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[使用高压液相色谱法测定血液胆汁酸:色谱法、酶法和免疫酶法的比较]

[Determination of blood bile acids using high pressure liquid chromatography: comparison of chromatographic, enzymatic and immunoenzymatic methods].

作者信息

Buratti M, Pellegrino O, Soardo A, Colombi A

机构信息

Istituti Clinici di Perfezionamento, Laboratorio di Ricerche Cliniche, Anatomia e Istologia Patologica, Milano.

出版信息

Med Lav. 1990 Jan-Feb;81(1):65-74.

PMID:2199808
Abstract

A method has been developed for measuring serum conjugated bile acids by HPLC. Serum samples, to which the internal standard is added, are purified by a solid phase procedure and injected on a reverse phase C18 column. Elution is accomplished by means of a flow gradient and peaks are detected at 198 nm. The detection limit ranges between 0.05 and 0.20 mumol/l for different analytes; between-day precision (CV 5.8%), working linear range (up to 50 mumol/l) and recovery (87%) were established. Comparison of the results obtained with HPLC, enzymatic and immuno-enzymatic methods gave high correlation coefficients. The method was applied for diagnostic purpose to a group of subjects suffering from various liver diseases. Also, 106 healthy workers, not occupationally exposed to known or potentially hepatotoxic agents, were studied in order to establish reference values for use in biological monitoring of chronic low level exposure to solvents. The method has the advantage of a more simple procedure compared to previously reported HPLC methods and appears to be well suited for routine use in toxicological and clinical test laboratories.

摘要

已开发出一种通过高效液相色谱法(HPLC)测量血清结合胆汁酸的方法。向血清样品中加入内标后,通过固相程序进行纯化,然后注入反相C18柱。通过流动梯度实现洗脱,并在198nm处检测峰。不同分析物的检测限在0.05至0.20μmol/L之间;确定了日间精密度(CV 5.8%)、工作线性范围(高达50μmol/L)和回收率(87%)。将HPLC法与酶法和免疫酶法所得结果进行比较,得到了较高的相关系数。该方法用于一组患有各种肝病的受试者的诊断。此外,对106名未职业接触已知或潜在肝毒性物质的健康工人进行了研究,以建立用于慢性低水平接触溶剂生物监测的参考值。与先前报道的HPLC方法相比,该方法具有程序更简单的优点,似乎非常适合毒理学和临床检测实验室的常规使用。

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