LC-MS/MS法在不明原因复发性流产患者血清10种氨基酸水平检测中的应用  

Application of LC-MS/MS method in detection of 10 kinds of amino acids in serum of patients with unexplained recurrent spontaneous abortion

作　　者：许红[1] 韩彦洁[1] 张瑶[1] 刘力[1] 

机构地区：[1]石家庄市第四医院石家庄市妇产医院,石家庄050000

出　　处：《山东医药》2017年第44期18-21,共4页Shandong Medical Journal

基　　金：河北省科技计划项目(122777100)

摘　　要：目的采用高效液相色谱串联质谱(LC-MS/MS)法检测不明原因复发性流产(URSA)患者血清10种氨基酸水平变化,并对该方法进行评价。方法选择URSA患者40例作为观察组,同期孕检正常的早孕(孕周<20周)妇女40例作为对照组。采用LC-MS/MS法检测两组血清鸟氨酸、瓜氨酸、甘氨酸、丙氨酸、缬氨酸、精氨酸、亮氨酸、蛋氨酸、苯丙氨酸、酪氨酸水平,并进行LC-MS/MS法检测的专属性、精密度及添加回收率、稳定性分析。结果观察组与对照组血清鸟氨酸分别为(89.3±8.9)、(33.6±5.4)μmol/L,瓜氨酸分别为(18.6±3.2)、(14.3±2.4)μmol/L,甘氨酸分别为(230.2±27.3)、(202.2±19.9)μmol/L,两组比较P均<0.01。两组血清丙氨酸、缬氨酸、精氨酸、亮氨酸、蛋氨酸、苯丙氨酸、酪氨酸水平比较差异均无统计学意义(P均>0.05)。专属性分析结果显示,观察组血清中出现相应氨基酸的质荷比信号图,空白甲醇溶液中只有添加内标的质荷比信号,未见相应的氨基酸信号。精密度及添加回收率分析结果显示,采用LC-MS/MS法检测血清10种氨基酸水平的添加回收率为82.1%~93.6%,日内标准偏差为1.5%~5.1%,日间标准偏差为2.5%~7.1%。稳定性分析结果显示,样本放置2~7天的平均回收率为95.3%~103.2%。结论 URSA患者血清鸟氨酸、瓜氨酸、甘氨酸水平均升高,血清丙氨酸、缬氨酸、精氨酸、亮氨酸、蛋氨酸、苯丙氨酸、酪氨酸水平无明显变化;LC-MS/MS法检测血清氨基酸的回收率、重复性和稳定性均较高。Objective To detect the changes of 10 kinds of amino acid levels in patients with unexplained recurrent spontaneous abortion （URSA） by high performance liquid chromatography-tandem mass spectrometry （LC-MS/MS） method, then to evaluate the performance of this method. Methods Forty URSA patients were used as the observation group, 40 normal early pregnancy women （ 〈 20 weeks） were arranged as the control group. LC-MS/MS method was used to detect ornithine, citrulline, glycine, alanine, valine, arginine, leucine, methionine, phenylalanine, and tyrosine levels in the two groups. The specificity, precision, recovery rate, and stability of LC-MS/MS method were also analyzed. Results The serum ornithine levels in the observation group and control group were （ 89.3 ± 8.9） and （33.6 ± 5.4 ）μmol/L, respectively; citrulline levels were separately （18.6 ±3.2） and （14.3 ±2.4） μmol/L, and glycine levels were separately （230.2 ± 27.3 ） and （ 202.2 ± 19.9 ） μmol/L（ all P 〈 0.01 ）. No significant difference was found in the levels of serum alanine, valine, arginine, leueine, methionine, phenylalanine, and tyrosine between the two groups （ all P 〉 0.05）. The specificity analysis showed that the amino acid m/z was observed in the serum of the observation group, and there was only internal standard m/z in methanol blank solution, no m/z of the corresponding amino acid. The precision and recovery rate analysis showed that the recovery rate of the method was 82.1%-93.6% , the within-day relative standard deviation （RSD） was 1.5%-5.1% and the between-days RSD was 2.5%-7.1%. The stability analysis showed that the average recovery rate was 95.3% -103.2% for 2-7 days. Conclusions The levels of serum ornithine, citrulline, and glyeine increase in URSA patients, but no significant changes are found in the serum alanine, valine, arginine, leucine, methionine, phenylalanine and tyrosine. The LC-MS/MS method has high recovery rate, repeatability, and stability in detection of serum

关 键 词：复发性自然流产 高效液相色谱串联质谱法 氨基酸 

分 类 号：R331[医药卫生—人体生理学]