Research Article Development and Validation of a ...downloads.hindawi.com/archive/2014/624635.pdfDevelopment and Validation of a Discriminating In Vitro Dissolution Method for Oral

Research ArticleDevelopment and Validation of a DiscriminatingIn Vitro Dissolution Method for Oral FormulationsContaining Satranidazole

Harshal Ashok Pawar and Pooja Rasiklal Joshi

Dr L H Hiranandani College of Pharmacy Smt CHM Campus Ulhasnagar Maharashtra 421003 India

Correspondence should be addressed to Harshal Ashok Pawar harshaldlhhcopgmailcom

Received 2 January 2014 Accepted 27 March 2014 Published 24 April 2014

Academic Editor Hakan Arslan

Copyright copy 2014 H A Pawar and P R Joshi This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The development of a meaningful dissolution procedure for drug products with limited water solubility has been a challenge tothe pharmaceutical industry Satranidazole (BCS Class II drug) is a new nitroimidazole derivative with potent antiamoebic actionThere is no official dissolution medium available in the literature In the present study parameters such as saturation solubilityin different pH medium dissolution behavior of formulations influence of sink conditions stability and discriminatory effect ofdissolution testing were studied for the selection of a proper dissolution medium Results of solubility data revealed that solubilityof Satranidazole decreases with an increase in pH Satranidazole showed better sink condition in 01 N HCl as compared to othermedia The drug and marketed formulations were stable in the dissolution media used An agitation speed of 75 rpm showed amore discriminating drug release profile than 50 rpm Using optimized dissolution parameters (paddle at 75 rpm 900mL 01 NHCl) greater than 80 of the label amount is released over 60 minutes UV-spectroscopic method used was validated for thespecificity linearity precision robustness and solution stabilityThemethod was successfully applied to granular formulations andalso to marketed tablets containing 300mg Satranidazole

1 Introduction

Satranidazole (STZ) is a new nitroimidazole derivative withpotent antiamoebic action It is used in the treatment ofintestinal and hepatic amoebiasis giardiasis trichomoniasisand anaerobic infections Its dose is 300mg twice daily for3ndash5 days in the treatment of amoebiasis and 600mg as asingle dose in the treatment of giardiasis and trichomoniasisIt is reported that Satranidazole exhibits significantly higherplasma concentrations than metronidazole and has a plasmaelimination half-life of 101 h which is significantly shorterthan the corresponding metronidazole half-life of 362 h [1]Also Satranidazole is having better tolerability absence ofneurological and disulfiram like reactions and it can bepreferred in patients with susceptible neurological symptoms[2]

Dissolution study is particularly important for insolubleor low solubility drugs where absorption is dissolution rate

limited The incorporation of adjuvants (eg diluents lubri-cants and surfactants) into the formulation of a solid oraldosage form can cause significant effects on the dissolutionrate of drugs especially those that are hydrophobic andpoorly soluble [3] In the case of Class 2 drugs in theBiopharmaceutics Classification System (BCS) dissolutionmay be the rate-limiting step for drug absorption so suit-able dissolution tests can be used to predict differences inbioavailability among different formulations [4] The choiceof formulation is often of critical importance in establishinga successful product for oral administration of this class ofdrugs [5] In this context the purpose of the present studywas to evaluate and compare the dissolution profiles of acompounded formulation to that of amarketed product usingSatranidazole as a model drug This drug is poorly solublein water and has high in vitro permeability it is thereforeclassified as BCS Class II [6] To date there is no publisheddissolution test for the evaluation of in vitro release profiles

Hindawi Publishing CorporationInternational Journal of SpectroscopyVolume 2014 Article ID 624635 7 pageshttpdxdoiorg1011552014624635

2 International Journal of Spectroscopy

of this drug from immediate-release solid oral dosage formsThe objective of the present study was to develop a validatedin vitro dissolution method for oral granular formulationcontaining Satranidazole

