A validated HPLC-FLD method for analysis of intestinal absorption and metabolism of capsaicin and dihydrocapsaicin in the rat

Mónika Kuzma, Krisztina Fodor, Gábor Maász, Péter Avar, G. Mózsik, Tibor Past, Emil Fischer, P. Perjési

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A sensitive and selective reverse-phase high performance liquid chromatographic method with fluorescence detection has been developed for determination of capsaicin (8-methyl-N-vanillyl-(trans)-6-nonenamid) and dihydrocapsaicin (8-methyl-N-vanillylnonanamide) in samples generated in rat small intestine luminal perfusion experiments. The experiments were designed to study the biotransformation of capsaicinoids in the small intestine in the rat. The chromatographic separation was performed at room temperature on a ZORBAX Eclipse® XDB-C8 column using isocratic elution with a mobile phase consisting 0.05M orthophosphoric acid solution and acetonitrile (60:40, v/v; pH 3.0) with a flow rate of 1.5mL/min. Fluorescence detection was performed at excitation and emission wavelengths of 230 and 323nm, respectively. The method was evaluated for a number of validation characteristics (accuracy, repeatability and intermediate precision, limit of detection, limit of quantification and calibration range). The limit of detection (LOD) was 50ng/mL and the limit of quantification (LOQ) was 100ng/mL for both capsaicin and dihydrocapsaicin reference standards dissolved in blank perfusate. The method was successfully applied for investigation of intestinal absorption of capsaicin and dihydrocapsaicin while 30μg/mL standardized Capsicum extract - containing capsaicin and dihydrocapsaicin - was luminally perfused for a 90min period. The structure of the glucuronide metabolites of capsaicin and dihydrocapsaicin appeared in the perfusate was identified by mass spectrometry.

Original languageEnglish
Pages (from-to)59-66
Number of pages8
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume103
DOIs
Publication statusPublished - Jan 5 2015

Fingerprint

Capsaicin
Intestinal Absorption
Metabolism
Rats
High Pressure Liquid Chromatography
Small Intestine
Limit of Detection
Fluorescence
Capsicum
Glucuronides
Biotransformation
Metabolites
Calibration
Mass spectrometry
Mass Spectrometry
Perfusion
Experiments
Flow rate
dihydrocapsaicin
Wavelength

Keywords

  • Capsaicin and dihydrocapsaicin
  • Fluorescent detection
  • Glucuronide metabolites
  • High performance liquid chromatography
  • Intestinal metabolism

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

Cite this

A validated HPLC-FLD method for analysis of intestinal absorption and metabolism of capsaicin and dihydrocapsaicin in the rat. / Kuzma, Mónika; Fodor, Krisztina; Maász, Gábor; Avar, Péter; Mózsik, G.; Past, Tibor; Fischer, Emil; Perjési, P.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 103, 05.01.2015, p. 59-66.

Research output: Contribution to journalArticle

@article{c2fd8c777b6f452ca6dafe9028518974,
title = "A validated HPLC-FLD method for analysis of intestinal absorption and metabolism of capsaicin and dihydrocapsaicin in the rat",
abstract = "A sensitive and selective reverse-phase high performance liquid chromatographic method with fluorescence detection has been developed for determination of capsaicin (8-methyl-N-vanillyl-(trans)-6-nonenamid) and dihydrocapsaicin (8-methyl-N-vanillylnonanamide) in samples generated in rat small intestine luminal perfusion experiments. The experiments were designed to study the biotransformation of capsaicinoids in the small intestine in the rat. The chromatographic separation was performed at room temperature on a ZORBAX Eclipse{\circledR} XDB-C8 column using isocratic elution with a mobile phase consisting 0.05M orthophosphoric acid solution and acetonitrile (60:40, v/v; pH 3.0) with a flow rate of 1.5mL/min. Fluorescence detection was performed at excitation and emission wavelengths of 230 and 323nm, respectively. The method was evaluated for a number of validation characteristics (accuracy, repeatability and intermediate precision, limit of detection, limit of quantification and calibration range). The limit of detection (LOD) was 50ng/mL and the limit of quantification (LOQ) was 100ng/mL for both capsaicin and dihydrocapsaicin reference standards dissolved in blank perfusate. The method was successfully applied for investigation of intestinal absorption of capsaicin and dihydrocapsaicin while 30μg/mL standardized Capsicum extract - containing capsaicin and dihydrocapsaicin - was luminally perfused for a 90min period. The structure of the glucuronide metabolites of capsaicin and dihydrocapsaicin appeared in the perfusate was identified by mass spectrometry.",
keywords = "Capsaicin and dihydrocapsaicin, Fluorescent detection, Glucuronide metabolites, High performance liquid chromatography, Intestinal metabolism",
author = "M{\'o}nika Kuzma and Krisztina Fodor and G{\'a}bor Ma{\'a}sz and P{\'e}ter Avar and G. M{\'o}zsik and Tibor Past and Emil Fischer and P. Perj{\'e}si",
year = "2015",
month = "1",
day = "5",
doi = "10.1016/j.jpba.2014.10.007",
language = "English",
volume = "103",
pages = "59--66",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
issn = "0731-7085",
publisher = "Elsevier",

