size from which repeated withdrawais may be made to dilute
or dissolve drugs for injection or the pFI is 5.7 (4-5 to 7'0).

40
Caution- Where rl'atcl ;. rr:ilirl'lrd fhi n;, r, : L

loruseirtncclttales,0llir;!ti.::,r'.'';.lrii.c.il.'
benzyl alchol is knou,n to induce tl-.;rr',,iir ')rii,,,
administration of bacteriostatic WFI \i'rii.r.'r.rt ;r .,r.rr,,r'
hemolysis.
Sterile water for inhalation
Sterile water fo:' inhalation cnt.itrtics ll i.r;i.1rri,)\litiL'.
antimicrobial agent or added buffer ancl is intende,-j ir;r irse ()rriv i{s
single-dose or short procedurc irrigatron. Wh.:n snrail':i i,rririlni':, aii
required the unused portion should be discarded. Sterle '*'ater il1
inhalation is Water for injection that is packaged enrl rerrr--1i.re,.1 sterilite
and is intended for use in inhalators and in the pre'p;rarion of inhal:riiori
solution [27]. It carries a less stringent spri'iiication lirr bacter-iii
endotoxins than sterile u'ater for inhalation. tli.'lei'lii,' l\ ilc.,t suitable ior
parenteral prepffation.
Sterile water for irrigation
Sterile water for irigation, USP is a sterile. drstilled, non
pyrogenic water for iniection intended onl;- for sterile irrigaticln,
washing, rinsing and dilution purposes. Sterile water fbr irrigations is
Water for injection packaged and sterilized in single dose containers oi
larger than I L in size that allows rapid delivery of its corrtents [2S]. I't
need not meet the requirement under small volume injection.
Sterile water for Irrigation, USP exerts a mechanical cleansing
action for sterile irrigation of body cavities, tissues or wounds.
indwelling wethral catheters and surgical drainage tubes, and for
washing, rinsing or soaking surgical dressings, instnunents and
laboratory specimens. It also serves as a diluent or vehicle for drugs used
for irrigation or other pharmaceutical preparations. It should have pH 5.5
(5.0 to 7.0). Examples of Sterile water tbr irrigation are Surgical
inigation solution, urologic irrigation solution, glycine solution, sorbitol
solution. Sterile Water for lrrigation, USP is indicated for all general
irrigation, washing, rinsing and dilution purposes which permit use of
sterile, nonpyrogenic, solute-free water.
Caulion-Possible adverse effects arising from the irrigation of body
cavities, tissues, or indrvelling catheters and tubes are completely
avoidable when proper procedures are followed. Excessive volume or
pressure during irrigation of closed cavities may cause undue distention
or disruption of tissues. Accidental contamination from careless
technique may transmit infection [29].
Water for hemodialysis
241
water for hemodialysis is used for hemodialysis applications. lt
may be packaged and stored in unreactive containers and preclude and
o'unreactive containers" implies that thc
bacterial- entt [30]. The term
container, .sp.Ciutty its water contact surfaces, are not changed in any
way by the water, such as by leaching of container-related compounds
intO ttre water or by any chemical reaction or corrosion caused by the
water [311. The rvater contains no added antimicrobial sans is not
intended for injection.
Pure steam: Pure steam iS also sometimes referred tO aS a "clean steam"'
It is generally used for following purposes:
. To remove any co-deposited impurity residue'
r For air humidification in controlled manufacturing environment.
r Used in steam sterilization of equipment and porous loads [32]
. For cleaning the places where condensates directly comes in contact
with officiai articles, product contact containers and surface.
Some water rniscible solvents are also used in parenterals.
principally to enhance dmg solubility, it is important to mentio,n_that also
."*" ut siabilizers for those drugs that degrade by hydrolysis [33]. Water
.rniscible polvent must be selected with grade care for it. must not be
irritating, toxic or sensitizing and it rilust not exert on the ingredients of
the forriulation. Solvents that are miscible with water are dioxolanes,
dimethylacetamide, burylenes glycol, polyethylene glycol 400 and 600'
propylene glycol. glycerin, ethyl alcohol etc' Various grades of water'
ih"i, ,rr"t in phurmaceuticals and preparation techniques are given in
Table l.
ahlc l.I f Jcr',( nnt renaratiott tcchniques of various es of
of water
Table
Grades of rvater Uses Preparation
iechnioues
. Cleaning ofortter surface ofihe factory Obtained from
Non potable
nahrral sources.
water Used in galden
".
Washing veittcles etc.
Potable rvater . As drinking rvater Obtained fronr
. Washing and extraction of the crude naflrral sources
drugs
. Prgparati;n of products to extemally
___ use 3tc
I Puritled s'aicr
Puritred u'"r o Foi tir,: pr,-,ductior, of non-parenlcral . Deionizatt
(Pw) on
I rni,'; pt cparation
r
I . For the cleal:trg of certain cqtriprrrcni Distillation
I l::;:d in non nat'enieral production r lon-
rl"-'i\i)1, : exchangc
I
| 'r';r'r-j].triii-,utl] rc;sr'
i
L__ __._ lI !l'" l:':-il' ' i ' Re
-- J
contact component osmosls
o For all types oftests and assay . Filtraticn
. For the preparation ofson:e bulk efc.
chemicals
. For the preparation ofniedia in
microbiology laboratories

