超高效液相色谱-串联质谱法测定燕窝中唾液酸的含量

Determination of Sialic Acid in Edible Bird’s Nest Using UPLC-MS/MS

摘要:

本研究建立了燕窝中唾液酸含量的超高效液相色谱-串联质谱（UPLC-MS/MS）测定方法。经磷酸水解后，燕窝中的唾液酸从与唾液酸糖蛋白的结合状态中游离出来，吸取上清液过Oasis HLB柱，用水淋洗固相萃取柱并弃去全部流出液，用真空泵在40～60 kPa负压下抽干Oasis HLB固相萃取柱，再用甲醇洗脱被测物，洗脱液40 ℃水浴中氮气吹干，用流动相溶液定容，采用负离子模式和多反应监测模式超高效液相色谱-串联质谱法检测。方法的线性范围在0.1~50 mg/L之间，相关系数0.999，检出限为0.02 mg/kg，定量限为0.1 mg/kg，样品平均加标回收率为93.5%，相对标准偏差1.52%。该方法前处理简单、重复性好、灵敏度高，可应用于燕窝中唾液酸含量的测定。

Abstract:

A simple and sensitive UPLC-MS/MS method was developed and validated for the quantification of sialic acid (N-Acetyl neuraminic acid) in edible bird’s nest. After acid hydrolysis, sialic acid in edible bird’s nest was released from the combination state of glycoprotein sialic acid. The supernate was further cleaned up using Oasis HLB SPE, followed by washed with water. The Oasis HLB SPE was drained by vacuum pump at 40~60 kPa prior to be eluted by methanol. The eluate was concentrated by nitrogen at 40 ℃ water bath, and then settled to permit by mobile phase. UPLC-MS/MS in negative-ion and multiple reaction monitor mode was used to detect the sialic acid. Validation results indicated that the limit of detection was 0.02 mg/kg and the limit of quantification was 0.1 mg/kg. The assay showed a linear range of 0.1-50 mg/L and gave a correlation coefficient (r) of 0.999. The average recovery was 93.5% with relative standard deviations (RSD) of 1.52% (n=6). The method showed good reproducibility and high sensitivity, suitable for analysis of sialic acid in edible bird’s nest.