| 摘要: |
| 目的 建立高效液相色谱-串联质谱（LC-MS/MS）法，用于测定原发性肝癌患者血清中7种甾类激素的含量。方法 血清样品采用乙酸乙酯液-液萃取，经丹磺酰氯衍生化后测定。色谱柱为Agilent Poroshell 120 EC-C18柱（2.1 mm×150 mm，2.7μm），流动相为乙腈-0.1%（V：V）甲酸水溶液，梯度洗脱，流速为0.3 mL/min，柱温为35℃，质谱检测采用动态多反应离子监测模式（MRM）。结果 7种甾类激素（雌二醇、雌三醇、雌酮、氢化可的松、睾酮、雄烯二酮和黄体酮）在8 min内达到基线分离。在相应浓度范围内线性关系良好，相关系数均>0.99，日内及日间精密度RSD均<15%，提取回收率为80%~119%，基质效应为85%~112%，样品在放置6 h后处理、处理后室温放置24 h、3个冻融循环以及-80℃保存30 d等测定过程中均稳定性良好。结论 该方法操作简便、结果准确、灵敏度高、重现性好，适用于原发性肝癌患者体内甾类激素水平检测。 |
| 关键词: 液相色谱-串联质谱 甾类激素 肝肿瘤 肝细胞癌 |
| DOI：10.16781/j.0258-879x.2016.07.0810 |
| 投稿时间：2016-02-19修订日期：2016-03-19 |
| 基金项目: |
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| LC-MS/MS for determination of seven steroid hormones in sera of primary hepatocellular carcinoma patients |
| QIAN Xian,LÜLei,ZHAN Qi,LI Yue-yue,LI Yun-qing,ZHANG Guo-qing,ZHAO Liang* |
(Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China
* Corresponding author) |
| Abstract: |
| Objective To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of seven steroid hormones in sera of patients with primary hepatocellular carcinoma. Methods The serum samples were extracted with ethyl acetate, and then derivatized with dansyl chloride. Chromatographic column was Agilent Poroshell 120 EC-C18 (2.1 mm×150 mm, 2.7 μm) column, mobile phase was acetonitrile and 0.1% formic acid-water solution, flow rate was 0.3 mL/min, the column temperature was 35℃, and the MS detection was selected in dynamic MRM mode. Results Seven steroid hormones (estradiol, estriol, estrone, hydrocortisone, testosterone, androstenedione and progesterone) were baseline separated within 8 min, which possessed good linear relationship (r>0.99) within the linear concentration range; the intra-day and inter-day precisions were less than 15%, the recoveries were ranged from 80% to 119%, the matrix effect was 85%-112%. The detection results were stable when the samples were examined 6 h after collection, 24 h after treatment at room temperature, after three freeze-thaw cycles, and 30 days when stored at -80℃. Conclusion The current LC-MS/MS method is simple, accurate, sensitive and reproducible, and it can be applied to determine steroid hormones in the sera of patients with primary hepatocellular carcinoma. |
| Key words: liquid chromatography-tandem mass spectrometry steroid hormone liver neoplasms hepatocellular carcinoma |
