QUANTITATION OF METFORMIN IN URINE BY RP-HPLC METHOD AND ITS APPLICATION IN PHARMACOKINETICS
Urine Metformin method
Objective: To develop and validate an easy and sensitive HPLC method for quantitation of metformin in urine.
Materials and Methods: The technique involved deproteinisation of urine sample with methanol and analysis of the supernatant the usage of Zorbax 300 – SCX, 4.6 X 150mm ID, 5 µm particle size and UV detection at a wavelength of 233 nm.
Results: The assay was specific for Metformin and linear from 1.25 to 50.0μg/ml. The relative well known deviation of intra- and inter-day assays was lesser than 7%. The recovery of Metformin from urine ranged from 97- 103%.
Conclusions: An easy and sensitive HPLC approach for quantitation of Metformin in urine has been developed. Due to its simplicity in sample preparation and instrumentation, this technique can be used for pharmacokinetic studies of metformin in urine samples.
1) Nelson R, Spann D, Elliott D, Brondos A, Vulliet R: Evaluation of the Oral Antihyperglycemic Drug Metformin in Normal and Diabetic Cats. J Vet Intern Med, 2004:18–24.
2) Scheen AJ: Clinical pharmacokinetics of metformin. Clin Pharmacokinet. 1996:30(5), 359–1.
3) Shah SA, Rathod IS, Savale SS, Patel BD: Development of a sensitive high-performance thin-layer chromatography method for estimation of ranitidine in urine and its application for bioequivalence decision for ranitidine tablet formulations. J Chromatogr B Analyt Technol Biomed Life Sci.2002:767(1), 83-91.
4) T?ma P: Large volume sample stacking for rapid and sensitive determination of antidiabetic drug metformin in human urine and serum by capillary electrophoresis with contactless conductivity detection. J Chromatogr A. 2014:1345, 207–211.
5) Skrzypek S, Mirceski V, Ciesielski W, Soko?owski A, Zakrzewski R: Direct determination of metformin in urine by adsorptive catalytic square-wave voltammetry. J Pharm Biomed Analysis.2007:45, 275–281.
6) Charles BG, Jacobsen NW, Ravenscroft PJ: Rapid liquid-chromatographic determination of metformin in plasma and urine. Clin Chem.1981:27, 434-6.
7) Benzi L, Marchetti P, Cecchetti P, Navalesi R: Determination of metformin and phenformin in human plasma and urine by reversed-phase high-performance liquid chromatography. J. Chromatogr. 1986:375, 184-9.
8) El-Gindy A, Nassar MW, El-Abasawy NM, Attia KA, Al-Shabrawi M: Optimization and Validation of an RP-HPLC Method for Direct Determination of Metformin Hydrochloride in Human Urine and in a Dosage Form. J AOAC Int. 2010:93(6), 1821-8.
9) Tong R, Qi X, Ding L, Song Q: Analysis of Urine Excretion of Metformin-Glipizide Tablet after Oral Administration in Healthy Human. Tianjin Med J. 2012:40(1), 7-11 (2012).
10) Zhang X, Wang X, Vernikovskaya DI, Fokina VM, Nanovskaya TN, Hankins GD et al: Quantitative determination of metformin, glyburide and its metabolites in plasma and urine of pregnant patients by LC-MS/MS. Biomed Chromatography. 2015:29(4),560-9.
11) Matin SB, Karam JH, Forshan PH: Simple electron capture gas chromatographic method for the determination of oral hypoglycemic biguanides in biological fluids. Anal Chem. 1975:47(3), 545–548.
12) Ross MSF: Determination of metformin in biological fluids by derivatization followed by high-performance liquid chromatography. J Chromatography. 1977:133, 408-411.
13) Lennard MS, Casey C, Tucker GT, Woods HF: Determination of metformin in biological samples. Br J Clinical Pharmacology. 1978: 6(2), 183–185.
14) Alamgir M, Hayat A, Majidano AA, Khuhawaret M.Y: Spectrophotometric Determination of Metformin in Pharmaceutical Preparations, Serum and Urine using Benzoin as Derivatizing Reagent. J Chem Soc Pak.2014:36(2), 344-349.
15) Deng B, Shi A, Kang Y, Li L: Determination of metformin hydrochloride using precolumn derivatization with acetaldehyde and capillary electrophoresis coupled with electrochemiluminescence. Luminescence. 2011:26(6), 592–597.
16) Mary Rebecca Y, Sudha V and Hemanth Kumar A K: A Validated High Performance Liquid Chromatography Method for the Determination of Metformin in Human Plasma and Its Application to Pharmacokinetic Study. Chromatography and Separation Techniques Journal. 2019:2(1):119.
17) Lennard MS, Casey C, Tucker GT, Woods HF: Determination of metformin in biological samples. Br J Clinical Pharmacology.1978:6(2), 183–185.
18) Tahara K, Yonemoto A, Yoshiyama Y, Nakamura T, Aizawa M et al.:Determination of antihyperglycemic biguanides in serum and urine using an ion-pair solid-phase extraction technique followed by HPLC-UV on a pentafluorophenylpropyl column and on an octadecyl column. Biomed Chromatography. 2006:20, 1200–1205.
19) Liu Y, Huang L, Wang J, Ding C, Xu Sh, Sun F,et al. Simultaneous Determination of Three Diabetic Drugs in Urine by High Performance Liquid Chromatography. J Instr Analysis, Fenxi Ceshi Xuebao. 2011:30(3), 293-297.
20) Nielsen F, Christensen Mette MH, Brøsen K: Quantitation of Metformin in Human Plasma and Urine by Hydrophilic Interaction Liquid Chromatography and Application to a Pharmacokinetic Study. Ther Drug Monitor.2014:36(2), 211–217.
21) El-Kimary Eman EI, Youssef RM, Allam: A High-Performance Thin-Layer Chromatographic Assay of Metformin in Urine Using Ion-Pair Solid-Phase Extraction: Application for Bioavailability and Bioequivalence Study of New Microbeads Controlled Release Formulation. J Planar Chromatogr – Mod TLC. 2014:27(5),377-384.
22) Garrett ER, Tsau J, Hinderling PH: Application of ion-pair methods to drug extraction from biological fluids. II. Quantitative determination of biguanides in biological fluids and comparison of protein binding estimates. J Pharm Sci. 1972:61(9),1411–1418.
23) Keal J, Somogyi A: Rapid and sensitive high-performance liquid chromatographic assay for metformin in plasma and urine using ion-pair extraction techniques. J Chromatography B Biomed Appl. 1986:378, 503-508.
24) Gabr RQ, Padwal RS, Brocks DR: Determination of metformin in human plasma an urine by high performance liquid chromatography using small sample volume and conventional octadecyl silane column. Pharm Sci. 2010:13, 486–494.
25) Eva Troja,Leonard Deda,Gezim Bocari: Ion pair HPLC method for the quantification of metformin in human urine.journal of applied bioanalysis. 2016:p.16-24.
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