色谱

色谱 ›› 2026, Vol. 44 ›› Issue (3): 296-301.DOI: 10.3724/SP.J.1123.2025.06017

• 研究论文 • 上一篇    下一篇

液相色谱-串联质谱法测定泪液中维生素A和维生素E及其与干眼的关系分析

罗保斌1, 焦婧然2, 白晶1, 刘向祎1,*()   

  1. 1.首都医科大学附属北京同仁医院，北京 100730
    2.首都医科大学附属北京佑安医院，北京 100069
  • 收稿日期:2025-06-16 出版日期:2026-03-08 发布日期:2026-03-12
  • 通讯作者: *Tel：010-58268695，E-mail：liuxiangyi@ccmu.edu.cn.
  • 基金资助:
    首都卫生发展科研专项(2020-1-4014)

Determination of vitamin A and vitamin E in human tear by liquid chromatography-tandem mass spectrometry and analysis of their relationship with dry eye

LUO Baobin1, JIAO Jingran2, BAI Jing1, LIU Xiangyi1,*()   

  1. 1.Beijing Tongren Hospital，Capital Medical University，Beijing 100730，China
    2.Beijing Youan Hospital，Capital Medical University，Beijing 100069，China
  • Received:2025-06-16 Online:2026-03-08 Published:2026-03-12
  • Supported by:
    Capital’s Funds for Health Improvement and Research(2020-1-4014)

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摘要：

泪液中维生素A（vitamin A， VA）和维生素E（vitamin E， VE）对于干眼的诊断和治疗具有潜在的临床预示价值，本研究建立了一种液相色谱-串联质谱法测定人泪液中VA和VE的分析方法。选用C18色谱柱（30 mm×2.1 mm， 2.6 μm），以0.1%甲酸水溶液-含2 mmol/L乙酸铵的0.1%甲酸甲醇溶液为流动相，采用大气压化学电离正离子模式，在0.4 mL/min的流速下进行梯度洗脱，4 min内可实现良好的色谱峰基线分离。经验证，该方法线性关系良好，VA的线性范围为5.00～300.00 ng/mL，检出限为2.00 ng/mL，加标回收率为98.3%～104.7%，日内精密度和日间精密度（以相对标准偏差（RSD）表示）为1.2%～7.0%；VE线性范围为25.00～1 000.00 ng/mL，检出限为6.00 ng/mL，加标回收率为97.9%～105.5%，日内精密度和日间精密度为3.0%～5.7%。使用本方法测定了5名干眼患者和9名健康志愿者的泪液标本，VA、VE质量浓度用中位数（四分位数间距）表示。干眼组泪液中VA含量为9.60（5.13～12.54） ng/mL，VE含量为42.00（31.75～128.00） ng/mL；健康组泪液中VA含量为18.10（12.46～21.69） ng/mL，VE含量为211.0（181.0～459.75） ng/mL。Wilcoxon Mann-Whitney两样本秩和检验干眼组和健康组泪液中VA、VE水平具有统计学差异，干眼组泪液中VA、VE浓度均低于健康组。本研究提供了一种简便可靠的泪液中VA、VE的液相色谱-串联质谱分析方法，探索了其与干眼的关系，为干眼相关因素的研究提供了新的可能的参考指标。

关键词: 液相色谱-串联质谱, 泪液, 干眼, 维生素A, 维生素E

Abstract:

Dry eye is characterized as a multifactorial disorder affecting the tear film and ocular surface， leading to symptoms of discomfort and visual disturbances， with the potential for ocular surface damage. It is among the most prevalent ocular surface disorders， with an estimated prevalence ranging from 5.0% to 50.0%， and an overall prevalence between 21.0% and 50.4% in China. Despite its high global prevalence， the fundamental mechanisms underlying the pathology of dry eye remain largely unexplored. Vitamins are crucial for maintaining ocular surface homeostasis， with vitamins A and E being physiologically present and essential for cell differentiation， development， and proper function. Deficiencies in these vitamins are frequently associated with dry eye， and systemic supplementation has been shown to benefit patients with low vitamin intake. Vitamins A and E may have potential clinical predictive value for the diagnosis and treatment of dry eye. In this study， a liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was developed to simultaneously measure the concentrations of vitamins A and E in human tears. A Phenomenex Kinetex C18 column was employed for gradient elution at a flow rate of 0.4 mL/min. The mobile phase A consisted of a 0.1% formic acid aqueous solution， while mobile phase B comprised a 2 mmol/L ammonium acetate and 0.1% formic acid methanol solution. Positive atmospheric pressure chemical ionization in multiple reaction monitoring mode was utilized for MS/MS detection. The analytical method for vitamin A demonstrated a validated linear range of 5.00-300.00 ng/mL， with a limit of detection at 2.00 ng/mL. The spiked recoveries were between 98.3% and 104.7%， and intra- and inter-day precision ranged from 1.2% to 7.0%. Similarly， the method for vitamin E exhibited a validated linear range of 25.00-1 000.00 ng/mL， with a limit of detection at 6.00 ng/mL. The spiked recoveries ranged from 97.9% to 105.5%， and intra- and inter-day precision varied between 3.0% and 5.7%. Tear samples from five patients with dry eye and nine healthy volunteers were analyzed using this method. In the dry eye group， the average concentrations of vitamin A and vitamin E were 9.60 （5.13-12.54） ng/mL and 42.00 （31.75-128.00） ng/mL， respectively. In the healthy group， the average concentrations of vitamin A and vitamin E were 18.10 （12.46-21.69） ng/mL and 211.00 （181.00-459.75） ng/mL， respectively. Statistical differences in vitamin A and vitamin E levels between the dry eye group and the healthy group were identified using the Wilcoxon Mann-Whitney test （P<0.05）. This study introduces a straightforward and reliable LC-MS/MS method for the detection of tear VA and VE levels， while also investigating their association with dry eye. This offers a novel reference for understanding the potential risk factors associated with dry eye.

Key words: liquid chromatography-tandem mass spectrometry （LC-MS/MS）, tears, dry eye, vitamin A, vitamin E

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