[1]刘 浩,彭 勇,尹一帆,等.基于UPLC-MS/MS 建立早产儿血清25-(OH)D3 和3-epi-25-(OH)D3 检测新方法及临床初步应用[J].现代检验医学杂志,2023,38(03):170-175.[doi:10.3969/j.issn.1671-7414.2023.03.031]
　LIU Hao,PENG Yong,YIN Yi-fan,et al.A Novel UPLC-MS/MS Method for Simultaneous Quantification of Serum 25- (OH)D3 and 3-epi-25(OH)D3 Followed by the Preliminary Application in Clinics[J].Journal of Modern Laboratory Medicine,2023,38(03):170-175.[doi:10.3969/j.issn.1671-7414.2023.03.031]
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基于UPLC-MS/MS 建立早产儿血清25-(OH)D3 和3-epi-25-(OH)D3 检测新方法及临床初步应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年03期
页码:
170-175
栏目:
研究简报·实验技术
出版日期:
2023-05-15

文章信息/Info

Title:
A Novel UPLC-MS/MS Method for Simultaneous Quantification of Serum 25- (OH)D3 and 3-epi-25(OH)D3 Followed by the Preliminary Application in Clinics
文章编号:
1671-7414（2023）03-170-06
作者:
刘 浩; 彭 勇; 尹一帆; 叶 亮; 陈 敏; 苗静琨
( 重庆市妇幼保健院/ 重庆医科大学附属妇女儿童医院儿科/ 新生儿筛查中心，重庆 401147)
Author(s):
LIU Hao;  PENG Yong;  YIN Yi-fan;  YE Liang;  CHEN Min;  MIAO Jing-kun
（Newborn Screening Center, Department of Pediatrics, Chongqing Health Center for Women and Children/Women and Children’s Hospital of Chongqing Medical University, Chongqing 401147, China）
关键词:
维生素D; 3-epi-25- 羟维生素D3; 超高效液相色谱- 串联质谱; 早产儿
分类号:
R714.21；R392.11
DOI:
10.3969/j.issn.1671-7414.2023.03.031
文献标志码:
A
摘要:
目的 建立血清25- 羟维生素D3[25-hydroxyvitamin D3, 25-(OH)D3] 及3-epi-25- 羟维生素D3[3-epi-25-hydroxyvitaminD3, 3-epi-25-(OH)D3] 超高效液相色谱- 串联质谱检测新方法（ultra-performance liquid chromatography-tandem massspectrometry，UPLC-MS/MS），并在早产儿中开展初步应用。方法 采用液- 液萃取法提取化合物，五氟苯基丙基（pentafluorophenylpropyl，PFPP）色谱柱分离25-(OH)D3 与3-epi-25-(OH)D3；从最低定量限、线性关系、精密度与准确度等四个方面对新建方法进行验证，并对134 例早产儿血清样本进行检测分析。结果 25-(OH)D3 及3-epi-25-(OH)D3 的最低定量限分别为6.05±0.78 nmol/L，1.48±0.20 nmol/L；线性范围3.78 ～ 480.00 nmol/L，1.00 ～ 128.00 nmol/L；相关系数r2 为0.992 2，0.992 8；25-(OH)D3 与3-epi-25-(OH)D3 低、中、高浓度质控品的日内精密度和日间精密度均小于15.00%；回收率109.92%，102.25%，98.76%；97.75%, 95.25%，99.80%。134 例早产儿血清25-(OH)D3 平均浓度22.35±13.28nmol/L，3-epi-25-(OH)D3 平均浓度5.43±4.35 nmol/L；3-epi-25-(OH)D3 占总25-(OH)D3 的平均比例为19.96%±12.08%，范围0.00% ～ 60.62%。以25-(OH)D3 计算，维生素D（Vitamin D, Vit D）缺乏率、不足率、充足率分别为78.36%，19.40%，2.24%；以总25-(OH)D3 计算，Vit D 缺乏率、不足率、充足率分别为65.67%，26.12%，8.21%；二者Vit D 缺乏率及充足率的比较, 差异均有统计学意义（χ2=5.351，1.719，4.823，均P ＜ 0.05）。结论 本研究建立的UPLC-MS/MS检测法性能良好且能精准检测早产儿血清25-(OH)D3 浓度，在早产儿Vit D 评估方面具有较好的应用价值。
Abstract:
Objective To establish a novel ultra-performance liquid chromatography-tandent mass spectrometry （UPLC-MS/ MS） method for quantification of serum 25-hydroxyvitamin D3 [25-(OH)D3] and 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH) D3], and to assess the Vitamin D (Vit D) levels in premature infants. Methods The liquid-liquid extraction was used to extract compounds after protein precipitation, and pentafluorophenyl propyl (PFPP) column was used to resolve 25-(OH)D3 and 3-epi- 25(OH)D3. Then used the established method for a clinical application in 134 premature infants after performance validation. Results For 25(OH)D3 and 3-epi-25(OH)D3, the low limit of quantification (LLOQ) were 6.05±0.78nmol/L, 1.48±0.20 nmol/ L. The linear ranges were 3.78~480.00nmol/L, 1.00~128.00nmol/L. The correlation coefficients (r2) were 0.992 2, 0.992 8. The relative standard deviations (RSD) of intra-day and inter-day for 25(OH)D3 and 3-epi-25(OH)D3 with low, middle, high concentrations were all less than 15.00%. The recovery rates were 109.92%, 102.25%, 98.76% and 97.75%, 95.25%, 99.80%. respectively. The average serum concentrations of 25-(OH)D3 and 3-epi-25-(OH)D3 in 134 premature infants were 22.35±13.28nmol/L and 5.43±4.35 nmol/L, respectively. The percentage of 3-epi-25-(OH)D3 to total 25-(OH)D3 was 19.96%±12.08%，ranging from 0.00% to 60.62%. Based on 25-(OH)D3, Vit D deficiency, insufficiency and sufficiency were 78.36%, 19.40% and 2.24%. While these values were 65.67%, 26.12% and 8.21%, respecitively based on total 25-(OH)D3. The P values of Vit D deficiency and Vit D sufficiency were both less than 0.05（χ2=5.351，1.719，4.823，all P ＜ 0.05）. Conclusion The novel UPLC-MS/MS method successfully detects the 25-(OH)D3 and 3-epi-25-(OH)D3 in premature infants, which has a great value in clinical applications for Vit D assessment of premature infants.

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备注/Memo

备注/Memo:
基金项目： 重庆市卫健委资助项目（2022WSJK018）: 基于UPLC-MS/MS 分析VD 代谢通路新方法的建立及其临床应用研究；重庆市妇幼保健院院级科研课题（2019YJMS08）：维生素A 通过肠道菌群影响妊娠期糖尿病及妊娠结局的研究。
作者简介：刘浩（1983-），男，医学硕士，主管技师，研究方向：遗传代谢病发病机制及诊断研究，E-mail：1320904@qq.com。
通讯作者：苗静琨（1979-），女，医学博士，副主任医师，研究方向：遗传代谢病发病机制及诊治研究，E-ma
更新日期/Last Update: 2023-05-15