人血清中牛磺酸的HPLC-荧光法测定方法学研究(1)
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[摘要] 目的：建立测定人血清中牛磺酸浓度的HPLC-荧光法。方法：以阿德福韦为内标，待测血清样品经乙腈沉淀，上清液加入荧光胺生成衍生化物，色谱柱为Phenomenex Kromasil NH2柱，乙腈-水(47∶53，磷酸调pH到4.0)为流动相，流速1.2 ml/min，荧光检测波长275 nm(激发波长)、420 nm(发射波长)。结果：每个样品分析时间约10 min，扣除血清中内源性牛磺酸，牛磺酸的线性范围为1.875～120.000 μg/ml，日内、日间精密度的RSD均小于15%，低、中、高浓度(3.750、15.000、60.000 μg/ml)的平均提取回收率分别为88.9%、96.0%和 98.7%。结论：该法操作快速、简单、准确，可用于临床药代动力学的研究。
[关键词] 牛磺酸；高效液相色谱-荧光法；血清药物浓度
[中图分类号] R969.1[文献标识码]B [文章编号]1673-7210(2009)01(b)-042-03
Study on the determination of taurine in human serum by HPLC-fluorescence
LIU Guo-bin1, LI Jian-chun2
(1.Pharmacy of the Affiliated Hospital of Bengbu Medical College, Bengbu233004, China; 2.Department of Pharmacy, Bengbu Medical College, Bengbu233030, China)
[Abstract] Objective: To establish a HPLC-fluorescence method for the determination of taurine in human serum. Methods: The adefovir was used as the internal standard, the serum samples to be determined were precipitated with acetonitrile, and fluorescamine was added into the supernate to form the derivation produce, the produce was analyzed on a Phenomenex Kromasil NH2 column, Acetonitrile-water (47∶53 V/V, adjusted to pH 4.0 with phosphonic acid) was used as the mobile phase with the flow rate of 1.2 ml/min. The λEx was set at 275 nm and λEm was set at 420 nm. Results: Each sample was analyzed within 10 minutes. The calibration curve was linear at the range of 1.875-120.000 μg/ml for taurine. The RSD of intra-day and inter-day were less than 15% respectively. The average extraction recoveries of taurine were 88.9%, 96.0% and 98.7% at concentrations of 3.750, 15.000 and 60.000 μg/ml respectively. Conclusion: A simple, rapid and specific HPLC-fluorescence method is developed to determine taurine in human serum.
[Key words] Taurine; HPLC-fluorescence; Serum concentration
牛磺酸(taurine)又名2-氨基乙磺酸，是一种人体必需的非蛋白质氨基酸，于1827年首次从牛胆汁中分离而得名，其没有遗传密码子，因此不能组成蛋白质和酶类。研究发现，牛磺酸具有多种生理功能，维持细胞内外渗透压平衡、清除氧自由基、抗脂质过氧化损伤、调节细胞钙稳态、直接的细胞膜稳定作用等。临床上，牛磺酸用来治疗支气管炎、急慢性肝炎、心力衰竭、子宫出血、高血压等[1，2]。血中牛磺酸含量测定方法有氨基酸自动分析仪[3]、电化学检测器[4]、薄层扫描法[5]，氨基酸自动分析仪主要用于测定正常人血浆，电化学检测器用于急性低血压时孤束核内牛磺酸含量变化，由于受到仪器检测范围的影响，缺乏血药浓度线性关系考察，而薄层扫描法操作费时，影响因素多，因此，目前的实验方法均不适于大批量样品的临床药代动力学研究 ......
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