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  

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

作　　者：韩润川 班艳娜 甄乾娜[3] 周莹 张晓清[1] 丁敏[1] HAN Runchuan;BAN Yanna;ZHEN Qianna;ZHOU Ying;ZHANG Xiaoqing;DING Min(Key Laboratory of Clinical Laboratory Dignostics(Ministry of Education of China),School of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China;Reproductive Health and Infertility Department the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Endocrinology Department,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[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

出　　处：《色谱》2019年第12期1337-1342,共6页Chinese Journal of Chromatography

基　　金：重庆市教育委员会科学技术研究青年项目(2018-9-76);重庆市研究生科研创新项目(CQCYS17137)~~

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, 4.6μm)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。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.

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 

分 类 号：O658[理学—分析化学]