Highly purified o HPW is intended for use in the a Deionizati


water (HPW) preparation ofproducts where water of on
high bioiogical quality is needed, t Reverse
except where WFI is required osmosis
a Ultrafiltrati
on etc.
Water for o For the production ofparenteral a Distillation
injection (WFI) preparation a Reverse
o For cleaning ofparenteral product osmosis
contact component a Membrane

Sterile WFI . For extemporaneous preparation rBy


(swFr) compounding distillation
. As a sterilize diluents for the parenteral of WFI
oroducts
Bacteriostatic r Used as a diluents in the preparation of . By using
WFI(BWFI) narenteral oroduct SWFI
Sterile water for a Preparation ofuse in inhalators r By using
inhalation a Preparation of inhalant solution SWFI
Sterile water for r To bath and moisten body tissue r Frorr
irrigations r Performing urologic procedure for water for
srrfgeon iniection
Sterile water for a Preparation ofuse in inhalators . By using
inhalation a Preoaration of inhalant solution SWFI
Sterile water for a To bath and moisten body tissue r From WFI
irrigation a Performing urologic procedure for
surseon
Water for o For the dilution of hemodialysis of r From safe
hemodialysis concentrate solution drinking
water

Specifications of various grades of water


USP has stipulated the specifications and definitions of the
various grades of water suitable for pharmaceutical use. It classifies
pharmaceutical water as (i) potable water (ii) purified water, used for the
manufacture of oral preparations and other formulations, and (iii) water
for injection (WFI) and (iv) sterile water for injection used for
injectables, parenterals and intravenous fluids. USP also specifies that

243

ll
purified water and WFI must adhere to the Environmental Protection
Agency's (EPA's) Part l4l, National Interim Primary Drinking Water
Regulations. Table 2 presents the principal specification for various
grades of water [34].

Table of waters as Der United State Pharm


Sr. Pararneter Potable water Purified Water for Sterile
No. water injection Water
for
iniection
I Appearance Clear, Clear, Clear, Clear,
colorless and colorless and colorless colorless
no visible no visible
uarticles particles
t Odor Odorless Odorless Odorless Odorless
3. Ph 6.5-8.5 5.0-7.0 5.0-7.0 5.0-7.0
4. Chloride NN{T 250 o ppm o Ppm o PPm
nnm
5. Aluminum 0.2 ms/L 0 ms/L 0 ms/L 0 me/L
6. Arsenic 0.01 ms/L 0 me/L 0 ms/L 0 me/L
7. Fluoride 1.5 me,{- 0 melL 0 mp/L 0 me/L
8. Boron 0.3 ms,'l- 0.3 rne,l- 0 me/L 0 ms/L
9. Sulfate NMT.lOO o ppm o ppm o ppm
nnm
10. Total NMT 500 o ppm o ppm o ppm
hardness Dnm
ll Mi crobial 500 cfir,'ml 100 chri n:l l0 cfu/100 l0 cfrr/
count ml ml
t2. Acidity or NMT0.l ml NMT0.l ml NMTO.I
alkalinity of 0.01M of 0-0lM ml of
NaOH NaOH 0.0tM
NaOH
t3. Ammonia 0.5 ppm 0.2 ppm 0 oom 0.2 oom
14. Heavy meta! 0.5 oom 0.1 oom 0 nom 0.1 opm
15. Conductivity NMT 0.3 ps NMT 0.1 ps NMT 0.1 ps NMT
0.05 us

