DEVELOPMENT AND VALIDATION OF A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR DETERMINING METFORMIN HYDROCHLORIDE IN DRIED BLOOD SPOTS


Yahdiana Harahap, Prastiwi Arum Sari, Catur Jatmika

Abstract


Objective: This study aimed to develop an analytical method for determining metformin concentrations in human dried blood spots (DBS) using
optimal 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 acetonitrile
phosphate 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 atorvastatin
calcium as an internal standard. Samples were prepared by protein precipitation with 60% methanol and then drying using nitrogen gas at 60°C for
15 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 was
also 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/mL
and can be applied to in vivo studies.


Keywords


Metformin, High-performance liquid chromatography, Optimization, Validation, Dried blood spot.

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About this article

Title

DEVELOPMENT AND VALIDATION OF A HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR DETERMINING METFORMIN HYDROCHLORIDE IN DRIED BLOOD SPOTS

Keywords

Metformin, High-performance liquid chromatography, Optimization, Validation, Dried blood spot.

DOI

10.22159/ijap.2018.v10s1.26

Date

20-12-2018

Additional Links

Manuscript Submission

Journal

International Journal of Applied Pharmaceutics
Vol. 10, Special Issue 1 (Dec.), 2018 Page: 126-129

Online ISSN

0975-7058

Authors & Affiliations

Yahdiana Harahap
Bioavailability and Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.

Prastiwi Arum Sari
Bioavailability and Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.

Catur Jatmika
Bioavailability and Bioequivalence Laboratory, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.


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