2 Materials and Methods

21 Materials Satranidazole was obtained as a gift sam-ple from Alkem Laboratories Mumbai Eudragit E100 wasobtained from Evonik Degussa Mumbai Satrogyl tablets(strength 300mg) were purchased from market All thechemicals and reagents used were of analytical grade

22 Formulation Development of Taste Masked STZ GranulesTwo different batches of taste masked granules of STZ wereformed by melt granulation technique using stearic acid andEudragit EPO using different ratio of drug to stearic acidThecomposition of the compounded formulation is mentionedbelowProduct A It is labeled to contain 300mg of the drug and thefollowing excipients stearic acid Eudragit EPO starch 1500magnesium oxide mannitol xylitol sodium carboxymethylcellulose hydroxypropyl cellulose vanilla aspartame andmagnesium stearate with drug to stearic acid ratio (1 2)

Product B It islabeled to contain 300mg of the drug andthe same above mentioned excipients with different drug tostearic acid ratio (1 15)

The assay of the above two products was performed usingpreviously developed and validated HPLC method and thecontents results were used to calculate the percentage releaseon each time of dissolution profile These two products wereused to study the discriminatory power of developedmethod

23 Instrumentation Dissolution test was performed in anElectrolab dissolution test system (TDT-08L) in accordancewith USP Pharmacopoeia general method A double-beamUV-Vis spectrophotometer (Shimadzu 1800 Japan) with10 cmquartz cells was used for all absorbancemeasurementsAll the absorbances were carried out at a UV wavelength of320 nm

24 Saturation Solubility Study The saturation solubility ofSatranidazole was determined in pH 12 (01 N HCl) 21(001N HCl) 45 (acetate buffer) 68 (phosphate buffer) 74(phosphate buffer) and distilled water at 37∘C Excess STZwas added to 100mL of dissolutionmedium in a conical flaskand agitated continuously at room temperature for 8 h on ashakerThe solutions were kept aside for 1 h until equilibriumwas achieved The solutions were then filtered through no 41Whatman filter paper and the filtrate was suitably diluted andanalyzed spectrophotometrically at 320 nm

25 Determination of Sink Conditions and Dissolution Con-ditions For poorly soluble drugs medium selection fordissolution tests is an important step in method validationdue to the difficulty to achieve sink condition [7] which isdefined as the volume of medium at least three times greater

than that required to dissolve the dose of the drug being tested[8]

Sink condition was determined using following equation

Maximum Dissolvable Dose = 119881 times119862119878

Sink (1)

where

119881 = Dissolution medium volume119862119878= Saturated solubility of the compound in the

mediumSink = Sink condition factor

From preselected 01 N HCl media dissolution testingwas performed on granules (119899 = 6) in compliance with USPlt1092gt using paddle (USP-II) The discriminatory power ofthe dissolution method was assessed by analyzing two in-house developed granular formulations of STZ (coded asproducts A and B) of 300mg strength prepared by using dif-ferent composition of excipients Product A contains 600mgstearic acid as a disintegrant whereas product B contains450mg stearic acid Other excipients in both products werethe same

The dissolution rate of STZ from granules of product Awas assessed at 50 and 75 rpm the recommended speeds forUSP apparatus II At 75 rpm product A exhibited a very rapiddissolution without coning

A calibrated dissolution apparatus (USP II) was used withpaddle at 75 rpm and bath temperature maintained at 37 plusmn05∘C Nine hundred millilitre freshly prepared 01 N HClsolution was used as the dissolution medium Dissolutionsamples were collected at 10 15 30 45 and 60min forimmediate-release granules (products A and B) and replacedwith an equal volume of the fresh medium to maintain aconstant total volume After the end of each test time samplesaliquots were filtered diluted in dissolution medium whennecessary and were analyzed by UV at 320 nm At each timepoint a 5mL sample was removed from each vessel andfiltered into labeled glass tubes diluted and analyzed by UVat 320 nm The dissolution of marketed formulation was alsocarried out in same conditions The cumulative release ofdrug was calculated using standard calibration curve of STZprepared in 01 N HCl

