J Korean Soc Clin Pharmacol Ther JKSCPT Journal of Korean Society for Clinical Pharmacology and Therapeutics 1225-5467 Korean Society for Clinical Pharmacology and Therapeutics 10.0000/jkscpt.2012.20.2.175 Original Article Targeted Plasma Metabolite Profiling of Metformin in Healthy Korean Volunteers Lihm Ho-Seob 1 † Cha Jaemin 2 3 † Seo Jeong Ju 2 3 Park Jeonghyeon 2 3 Lee Joomi 2 3 Lee Hae Won 3 Bae Kyun Seop 4 Kim Woomi 5 Yoon Young-Ran 2 3 Department of Family Medicine, Kosin University College of Medicine, Busan, Korea. Department of Biomedical Science, Kyungpook National University Graduate School, Daegu, Korea. Clinical Trial Center, Kyungpook National University Hospital, Daegu, Korea. Department of Clinical Pharmacology & Therapeutics, Asan Medical Center, Seoul, Korea. Department of Pharmacology, Kosin University College of Medicine, Busan, Korea. Corresponding author (yry@knu.ac.kr)

†These authors equally contributed to this work.

 12 2012 31 12 2012 20 2 175 181 20 11 2012 11 12 2012 15 12 2012 Copyright © 2012 Korean Society for Clinical Pharmacology and Therapeutics 2012 Background

Metformin is an effective oral antihyperglycaemic agent for type 2 diabetes mellitus, with a variety of metabolic effects. In addition to controlling blood glucose level, it has been appeared to decrease the long-period complications of diabetes, including macrovascular disease. Few reports have addressed the metabolite profiling of metformin. The study was to evaluate if targeted metabolic profiling approach is sensitive enough to predict the therapeutic effects of metformin after a single oral dose.

 Methods

A randomized, open-label, single-dose study was conducted in twenty eight healthy Korean male volunteers. To determine the concentrations of endogenous metabolites in their pre-dose and post-dose plasma samples, blood samples were collected before and at 2 and 6 h after a single oral dose of 500 mg metformin. Both Modular P/Modular D analyzer and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS)-based metabolic profiling was performed.

 Results

We quantified pre-dose and post-dose creatinine, blood urea nitrogen (BUN), lactic acid, 7 amino acids (lysine, glutamic acid, alanine, valine, leucine, phenylalanine, tryptophan), and 5 lysophosphatidylcholines (14:0, 16:0, 17:0, 18:0, and 18:1) using autoanalyser and UPLC-MS/MS. The postdose levels of alanine, lactic acid, glutamic acid, lysine, valine, leucine, phenylalanine, tryptophan, and lysoPC (18:1) were slightly decreased with statistical significance, but there is no clinical significance.

 Conclusion

In order to explore the potential endogenous metabolites associated with the therapeutic effects of metformin, further study including non-targeted (global) metabolite profiling is needed.

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Chemical structure of metformin.

Comparison of targeted metabolite concentrations at 0 (predose), 2h and 6h after a single 500-mg oral dose of metformin. Boxes indicate interquantile range and whisker bars indicate 10th and 90th percentiles. Horizontal bars located in the middle of the boxes represent the median values. *P-value < 0.05, compared between baseline (predose) and 2h or 6h values by repeated measures ANOVA test.

Analytical method for UPLC-MS/MS

*A buffer: 0.1% formic acid in DW; B buffer: 0.1% formic acid in acetonitrile.

Analytical condition of the MRM transition of UPLC-MS/MS