Development and validation of RP-HPLC method for determination of acetazolamide,furosemide and phenytoin extemporaneous suspensions

Sirilk Rkrod Srote Ornlor

aUbon Ratchathani University,Ubon Ratchathani 34190,Thailand

bDivision of Pharmacy,Sappasit Prasong Hospital,Ubon Ratchathani,Thailand

Development and validation of RP-HPLC method for determination of acetazolamide,furosemide and phenytoin extemporaneous suspensions

Warisada Sila-ona,*,Salinee Na-Ranonga,Sirilak Rakroda, Sarote Ornlaorb,Zongporn Joungmunkonga

aUbon Ratchathani University,Ubon Ratchathani 34190,Thailand

bDivision of Pharmacy,Sappasit Prasong Hospital,Ubon Ratchathani,Thailand

A R T I C L E I N F O

Article history:

Available online 25 November 2015

Development and validation of RP

HPLC method

Extemporaneous suspensions

The development of the extemporaneous preparations allows physicians to adjust the dose for pediatric patients and provides for a more convenient dosage vehicle for those patients with diffculty swallowing tablets[1].As such,the production unit of pharmacy division,Sappasit Prasong Hospital,Ubon Ratchathani province,prepared the extemporaneous formulations such as Acetazolamide(AM),Furosemide(FM)and Phenytoin(PT)powder for suspensions.The extemporaneous suspensions of 10 mg/mL AM,2 mg/mL FM and 10 mg/mL PT were prepared from 250 mg Diamox?,40 mg Lasix?and 50 mg Dilantin?tablets,respectively and diluted with syrup vehicle. The syrup vehicle is composed of appropriate sweetening agents,viscosity inducing agent,favoring agent and preservative.To quantify drugs in the multicomponent formulas,high performance liquid chromatography(HPLC)was performed since it can be selective and sensitive[2].Thus,the objective of this study was to develop and validate a stability-indicating HPLC method for AM,FM and PT in their extemporaneous suspensions.The obtained data would be used to further study of the stability assessment of these 3 extemporaneous suspensions.

The optimal chromatographic conditions for each analysis were modifed from USP[3].The chromatographic separation of AM/FM and PT was performed on a BDS Hypersil column (C18Thermo,4.6×250 mm)and Symmetry column(C8Waters, 3.9×150 mm),respectively.The solution of 0.5 N NaOH was the solvent extraction of AM from suspended matrix,while FM and PT were extracted by using the mixture of acetonitrile/water/ glacial acetic acid(22:22:1 ratio)and methanol,respectively.The mobile phase for AM,FM and PT quantifcation composted correspondingly of 0.05 M sodium acetate anhydrous,acetonitrile and methanol(475:30:20 ratio);acetonitrile and 1/99 glacial acetic acid in water(38:62 ratio);and water,methanol,acetonitrile,triethylamine solution and acetic acid(500:270:230:5:1 ratio).All mobile phases were performed under isocratic conditions,at a fow rate of 1 mL/min with an injection volume of 20 μL.All samples were analyzed using UV detector at the wavelength of 254 nm.

The validation method was carried out assessing parameters such as precision,accuracy,limit of detection(LOD),limit of quantifcation(LOQ)and linearity.The precision was evaluated by repeatability of three concentrations of each drug and the results were expressed as relative standard deviation(RSD). The%RSD of AM,FM and PT were 1.45,0.74 and 0.98,respectively.These indicated that the method proposed presents good precision for the analysis of these three drugs in syrup vehicle regarding the lowering of%RSD(less than 2%).The accuracy was evaluated by means of recovery assessments at three concentration levels and expressed as%recoveries.The%recoveries and%RSD obtained for AM(112.13%,1.69%),FM(1.56%)and PT(1.68%)were within acceptable limits(80–110%)and revealed that the chromatographic method has good accuracy in the analysis of drugs in syrup.The LOD and LOQ of AM (0.0225,0.0684 mg/mL),FM(0.0071,0.0215 mg/mL)and PT(0.0285, 0.0866 mg/mL)indicated that the optimized method is desirable for determination at low concentration of these three drugs in syrup vehicle.Linearity was evaluated on a calibration curve constructed with standard solutions of drugs in the range of 0.050–0.250 mg/mL(AM),0.1–0.5 mg/mL(FM)and 0.25–1.5 mg/mL (PT).Correlation coeffcients for all drugs were greater than 0.990.The data obtained revealed that the HPLC conditions of all 3 drugs were reasonable and led to be a condition for the further stability study of extemporaneous suspensions.

Acknowledgments

This work was fully supported by the grant of Faculty of Pharmaceutical Sciences,Ubon Ratchathani University and Sappasit Prasong Hospital for the extemporaneous suspensions.

R E F E R E N C E S

[1]Viriyarote A.Extemporaneous oral liquid preparations.TJPS 2007;2(3):320–326.

[2]Sila-on W,Sungkapat S,Anansuchartkul J,et al.Physical and chemical stability study of morphine sulfate syrup.IJPS 2008;4(2):134–139.

[3]The United States Pharmacopeia 32 and NF 27.Rockville,MD: The United States Pharmacopeial Convention,Inc.;2009.

*E-mail address:warisada.s@ubu.ac.th.

Peer review under responsibility of Shenyang Pharmaceutical University.

http://dx.doi.org/10.1016/j.ajps.2015.11.087

1818-0876/?2016 Production and hosting by Elsevier B.V.on behalf of Shenyang Pharmaceutical University.This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/).