羊水样本卵磷脂和鞘磷脂检测超高效液相色谱串联质谱方法的建立及应用

doi:10.3969/j.issn.1673-8640.2020.03.009

检验医学 ›› 2020, Vol. 35 ›› Issue (3): 233-237.DOI: 10.3969/j.issn.1673-8640.2020.03.009

羊水样本卵磷脂和鞘磷脂检测超高效液相色谱串联质谱方法的建立及应用

曹正¹, 刘京瑞¹, 谢鑫¹, 刘一林², 董莹¹, 唐国栋¹, 翟燕红¹

1.首都医科大学附属北京妇产医院检验科,北京 100026
2.北京奥米科生物科技有限公司实验室,北京 100094

收稿日期:2019-06-14 出版日期:2020-03-30 发布日期:2020-04-17
通讯作者: 翟燕红
作者简介:通信作者：翟燕红,联系电话：010-52276406。
作者简介：曹正,男,1982年生,博士,副主任技师,主要从事临床质谱的应用研究。
基金资助:
北京市科技计划课题“首都临床特色项目研究”基金项目（Z161100000516073）

Determination of lecithin and sphingomyelin in amniotic fluid by ultra-performance liquid chromatography-tandem mass spectrometry

CAO Zheng¹, LIU Jingrui¹, XIE Xin¹, LIU Yilin², DONG Ying¹, TANG Guodong¹, ZHAI Yanhong¹

1. Department of Clinical Laboratory,Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing 100026,China
2. Laboratory of Beijing Omics Biological Technology Co.,Ltd.,Beijing 100094,China

Received:2019-06-14 Online:2020-03-30 Published:2020-04-17
Contact: ZHAI Yanhong

摘要/Abstract

摘要：

目的建立检测羊水中卵磷脂和鞘磷脂（L/S）的超高效液相色谱串联质谱（UPLC-MS/MS）方法,以便准确、高效地预测胎儿肺成熟度。方法收集孕32~39周孕妇分娩时的羊水样本23份。依据新生儿Apgar评分标准,有3例胎儿胎肺未成熟、20例胎儿胎肺成熟。另收集孕18周孕妇羊水样本7份作为基线对照,建立检测羊水中卵磷脂和鞘磷脂水平的UPLC-MS/MS方法,计算卵磷脂/鞘磷脂（L/S）比值,同时采用板层小体计数（LBC）法检测板层小体（LB）,评价2种方法在预测胎肺成熟度中的价值。结果建立的检测羊水中卵磷脂和鞘磷脂的UPLC-MS/MS方法精密度良好,离子峰强度和保留时间均在可检测范围内,主成分分析（PCA）显示6个质控样本聚类良好。以L/S比值=10作为判断胎肺成熟度与不成熟的临界值,UPLC-MS/MS的敏感性和特异性均为100%。以LB=50×10⁹/L作为判断胎肺成熟度与不成熟的临界值,LBC法的敏感性和特异性分别为80%和95%。结论建立了检测羊水中卵磷脂和鞘磷脂水平的UPLC-MS/MS方法,其结果可靠,可以准确、高效地预测胎肺成熟度。

关键词: 卵磷脂/鞘磷脂比值, 超高效液相色谱串联质谱, 胎肺成熟度

Abstract:

Objective To establish a method based on ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） to measure the ratio of lecithin/sphingomyelin（L/S） in amniotic fluid to accurately and efficiently predict fetal lung maturity. Methods According to the Apgar scoring standard for newborns,23 amniotic fluid specimens obtained at 32-39 gestational weeks were classified into 3 fetal lung immature cases and 20 fetal lung mature cases. Totally,7 amniotic fluid specimens obtained at 18 gestational week were used as baseline controls. UPLC-MS/MS for determining the ratio of L/S in amniotic fluid was established. Lamellar body （LB） was determined by lamellar body count （LBC） method. The roles of the 2 methods for predicting fetal lung maturity were evaluated. Results The UPLC-MS/MS for the determination of L/S had good precision. Both ion peak intensity and retention time were within the determinable range,and principal component analysis （PCA） showed good clustering of 6 quality control samples. When L/S=10 was used as the cut-off value to discriminate maturity and immaturity,the sensitivity and specificity reached 100%. When LB=50×10⁹ /L was used as the cut-off value,2 cases of positive specimens and 1 case of negative specimen were incorrectly identified,with a sensitivity of 80% and a specificity of 95%. Conclusions The measurement of L/S in amniotic fluid by UPLC-MS/MS is reliable and can predict fetal lung maturity accurately and efficiently.

Key words: Lecithin/Sphingomyelin ratio, Ultra-performance liquid chromatography-tandem mass spectrometry, Fetal lung maturity

中图分类号:

R446.1

曹正, 刘京瑞, 谢鑫, 刘一林, 董莹, 唐国栋, 翟燕红. 羊水样本卵磷脂和鞘磷脂检测超高效液相色谱串联质谱方法的建立及应用[J]. 检验医学, 2020, 35(3): 233-237.

CAO Zheng, LIU Jingrui, XIE Xin, LIU Yilin, DONG Ying, TANG Guodong, ZHAI Yanhong. Determination of lecithin and sphingomyelin in amniotic fluid by ultra-performance liquid chromatography-tandem mass spectrometry[J]. Laboratory Medicine, 2020, 35(3): 233-237.

图/表 3

参考文献 18

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组别	例数	L/S比值	LB/×10⁹/L
基线组	7	1.73±0.32	1.71±0.49
胎肺不成熟组	3	3.06±1.73	59.67±13.87
胎肺成熟组	20	60.51±25.61	56.60±6.60

组别	例数	L/S比值	LB/×10⁹/L
基线组	7	1.73±0.32	1.71±0.49
胎肺不成熟组	3	3.06±1.73	59.67±13.87
胎肺成熟组	20	60.51±25.61	56.60±6.60

羊水样本卵磷脂和鞘磷脂检测超高效液相色谱串联质谱方法的建立及应用

Determination of lecithin and sphingomyelin in amniotic fluid by ultra-performance liquid chromatography-tandem mass spectrometry

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