Quantification of hydroxyproline in small amounts of skin tissue using isocratic high performance liquid chromatography with NBD-F as fluorogenic reagent

In order to determine the collagen content of small amounts of skin tissue, we developed a new, simple and highly sensitive method of measuring the quantity of hydroxyproline (Hyp) using isocratic high performance liquid chromatography (HPLC) with a fluorogenic agent, 7-fluoro-4-nitrobenzo-2-oxa-1,3...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-09, Vol.824 (1), p.161-165
Main Authors: Kakinuma, Miwa, Watanabe, Yasuo, Hori, Yusuke, Oh-i, Tsunao, Tsuboi, Ryoji
Format: Article
Language:English
Subjects:
4-Chloro-7-nitrobenzofurazan - analogs & derivatives
4-Chloro-7-nitrobenzofurazan - chemistry
Amino Acids - analysis
Amino Acids - isolation & purification
Analysis
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Collagen
Diaphragm - chemistry
Fluorescent Dyes - chemistry
Fundamental and applied biological sciences. Psychology
General pharmacology
High performance liquid chromatography (HPLC)
Hydroxyproline
Hydroxyproline - analysis
Kidney - chemistry
Liver - chemistry
Male
Medical sciences
Muscles - chemistry
Myocardium - chemistry
NBD-F
Pharmacology. Drug treatments
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Skin
Skin - chemistry
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Summary:In order to determine the collagen content of small amounts of skin tissue, we developed a new, simple and highly sensitive method of measuring the quantity of hydroxyproline (Hyp) using isocratic high performance liquid chromatography (HPLC) with a fluorogenic agent, 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-F). The recovery rate of Hyp and reproducibility of the assay were high, and the test was sensitive enough to detect Hyp in less than 1 mg of skin tissue. This method is clinically useful for ensuring accurate diagnosis and for monitoring specific skin conditions using small human skin samples collected in biopsies.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2005.07.019