Maximum allowable concentrations of toxic substances according to the


international standard are given in 'fable 3 [35].
Table 3: Maximum allowable concentration of toxic substances
f r*i*u.iunces I lr{ aximum allowable concenf ration

i Cr.'aa!cl,; l('r'J i i) :.

!+4

lt
i . 3,:illr:.::) : . j.:,)
..,. _ .., .

LEgtYslEo---Li () ---- ----l

Quality Control of Pharmaceutical Water SSstem


In the pharmaceutical quaiity contrrri q,rte:: anal'-,sis is of vital
importance to ensure the prodtrct's sterilrty. Orice water for
pharmaceutical use has been cbiained. it must be siored and distributed
to the points of use; there is nc poiut in producing quelity water unless it
is correctly stored and distributed. This stage is of viral importance to
minimize possible contamination of the water and the proliferation of
micro-organisms. Systems must be sealed with continuous recirculation
and must have a sanitization system.
Location of sampling points:
Samples must be taken from locations that are representative of
the water source, fieatment piant, storage facilities, distribution network,
points at which water is delivered to the consumer, and points of use. In
selecting sampling points, each localiry should bre considei-ed
individually; however, the following general criteria are t:s:ialiv
applicable [36]:
. Sampling points should be selected such that the samples taken are
representative of the different sources from which water is obtained
by the public or enters the system.
o These points should include those that yield samples representative of
the conditions at the most unfavorable sources or places in the sr.rpply
system, particularly points of possible contamination such as
unprotected sources, loops, reservoirs, lorv-pressure zones, ends ofthe
system, etc.
. Sampling points should be uniformly distributed throughout a piped
distribution system, taking population distribution into account; the
number of sampling points should be proportional to the number of
links or branches. t

. The points chosen shouid generally yield samples that are


representative of the system as a whole and of its main components
[3 7].
Sampling frequency
The most important tests used in water-qualily surveillance or
quality control in small communities are those for microbiological
quality and turbidity, and for free chlorine residual and pH where
chlorination is used. The recommended minimum frequencies for these

245

ll
critical nteasurements in unpiped water supplies are summarized, in Table
4.

Table 4: Mini mum ol sam and anal


Source and mode of Minimum frequencl of sampling and analvsis
supnlv Bacieriolonical Phvsical/Chemical
Open wells for Sanitary protection Once initially, thereafter
conrmunity supply measures: bacteriological as
testing only if situation demands
situation demands
Covered dug wells and Sanitary protection Once initially, thereafter
shallorv tubewells with measures: bacteriological as
hand-pumps tesling only if situation demands
situation demands
Deep nrbewells with Once initially thereafter as Once initially, thereafter
hand-purnps situation demands as
situation demands
Protected springs Once inirially thereafter as Periodically for residual
situation demands chlorine. if water is
chlorinated
Commrtniry . rainwater Sanitary protection Not needed
coliection systems measures: bacteriological
testing only if
situation demands

