Transl Clin Pharmacol.  2017 Jun;25(2):67-73. 10.12793/tcp.2017.25.2.67.

Metabolomic analysis of healthy human urine following administration of glimepiride using a liquid chromatography-tandem mass spectrometry

• Do EY
• Gwon MR
• Kim BK
• Ohk B
• Lee HW
• Kang WY
• Seong SJ
• Kim HJ
• Yoon YR

• KMID: 2386763
• DOI: http://doi.org/10.12793/tcp.2017.25.2.67

Abstract

Glimepiride, a third generation sulfonylurea, is an antihyperglycemic agent widely used to treat type 2 diabetes mellitus. In this study, an untargeted urinary metabolomic analysis was performed to identify endogenous metabolites affected by glimepiride administration. Urine samples of twelve healthy male volunteers were collected before and after administration of 2 mg glimepiride. These samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and then subjected to multivariate data analysis including principal component analysis and orthogonal partial least squares discriminant analysis. Through this metabolomic profiling, we identified several endogenous metabolites such as adenosine 3"², 5"²-cyclic monophosphate (cAMP), quercetin, tyramine, and urocanic acid, which exhibit significant metabolomic changes between pre- and posturine samples. Among these, cAMP, which is known to be related to insulin secretion, was the most significantly altered metabolite following glimepiride administration. In addition, the pathway analysis showed that purine, tyrosine, and histidine metabolism was affected by pharmacological responses to glimepiride. Together, the results suggest that the pharmacometabolomic approach, based on LC-MS/MS, is useful in understanding the alterations in biochemical pathways associated with glimepiride action.

Keyword

metabolomic profiling; glimepiride; liquid chromatographytandem mass spectrometry

MeSH Terms

Adenosine
Diabetes Mellitus, Type 2
Histidine
Humans*
Insulin
Least-Squares Analysis
Male
Mass Spectrometry*
Metabolism
Metabolomics*
Principal Component Analysis
Quercetin
Statistics as Topic
Tyramine
Tyrosine
Urocanic Acid
Volunteers
Adenosine
Histidine
Insulin
Quercetin
Tyramine
Tyrosine
Urocanic Acid

Figure

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  Figure 1 Representative base peak chromatogram (m/z 50–1000; 0–35 min) of urine samples obtained from healthy volunteers before (A) and after (B) 2 mg glimepiride administration in a positive ion mode.

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  Figure 2 PCA score plot based on LC-MS/MS chromatographic data. The triangles designate the quality control (QC) samples (Δ, n=8). Pre- (●, n=12) and post-dose groups (□, n=12) were separated on PCA score

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  Figure 3 OPLS-DA plots obtained from untargeted metabolomics analysis data. (A) Score plot, (B) Loading plot, (C) permutation test plot for internal validation of OPLS-DA model.

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  Figure 4 S-plot of urinary metabolites. The variables labeled with colors are four identified metabolites including cAMP (blue), quercetin (red), tyramine (yellow), and urocanic acid (green).

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  Figure 5 Box plots of four identified metabolites from pre- and post-dose urine samples obtained from urinary metabolomic analysis. (A) cAMP, (B) quercetin, (C) tyramine, and (D) urocanic acid. Statistical significance was compared with pre-dose samples (*p<0.05; **p<0.01).

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