26 Comparison of Dissolution Profiles by a Model-Inde-pendent Method The in vitro dissolution data of products Aand B was compared by two-tailed Studentrsquos 119905-test Mooreand Flanner proposed a model-independent mathematicalapproach to compare the dissolution profile using two factors1198911and 119891

2[9]

1198911=

[sum

119899

119905=1

1003816100381610038161003816

119877119905minus 119879119905

1003816100381610038161003816

]

[sum

119899

119905=1

119877119905]

times 100

1198912= 50 times log[1 + (1

119899

)

119899

sum

119905=1

(119877119905minus 119879119905)

2

]

minus05

times 100

(2)

where 119877119905and 119879

119905are the cumulative percentage dissolved

at each of the selected 119899 time points of the reference and

International Journal of Spectroscopy 3

test product respectively The factor 1198911is proportional to

the average difference between the two profiles whereasfactor 119891

2is inversely proportional to the average squared

difference between the two profiles with emphasis on thelarger difference among all the time points The factor 119891

2

measures the closeness between the two profiles Because ofthe nature of measurement 119891

1was described as difference

factor and 1198912as similarity factor [10] FDA has set a public

standard of1198912value between 50 and 100 to indicate similarity

between two dissolution profiles

27 AnalyticalMethodValidation UV-spectroscopicmethodused for estimation of Satranidazole in dissolution sampleswas validated by determining the specificity linearity pre-cision robustness and solution stability according to USPand ICH guidelines [11ndash13]The standard solution containing25 120583gmL of Satranidazole in 01 NHCl was scanned between200 and 400 nm to determine the 120582 max using 01 N HCl asblank in UV spectrophotometer (Shimadzu 1800)

271 Specificity Placebo batch of the granules was preparedin its usual concentration Dissolution was performed sim-ilarly as that of the STZ granules in 900mL of 01 N HCLas dissolution medium using USP apparatus II at 37 plusmn 05∘Cat 75 rpm for 1 hr Aliquots of this solution were filtereddiluted appropriately and analyzed by UV spectrophotomet-ric method

272 Linearity Stock solution of 1000 ppm was preparedby dissolving 50mg drug in 50mL methanol From this2nd stock solution of 100 ppm was prepared in 01 N HCLFurther dilutions were carried out to give solutions of 2 46 8 10 15 25 and 30 120583gmL 01 N HCL was used as blankand absorbances of prepared solutions were noted at 320 nmand calibration curve was constructed Each solution wasprepared in triplicate

273 RecoveryAccuracy A recovery studywas carried out byadding granules equivalent to 80 100 and 120 of drug in thedissolution vessel The dissolution test was done for 60minusing 900mL of dissolution 01 N HCL apparatus 2 rotatingat 75 rpm Aliquots of 5mL were filtered with quantitativefilter and analyzed by UV spectrophotometric method at thefinal concentrations 133 166 and 20120583gmL respectivelyEach concentration was prepared in duplicate and each onewas analyzed in triplicate [14]

274 Method PrecisionRepeatability It was performed on 6doses of granules from same batch and sampleswere analyzedaccording to the test method and aliquots were taken at theend of 60 minutes [15]

Intermediate precision was evaluated to determine theeffects of random events on the precision of the analyticalprocedure This was done by performing the dissolution onsame batch of granules on different day by changing theanalyst

Table 1 pH dependent solubility of Satranidazole in different buffersolutions

pH Solubility (mgmL)lowast Sink condition12 (01 N HCl) 15125 4537521 (001N HCl) 11985 3595545 (acetate buffer) 09813 2943968 (phosphate buffer) 08251 2475374 (phosphate buffer) 09825 29475Distilled water 08133 24399lowastMean of 3 determinations

275 Robustness Robustness was studied by changing thewavelength of UV spectrophotometer at 320 plusmn 2 nm and alsoby changing agitation rate of dissolution apparatus at 75 plusmn2 rpm

276 Solutions Stability To evaluate solution stability thesample solution was stored at room temperature and wasanalyzed by UV spectrophotometer for 24 hrs at various timeintervals

