色谱

色谱 ›› 2019, Vol. 37 ›› Issue (12): 1337-1342.DOI: 10.3724/SP.J.1123.2019.05039

• 研究论文 • 上一篇    下一篇

高效液相色谱-荧光法测定多囊卵巢综合征患者血浆中吲哚类物质

韩润川1, 班艳娜2, 甄乾娜3, 周莹1, 张晓清1, 丁敏1   

  1. 1.重庆医科大学检验医学院, 临床检验诊断学教育部重点实验室, 重庆 400016;
    2.重庆医科大学附属第一医院生殖健康与不孕症专科, 重庆 400016;
    3.重庆医科大学附属第一医院内分泌科, 重庆 400016
  • 收稿日期:2019-05-23 出版日期:2019-12-08 发布日期:2020-12-11
  • 通讯作者: 丁敏.Tel:(023)68485239,E-mail:dingmin@cqmu.edu.cn.
  • 基金资助:
    重庆市教育委员会科学技术研究青-项目（2018-9-76）；重庆市研究生科研创新项目（CQCYS17137）.

Determination of indoles in plasma of polycystic ovarian syndrome patients by high performance liquid chromatography-fluorescence detection

HAN Runchuan1, BAN Yanna2, ZHEN Qianna3, ZHOU Ying1, ZHANG Xiaoqing1, DING Min1   

  1. 1. Key Laboratory of Clinical Laboratory Dignostics(Ministry of Education of China), School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China;
    2. Reproductive Health and Infertility Department the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;
    3. Endocrinology Department, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Received:2019-05-23 Online:2019-12-08 Published:2020-12-11
  • Supported by:
    Youth Project of Science and Technology Research Program of Chongqing Education Commission of China (No. 2018-9-76); Graduate Innovation Foundation of Chongqing (No. CQCYS17137).

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摘要： 建立了一种同时测定血浆吲哚类物质（吲哚丙酸、吲哚乙酸、吲哚与3-甲基吲哚）的高效液相色谱-荧光法（HPLC-FLD）。以液-液萃取法进行样本预处理，采用Shim-pack VP-ODS色谱柱（150 mm×4.6 mm，4.6 μm）分离，流动相为15 mmol/L磷酸二氢钠溶液-甲醇（42：58，v/v），柱温为30℃，流速为0.8 mL/min。吲哚丙酸、吲哚乙酸、吲哚与3-甲基吲哚在各自范围内线性关系良好。该方法的日内与日间相对标准偏差均小于6.31%，平均回收率为97.5%~107.0%。利用该方法对多囊卵巢综合征（PCOS）患者（n=61）与作为对照的正常育龄期女性（n=25）的血浆样本进行测定，结果发现PCOS患者血浆吲哚类物质显著高于对照组，其中吲哚是PCOS的危险因子和潜在的诊断标志物。该方法操作简便，灵敏度高，适用于临床检测和实验室研究。

关键词: 高效液相色谱-荧光法, 多囊卵巢综合征, 吲哚类物质, 血浆

Abstract: A new method was established for the determination of indoles (indole (IND), 3-methylindole (3-MI), indolyl-3-acetic acid (IAA) and indolyl-3-propionic acid (IPA)) in plasma by high performance liquid chromatography-fluorescence detection (HPLC-FLD). The analytes were separated simultaneously on a Shim-pack VP-ODS column (150 mm×4.6 mm, 4.6 μm) using 15 mmol/L sodium dihydrogen phosphate solution and methanol (48:52, v/v) as the mobile phases. The column temperature was 30 ℃, and the flow rate was 0.8 mL/min. The calibration curves of IND, 3-MI, IPA and IAA showed good linear relationships. The intra-day and inter-day relative standard deviations (RSDs) for the analytes were both less than 6.31%. The average recoveries of the analytes in plasma were 97.5%-107.0%. The method was successfully applied to the analysis of indoles in the plasma of healthy women of reproductive age (n=25) as controls and patients with polycystic ovarian syndrome (n=61). The results showed that the concentrations of indoles in the plasma were significantly different between the two groups, and IND was found to be a risk factor and a potential diagnostic biomarker for polycystic ovarian syndrome (PCOS). The method is simple, sensitive and suitable for use in clinical testing and laboratory research.

Key words: high performance liquid chromatography-fluorescence detection (HPLC-FLD), polycystic ovarian syndrome (PCOS), indoles, plasma

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