目的：对女贞子中主要药用成分进行定性分析,用电喷雾质谱法探究人血清白蛋白与齐墩果酸相互作用。方法：定性分析采用高效液相色谱电喷雾高分辨质谱（HPLC-ESI-MS）法,色谱柱使用Agilent ZORBAX Eclipse XDB-C18柱（2.1×150 mm,5-Micron）,流动相为甲醇（A）-0.1%乙酸水溶液（B）,梯度洗脱：0-30 min 10%-90%A,30-40 min 90%A,40-60 min 90%-10%A,柱温：30℃,进样量：1μL,流速：0.2 m L·min^-1,女贞子乙醇提取物采用HPLC-ESI-MS负离子检测模式。结果：采用HPLC-ESI-MS定性地检测了女贞子乙醇超声提取液,鉴定出9种有效成分（齐墩果酸、红景天苷、特女贞苷、橄榄苦苷、芹菜素、委陵菜酸、2α-羟基熊果酸、2α-羟基齐墩果酸和19α-羟基-3-乙酰乌索酸）。电喷雾质谱观测到齐墩果酸能与人血清白蛋白形成较为稳定的1：1蛋白质复合物,其结合常数为1.63×10^4L·mol^-1。结论：该方法定性分析了女贞子乙醇提取物中主要药用成分,首次用电喷雾质谱法研究了人血清白蛋白与齐墩果酸的相互作用,从分子水平上揭示了齐墩果酸与人血清白蛋白相互作用的质谱信息。
A sensitive and rapid high performance liquid chromatography-mass spectrometry(HPLC-MS)method was developed and validated for simultaneous quantifcation of ten steroid hormones,including estrogens,androgens,progesterones,and corticosteroids four classes of steroids.The following ten steroid hormones were analyzed:progesterone,21-deoxycortisol,estrone,4-androstenedione,testosterone,dihydro-testosterone,androstenone,dehydroepiandrosterone,corticosterone and cortisone.Stable deuterated isotopes were used as internal standards for quantifcation.Sample preparation with and without derivatization were performed after liquid-liquid extraction,and the corresponding results were compared according to sensitivity and selectivity.Hydroxylamine derivatization was found to improve the ionization efciency of the analytes for electrospray ionization MS analysis.The gradient of mobile phase and experimental parameters for HPLC separation were optimized.The lower limits of quantifcation were in the range of 0.05-5 ng mL^(−1) with wide linear range for the ten steroid hormones.The intra-day precision<11.1%and recovery of 84.5-120% with negligible matrix efect were achieved,where within the acceptance limits of the FDA guideline.Total HPLC-MS analysis time was 6 min.This method enables simultaneous quantifcation of steroids in human serum.It will be helpful for the serum steroid profling in order to understand various endocrinology diseases.
