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Quantitation of underivatized branched-chain amino acids in sport nutritional supplements by capillary electrophoresis with direct or indirect UV absorbance detection

机译：运动营养补品中未衍生的支链氨基酸的定量分析采用毛细管电泳直接或间接紫外吸收检测

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The branched-chain amino acids (BCAAs) including leucine (Leu), isoleucine (Ile) and valine (Val) play a pivotal role in the human body. Herein, we developed capillary electrophoresis (CE) coupled with conventional UV detector to quantify underivatized BCAAs in two kinds of sport nutritional supplements. For direct UV detection at 195 nm, the BCAAs (Leu, two enantiomers of Ile and Val) were separated in a background electrolyte (BGE) consisting of 40.0 mmol/L sodium tetraborate, and 40.0 mmol/L β-cyclodextrin (β-CD) at pH 10.2. In addition, the indirect UV detection at 264 nm was achieved in a BGE of 2.0 mmol/L Na2HPO4, 10.0 mmol/L p-aminosalicylic acid (PAS) as UV absorbing probe, and 40.0 mmol/L β-CD at pH 12.2. The β-CD significantly benefited the isomeric separation of Leu, L- and D-Ile. The optimal conditions allowed the LODs (limit of detections) of direct and indirect UV absorption detection to be tens μmol/L level, which was comparable to the reported CE inline derivatization method. The RSDs (relative standard deviations) of migration time and peak area were less than 0.91% and 3.66% (n = 6). Finally, CE with indirect UV detection method was applied for the quantitation of BCAAs in two commercial sport nutritional supplements, and good recovery and precision were obtained. Such simple CE method without tedious derivatization process is feasible of quality control and efficacy evaluation of the supplemental proteins.
机译：包括亮氨酸（Leu），异亮氨酸（Ile）和缬氨酸（Val）在内的支链氨基酸（BCAA）在人体中起关键作用。在这里，我们开发了毛细管电泳（CE）结合常规的UV检测器，以定量两种运动营养补品中未衍生的BCAA。为了在195 nm处进行直接UV检测，将BCAAs（Leu，Ile和Val的两个对映体）在背景电解质（BGE）中分离，该电解质由40.0 mmol / L四硼酸钠和40.0 mmol / Lβ-环糊精（β-CD）组成）在pH 10.2下。另外，在BGE为2.0 mmol / L Na2HPO4、10.0 mmol / L对氨基水杨酸（PAS）作为UV吸收探针，40.0 mmol / Lβ-CD在pH 12.2的BGE下，实现了在264 nm处的间接UV检测。 β-CD显着有利于Leu，L-和D-Ile的异构体分离。最佳条件使得直接和间接UV吸收检测的LOD（检测限）达到数十μmol/ L，与报道的CE在线衍生化方法相当。迁移时间和峰面积的RSD（相对标准偏差）分别小于0.91％和3.66％（n = 6）。最后，采用间接紫外检测法对两种商业商业营养补品中的BCAA进行定量分析，获得了良好的回收率和精密度。这种简单的没有繁琐的衍生化过程的CE方法对于补充蛋白的质量控制和功效评估是可行的。

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