6-oxy-(acetyl piperazine) fluorescein as a new fluorescent labeling reagent for free fatty acids in serum using high-performance liquid chromatography.
A new fluorescein-based fluorescent derivatizating reagent, 6-oxy-(acetyl piperazine) fluorescein (APF), has been designed, synthesized and developed for carboxylic acid labeling. It was used as a pre-column derivatizing reagent for the determination of seven free fatty acids (lauric acid, myristic acid, arachidonic acid, linoleic acid, palmitic acid, oleic acid, and stearic acid) with high-performance liquid chromatography (HPLC). The derivatization reaction of APF with seven fatty acids was completed at 60 degrees C for 1 h using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as the condensing reagent. On a C18 column, the derivatives of APF with seven free fatty acids could be separated completely in 22 min using a mobile phase of methanol-water (88:12, v/v) containing 7 mmol L(-1) pH 6.5 Na2HPO4-H3Cit3 buffer with fluorescence detection at lambdaex/lambdaem=467/512 nm. The detection limits could reach 0.1-6.4 nmol L(-1) (signal-to-noise=3). This reagent was applied to the determination of the free fatty acids in human serum samples with satisfying recovery efficiencies varying from 93 to 105%.
Du
Xiao-Lan
XL
Department of Chemistry, Wuhan University, Wuhan 430072, China.
Zhang
Hua-Shan
HS
Guo
Xiao-Feng
XF
Deng
Ying-Hua
YH
Wang
Hong
H
D002264:Carboxylic Acids / Q000097:blood / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005230:Fatty Acids, Nonesterified / Q000737:chemistry / Q000302:isolation & purification*; D005452:Fluoresceins; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D010879:Piperazines; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*
C524827:6-oxy-(acetyl piperazine) fluorescein; D002264:Carboxylic Acids; D005230:Fatty Acids, Nonesterified; D005452:Fluoresceins; D005456:Fluorescent Dyes; D010879:Piperazines