Simultaneous Determination of Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate and High-Performance Liquid Chromatography

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Effect of Dietary Supplementation of Sypjeondaebotang or Jahyulyanggeuntang on Iron Bioavailability in Rats

The effect of dietary supplementation of the two oriental medicinal prescriptions, Sypjeondaebotang or Jahyulyanggeuntang, on iron bioavailability was evaluated in rats which were depleted of iron by being fed an irondeficient diet for 4 weeks. Seventy two iron-depleted rats were randomly divided into 3 groups (n = 24) , and fld one of the following experimental diets for 4 (n=8), 12 (n=8), and 25 days (n=8) : Control diet (CD), Sypjeondaebotang-supplemented diet (SD), Jahyulyanggeuntang -supplemented diet (JD). The CD contained 20 mg Fe/kg diet as FeSO4, and the SD or JD was identical except that the respective oriental medicinal prescription extract was included at the level of 4 g/kg diet. Animals fed the SD and JD for 25 days consumed significantly less food (p0.05) were lower in rats fed the SD compared to those fed the CD. Dietary supplementation of Sypjeondaebotang during the period of iron repletion significantly increased blood levels of hemoglobin (p<0.05) and hematocrit (p<0.01) measured at day 12, and increased mean corpuscular volume (p<0.05) measured at day 25, compared to the values for the CD rats. Regression analyses of hemoglobin-repletion bioassay data from rats fed the SD and JD showed the relative biological values of 123% and 99%, respectively, calculated against the slope for the CD rats. Apparent iron absorption and retention values were also significantly lower (p<0.05) in rats fed the SD for 25 days than those for the CD rats. Based on the results from diverse biochemical indices of iron status and the chemical balance study, the effect of Jahyulyanggeuntang on iron bioavailability appears to be less prominent than that for Sypjeondaebotang. Taken together, these results indicate that Sypjeondaebotang has a positive effect in restoring iron depletion by increasing the iron bioavailability in rats.