Simultaneous determination of l-citrulline and l-arginine in plasma by high performance liquid chromatography.
OBJECTIVE
To develop a method for simultaneously determining l-citrulline and L-arginine levels in plasma using RP-HPLC with ultraviolet detection.


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.


CONCLUSIONS
This approach offers a reliable, efficient analytical platform for the simultaneous determination of citrulline and arginine levels in plasma.
Mao
Hui-ming
HM
Department of Central Laboratory, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
Wei
Wei
W
Xiong
Wu-jun
WJ
Lu
Ying
Y
Chen
Bing-guan
BG
Liu
Zhongmin
Z
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001120:Arginine / Q000097:blood*; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D002956:Citrulline / Q000097:blood*; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D057230:Limit of Detection; D008297:Male; D008722:Methods; D008875:Middle Aged; D009369:Neoplasms / Q000097:blood
D007202:Indicators and Reagents; D002956:Citrulline; D001120:Arginine