3 Results and Discussions

For immediate-release formulations apparatus 1 (baskets) at100 rpm or apparatus 2 (paddles) at 50 or 75 rpm is mostcommonly used Other agitation speeds and apparatus areacceptable with appropriate justification For dosage formsthat exhibit coning (mounding) under the paddle at 50 rpmthe coning can be reduced by increasing the paddle speed to75 rpm thus reducing the artifact and improving the data

Reference compendia and guidelines of Food DrugAdministration United States Pharmacopeia FederationInternational Pharmaceutique World Health OrganizationEuropean Pharmacopoeia and Japanese Pharmacopoeia rec-ommend the use of rotating paddle between 50 and 100 rpmwith volume of 500 to 1000mL along with surfactant toprovide sink condition for insoluble drug products [16]

The most common way to check the discriminatorypower of the method is to test formulations with differencesresulting forms changes in the characteristics of the APIdrug product composition product manufacturing processand stability conditions [10 17ndash19]

Drug solubility and solution stability are importantproperties to be considered when selecting the dissolutionmedium From the saturation solubility data and establishedsink conditions (Table 1) it was found that pH 12 (01 NHCL)was better dissolution medium for Satranidazole

The solution stability data is represented in Table 2 Thesolution of drug was found to be stable in 01 N HCL for 24hours The cumulative RSD obtained was less than 2 at theend of 24 hrs

The stirring speeds of 50 rpm and 75 rpm for productA were tested The statistical Student 119905-test was applied tocompare the percent drug release using 50 or 75 rpm forgranules (Table 3) The 119875 value for granules was smallerthan the delineated significance level indicating that thereis statistically significant difference between the drug release


Table 2 Stability data in 01 N HCL

Time Absorbance Mean SD RSD30 minutes 0757 mdash mdash mdash1hr 0758 07575 000071 00932 hrs 0752 075567 000321 04254 hrs 0759 07565 000311 04118 hrs 0745 07542 000581 077016 hrs 0734 075083 000975 129824 hrs 0745 075 000917 1222

Table 3 Product A dissolution tests results (119899 = 12) using differentstirring speeds and HCl 01 N as dissolution medium

Time in minutes Cumulative release119905-test 119875

50 rpm 75 rpm0 0 0

44858 00065

5 591 86510 1757 331515 3962 452930 4588 666745 6043 764860 7138 8759

Table 4 Comparison of dissolution profile of two products (A andB) with different ratio of drug stearic acid

Time (minutes)Product A

Drug stearic acid(1 2)

Product BDrug stearic acid

(1 15)0 0 05 871 plusmn 0426 802 plusmn 051210 339 plusmn 0299 5299 plusmn 06915 4605 plusmn 0343 6268 plusmn 048930 6583 plusmn 0378 7340 plusmn 039745 7857 plusmn 0418 8166 plusmn 043460 8765 plusmn 0404 9001 plusmn 0546Similarity factor 119891

2

46Difference factor 119891

1

17

Table 5 Linearity data

Concentration (in mcgmL) Absorbance2 008224 014336 021758 0279310 0339815 0505720 0630125 0751830 09204

percent and suggested that the stirring speed of 75 rpm isbetter than 50 rpm

04

03

02

01

0

minus01

Abs

200 250 300 350 400

Wavelength (nm)

Figure 1 UV spectrum of STZ in 01 N HCl

Table 6 Results of accuracy

Addedamount (120583g)

Recoveredamount (120583g) Recoverylowast () Mean () RSD

133 (80) 1293 9721 plusmn 12430991133 190166 (100) 1646 9915 plusmn 13256

20 (120) 20196 10098 plusmn 15234lowastEach value is the mean of 3 determinations

On comparison of dissolution of products A and B the 1198911

value was found to be 46 and 1198912value was found to be 17 as

shown in Table 4 indicating dissimilarity between productsA and B It can be concluded that the drug release profile at75 rpm detected small changes in drug product composition

The 120582maxwas found to be 320 nm in dissolutionmediumand hence it was taken as the analytical wavelength The UVspectrum of STZ in 01 N HCl is shown in Figure 1

