Quantitative analysis of valiolamine through pre-column derivatization with phenylisocyanate using high-performance liquid chromatography with UV detection: selection of reagent, identification of derivative and optimization of derivatization conditions.
This report describes the improved quantitative determination of valiolamine in a medium for microbial culture using high-performance liquid chromatography with UV detection. Valiolamine aqueous solution was dried, dissolved in dimethyl sulfoxide and derivatization performances of phenylisocyanate (PHI), 1-fluoro-2,4-dinitrobenznene and 1-naphthylisothiocyanate were compared in the presence of triethylamine. The PHI was chosen as the most suitable derivatization reagent and the valiolamine-PHI derivative was identified by thin-layer chromatography and electrospray ionization mass spectrometry. The derivative eluted at 10.5 min on a reverse-phase column using a mobile phase composed of 10% acetonitrile in water containing 0.5 mM sodium octyl sulfate (pH 3.0), at a column flow rate of 1.0 mL/min with UV detection at 240 nm. The optimum conditions for derivatization were a reaction temperature of 30 degrees C, reaction time of 30 min, and PHI concentration higher than 33.6 mM. Calibration curves were linear in the range of 0.99-19.95 microg/mL for the standard solutions and 24.9-99.7 microg/mL for the spiked sample. The proposed method was validated and proven to be selective, accurate and precise and suitable for the quantitative analysis of valiolamine in medium for microbial cultures.
Li
He
H
Department of Chemical & Biological Engineering and ERI, GyeongSang National University, Jinju, Gyeongnam, Republic of Korea.
Lee
Jae Ran
JR
Nguyen
Do Quynh Anh
do QA
Kim
Sung Bae
SB
Seo
Yang-Gon
YG
Chang
Yong Keun
YK
Hong
Soon-Kwang
SK
Moon
Myung Hee
MH
Chung
In-Young
IY
Kim
Chang-Joon
CJ
D000704:Analysis of Variance; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D001681:Biological Assay; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D007202:Indicators and Reagents; D007294:Inositol / Q000031:analogs & derivatives / Q000032:analysis / Q000494:pharmacology; D017953:Isocyanates / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D050111:Imino Sugars
D000900:Anti-Bacterial Agents; D007202:Indicators and Reagents; D007294:Inositol; D017953:Isocyanates; D012997:Solvents; C043270:valiolamine; D050111:Imino Sugars