中国临床药理学杂志2025，Vol.41Issue(6)：866-871,6.DOI:10.13699/j.cnki.1001-6821.2025.06.022
用LC-MS/MS法检测血清中维生素D的含量
Determination the contents of vitamin D in serum by LC-MS/MS
摘要
Abstract
Objective To establish a quantitative analysis method for serum vitamin D(VD)based on liquid chromatography-tandem mass spectrometry(LC-MS/MS)and evaluate its clinical application value in pediatric nutritional support.Methods Serum samples were processed with acetonitrile,using 25-hydroxyvitamin D2-d6(25-OH-VD2-d6)and 25-hydroxyvitamin D3-d6(25-OH-VD3-d6)as internal standards.The chromatographic column was Kinetex C18(50.0 mm×3.0 mm,2.6 μm).The mobile phase consisted of 0.1％formic acid aqueous solution and 0.1％formic acid methanol solution,with gradient elution at a flow rate of 0.5 mL·min-1 and an injection volume of 10 μL.The column temperature was set to 30 ℃.Electrospray ionization was used in positive ion mode,with multiple reaction monitoring.The LC-MS/MS method was evaluated for specificity,calibration curves,lower limits of quantification,precision,recovery rate,stability,and matrix effects.The method was applied to detect VD levels in the serum of infants from the external group(oral 10 μg·kg-1 VD drops,qd)and the parenteral group[parenteral nutrition solution with injectable fat-soluble vitamin(Ⅰ)administered intravenously,it was equivalent to 10 μg·mL-1 VD,qd].Results Both 25-OH-VD2 and 25-OH-VD3 showed good linearity in the range of 5-100 ng·mL-1,with the 25-OH-VD2 standard curve of y=1.00x+2.34×10-2(R=0.9998),and a lower limit of quantification(LLOQ)of 0.5 ng·mL-1;25-OH-VD3 standard curve of y=1.00x+5.6×10-3(R=0.9998),and LLOQ of 1 ng·mL-1.The recovery rate of 25-OH-VD2 quality control samples was 95.59％-105.38％,with intra-and inter-day precision≤4.75％and ≤ 4.68％,respectively.The recovery rate of 25-OH-VD3 was 98.38％-113.63％,with intra-and inter-batch precision ≤5.64％and ≤7.83％,respectively.Stability of 25-OH-VD2 and 25-OH-VD3 samples at different concentrations was good under various conditions,with matrix effects ranging from 85％-110％and 88％-112％,respectively.After enteral or parenteral VD supplementation,the serum levels of 25-OH-VD2 and 25-OH-VD3 in both groups of preterm infants significantly increased,with the parenteral group showing significantly higher concentrations of 25-OH-VD2 and 25-OH-VD3 compared to the enteral group(P＜0.05).The daily weight gain rate of preterm infants in the parenteral group was also significantly higher than those in the enteral group(all P＜0.05).Conclusion The quantitative analysis method of serum VD based on LC-MS/MS has strong specificity and high sensitivity,which is suitable for the monitoring of VD level in premature infants.关键词
维生素D/液相色谱-串联质谱/定量分析/早产儿/营养支持Key words
vitamin D/liquid chromatography tandem mass spectrometry/quantitative analysis/preterm infants/nutritional support分类
医药卫生引用本文复制引用
施卉,李海娟,顾玲琪,张小华,金霞,顾锦华..用LC-MS/MS法检测血清中维生素D的含量[J].中国临床药理学杂志,2025,41(6):866-871,6.基金项目
南通市药物政策与药学服务研究基金资助项目(2023NTPA11) （2023NTPA11）
