International Journal of Pharmacy and Pharmaceutical Sciences

ISSN- 0975-1491 Vol 2, Issue 4, 2010

ResearchArticle

QUANTITATIVEDETERMINATIONOFBORONCONTENTINTAMSULOSINHYDROCHLORIDE USINGINDUCTIVELYCOUPLEDPLASMAOPTICALEMISSIONSPECTROSCOPY
MITHLESHRAJPUT,VINAYKUMARJAIN,DHARAMPALJAIN,MANJEETAGGARWALANDRAKESHKUMAR KHANDAL
Director,ShriramInstituteforIndustrialResearch,Delhi110007India,Email:rkhandal@shriraminstitute.org,sridlhi@vsnl.com Received:24May2010,RevisedandAccepted:26Jun2010 ABSTRACT Aprecise,accurate,sensitiveandselectiveanalyticalmethodusingInductivelyCoupledPlasmaOpticalEmissionSpectroscopy(ICPOES)hasbeen developedandvalidatedforthedeterminationoftracelevelsofboron,presentasanimpurityinTamsulosinhydrochloride,analphablockerdrug. Boron was suitably extracted from Tamsulosin hydrochloride and brought into the solution using ashing technique followed by quantitative determinationbyICPOES. Thelimitofdetectionofthevalidatedmethodwasfoundas15 g/landthelimitofquantitationwascalculatedas25g/l.Themethodwasfoundto belinearinthewideworkingrangeof25 g/lto800 g/lwithcorrelationcoefficientof0.99978.Therecoveriesofboronfromthespikedsamples ofTamsulosinhydrochloridewerefoundintheacceptablerangeof90to98%atthreedifferentspikinglevels.Themethodcanroutinelybeused forthequantitativedeterminationofborontoensurethequalityofTamsulosinhydrochloride Keywords:ICPOES,Boron,Tamsulosinhydrochloride,Methoddevelopment INTRODUCTION Tamsulosin hydrochloride, 5[(2R) 2 [[2 (2Ethoxyphenoxy) ethyl] amino] propyl] 2 methoxy benzene sulfonamide hydrochloride1,2,3, is an alpha blocker drug . It is used for the treatment of urinary problems caused by an enlarged prostate. Its intakerelaxesthemusclesoftheprostateandbladder,whichhelps freeflowofurine2,3. absorptiontechnique(FlameAAS)hasunsatisfyingdetectionlimits for boron46, 8. Similarly, other analytical techniques for the determinationofboronaretimeconsumingandnotsosensitiveand precise8. But the use of Inductively Coupled Plasma Optical Emission Spectroscopy(ICPOES),provideshighersensitivity,lowerdetection limits, less chemical interferences and is less time consuming as comparedtoalltheabovementionedtechniques. Thepresentpaperdescribesthedeterminationofresiduesofboron in Tamsulosin hydrochloride by ICPOES using radial torch, which caneasilybeadoptedbyanyanalyticallaboratory.Themethodwas validated for various parameters in accordance with International Conference on Harmonization (ICH) and Eurachem guidelines. Method was also optimized for sample preparation using ashing procedurewherebycalciumhydroxidewasaddedtotrapboron. MATERIALSANDMETHODS Chemicalsandreagents SamplesofTamsulosinhydrochloriderawmaterial(fiveinnumber coded as A, B, C, D and E) were procured from M/S Ranbaxy Research Laboratories. HPLC grade water used throughout the experimental analysis was procured from s.d.finechem limited. CalciumhydroxideandHydrochloricAcid,ARgradewereprocured fromMerckSpecialtiesChemicalLimited.Alltheglasswareusedwas TypeAandBorosilmake.Calibratedmicropipettewithrange100 l1000lwas used.Standardreferencesolutionsof1000g/ml boron, nickel, platinum and palladium (traceable to NIST) were procuredfromScharlauChemie,Spain.Whatmanfilterpaperno.41 wasused. Saturatedsolutionofcalciumhydroxide Saturatedsolutionofcalciumhydroxidewaspreparedbydissolving anexcessamountofcalciumhydroxideinwarmHPLCgradewater till the point where no further dissolution of calcium hydroxide occursevenonwarmingthesolution.Theexcesscalciumhydroxide wasfilteredpriortouse. Instrumentation Varian (Australia) Vista MPX Inductively Coupled Plasma Optical Emission Spectrometer (ICPOES) equipped with argon saturation assembly, CCD detector and 21 CFR 11 version 4.1.0 software for data acquisition and processing was used. Muffle furnace:

