OBJECTIVE: To develop a method for simultaneously determining l-citrulline and L-arginine levels in plasma using RP-HPLC with ultraviolet detection. DESIGN AND METHODS: Plasma samples were deproteinized by trichloroacetic acid and heat. Phenyl-isothiocyanate (PITC) solution was used as derivatization reagent and a gradient elution was carried out. RESULTS: The linearity for L-citrulline and L-arginine ranged from 0 to at least 1000 micromol/L. R(2) values were above 0.9999 for both. LODs for L-citrulline and L-arginine were 0.0201 micromol/L and 0.0476 micromol/L, respectively, while LOQs were 0.240 micromol/L and 0.448 micromol/L, respectively. Intra- and inter-day CVs were less than 3.40% and 7.2%, respectively. The average recovery was from 86.22% to 118.9%. L-citrulline and L-arginine concentrations in healthy controls were 60.77+/-9.18 micromol/L and 58.19+/-16.43 micromol/L, respectively. CONCLUSION: This approach offers a reliable, efficient analytical platform for the simultaneous determination of citrulline and arginine levels in plasma. Copyright (c) 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
OBJECTIVE: To develop a method for simultaneously determining l-citrulline and L-arginine levels in plasma using RP-HPLC with ultraviolet detection. DESIGN AND METHODS: Plasma samples were deproteinized by trichloroacetic acid and heat. Phenyl-isothiocyanate (PITC) solution was used as derivatization reagent and a gradient elution was carried out. RESULTS: The linearity for L-citrulline and L-arginine ranged from 0 to at least 1000 micromol/L. R(2) values were above 0.9999 for both. LODs for L-citrulline and L-arginine were 0.0201 micromol/L and 0.0476 micromol/L, respectively, while LOQs were 0.240 micromol/L and 0.448 micromol/L, respectively. Intra- and inter-day CVs were less than 3.40% and 7.2%, respectively. The average recovery was from 86.22% to 118.9%. L-citrulline and L-arginine concentrations in healthy controls were 60.77+/-9.18 micromol/L and 58.19+/-16.43 micromol/L, respectively. CONCLUSION: This approach offers a reliable, efficient analytical platform for the simultaneous determination of citrulline and arginine levels in plasma. Copyright (c) 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
Authors: Crystal L Richards; Sandra J Raffel; Sébastien Bontemps-Gallo; Daniel P Dulebohn; Tessa C Herbert; Frank C Gherardini Journal: PLoS Pathog Date: 2022-03-14 Impact factor: 6.823