Determination of sterols in Candida albicans by GC-SIM-MS
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摘要: 目的 建立气质联用法(GC-SIM-MS)测定白念珠菌中羊毛甾醇和麦角甾醇的含量。 方法 白念珠菌经离心、皂化、提取出甾醇部分后溶解于环己烷中,采用HP-5毛细管柱进行气相色谱分析,柱温:程序升温150~320℃,起始温度150℃,按10℃/min 升至300℃,再按20℃/min升至320℃;检测器温度270℃;进样口温度270℃;载气:氦气1 ml/min;质谱检测器:EI离子源,电力电压 70 eV;SIM Scan; the selected ions:3.00~17.50 min, 275,386;17.50~19.00 min, 363,396;19.00~30.00 min,411,426;进样量:1 μl;无分流比。 结果 羊毛甾醇和麦角甾醇线性关系良好(r分别为0.999 8和0.999 5),回收率在91.2%~117.3%(RSD<2.5%),最低检测限分别为18.29 ng/ml和68.13 ng/ml,日内及日间精密度良好(RSD均小于1.2%)。 结论 本方法准确、灵敏、专属性强,结果可靠,为抗真菌药物研究提供了有力的工具。Abstract: Objective To establish a method of GC-SIM-MS for the determination of sterols in Candida albicans, including lanosterol and ergosterol. Methods The sterols extracted from Candida albicans were analyzed by gas chromatography with a HP 50 capillary column, temperature from 150℃ to 300℃, 10℃/min, then, increased from 300℃ to 320℃, 20℃/min;detected temperature was 270℃; injection temperature was 270℃; carrier gas was Helium, flow rate was 1 ml/min; no splic.Mass spectrometry:EI Ion source, Ionization voltage:70 eV; SIM Scan; the selected ions:3.00~17.50 min, 275, 386; 17.50~19.00min, 363, 396;19.00~30.00 min 411, 426. Results Linear relation of lanosterol and ergosterol were good(r=0.999 8 and 0.999 5 respectively). The average recoveries were between 91.2%~117.3%, RSD<2.5%. The lowest limit of detect was 18.29 ng/ml and 68.13 ng/ml respectively, the precision within day were good (RSD<1.2%). Conclusion This method was accurate, sensitive and specific. The results were reliable, which provided a powerful tool for the study of antifungal agents.
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Key words:
- gas chromatography mass spectrometry /
- lanosterol /
- ergosterol /
- determination /
- Candida albicans
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