Fig.1:StructureofTamsulosinHydrochloride During the synthesis of Tamsulosin hydrochloride, which involves several steps, sodium borohydrideiodine or sodium borohydride sulfate is used as a reducing agent in one of the steps2, 3. As a result, trace amount of boronmay remaininthe final product,whichisnot desirableasresidualimpuritiesmaycauseadverseeffectsonhealth.It is therefore imminent that the boron used as catalyst during the manufacturingprocessisremovedfromthefinalproductbyadopting suitablepurificationprocesses.Inordertoensurethattheproductis freefromimpuritiesofboron,itisutmostessentialthatthevalidated methodisusedfordeterminingthecontentofresidualimpurities.The methodshouldbesuchthatitdoesnotsufferfromanyinterferences fromtheresidualimpuritiesofothermetalsusedascatalysts(suchas nickel,palladiumandrhodium2,3).Thepresentstudywasundertaken to develop and validate an analytical method to detect boron at low concentrationlevels;wellbelowacceptablelimits. Anumber of methods have beenreported for the determinationof boron, most of them being colorimetric methods4,5,7. For the quantitative determination of boron by colorimetry, a number of reagents are available such as curcumin4,5,7, carminic acid4,5,7 or carmine4,5,7, quinalizarin4,5,7 , azomethineH4,5 etc. The colorimetric methods are not so sensitive and moreover suffer due to the interference form other metallic impurities as well as from the components of matrix10. The fluorimetric methods are more sensitive than colorimetric methods but are susceptible to interferencesfromcertainchemicalspeciesandarealsosensitiveto pHandthetemperature7,8.Therefore,fluorimetricmethodsare not widely used for the determination of boron. The Flame Atomic

Khandaletal. IntJPharmPharmSci,Vol2,Issue4,182185 Ambassador with least count 1 C was used. Electronic analytical balance:Afcoset3200,Mettlertoddlerwithreadability0.01mgwas used. Preparationofcalibrationstandardsofboron Preparationofstocksolutionofboron(5g/ml) 5 ml of standard reference solution of boron (1000g/ml) was pipettedintoa 100mlvolumetricflaskanddilutedtovolume with HPLC grade water. This gave a solution with concentration of 50 g/ml (solution A). From solution A, 10 ml was further diluted to 100 ml to give a solution with boron concentration of 5 g/ml (solutionB).Thiswasthenusedasastocksolutionforpreparation ofcalibrationstandardsolutionsofboron. Preparationofcalibrationsolutionsofboron From the solution B, aliquots of 0.125 ml, 0.25 ml, 0.50 ml, 1.00 ml, 2.00mland4.00mlwerepipettedintosixdifferentvolumetricflasks of 25 ml and diluted to mark using HPLC grade water. This gave a series of standard calibration solutions having concentration of 25 g/l,50g/l,100g/l,200g/l,400g/land800g/lrespectively. Sampletreatment About 4.00 0.01 gram sample of Tamsulosin hydrochloride was weighed accurately in a silica crucible, and 10 ml of saturated solution of calcium hydroxide was added. The crucible was then heatedonahotplatetodrynessandthemixturewasashedat550 25C in a muffle furnace for 4 hours. The contents were cooled and leached in 2 ml hydrochloric acid and transferred to a 25 ml volumetricflaskandmadetovolumeusingHPLCgradewater.All sample preparation required for studies of various validation parameterswasdoneaspertheaboveprocedure. Preparationofreagentblank 10mlofsaturatedsolutionofcalciumhydroxidewastakeninsilica crucible and heated on a hot plate to dryness and ashed at 550 25Cinamufflefurnacefor4hours.Aftercooling,thecontentswere leached using 2 ml hydrochloric acid and transferred to 25 ml volumetricflaskandmadetovolumeusingHPLCgradewater.Five replicatesofreagentblankwereprepared.

