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Determination of Triterpenic Acids in Human Serum by High-Performance Liquid Chromatography: Triterpenoid Interaction with Serum Protein
- Source :
- Digital.CSIC. Repositorio Institucional del CSIC, instname
- Publication Year :
- 2011
- Publisher :
- American Chemical Society (ACS), 2011.
-
Abstract
- 6 páginas, 4 figuras, 1 tabla.<br />Terpenic 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.<br />This work was supported in part by grants from Comision Interministerial de Ciencia y Tecnologia (CICYT, AGL2008-02258), Instituto de Salud Carlos III (RETICS, RD06/0045), and Exxentia Group (110101080017 21).
- Subjects :
- Terpenic acids
High-performance liquid chromatography
Terpenoid
Triterpene
Spectrophotometry
medicine
Humans
Chromatography, High Pressure Liquid
chemistry.chemical_classification
Detection limit
Chromatography
medicine.diagnostic_test
Chemistry
Elution
Extraction (chemistry)
Human serum
Blood Proteins
General Chemistry
Repeatability
Triterpenes
Spectrophotometry, Ultraviolet
HPLC
General Agricultural and Biological Sciences
Protein Binding
Subjects
Details
- ISSN :
- 15205118 and 00218561
- Volume :
- 59
- Database :
- OpenAIRE
- Journal :
- Journal of Agricultural and Food Chemistry
- Accession number :
- edsair.doi.dedup.....02b86adba6907a51a276f14323931fdb