Storage of samples for microbiological analysis


'l'he time betrveen sample collection and analysis should,
in
general, not exceed 6 hours, and 24 hours is considered. It is assumed
that the samples are immediately placed in a lighproof insulated box
contalning melting ice or ice-packs ''vith water to ensure rapid cooling. If
ice is not available, the transportation time must not exceed 2 hours. It is
impcrative that samples are kepr in the dark and that cooling is rapid. If
these r:onditions are not met, the sampies should be discarded. When
water that contains or may contain even traces of chlorine is sampled, the
chlorin.' n:ust be inactivared. lf it is not, microbes may be killed during
iransit and an enoneous result rvill be obtained. The bottles in which the
samplcs are placed should therefore contain sodium thiosulfate to
neutraiize any chlorine presrrrt [38].
Sam p ling methods for ph.v-sicochernical analysis
fhis has important qonsequence:; for sampling regimes, sampling
prot:r;js6s5. etnd nrcrhcds oi sanrple preservation and storage. In general,
iilc tirirt between -iainDling iiiiri arraiysi.s should be kept to a minimum.
Sior-:l'-.'irr r.la',.; r:; r,iilvsii:,,'i,-.:t,, Lri,lii,:s;li a lclw telnp,Jrature (e.g.4.C) in

lt
the dark is recommended. Sample bottles must be clean but need not be
sterile. Special preservatives may be required for some analyes.
Residual chlorine, pH, and turbidity should be tested immediately after
sampling as they will change during storage and transport.
Microbiological analysis
The microbiological examination of drinking-water emphasizes
assessment of the hygienic quality of the supply. This requires the
isolation and enumeration of organisms that indicate the presence of
faecal contamination. In certain circumstances, the same indicator
organisms may also be used to assess the efficiency of drinking-water
treatment plants, which is an important element of quality control [39].
In the microbiological examination of water there are four different
cultural methods routinely employed and recommended in standards.
These include:
r Aerobic or Heterohophic Plate Count (IfC),
r Presence -Absence (P-A) testing,
o Most Probable Number (MPN) method and
o Membrane Filtration (MF) method.

LHeterotrophic Plate Count (HPC)


Heterotrophic Plate Count is commonly deterrrined using the
pour plate technique. One ml of a water sample or a decimal dilution
series is transferred to separate Petri dishes.l5ml of liquified agar
medium is then added to each Petri dish. The sample is thoroughly mixed
by rotation (three times left, three times right and once through the
centre). The agar is left to solidifu on a flat level, preferably cool,
surface. After complete solidification the plates are inverted and
incubated. Plates showing 25 to 250 colonies should be considered in
determining the standard plate count. A count is designated as standard
plate count at temperature of incubation. The incubation temperature can
either be 20,30 or 35-37"C. Depending on incubation temperature and
afinosphere, the counts are termed psychrotrophic aerobic or anaerobic
standard count (20oC) or mesophilic aerobic or anaerobic plate counts
(30 or 3s-37'C) [40].
ilPresence-Absence (P-A) testing
Presence-Absence testing is to obtain qualitative information on
the presence and absence of the target organism or group of organisms.
For the Presence-Absence test commonly a 100 ml sample is transferred
to a single flask. A double or triple strength liquid culture medium is then

247

lt
added. The use of a double or triple strength medium prevents that the
sample dilutes in the culture medium and thereby reduces its selectivity.
iil Most Probable Number (MPN) or Maltiple tube technique
The multiple tube testing is a modification of the Presence-
Absence testing. lnstead of adding the sample to a single tube, the
sample is divided in portions and multiple tubes are inoculated with
variable volumes of the same water sample. The reference methods
employ different volumes and replicates for the Multiple tube testing.
For sample volumes less than 5 ml or l0 ml the sample is added to an
equal volume of single strength media. For volumes of 10-100 ml,
double strength media are commonly used. Standard methods, however,
recommend the use of triple strength media for volumes of 100 ml and
higher [al]. The positive and negative test results of each tube can be
used for a calculation of the estimated number of counts of
microorganisms.
iv.Membrane Filtarton (M F)
Membrane Filtration technique is relatively simple. At the
beginning of each filtration series, the filtration units are sterilised to
avoid contamination. Using sterile tweezers, a sterile membrane filter is
placed over the porous plate of the filtration unit, grid side up. The
matched funnel unit is careftrlly placed over the receptacle and locked in
place. A volume of water is then filtered through the membrane filter
under partial vacuum. The membrane filter retains the organisms on the
surface. The filter may be rinsed with three 30 ml portions of sterile
buffered water or membrane rinsing fluid. It is advised to validate the
number of washes.
After the filtration, the funnel is unlocked and removed. The
membrane filter is removed with sterile tweezers and placed on the agar
meciium with a rolling motion to avoid the entrapment of air. Agar plates
used, should have a visually dry surface. The agar plate is incubated
inverted. After the recommended incubation period the colonies are
counted. The counts of typical colonies on a selective medium are
presumptive counts. A confirmation of a square root of typical colonies
gives the confirnrative count. Counts are expressed as cfu per ml for a
fiitered sampb |"42].
Rapid testing nrethods
(i) Fluorogenic utrd chromogenic culture media
On a medium containing fluorogenic and/or chromogenic
substrates a. iai.gct o:'i:anism is identified by the activity of an en-zyme
spclir-i; i.o ':irrt .rr':I:,'l;:irir. in fltrorogenic culttrc media the enzymc
1io