ICPOESconditions Inductivelycoupledplasmaopticalemissionspectrometery(ICPOES) withradialtorchequippedwithargonsaturationassemblywasused for the determination of boron in Tamsulosin hydrochloride. High purity (99.99%) argon was used as plasma, auxiliary and nebulizer gas. The gas flows were kept at 15.0 l/minute for plasma, 1.50 l/ minuteforauxiliaryand0.56l/minutefornebulizer.Radiofrequency (R.F.)poweroftheplasmageneratorwas1.20kW.Verticalheightof theplasmawasfixedat7mm.Sampleuptaketimeof30.0sec,delay timeof5sec,rinsetimeof5sec,initialstabilizationtimeof5secand time between replicate analysis of 5 sec was maintained throughout the studies for ICPOES. All the observations of emission were recorded at 249.772nm, which corresponds to the most sensitive emission wavelength of boron. The instrument was calibrated for variousparametersbeforethestudies. RESULTSANDDISCUSSION Analyticalmethoddevelopment Sampledigestion One of the challenges of method development was the efficient extraction of boron from Tamsulosin hydrochloride. For this purpose, generally, the digestion using acids is recommended. During the initial development work, it was noted that the acids usedforthispurposewerecontaminatedwithimpuritiesofboronto the extent beyond the residual impurity of boron in the drug. The methodusedhereforthispurposetherefore,wasashingofthedrug inthepresenceofcalciumhydroxide.Theuseofcalciumhydroxide prevents volatilization of boron since calcium hydroxide helps in formationofcalciumboride4,5,whichisahighlystablecompoundas perthereaction: 2Ca(OH)2+12B2CaB6+2H2O Optimization of requirement of saturated solution of calcium hydroxide For the purpose of complete extraction of boron, the studies for optimization of the requirements of calcium hydroxide for the purpose were conducted. Tamsulosin hydrochloride was spiked withdifferentconcentrationofstandardsolutionofboronandeach spiked sample was ashed in the presence of varying amount of calciumhydroxide.TheresultsofthestudyarepresentedinTable 1.

Table1:Optimizationofsaturatedsolutionofcalciumhydroxide Volumeofsaturatedsolutionof Ca(OH)2,ml Spikedconcentrationofboron,g/l (n=3) %Recovery 5 25 60 200 55 800 52 8 25 75 200 73 800 73 10 25 92 200 94 800 97 12 25 91 200 94 800 95

14000 Emission intensity (c/s) 12000 10000 8000 6000 4000 2000 0 0 100 200 300 400 500 600 700 800 900 Concentration of boron (mcg/l) R = 0.9996
2

Fig.2:Calibrationcurveforconcentrationofboron(g/l)vsemissionintensity(c/s)

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Khandaletal. IntJPharmPharmSci,Vol2,Issue4,182185 Table2:BoroncontentinsamplesofTamsulosinhydrochloride(Resultsarethemeanofthefivereplicates) Sample code Concentrationof boroninreagent blank,g/l 62.79 62.79 62.79 62.79 62.79 Totalconcentration ofboroninthe samplesolution,g/l 78.54 82.13 84.73 81.97 83.69 Observations Concentrationofboron inthesamplesolution duetoTamsulosinHCl, g/l 15.75 19.34 21.94 19.18 20.90 Weightof TamsulosinHCl taken,g 4.00 4.00 4.00 4.00 4.00 Dilutionfactor Boroncontentin thesampleof TamsulosinHCl, ppb 98.44 120.88 137.13 119.875 130.625

