毛细管电泳激光诱导荧光检测肌腱和肌腱细胞中的羟脯氨酸
羟脯氨酸,激光诱导荧光检测,毛细管电泳,肌腱,细胞,,羟脯氨酸，激光诱导荧光检测，毛细管电泳，肌腱，细胞,1引言,2实验部分,3结果与讨论
摘 要 建立毛细管电泳激光诱导荧光检测(CELIF)分析羟脯氨酸的方法。肌腱和肌腱细胞中的胶原蛋白碱水解生成氨基酸，经异硫氰酸荧光素(FITC)衍生，采用LIFCE分离测定胶原蛋白特异性氨基酸羟脯氨酸。羟脯氨酸在0.5 μg/L~8×103 μg/L浓度范围内线性关系良好；检出限为0.5 μg/L。相对迁移率和相对峰高的相对标准偏差(RSD)分别为5.0%和6.1%。测定了60份肌腱和9份细胞样品，加标回收率为95%~110%。将所建立的毛细管电泳方法与高效液相色谱法(HPLC)进行比较，两者测定结果相对误差为-1.9%~2.0%。本法仅需一次荧光标记，操作简单、快速灵敏，12min内完成一个分析周期，适于测定肌腱和细胞样品。关键词 羟脯氨酸，激光诱导荧光检测，毛细管电泳，肌腱，细胞
Dertermination of Hydroxyproline in Tendon and Cell by Laser Induced FluorescenceCapillary Electrophoresis
Zou Xiaoli，Zhou Chunyan，Li Yuanqian， Zeng Hongyan
(Department of Sanitary Technology, West China School of Public Health, Sichuan University, Chengdu 610041)
Abstract The methods for analysis of hydroxyproline by capillary electrophoresis (CE) with laser induced fluorescence (LIF) were developed. The collagen in tendons and cells can be decomposed to amino acid and derivatizated with fluorescein isothiocyanate (FITC). The content of its special amino acid hydroxyproline was determined by LIFCE method. The linear range was 0.5-8×103 μg/L and the detection limit of the method was 0.5 μg/L. The RSD for relative migration time and relative fluorescence intensity ratio were 5.0 % and 6.1%. The proposed method was used in analysis of 60 tendons and 9 cells sample to study the decompositon of tendons and the change of cells growth after culture. The average recoveries were 95%-110%. And compared with high performance liquid chromatography method, the relative error of determination results between the two methods was -1.9%-2.0%. Only one fluorescent labeling was needed. One analysis can be completed within 12 min. This method was available for determination of hydroxyproline content in tendons and cells. ......
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