}

TY - JOUR

T1 - A validated HPLC-FLD method for analysis of intestinal absorption and metabolism of capsaicin and dihydrocapsaicin in the rat

AU - Kuzma, Mónika

AU - Fodor, Krisztina

AU - Maász, Gábor

AU - Avar, Péter

AU - Mózsik, G.

AU - Past, Tibor

AU - Fischer, Emil

AU - Perjési, P.

PY - 2015/1/5

Y1 - 2015/1/5

N2 - A sensitive and selective reverse-phase high performance liquid chromatographic method with fluorescence detection has been developed for determination of capsaicin (8-methyl-N-vanillyl-(trans)-6-nonenamid) and dihydrocapsaicin (8-methyl-N-vanillylnonanamide) in samples generated in rat small intestine luminal perfusion experiments. The experiments were designed to study the biotransformation of capsaicinoids in the small intestine in the rat. The chromatographic separation was performed at room temperature on a ZORBAX Eclipse® XDB-C8 column using isocratic elution with a mobile phase consisting 0.05M orthophosphoric acid solution and acetonitrile (60:40, v/v; pH 3.0) with a flow rate of 1.5mL/min. Fluorescence detection was performed at excitation and emission wavelengths of 230 and 323nm, respectively. The method was evaluated for a number of validation characteristics (accuracy, repeatability and intermediate precision, limit of detection, limit of quantification and calibration range). The limit of detection (LOD) was 50ng/mL and the limit of quantification (LOQ) was 100ng/mL for both capsaicin and dihydrocapsaicin reference standards dissolved in blank perfusate. The method was successfully applied for investigation of intestinal absorption of capsaicin and dihydrocapsaicin while 30μg/mL standardized Capsicum extract - containing capsaicin and dihydrocapsaicin - was luminally perfused for a 90min period. The structure of the glucuronide metabolites of capsaicin and dihydrocapsaicin appeared in the perfusate was identified by mass spectrometry.

AB - A sensitive and selective reverse-phase high performance liquid chromatographic method with fluorescence detection has been developed for determination of capsaicin (8-methyl-N-vanillyl-(trans)-6-nonenamid) and dihydrocapsaicin (8-methyl-N-vanillylnonanamide) in samples generated in rat small intestine luminal perfusion experiments. The experiments were designed to study the biotransformation of capsaicinoids in the small intestine in the rat. The chromatographic separation was performed at room temperature on a ZORBAX Eclipse® XDB-C8 column using isocratic elution with a mobile phase consisting 0.05M orthophosphoric acid solution and acetonitrile (60:40, v/v; pH 3.0) with a flow rate of 1.5mL/min. Fluorescence detection was performed at excitation and emission wavelengths of 230 and 323nm, respectively. The method was evaluated for a number of validation characteristics (accuracy, repeatability and intermediate precision, limit of detection, limit of quantification and calibration range). The limit of detection (LOD) was 50ng/mL and the limit of quantification (LOQ) was 100ng/mL for both capsaicin and dihydrocapsaicin reference standards dissolved in blank perfusate. The method was successfully applied for investigation of intestinal absorption of capsaicin and dihydrocapsaicin while 30μg/mL standardized Capsicum extract - containing capsaicin and dihydrocapsaicin - was luminally perfused for a 90min period. The structure of the glucuronide metabolites of capsaicin and dihydrocapsaicin appeared in the perfusate was identified by mass spectrometry.

KW - Capsaicin and dihydrocapsaicin

KW - Fluorescent detection

KW - Glucuronide metabolites

KW - High performance liquid chromatography

KW - Intestinal metabolism

UR - http://www.scopus.com/inward/record.url?scp=84914129355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84914129355&partnerID=8YFLogxK

U2 - 10.1016/j.jpba.2014.10.007

DO - 10.1016/j.jpba.2014.10.007

M3 - Article

C2 - 25462121

AN - SCOPUS:84914129355

VL - 103

SP - 59

EP - 66

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

ER -