A B C D E

25 25 25 25 25

Fromtheresults,itbecomeevidentthat10mlofsaturatedsolution of calcium hydroxide was sufficient enough for more than 90 % recovery of boron at concentration levels between 25 g/l to 800 g/l.Whilestudyingtheeffectofdifferentconcentrationofcalcium on the intensity of boron signal, no significant signal decrease was observedforboron. Analyticaldata For the quantitative determination of boron using ICPOES, conditions were optimized as detailed above so as to get the maximum signal intensity. Boron content in both i.e. sample of Tamsulosin hydrochloride and the reagent blank was determined against a six point calibration curve plotted for the standard solutions of boron ranging from 25 g/l to 800 g/l vs their emission intensity (Figure 2). Results for boron content in five samplesofTamsulosinhydrochloridearegiveninTable2. Methodperformancecharacteristics The method was validated for various parameters as per the InternationalConferenceonHarmonization(ICH)andtheEurachem guidelines. Linearity Asixpointcalibrationcurve(Figure2)forthestandardsolutionsof boronrangingfrom25 g/lto800 g/lvsemissionintensity(c/s) was found to be linear and showed a correlation coefficient of 0.99978. Precision Precisionstudiesofthemethodwerecarriedoutforbothintraday and interday repeatability and reproducibility using replicates of

standard solutions of boron at three different concentrations (25g/l, 200g/l and 400g/l) and using digested samples of Tamsulosin hydrochloride spiked with standard solutions of boron at three different concentrations (25g/l, 200g/l and 400g/l). Data for the precision studies are given in Table 3. Precision (% RSD)of4.9%and4.4%at 25 g/l;4.3%and3.1%at200 g/land 3.2% and 2.5% at 400 g/l was obtained for analysis on the same dayorintradayforthestandardsolutionsofboronandthespiked solutionsrespectively. Precisionfortheinterdayanalysisforthreesubsequentdayswas foundtobe5.0%and4.8%at25 g/l;4.2%and3.6%at200 g/l and3.6%and2.4%at400 g/lforthestandardsolutionsofboron andthespikedsolutionsrespectively.Precisionobtainedforboth interday&intradaywaswellwithintheacceptablelimits. Accuracy For accuracy (recovery) studies standard solutions of boron were spikedinthreesampleofTamsulosinhydrochloride(markedasA,B and C) at the levels of 25g/l, 50g/l and 100g/l. The spiked samplesweredigestedastheproceduredescribedabove. Theboroncontentinreagentblankandthespikedsamplesolutions was determined against the standard calibration curve. Percent recoverywasevaluatedonthebasisofthecomparisonoftheactual concentrationlevelofthespikedsolutionswiththatoftheobserved values of the concentration of boron. Recoveries of more than 90%were obtainedforallthethreespikedsamplesateachspiked level, which is well within the acceptable criteria at trace concentration levels. Data for the accuracy studies are given in Table4.

Table3:IntradayandInterdayprecisionstudies Solutiontype Standardsolutionof Boron TamsulosinHClspiked withstandard solutionsofboron Boronconcentrationin solution,g/l 25 200 400 25 200 400 Intradayprecision(n=6) meanobserved concentration,g/l 24.7 195.6 393.4 24.5 194.3 391.5 %RSD 4.9 4.3 3.2 4.4 3.1 2.5 Interdayprecision(n=6) meanobserved concentration,g/l 24.5 196.2 394.7 24.6 193.6 390.2 %RSD 5.0 4.2 3.6 4.8 3.6 2.4

Table4:RecoverystudiesfordeterminationofboroninTamsulosinhydrochloride Sample code A B C Original concentrationof boroninsample solution,g/l 15.75 19.34 21.94 Spiked concentration,g/l 25 50 100 25 50 100 25 50 100 Totalconcentration, g/l 40.75 65.75 115.75 44.34 69.34 119.34 46.94 71.94 121.94 Meanobserved concentration,g/l (n=3) 38.47 62.73 110.53 42.18 65.52 113.47 44.98 67.33 115.61 %Recovery %RSD

