[Synthesis of fluorescent derivatization reagent 2-(4-carboxyphenyl)-naphthoimidazole and its application in the determination of alcohols by high performance liquid chromatography].
A new sensitive pre-column fluorescent derivatization method, which employs high performance liquid chromatography(HPLC) with fluorometric detection, for the determination of alcohols is described. The alcohols were transformed into the corresponding fluorescent esters by treating with 2-(4-carboxyphenyl)-naphthoimidazole in dichloromethane in the presence of 4-piperidinopyridine (PIP) and 1-isopropyl-3-(3-dimethylaminopropyl)-1-carbodiimide perchlorate(IDC) at 80 degrees C for 10 min. The resultant mixture was analyzed by HPLC on a reversed phase column(Zorbax Bp C8, 4.6 mm i.d. x 250 mm, 5 microns) with acetonitrile-methanol (volume ratio was 90:10) as mobile phase, and the sample volume was 20 microL. Monitored excitation and emission wavelengths were 345 nm and 485 nm respectively in determination of the alcohol. The calibration curve of cholesterol is linear between 0 and 2.5 g/L with correlation coefficient of 0.9992. The average recovery was 96.15% with RSD of 1.39%. The lowest detectable mass concentration of cholesterol was 1.0 microgram/L (S/N = 3).
Zhou
H B
HB
Department of Pharmacy, Guangdong College of Pharmacy, Guangzhou 510224, China.
Mukai
Y
Y
D000438:Alcohols / Q000032:analysis*; D002784:Cholesterol / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes; D007093:Imidazoles / Q000138:chemical synthesis*; D007202:Indicators and Reagents / Q000737:chemistry*; D009281:Naphthalenes / Q000138:chemical synthesis*
C476150:2-(4-carboxyphenyl)naphthoimidazole; D000438:Alcohols; D005456:Fluorescent Dyes; D007093:Imidazoles; D007202:Indicators and Reagents; D009281:Naphthalenes; D002784:Cholesterol