液相色谱-串联质谱法检测血清皮质醇方法的建立及性能评价

doi:10.3969/j.issn.1673-8640.2017.06.018

摘要/Abstract

摘要：

目的建立同位素稀释液相色谱-串联质谱法（ID-LC-MS/MS）检测血清皮质醇的方法并对其进行性能评价。方法采用ABSCIEX TRIPLE QUAD 5500液相色谱-串联质谱联用系统定量检测皮质醇,参照美国临床实验室标准化协会（CLSI）相关文件对建立的方法进行线性、精密度、提取回收率和准确性等基本性能验证。结果内标平衡时间为室温下1 h。ID-LC-MS/MS检测皮质醇的线性回归方程为Y=0.018 7X+0.043 4,r=0.999 8;线性范围为25~500 ng/mL。检出限为0.05 ng/mL。批内、批间变异系数（CV）分别为<4%、<6%。提取回收率为84.5%~90.4%。国际临床化学和检验医学联合会参考实验室外部质量评价计划（RELA）-A和RELA-B样本测定结果的偏移分别为3.19%和2.84%。结论建立的ID-LC-MS/MS方法的精密度、准确度、重现性均较好,结果准确,可用于临床实验室检测血清皮质醇。

关键词: 同位素稀释液相色谱-串联质谱, 性能验证, 皮质醇

Abstract:

Objective To establish isotope-dilution liquid-chromatography tandem-mass spectrometry（ID-LC-MS/MS） for determining serum cortisone,and to evaluate its performance. Methods ABSCIEX TRIPLE QUAD 5500 liquid-chromatography tandem-mass spectrometry was used to determine serum cortisone quantitatively. According to the Clinical and Laboratory Standards Institute（CLSI） document,the performance,including linearity,precision,extraction and recycling rates and accuracy,was verified. Results Cortisone-d4 was added to analytes, and they were equilibrated at room temperature for 1 h. ID-LC-MS/MS for determining serum cortisone had linear regression equation Y=0.018 7X+0.043 4 （r=0.999 8）. The linear range was 25-500 ng/mL. The determination limit was 0.05 ng/mL for serum cortisone. The within-run and between-run coefficients of variation（CV） were <4% and <6%. The extraction and recycling rates were 84.5%-90.4%. The biases of the International Federation of Clinical Chemistry and Laboratory Medicine External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine（RELA）-A and RELA-B were 3.19% and 2.84%. Conclusions The established ID-LC-MS/MS has good precision,accuracy and repeatability,which is suitable for determining serum cortisone.

Key words: Isotope-dilution liquid-chromatography tandem-mass spectrometry, Performance verification, Cortisone

中图分类号:

R446.1

朱宇清, 朱岭峰, 赵晓君, 徐翀, 邵维杰, 曹丹如, 陆银华. 液相色谱-串联质谱法检测血清皮质醇方法的建立及性能评价[J]. 检验医学, 2017, 32(6): 524-530.

ZHU Yuqing, ZHU Lingfeng, ZHAO Xiaojun, XU Chong, SHAO Weijie, CAO Danru, LU Yinhua. Establishment and performance evaluation of liquid-chromatography tandem-mass spectrometry for determining serum cortisone[J]. Laboratory Medicine, 2017, 32(6): 524-530.

图/表 9

参考文献 13

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物质	母离子（m/z）	子离子（m/z）	裂解电压（V）	撞击能（V）
皮质醇	363.1	120.9,183.0	150	32.10,39.80
皮质醇-d4	367.1	121.3	150	25.49

物质	母离子（m/z）	子离子（m/z）	裂解电压（V）	撞击能（V）
皮质醇	363.1	120.9,183.0	150	32.10,39.80
皮质醇-d4	367.1	121.3	150	25.49

理论值（ng/mL）	测定值1（ng/mL）	测定值2（ng/mL）	x（ng/mL）	s（ng/mL）	CV（%）	偏移（%）
25	24.5	23.5	24.00	0.71	2.95	4.00
50	47.8	52.0	49.90	2.97	5.95	0.20
100	103.0	97.0	100.00	4.24	4.24	0.00
250	265.0	271.0	268.00	4.24	1.58	7.20
500	484.0	481.0	482.50	2.12	0.44	3.50

理论值（ng/mL）	测定值1（ng/mL）	测定值2（ng/mL）	x（ng/mL）	s（ng/mL）	CV（%）	偏移（%）
25	24.5	23.5	24.00	0.71	2.95	4.00
50	47.8	52.0	49.90	2.97	5.95	0.20
100	103.0	97.0	100.00	4.24	4.24	0.00
250	265.0	271.0	268.00	4.24	1.58	7.20
500	484.0	481.0	482.50	2.12	0.44	3.50

样本	批内			批间
样本	x（ng/mL）	s（ng/mL）	CV（%）	x（ng/mL）	s（ng/mL）	CV（%）
低值	109.56	3.21	2.93	108.15	5.36	4.96
高值	330.50	11.63	3.52	335.02	18.49	5.52