90.88 93.96 94.78 91.36 92.36 94.13 92.16 90.78 93.67

5.0 4.9 3.2 4.8 4.7 3.1 4.9 4.7 2.9

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Specificity Themethodwasevaluatedforspecificitytowardsdetermination of boron in presence of other interferences(suchas nickel,palladium and rhodium), which may also be present as impurities in the sample matrix at different concentration levels. Data for the specificitystudiesaregiveninTable5. In order to determine the specificity of the method, three boron solutionswerepreparedatconcentrationlevelsof25g/l,100 g/l and 800 g/l the concentration of boron was measured using calibration curve. Boron solutions with different concentrations

were then subsequently spiked with nickel (1000 g/l), palladium (1000 g/l) and rhodium (1000 g/l) and the concentration ofthe boron was determined after addition of each of these interfering elements. It was found that impurities of nickel, palladium and rhodiumevenifpresentathighconcentrationsof1000 g/ldidnot interferewiththedeterminationofboronpresentatextremely low concentration (the lowest concentration being 25 g/l) and the recoveriesof boron obtained in all the cases were between 98.3to 102 %, thus, indicating that the method is specific to the determination of trace amount of boron in the presence of nickel, palladiumandrhodium.

Table5:SpecificityofBoroninpresenceofNickel,PalladiumandRhodium Solutioncomposition,g/l B:Boron;Ni:Nickel;Pd:Palladium;Rh: Rhodium B:25 B:25+Ni:1000 B:25+Ni:1000+Pd:1000 B:25+Ni:1000+Pd:1000+Rh:1000 B:100 B:100+Ni:1000 B:100+Ni:1000+Pd:1000 B:100+Ni:1000+Pd:1000+Rh:1000 B:800 B:800+Ni:1000 B:800+Ni:1000+Pd:1000 B:800+Ni:1000+Pd:1000+Rh:1000 Limitofdetection(LOD)andLimitofquantification(LOQ) Limit of detection (LOD) was determined on the basis of signal to noise ratio (S:N) of 3:1 and was calculated as 15 g/l. Limit of quantification(LOQ)wasobtainedas25 g/levaluatedonthebasis of minimum concentration for which a reproducible signal was obtainedwith%RSDlessthan5forfivereplicates. CONCLUSION The experimental data demonstrates that the proposed analytical method is precise, accurate, sensitive and selective for the quantitative determination of boron in raw material of Tamsulosin hydrochloride using Inductively coupled plasma optical emission spectroscopy (ICPOES). The method provides wide linearity, specificity without interference from endogenous impurities like nickel, palladium and rhodium. The developed analytical method also provides excellent recovery at various concentration levels. In additiontoallthesefeatures,oneoftheimportantadvantagesofthe developed method is the simplicity and fast sample preparation procedure.Therefore,themethodcanbeeasilyadoptedforroutine quantitative analysis of boron, present as residual impurity in qualityofrawmaterialsofTamsulosinhydrochloride ACKNOWLEDGEMENTS TheauthorsarethankfultothemanagementofShriramInstitutefor Industrial Research for the guidance and support provided for undertaking the research study. The authors also thank M/S Ranbaxy Research Laboratory for providing the samples of Tamsulosinhydrochlorideforthepurposeofstudy. REFERENCES 1. British Pharmacopoeia 2009, The Stationary Office, British PharmacopoeiaCommission,London,UK:VolumeII,19721974. MeanobservedBoroncontent,g/l (n=3) 25.16 24.70 24.57 25.28 101.10 100.99 98.70 102.00 786.11 806.36 811.42 788.19 2. %RSD 4.48 0.23 1.95 0.77 0.01 0.07 0.48 0.79 0.21 0.04 0.77 0.86 %Recovery 100.6 98.8 98.3 101.1 101.1 101 98.7 102.0 98.3 100.8 101.4 98.5

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