lt
characterizing the target organism splits a fluorogenic substrate), into
two separate components, a sugar or amino acid and a fluorogen- The
mentioned fluorogen converts UV-light to visible light. The fluorescence
produced is read in the dark under exposure to a UV light at 366 nm. An
agar, containing a fluorogenic, should be read aftet 24 hours as these
substrates have the tendency to migrate into the agar making it
impossible to identiff a single colony of the target organism- It is also
important to mention that fluorogenic.substrates are heat sensitive and
pH dependent. Tests have shown that the fluorescence is most intense at
a pH above 8.5.
In chromogenic culture media, indolyl derivatives are quite
commonly used. These chromogenic substrates are water soluble, heat
stable and pH independent. As with fluorogenic culture media the
enzyme that characterizes the target organism splits the chromogenic dye
into two components. The chromogenic colors the broth and/or the
colony. The color does not diffuse into the agar, therefore, only the target
colonies are colored [43].
The presence of the target organism(s) in fluorogenic and chromogenic
culture media is identified by the color and/or fluorescence. The high
specificity of the differential system eliminates the need for subculturing
and further biochemical tests and greatly improves identification'
ii. Total coliform bacteria
When using FluoroCult Broth media with the Presence-Absence
or the multiple tube (MPl$ method, results of total coliform bacteria are
available within 24 hours. A positive total coliform test is indicated by a
blue-green color. The distinct blue-green color of a positive test sample
is easily distinguished even with colored water samples [44].

Validation and Qualification


Validation and qualification of water purification, storage and
distribution systenN are a fi.rndamental part of Good Manufactqring
Process (GMP) and form an integral part of the GMP inspection' The
grade of water used at different stages in the manufacture of the active
pharmaceutical ingredients and pharmaceutical products should be
discussed in the'pharmaceutical dossier. The grade of water used should
take account ofthe nature and intended uses ofthe finished product and
the stage at which the water is used [45].
In the past two decades the validation approach in
pharmaceutical industries has been discussed due to its real.importance
within a productive process in relation to the products quality attributes,
249

lt
like purity, safety and effectiveness that constitr.ltes the base of the
thought and working of professionals involved to the process validation.
The U.S. Food and Drug Administration (FDA) in l9g7 in its most
recently proposed 'Guidelines on General principles of process
Yalidation' has offered a definition to process validation, after a series of
incidents involved cross-contamination problems by some
pharmaceutical manufacturing establishments [46] :
"Process validation is establishing documented evidence which
provides a high degree of assurance that a specific process will
consistently produce a product meeting its predetermined specifications
and quality characteristics".
validation is legible through the documentation establishment;
the guarantee of that manufacturing process assures the product quality
with homogeneity. The validation concept applies to the end product,
and all the previous stages are called qualification. So, the validation
consists of a series of qualifications that involves tests, systems
verifications and also critical process parameters that are its regulating
keys, and can vary within an acceptance limit. The different
qualifications form stages of validation:
r Design Qualification,
I.i,. Installation Qualification,
r Operation Qualification and
o Performance Qualification.
Each one must follow its own scheduie and protocol. The water,
in a pharmaceutical industry, is the most important raw material for the
accomplishment of any process. Therefore, it is essential to not only
assure that water, but also installations and procedures are validated. So,
it is necessary to choose a well-designed water system, by using a
combination of methods that allows reaching the quality levels of water
to a determinate application, by optimizing its particular capacity of
remove contaminants [a7]. Successful accomplishn'rent of validation is
ensured by various testing phases. Usually, a three-phase testing
approach is recommended over an extended period to prove reliability
and robustness of the system for producing water of specified quality
with a high degree of assurance.
Guidelines
The primary purpose of the 'Guideline.r _for y'l/ater euality, is the
protection of public health. The Guidelines provide the recommendations
of the World Health Organization (WHO) for managing the risk from
hazards that ma)' compromise the safety of water quality. The
250