31 Specificity It was found that dissolution of placebo washaving no effect on dissolution of Satranidazole granulesAlso the excipients used for the formulation of granules werenot showing interference with the maximum absorption ofdrug

32 Linearity The calibration curve of STZ in 01 NHCLwasplotted as shown in Figure 2 The correlation coefficient wasfound to be 0997 These data indicated good linearity of STZin the range of 2ndash30120583gmL as shown in Table 5

Limit of quantitation (LOQ) is the lowest amount ofanalyte in a sample that can be determined with acceptableprecision and accuracy under stated experimental conditionsThe quantitation limit is expressed as the concentration ofanalyte in the sample The standard deviation and relatedstandard deviation for the limit of quantitation were wellwithin the desirable limit of no more than 20 The lowestquantifiable concentration was 522mcgmL and this param-eter can be used for predicting the drug release in low doseformulation


Table 7 Method precisionrepeatability

Sample number cumulative release at 60min

Day 1 Day 2Analyst 1 Analyst 2

1 8709 88342 8623 89233 8724 87894 8878 86215 8765 87476 8894 8701Mean 87655 87691Standard deviation (SD) 1042 1052Relative standard deviation ( RSD) 1189 1199Mean 87673Standard deviation (SD) 0999Relative standard deviation ( RSD) 1139

Table 8 Change in wavelength

Time (minutes) At 318 At 320 At 322Average release plusmn SD (119899 = 6)

0 0 05 895 plusmn 05456 8512 plusmn 07678 8398 plusmn 0678910 3401 plusmn 04345 9823 plusmn 06678 9731 plusmn 0801215 4425 plusmn 05987 9981 plusmn 08978 9812 plusmn 0561230 6687 plusmn 06745 10129 plusmn 07809 9924 plusmn 0676945 7712 plusmn 07568 7684 plusmn 05982 7746 plusmn 0814560 8593 plusmn 04768 8709 plusmn 03780 8576 plusmn 04568Average at 60min 8626 plusmn 06744 RSD at 60min 0839

0 10 20 30 40

AbsorbanceLinear (absorbance)

0

01

02

03

04

05

06

07

08

09

1

y = 00294x + 00375

R2 = 09975

Figure 2 Linearity curve of STZ in 01 N HCl

33 Accuracy According to USP guidelines the recoveryfor a dissolution test must be in the range of 95ndash105 Thepercent recovery was from 9721 to 10098 The accuracyof the method is acceptable from the results seen in Table 6

34 Method Precision and Intermediate Precision The per-cent RSD did not exceed 2 for the repeatability andintermediate precision demonstrating suitable precisionTheresults are represented in Table 7

35 Robustness The percent RSD values were within thespecified limit of 2 indicating the robustness of dissolutionmethodon changing the agitation rate and also ofUVmethodby changing the wavelength The results are shown in Tables8 and 9

36 Evaluation of Marketed Formulation By using aboveoptimized dissolution conditions the dissolution ofmarketedSTZ film coated tablet was performed The satisfactory drug releasewas obtained at the endof 60minutesThe resultsare shown in Table 10


Table 9 Change in agitation rate

Time (minutes) At 73 rpm At 75 rpm At 77 rpmAverage release plusmn SD (119899 = 6)

0 0 0 05 898 plusmn 03987 817 plusmn 05679 821 plusmn 0501210 3518 plusmn 05671 3693 plusmn 05712 3394 plusmn 0678215 4445 plusmn 06745 4281 plusmn 07986 4524 plusmn 0586730 6623 plusmn 07681 6981 plusmn 06897 6687 plusmn 0613945 7656 plusmn 08142 7426 plusmn 05893 7843 plusmn 0783160 8732 plusmn 04587 8823 plusmn 06897 8842 plusmn 04879Average at 60min 8799 plusmn 07633 RSD at 60min 0668

Table 10 Dissolution of marketed tablet

Time (minutes) cumulative release0 05 934 plusmn 043210 2887 plusmn 047115 391 plusmn 057230 6543 plusmn 050345 8366 plusmn 06660 881 plusmn 055

