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采用高效液相色谱荧光检测法测定生物样品和分离的肝细胞中的牛磺酸。

Determination of taurine in biological samples and isolated hepatocytes by high-performance liquid chromatography with fluorimetric detection.

作者信息

Waterfield C J

机构信息

Department of Toxicology, School of Pharmacy, University of London, UK.

出版信息

J Chromatogr B Biomed Appl. 1994 Jul 1;657(1):37-45. doi: 10.1016/0378-4347(94)80067-7.

Abstract

A high-performance liquid chromatographic method with fluorimetric detection is described for the routine and selective determination of taurine in urine, serum, tissues and isolated hepatocytes. The preparation and use of ion-exchange resins to extract taurine from biological samples is included. Taurine was derivatised with o-phthalaldehyde/2-mercaptoethanol prior to injection onto a C18 column (LiChrospherR 100 RP-18, 5 microns, 125 x 4 mm I.D.). Isocratic elution of the adduct was carried out using NaH2PO4 (0.05 M, pH 5.4) in methanol and water (43:57, v/v). Homoserine was used as an internal standard to facilitate the standardisation and quantitation of samples and analysis was completed in 6 min with homoserine and taurine eluting after 3 and 4 min, respectively. The method will detect 0.5 pmol of taurine on the column. Appropriate dilutions of these biological samples enable these samples to be assayed on an autosampler, using the same standard curve. Concentrations of taurine in human, dog and rat urine, rat liver, serum and isolated hepatocytes are reported.

摘要

本文描述了一种采用荧光检测的高效液相色谱法,用于尿液、血清、组织和分离的肝细胞中牛磺酸的常规选择性测定。其中包括离子交换树脂的制备及其用于从生物样品中提取牛磺酸的方法。牛磺酸在注入C18柱(LiChrospherR 100 RP - 18,5微米,内径125×4毫米)之前,先用邻苯二甲醛/2 - 巯基乙醇进行衍生化。加合物的等度洗脱使用甲醇和水(43:57,v/v)中的NaH2PO4(0.05 M,pH 5.4)进行。使用高丝氨酸作为内标以促进样品的标准化和定量,分析在6分钟内完成,高丝氨酸和牛磺酸分别在3分钟和4分钟后洗脱。该方法在柱上可检测到0.5皮摩尔的牛磺酸。对这些生物样品进行适当稀释后,可使用相同的标准曲线在自动进样器上对这些样品进行测定。文中报道了人、狗和大鼠尿液、大鼠肝脏、血清及分离的肝细胞中牛磺酸的浓度。

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