lt
recommendations should be considered in the context of managing the
risk from other sources of exposure to these hazards, such as rvaste, air,
food and consumer products [48]. Different grades of water are produced
according to USP and EP requirements, usually by: distillation; reverse
osmosis; deionisation; or ultrafiltration. All of these processes need to be
validated and recorded according to specific standards [49].
Each set of guidelines outline legal requirements for the chemical content
of each grade of water, including a three stage conductivity test for
inorganic compounds that will determine pH and total organic oxygen
(TOC) levels. The FDA states that implicit in the term "Purified Water"
is that it has some reasonable, objective level of purity. TOC testing
allows for evaluating impurities in water besides those which are
inorganic anions and cations.
Conclusion
In the pharmaceutical companies, different grades of water are
used for different puposes. So it is necessary to know all the grades of
pharmaceutical water, its preparation, specifications and uses to follow
the guidelines for the maintenance of quality atfribules. Water treatment
systems must be operated within regulatoryr :'guidelines .for
pharmaceutical production facilities by performing various quality
checks for physicochemical and microbiological impurities. Furthermore
documented evidence is necessary to ensure and validate system
operation as per desired specifications. Validation is therefore an
essential tool for process optimization and total quality management for
safety, efficacy and assurance of water quality for pharmaceutical
purpose.

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I

Prof. (Dr.) S.S. Gill


After doing MS (Orthopedics) from PGIMER. Chandigarh in 1976, he was specially trained in Orthopedics
from University College. Los Angles, U.S.A. Belorc assuming the o{fice ofthe Vice Chancellor ofBaba Farid
Uliversity of Health Sciences, Faridkot. he serued as Prof'essor & Head Dept of Orthopedics at PGIMER.
Chandigarh. Profl Gill was Member ofsenate and Syndicate, Dean, Faculty ofMedical Sciences and Dean.
Faculty ofDesign & Fine Arts at Panjab University, Chandigarh and on several other high profile committees o1'
nany National Institutes. He has been an active researcher. teacher and orthopeadician during his professional
career and contributed 2 books. 5 chapters in books and published more than 1 00 research papers including 25 in
sT
I
intemationaljoumals ofrepute. Prof. Gill has visited various corurtries like England, USA. Canada. Hongkong.
China, Australia and many more . He has becn decorated with numerous presligious awards including SCROL L
G
O F HONOUR by North Zone Indian Orthopeadician Association, Jammu in 2002, Bhagat Puran Singh Award
in 2007, lor social services by Nagrik Manch and Baba Farid Award fbr Honesty 20 I 2 by Baba Farid Society.
g
I
-
L
Dr. Rajeev Manhas
Ph.D. (LISc), AMSPI, FUWAI, is Head of
o
5
Department of Health Sciences Library &
Infonnation System and University Library &
L
Informatics Division, BFUHS, Faridkot. He has
morc lhan l7 years work expcrience in g
T
librarianship. He has attended morc than 50
conferences including an intemational conlercnce
at Canada. He has organized more than J0
conferences and workshops at national and
intcmational level. He has published more than 50
research papers in national and intcmational
G
joumals, 3 books, and 7 chapters in books. Hc has
Young Scientist Award-2O11 to his credit lbr his
contribution in development of Health Sciences
Library Network in Punjab. He has setup
Department of Health Sciences Library &
Information System in the University for teaching
& research in health sciences librarianship. He is on
=
the editorial board of5 intemational and 4 national
journals. He is Board Member of Informing
Science lnstitute. USA.

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