Open AccessDEVELOPMENT AND VALIDATION OF A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR DETERMINING METFORMIN HYDROCHLORIDE IN DRIED BLOOD SPOTS
Author(s) -
Yahdiana Harahap,
Prastiwi Arum Sari,
Catur Jatmika
Publication year - 2018
Publication title -
international journal of applied pharmaceutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.238
H-Index - 15
ISSN - 0975-7058
DOI - 10.22159/ijap.2018.v10s1.26
Subject(s) - chromatography , bioanalysis , protein precipitation , chemistry , metformin hydrochloride , dried blood , precipitation , high performance liquid chromatography , analytical chemistry (journal) , metformin , medicine , physics , meteorology , diabetes mellitus , endocrinology
Objective: This study aimed to develop an analytical method for determining metformin concentrations in human dried blood spots (DBS) usingoptimal chromatographic conditions and DBS preparation methods and validate the analytical methods.Methods: Analysis metformin HCl performed using C-18 columns (250 mm × 4.6 mm) at a column temperature of 40°C with a mobile phase acetonitrilephosphate buffer (40:60% v/v) at pH 7.0 using a flow rate of 0.8 mL/min, a photodiode array detector at a wavelength of 234 nm, and atorvastatincalcium as an internal standard. Samples were prepared by protein precipitation with 60% methanol and then drying using nitrogen gas at 60°C for15 min before reconstituting in 200 μL of the mobile phase.Results: Invalidation procedures, the present methods met the acceptance criteria stated in the EMEA Bioanalytical Guideline 2011. The method wasalso linear at the concentration range of 25.0–5000.0 ng/mL (r=0.9997).Conclusion: The developed method for determination of metformin HCl in DBS sample was valid with a concentration range of 25.0–5000.0 ng/mLand can be applied to in vivo studies.