4 Conclusion

Thedissolution test developed and validated for STZ granuleswas considered satisfactory The conditions that allowed thedissolution determination were USP apparatus II (paddle)with 01 N HCl dissolution medium at 370 plusmn 05∘C rotatingat a speed of 75 rpm STZ was found to be stable for24 hrs indicating good stability of the drug in dissolutionmedium The validated method was found to be specificlinear precise and accurate The stated analytical methodcan be successfully used for in vitro dissolution and routineanalysis of samples for STZ granules and also marketed STZtablets

Conflict of InterestsThe authors declare that there is no conflict of interestsregarding the publication of this paper

AcknowledgmentAuthors are very much thankful to Dr P S Gide Principal ofHyderabad Sindh National Collegiate Boards (HSNCB) DrL H Hiranandani College of Pharmacy Ulhasnagar for hiscontinuous support guidance and encouragement

References

[1] A Pargal C Rao K K Bhopale K S Pradhan K BMasani and C L Kaul ldquoComparative pharmacokinetics andamoebicidal activity of metronidazole and satranidazole in the

golden hamsterMesocricetus auratusrdquo Journal of AntimicrobialChemotherapy vol 32 no 3 pp 483ndash489 1993

[2] D M Parmar and S P Jadav ldquoThe concept of personal drugs inthe undergraduate pharmacology practical curriculumrdquo IndianJournal of Pharmacology vol 39 no 3 pp 165ndash167 2007

[3] M E Aulton Pharmaceutics The Science of Dosage FormDesign Churchill LivingstoneNewYorkNYUSA 2nd edition2001

[4] US Department of Health and Human Services Food andDrug Administration Dissolution Testing of Immediate ReleaseSolidOralDosage Forms Guidance for Industry Center forDrugEvaluation and Research (CDER) US Government PrintingOffice Washington DC USA 1997

[5] C W Pouton ldquoFormulation of poorly water-soluble drugs fororal administration physicochemical and physiological issuesand the lipid formulation classification systemrdquo European Jour-nal of Pharmaceutical Sciences vol 29 no 3-4 pp 278ndash2872006

[6] EPAR ldquoAcomplia scientific discussionrdquo European PublicAssessment Report EMEAHC666 European MedicinesAgency London UK 2006

[7] M Zahirul and I Khan ldquoDissolution testing for sustained orcontrolled release oral dosage forms and correlation with invivo data challenges and opportunitiesrdquo International Journalof Pharmaceutics vol 140 no 2 pp 131ndash143 1996

[8] ldquoPharmacopeial previewsrdquo Pharmacopeial Forum vol 30 pp351ndash363 2004

[9] J W Moore and H H Flanner ldquoMathematical comparison ofcurves with an emphasis on in vitro dissolution profilesrdquo Phar-maceutical Technology vol 20 no 6 pp 64ndash74 1996

[10] V P Shah Y Tsong P Sathe and J Liu ldquoIn vitro dissolutionprofile comparisonmdashstatistics and analysis of the similarityfactor f2rdquo Pharmaceutical Research vol 15 no 6 pp 889ndash8961998

[11] ldquoThe United States Pharmacopeiardquo Pharmacopeial ForumUSP-32 NF-27 31th edition 2009

[12] ldquoICH harmonized tripartite guideline validation of analyticalprocedure Q2Ardquo in Proceedings of the International Conferenceon Harmonisation (ICH rsquo94) March 1994

[13] ldquoICH harmonized tripartite guideline validation of analyticalprocedure Q2Brdquo in Proceedings of the International Conferenceon Harmonisation (ICH rsquo94) March 1994

[14] L C Vaucher C S Palm A D Lange and E E S Schapo-val ldquoDevelopment and validation of a dissolution test for


telithromycin in coated tabletsrdquo Quimica Nova vol 32 no 5pp 1329ndash1333 2009

[15] A P Kulkarni S Mohd Z Zaheer and M H G DehghanldquoDevelopment and validation of a dissolution method forpioglitazonetabletsrdquo Dissolution Technologies In press

