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

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=50×10^9/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。

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

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=50×10^9/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.