用液相色谱串联质谱法检测脐带血血清25-羟基维生素D2和25-羟基维生素D3浓度  

Determinations of serum 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 concentrations in cord blood by liquid chromatography tandem-mass spectrometry

作　　者：赵丽[1] 王伟业[1] 

机构地区：[1]上海交通大学医学院附属新华医院环境与儿童健康教育部和上海市重点实验室,上海200092

出　　处：《检验医学》2015年第8期825-829,共5页Laboratory Medicine

基　　金：上海市科学技术委员会资助项目(13430710300)

摘　　要：目的建立一种测定脐带血25-羟基维生素D2[25（OH）D2]和25-羟基维生素D3[25（OH）D3]浓度的方法。方法脐带血血清样本前处理采用饱和硫酸锌和乙腈的蛋白沉淀法。采用液相色谱串联质谱法（LC．MS／MS）测定25（OH）D2和25（OH）D3浓度。固定相为Poroshell@120EC-C18正向柱；电喷雾离子源（ESI）；多反应监测模式（MRM）；喷雾电压35．0psi；喷雾温度180℃；鞘气温度350℃；鞘气流速11．0L／min。以甲醇（80％）、超纯水（20％）为流动相进行梯度洗脱，流速为0．5mL／min，柱温为50℃；质谱采用高温ESI，以选择反应监测模式进行定量分析。结果25（OH）D2和25（OH）D3在3．125～100μg/L范围内呈良好的线性关系（R^2=0．9987、0．9991）。血样冻融3次进行检测，日内、日间相对标准偏差（RSD）均〈15％。对血清25．（OH）D2和25（OH）D3质控品进行检测，结果均符合要求。结论LC—MS／MS敏感性高，分析时间短，适用于脐带血1．25（OH）D2和25（OH）D3浓度检测。Objective To establish a method for determining the concentrations of 25-hydroxyvitamin D2 [ 25 （OH） D2 ] and 25-hydroxyvitamin D1 [ 25 （OH） D3 ] in cord blood. Methods The serum samples from cord blood were pretreated by acetonitrile and ZnSO4. High performance liquid chromatography tandem-mass spectrometry （LC-MS/MS） was used for the determinations of 25 （OH） D2 and 25 （OH） D3 concentrations. The analysis was carried on a Poroshell 120 EC-C18 column. Electrospray ionization （ ESI）, muhiple reaction monitor （ MRM ）, nebulizer 35.0 psi, gas temperature 180℃, sheath gas temperature 550℃ and sheath gas flow 11.0 L/min were performed. The column temperature was 50℃ with linear gradient elution using methanol （ 80% ） and water （ 20% ）, and the flow rate was 0.5 mL/min. The quantitative analysis was performed by selective reaction monitoring mode of heater ESI. Results A good linearity was detected in the hnear range of 3. 125-100 μg/L for 25 （OH） 132 and 25 （OH） 133 （R2 = 0. 998 7 and 0. 999 1 ）. Blood samples had frozen and thawed for 3 times. Both intra-day and inter-day relative standard deviations （RSD） were 〈 15%. The quality control materials of serum 25 （OH） D2 and 25 （OH） D3 were analyzed, and the results met the requirements of quality control. Conclusions LC-MS/MS is sensitive, rapid and suitable for the determinations of serum 25 （OH） D2 and 25 （OH） D3 concentrations in cord blood.

关 键 词：25-羟基维生素D2 25-羟基维生素D3 液相色谱串联质谱 脐带血 

分 类 号：R446.1[医药卫生—诊断学]