华西口腔医学杂志

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紫外检测-反相高效液相色谱法测定唾液富组蛋白5的实验研究

李岩1,2 曾红燕1 邹晓莉1 李庆福3 周红梅3   

  1. 1.四川大学华西公共卫生学院卫生检验教研室, 四川成都610041;2.四川安全生产检测检验技术研究院职业危害检测所, 四川成都610000;3.四川大学华西口腔医院黏膜科, 四川成都610041
  • 收稿日期:2010-12-25 修回日期:2010-12-25 出版日期:2010-12-20 发布日期:2010-12-20
  • 通讯作者: 邹晓莉,Tel:028-85501301
  • 作者简介:李岩(1980—),男,新疆人,检验师,学士
  • 基金资助:

    教育部新世纪优秀人才支持计划基金资助项目(NCET07-0591)

Determination of histatins 5 in salivary by reversed-phase high performance liquid chromatography method with ultraviolet detection

LI Yan1,2, ZENG Hong-yan1, ZOU Xiao-li1, LI Qing-fu3, ZHOU Hong-mei3   

  1. 1. Dept. of Sanitary Technology, West China School of Public Health of Sichuan University, Chengdu 610041, China; 2. Examination Institute of Occupational Hazards, Sichuan Academy of Inspecting and Testing Technology for Safety Production, Chengdu 610000, China;3. Dept. of Oral Medicine, West China College of Stomatology, Sichuan University,Chengdu 610041, China
  • Received:2010-12-25 Revised:2010-12-25 Online:2010-12-20 Published:2010-12-20
  • Contact: ZOU Xiao-li,Tel:028-85501301

摘要:

目的建立测定唾液中富组蛋白5的紫外检测-反相高效液相色谱法。方法将收集的唾液样品用pH2.5的磷酸盐缓冲液适度稀释后,离心,取上清液进样,于0.01 μg·mL-1磷酸盐缓冲液(pH3.5)中洗脱,经反相C18柱分离,紫外检测器在276 nm波长处测定其中的富组蛋白5含量。结果方法的线性范围为1.0~50.0 μg·mL-1,检出限为0.12μg·mL-1,标准溶液测定保留时间和峰面积的相对标准偏差分别为0.68%和4.13%。将所建立的方法用于唾液样品的分析,获得了较为满意的结果,样品测定的相对标准偏差为4.41%,加标回收率为88.4%~109.0%。结论紫外检测-反相高效液相色谱法准确快速,样品处理简单,15 min内可完成1次分析周期,适用于唾液中富组蛋白5的快速检测。

关键词: 富组蛋白5, 唾液, 高效液相色谱法

Abstract:

Objective The determination method of histatins 5 in human saliva with reversed-phase high performance liquid chromatography(HPLC) was developed. Methods Salivary samples were collected and diluted with phosphate buffer(pH2.5). The upper solution was determined with HPLC. Phosphate buffer(pH 3.5) of the mobile phase and C18 column was used throughout the experiment. The detection wavelength was 276 nm. Results The linear ranges were 1.0-50.0 μg·mL-1. The detection limit was 0.12 μg·mL-1. The relative standard derivations(RSD) of standard solution for reserved time and peak area were 0.68% and 4.13% respectively. The proposed method was applied for analysis of salivary samples and the satisfactory results were obtained. RSD for sample determination was 4.41% and the average recoveries were 88.4%-109.0%. Conclusion The method was quick, simple and accurate. Analytical time was less than 15 min. It was adapted for analysis of salivary histatins 5 in salivary samples.

Key words: histatins 5, salivary, reversed-phase high performance liquid chromatography