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Title

Determination of Triterpenic Acids in Human Serum by High-Performance Liquid Chromatography: Triterpenoid Interaction with Serum Protein

AuthorsRada, Mirella ; Ruiz-Gutiérrez, Valentina ; Guinda Garín, Mª Ángeles
KeywordsTerpenic acids
HPLC
Terpenoid
Human serum
Issue Date23-Feb-2011
PublisherAmerican Chemical Society
CitationJournal of Agricultural and Food Chemistry 59(6): 2308-2313 (2011)
AbstractTerpenic acids are under development as therapeutic agents in numerous treatments. In support of pharmacokinetic and toxicological evaluations, a robust assay based on high-performance liquid chromatography (HPLC) was developed for the analysis of the terpenoids in human serum. For a clear understanding of the differences in biological activity of these compounds, the interactions between oleanolic or betulinic acids and human serum protein have been studied by ultraviolet−visible (UV−vis) absorption under physiological conditions. A combination of liquid/liquid extraction, centrifugation, and consecutive HPLC resulted in simultaneous separation, identification, and quantification of the oleanolic, betulinic, and ursolic acids. The validity of the developed method was established by determining linearity, recovery, precision, accuracy, limit of detection, and quantification. Detection limits were in the range of 3.3−4.3 ng/mL, and linearity values ranged up to 1 μg/mL. The repeatability of the method was good. All compounds can be well-distinguished by order of elution during liquid chromatography. The pentacyclic triterpenoids have been identified by retention time comparison to pure standards and quantified by an internal standard. The results by UV−vis absorption spectra experiments (240−340 nm) indicate that protein structures have been perturbed in the presence of oleanolic and betulinic acids.
Description6 páginas, 4 figuras, 1 tabla.
Publisher version (URL)http://dx.doi.org/10.1021/jf1039537
URIhttp://hdl.handle.net/10261/42286
DOI10.1021/jf1039537
ISSN0021-8561
E-ISSN1520-5118
Appears in Collections:(IG) Artículos
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