检验医学与临床2017，Vol.14Issue(2)：180-183,4.DOI:10.3969/j.issn.1672-9455.2017.02.011
HPLC-MS／MS检测血清25-羟维生素D方法的建立和基本性能评价
Establishment and evaluation of a high performance liquid chromatography-tandem mass spectrometry method for the determination of 25-hydroxyvitamin D in serum
摘要
Abstract
Objective To establish a high performance liquid chromatography‐tandem mass spectrometry method (HPLC‐MC/MC) for the quantitation determination of serum 25‐hydroxyvitamin D2 [25(OH)D2] and 25‐hydroxyvitamin D3 [25(OH)D3] and to be used for routine clinical evaluation of Vitamin D levels .Methods Serum samples and standard samples were mixed respective‐ly with 25(OH)D2‐d3 and 25(OH)D3‐d3 (internal standard) and treated with ZnS04 solution and methanol to precipitate .The re‐siduals were then analyzed by HPLC‐MS/MS system carried on a Poroshell 120 EC‐C18 column .Electrospray ionization (ESI) , multiple reaction monitor (MRM) ,dry gas temperature 275 ℃ ,dry gas flow 10 L/min ,nebulizer 50 psi capillary voltage 5 000 V 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 The correlation coefficients between the peak area ratios and 25(OH)D2 as well as 25(OH)D3 concentrations were higher than 0 .995 .The limit of detection (LOD) of 25(OH)D2 and 25(OH)D3 both were 1 .0 ng/mL and limit of quantification (LOQ) were 2 .0 ng/mL and 1 .5 ng/mL respectively .The within‐run and total coefficients of variation of 25(OH)D2 were 3 .01% -5 .23% and 3 .45% -8 .72% ;within‐run and total coefficients of variation of 25(OH)D3 were 1 .78% -3 .43% and 2 .19% -4 .16% .The accu‐racy was 105 .4 for 25(OH)D2 and it ranged from 98 .0% to 104 .9% for 25(OH)D3 .The recovery rates of 25(OH)D2 were 99 .67% -103 .92% ,which of 25(OH)D3 were 98 .72% -104 .04% .Conclusion A HPLC‐MS/MS method for serum 25(OH)D2 and 25(OH)D3 has been established .It is sensitive and accurate ,and can be used in the evaluation of vitamin D in routine clinical la‐boratory .关键词
25-羟维生素D/液相色谱法/串联质谱法Key words
25-hydroxy vitamin D/liquid chromatography/tandem mass spectrometry引用本文复制引用
赵平,周慧,张曼..HPLC-MS／MS检测血清25-羟维生素D方法的建立和基本性能评价[J].检验医学与临床,2017,14(2):180-183,4.基金项目
国家高技术研究发展计划（2014AA022304）。 （）