[16] C K Brown H P Chokshi B Nickerson R A Reed B RRohrs and P A Shah ldquoAcceptable analytical practices for dis-solution testing of poorly soluble compoundsrdquo PharmaceuticalTechnology vol 28 no 12 pp 56ndash65 2004

[17] C Noory N Tran L Ouderkirk and V Shah ldquoSteps for devel-opment of a dissolution test for sparingly water-soluble drugproductsrdquoDissolution Technologies vol 7 no 1 pp 16ndash18 2000

[18] M Siewert ldquoFIP guidelines for dissolution testing of solid oralproductsrdquoPharmazeutische Industrie vol 57 no 5 pp 362ndash3691995

[19] ldquoInternational Pharmaceutical Federation (FIP) guidelines fordissolution testing of solid oral productsrdquo Drug InformationJournal vol 30 pp 1071ndash1084 1996

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Analytical Methods in Chemistry

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Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Applied ChemistryJournal of



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Spectroscopy

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Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


of this drug from immediate-release solid oral dosage formsThe objective of the present study was to develop a validatedin vitro dissolution method for oral granular formulationcontaining Satranidazole

2 Materials and Methods

21 Materials Satranidazole was obtained as a gift sam-ple from Alkem Laboratories Mumbai Eudragit E100 wasobtained from Evonik Degussa Mumbai Satrogyl tablets(strength 300mg) were purchased from market All thechemicals and reagents used were of analytical grade

22 Formulation Development of Taste Masked STZ GranulesTwo different batches of taste masked granules of STZ wereformed by melt granulation technique using stearic acid andEudragit EPO using different ratio of drug to stearic acidThecomposition of the compounded formulation is mentionedbelowProduct A It is labeled to contain 300mg of the drug and thefollowing excipients stearic acid Eudragit EPO starch 1500magnesium oxide mannitol xylitol sodium carboxymethylcellulose hydroxypropyl cellulose vanilla aspartame andmagnesium stearate with drug to stearic acid ratio (1 2)

Product B It islabeled to contain 300mg of the drug andthe same above mentioned excipients with different drug tostearic acid ratio (1 15)

The assay of the above two products was performed usingpreviously developed and validated HPLC method and thecontents results were used to calculate the percentage releaseon each time of dissolution profile These two products wereused to study the discriminatory power of developedmethod

23 Instrumentation Dissolution test was performed in anElectrolab dissolution test system (TDT-08L) in accordancewith USP Pharmacopoeia general method A double-beamUV-Vis spectrophotometer (Shimadzu 1800 Japan) with10 cmquartz cells was used for all absorbancemeasurementsAll the absorbances were carried out at a UV wavelength of320 nm

24 Saturation Solubility Study The saturation solubility ofSatranidazole was determined in pH 12 (01 N HCl) 21(001N HCl) 45 (acetate buffer) 68 (phosphate buffer) 74(phosphate buffer) and distilled water at 37∘C Excess STZwas added to 100mL of dissolutionmedium in a conical flaskand agitated continuously at room temperature for 8 h on ashakerThe solutions were kept aside for 1 h until equilibriumwas achieved The solutions were then filtered through no 41Whatman filter paper and the filtrate was suitably diluted andanalyzed spectrophotometrically at 320 nm

25 Determination of Sink Conditions and Dissolution Con-ditions For poorly soluble drugs medium selection fordissolution tests is an important step in method validationdue to the difficulty to achieve sink condition [7] which isdefined as the volume of medium at least three times greater

than that required to dissolve the dose of the drug being tested[8]

Sink condition was determined using following equation

Maximum Dissolvable Dose = 119881 times119862119878

Sink (1)

where

119881 = Dissolution medium volume119862119878= Saturated solubility of the compound in the

mediumSink = Sink condition factor

From preselected 01 N HCl media dissolution testingwas performed on granules (119899 = 6) in compliance with USPlt1092gt using paddle (USP-II) The discriminatory power ofthe dissolution method was assessed by analyzing two in-house developed granular formulations of STZ (coded asproducts A and B) of 300mg strength prepared by using dif-ferent composition of excipients Product A contains 600mgstearic acid as a disintegrant whereas product B contains450mg stearic acid Other excipients in both products werethe same

