Determination of metformin bio-distribution by LC-MS/MS in mice treated with a clinically relevant paradigm

Abstract

Metformin, an anti-diabetes drug, has been recently emerging as a potential "anti-aging" intervention based on its reported beneficial actions against aging in preclinical studies. Nonetheless, very few metformin studies using mice have determined metformin concentrations and many effects of metformin have been observed in preclinical studies using doses/ concentrations that were not relevant to therapeutic levels in human. We developed a liquid chromatography-tandem mass spectrometry protocol for metformin measurement in plasma, liver, brain, kidney, and muscle of mice. Young adult male and female C57BL/6 mice were voluntarily treated with metformin of 4 mg/ml in drinking water which translated to the maximum dose of 2.5 g/day in humans. A clinically relevant steady-state plasma metformin concentrations were achieved at 7 and 30 days after treatment in male and female mice. Metformin concentrations were slightly higher in muscle than in plasma, while,~3 and 6-fold higher in the liver and kidney than in plasma, respectively. Low metformin concentration was found in the brain at~20% of the plasma level. Furthermore, gender difference in steady-state metformin bio-distribution was observed. Our study established steady-state metformin levels in plasma, liver, muscle, kidney, and brain of normoglycemic mice treated with a clinically relevant dose, providing insight into future metformin preclinical studies for potential clinical translation.

Meta-Assessment of Metformin Absorption and Disposition Pharmacokinetics in Nine Species

“…The marked difference between V SS (6-10 L/kg for dogs and 0.432-0.856 L/kg for man) and V d,area (44.8 L/kg for dogs and 0.859-3.83 L/kg for man) [36,37,40,71] is likely related to differences in transporters and related tissue uptake. From the collected literature information (Supplementary Table S3), tissue K p values at early times (e.g., <24 h) ranged 0.359 to 0.771 for muscle and 0.0354 for brain in mouse [55,130,131], while the prolonged treatment of metformin (i.e., from 7 to 30 days) produced K p values of 1.00-2.06 for muscle and 0.174-0.257 for the brain [132]. Similarly in rats, the brain K p increased from 0.2 (at 1 h) to 1.48 (at 24 h) [133], while the K p values for muscle were 0.455 at 2 h of day 1 and 0.738 at 2 h of day 7 [134].…”

Development and Validation of a Simple and Sensitive LC-MS/MS Method for Quantification of Metformin in Dried Blood Spot Its Application as an Indicator for Medication Adherence

“…In this analysis, the retention time was the shortest, which was comparable to the reported data for MET. It is reported to be at 0.89 min, 15 , 26 at 1.26 min, 41 at 1.4 min, 27 at 1.45 min, 42 1.8 min, 14 at 2.65 min 25 and at 3.1 min. 28 …”

“…Recently, few sensitive methods were reported for individual estimation of MET using LC-MS/MS. 6,[12][13][14][15][25][26][27][28] Only a few studies published the analysis of MET from DBS utilizing LC-MS/MS. 1,4,[29][30][31][32] This sampling technique (DBS) is also acceptable for infants, which reduces the risk of HIV infection and other infectious pathogens to a minimum.…”

Smart UV Derivative Spectrophotometric Methods for Simultaneous Determination of Metformin and Remogliflozin: Development, Validation and Application to the Formulation

“…In addition, REM helps in reducing the body weight and blood pressure in diabetes mellitus patients. [8][9][10] Many analytical methods were illustrated for the estimation of MET from pharmaceutical preparations and plasma by spectroscopic, 11,12 HPLC, [13][14][15] HPLTC, 16 LC-MS, 17,18 and CZE, 19,20 alone and with other drugs. Few analytical procedures were also depicted in the texts for the analysis of REM.…”