液相色谱-串联质谱法测定健康人唾液中睾酮和皮质醇的含量
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摘要
目的建立一种方便、可靠和灵敏的液相色谱-串联质谱(LC-MS/MS)方法用于检测健康人唾液中2种内源性激素(睾酮和皮质醇)的水平。方法在Agilent6410A串联质谱仪上采用正离子检测模式,以卡马西平为内标,唾液样品经OMGEA NANOSEP 10K超滤管21 912.8×g超滤10 min后,在Agilent ZORBAX SB-C18(3.5μm,2.1 mm×100 mm)色谱柱上采用等度洗脱进行色谱分离。流动相为乙腈∶0.1%甲酸水溶液(60∶40);流速为0.3 mL/min;进样量为10μL;柱温为25℃;每个样品的分析周期为3 min。结果睾酮和皮质醇线性关系良好,其相关系数r>0.990,日内与日间精密度均<15%。结论该方法可以用于测定唾液中睾酮和皮质醇的含量,为迅速简便地检测体内激素水平提供参考。
Objective To establish a convenient, reliable and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS) method for the determination of two kinds of endogenous hormones(testosterone, cortisol) in the saliva of healthy human. Methods A positive ion mode was performed on the Agilent 6410 A mass spectrometer. Carbamazepine was used as the internal standard. The saliva samples were centrifuged at 21 912.8×g for 10 min with OMGEA NANOSEP 10 K ultrafiltration tubes, and chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column with isocratic elution. The mobile phase was acetonitrile∶0.1% formic acid awter solution(60∶40); flow rate was 0.3 mL/min; injection volume was 10 μL; column temperature was 25 ℃; and the analysis period of each sample was 3 min.Results Testosterone and cortisol had good linear relationship with correlation coefficient(r) being both >0.990. Inter-and intra-day precision was both less than 15%. Conclusion This method can be used to determine the contents of testosterone and cortisol in saliva, which provides a reference for rapid and simple detection of hormone levels in vivo.
Objective To establish a convenient, reliable and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS) method for the determination of two kinds of endogenous hormones(testosterone, cortisol) in the saliva of healthy human. Methods A positive ion mode was performed on the Agilent 6410 A mass spectrometer. Carbamazepine was used as the internal standard. The saliva samples were centrifuged at 21 912.8×g for 10 min with OMGEA NANOSEP 10 K ultrafiltration tubes, and chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column with isocratic elution. The mobile phase was acetonitrile∶0.1% formic acid awter solution(60∶40); flow rate was 0.3 mL/min; injection volume was 10 μL; column temperature was 25 ℃; and the analysis period of each sample was 3 min.Results Testosterone and cortisol had good linear relationship with correlation coefficient(r) being both >0.990. Inter-and intra-day precision was both less than 15%. Conclusion This method can be used to determine the contents of testosterone and cortisol in saliva, which provides a reference for rapid and simple detection of hormone levels in vivo.
引文
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[11]马晓斐,吕品,李挥,张岩.液相色谱-串联质谱法同时测定人血清中皮质酮与皮质醇[J].分析测试学报,2013,32:886-889.
[12]张彤杰,王建跃,张乾通,王恒.婴儿尿液中睾酮和雄烯二酮的超高效液相色谱-串联质谱法测定[J].浙江预防医学,2015,27:646-648.
[2]王林,张云丽.低氧训练对睾酮、皮质醇影响的研究进展[J].辽宁体育科技,2004,26:46-47.
[3]干伟,张林,安振梅.唾液皮质醇的研究进展[J].中国实验诊断学,2009,13:1304-1306.
[4]KOBAN L,POURTOIS G.Brain systems underlying the affective and social monitoring of actions:an integrative review[J].Neurosci Biobehav Rev,2014,46(Pt 1):71-84.
[5]周志芳,夏运成.唾液分析的临床应用价值及研究进展[J].实用预防医学,2010,17:413-416.
[6]张东春.唾液类固醇激素检测的临床意义[J].中国保健营养,2013(4):980-981.
[7]姚剑婷,许红峰,吴梦如.唾液皮质醇分泌节律影响因素研究进展[J].新疆中医药,2017,35:78-81.
[8]金伟,黄斌,王彬,王道玮,赵世民,潘学军.气相色谱/质谱法测定水和沉积物中雄激素与孕激素[J].分析化学,2013,41:205-209.
[9]刘齐,张华珺,郭洁,孙亮,马长华.HPLC-MS/MS法同时测定育发液中的9种禁限用物质[J].药物分析杂志,2016,36:1660-1666.
[10]黄河花,刘东阳,胡蓓,江骥.高效液相色谱串联质谱法同时定量测定人血清中脱氢表雄酮、睾酮及雄酮[J].药物分析杂志,2012,32:210-216.
[11]马晓斐,吕品,李挥,张岩.液相色谱-串联质谱法同时测定人血清中皮质酮与皮质醇[J].分析测试学报,2013,32:886-889.
[12]张彤杰,王建跃,张乾通,王恒.婴儿尿液中睾酮和雄烯二酮的超高效液相色谱-串联质谱法测定[J].浙江预防医学,2015,27:646-648.