Meng Yong, Liu Jin-Feng, Yang Shi-Zhong, Ye Ru-Qiang, Mu Bo-Zhong
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology.
Anal Sci. 2015;31(5):377-82. doi: 10.2116/analsci.31.377.
A highly sensitive and selective high-performance liquid chromatographic (HPLC) method has been developed for the determination of microbial lipopeptides of fluorescent derivatization with 1-bromoacetylpyrene to overcome the limitations of trace detection of lipopeptides in aqueous solutions. The derivatization of lipopeptides with 1-bromoacetylpyrene was conducted at 60°C for 20 min under catalysis of triethylamine. The resulting derivative products were separated by HPLC and determined by a fluorescence detector. Each homolog of lipopeptides in samples was identified by HPLC-MS and the detection limit after derivatization in an aqueous solution was 2.5 μg/mL (S/N = 3). The calibration curve for lipopeptides was linear in the concentration range of 0.250 - 4.00 mg/mL. This method has adequate sensitivity and selectivity for microdetection of lipopeptides in aqueous solutions in mild reaction conditions, which allows this method to be used in the determination of trace lipopeptides in environmental samples and complex samples.
已开发出一种高灵敏度和高选择性的高效液相色谱（HPLC）方法，用于测定经1-溴乙酰芘荧光衍生化的微生物脂肽，以克服水溶液中脂肽痕量检测的局限性。脂肽与1-溴乙酰芘的衍生化反应在三乙胺催化下于60°C进行20分钟。所得衍生产品通过HPLC分离，并用荧光检测器测定。通过HPLC-MS鉴定样品中脂肽的每个同系物，水溶液中衍生化后的检测限为2.5μg/mL（S/N = 3）。脂肽的校准曲线在0.250 - 4.00mg/mL浓度范围内呈线性。该方法在温和的反应条件下对水溶液中脂肽的微量检测具有足够的灵敏度和选择性，这使得该方法可用于环境样品和复杂样品中痕量脂肽的测定。
