High Pressure Liquid Chromatographic Assay for Salicylic Acid and its Metabolites in Rat Urine
作者:
BhaskerV. Shetty,
Srikumaran Melethil,
期刊:
Analytical Letters
(Taylor Available online 1988)
卷期:
Volume 21,
issue 3
页码: 395-410
ISSN:0003-2719
年代: 1988
DOI:10.1080/00032718808066333
出版商: Taylor & Francis Group
关键词: Salicylic acid;salicyluric acid;gentisic acid;salicyl acyl glucuronide;salicyl phenolic glucuronide;analysis;rat urine;HPLC
数据来源: Taylor
摘要:
An HPLC method for the determination of salicylic acid (SA), gentisic acid (GA), salicyluric acid (SU), and salicyl acyl glucuronide (SAG) in rat urine was developed. The method consisted of extracting SA, GA, and SU from acidified urine into 50:50 mixture of ethyl acetate and butyl chloride. Salicyl acyl glucuronide was extracted from neutral urine after conversion to salicyl hydroxamic acid with hydroxylamine. Salicyl phenolic glucuronide was estimated indirectly as the difference between total salicylate and sum of the four constituents mentioned above. Chromatographic separation was done on a C18column with U.V. detection at 310 nm using a mobile phase consisting of 5–10% acetonitrile in 3% glacial acetic acid. The extraction recovery of these compounds from spiked urine ranged from 90–108%. The detection limits were 10 μg/ml for GA, SU and SA, and 2.5 μg/ml for SHA. The method was applied to the study of salicylic acid metabolism in the rat.
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