The dissolution rate of STZ from granules of product Awas assessed at 50 and 75 rpm the recommended speeds forUSP apparatus II At 75 rpm product A exhibited a very rapiddissolution without coning

A calibrated dissolution apparatus (USP II) was used withpaddle at 75 rpm and bath temperature maintained at 37 plusmn05∘C Nine hundred millilitre freshly prepared 01 N HClsolution was used as the dissolution medium Dissolutionsamples were collected at 10 15 30 45 and 60min forimmediate-release granules (products A and B) and replacedwith an equal volume of the fresh medium to maintain aconstant total volume After the end of each test time samplesaliquots were filtered diluted in dissolution medium whennecessary and were analyzed by UV at 320 nm At each timepoint a 5mL sample was removed from each vessel andfiltered into labeled glass tubes diluted and analyzed by UVat 320 nm The dissolution of marketed formulation was alsocarried out in same conditions The cumulative release ofdrug was calculated using standard calibration curve of STZprepared in 01 N HCl

26 Comparison of Dissolution Profiles by a Model-Inde-pendent Method The in vitro dissolution data of products Aand B was compared by two-tailed Studentrsquos 119905-test Mooreand Flanner proposed a model-independent mathematicalapproach to compare the dissolution profile using two factors1198911and 119891

2[9]

1198911=

[sum

119899

119905=1

1003816100381610038161003816

119877119905minus 119879119905

1003816100381610038161003816

]

[sum

119899

119905=1

119877119905]

times 100

1198912= 50 times log[1 + (1

119899

)

119899

sum

119905=1

(119877119905minus 119879119905)

2

]

minus05

times 100

(2)

where 119877119905and 119879

119905are the cumulative percentage dissolved

at each of the selected 119899 time points of the reference and






2




























50 rpm 75 rpm0 0 0

44858 00065

5 591 86510 1757 331515 3962 452930 4588 666745 6043 764860 7138 8759






2


1

17




04

03

02

01

0

minus01

Abs

200 250 300 350 400

Wavelength (nm)





133 (80) 1293 9721 plusmn 12430991133 190166 (100) 1646 9915 plusmn 13256

















0 10 20 30 40


0

01

02

03

04

05

06

07

08

09

1

y = 00294x + 00375

R2 = 09975












4 Conclusion




References
































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of






2




























50 rpm 75 rpm0 0 0

44858 00065

5 591 86510 1757 331515 3962 452930 4588 666745 6043 764860 7138 8759






2


1

17




04

03

02

01

0

minus01

Abs

200 250 300 350 400

Wavelength (nm)





133 (80) 1293 9721 plusmn 12430991133 190166 (100) 1646 9915 plusmn 13256

















0 10 20 30 40


0

01

02

03

04

05

06

07

08

09

1

y = 00294x + 00375

R2 = 09975












4 Conclusion




References
































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of






50 rpm 75 rpm0 0 0

44858 00065

5 591 86510 1757 331515 3962 452930 4588 666745 6043 764860 7138 8759






2


1

17




04

03

02

01

0

minus01

Abs

200 250 300 350 400

Wavelength (nm)





133 (80) 1293 9721 plusmn 12430991133 190166 (100) 1646 9915 plusmn 13256

















0 10 20 30 40


0

01

02

03

04

05

06

07

08

09

1

y = 00294x + 00375

R2 = 09975












4 Conclusion




References
































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of









0 10 20 30 40


0

01

02

03

04

05

06

07

08

09

1

y = 00294x + 00375

R2 = 09975












4 Conclusion




References
































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of







4 Conclusion




References
































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

















Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of










Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

Documents

Research Article Development and Validation of a ...downloads.hindawi.com/archive/2014/624635.pdfDevelopment and Validation of a Discriminating In Vitro Dissolution Method for Oral