EurekaMag.com logo
+ Site Statistics
References:
53,214,146
Abstracts:
29,074,682
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

2-(4-Hydrazinocarbonylphenyl)-4,5-diphenylimidazole as a versatile fluorescent derivatization reagent for the high-performance liquid chromatographic analysis of free fatty acids






Journal of Chromatography, Biomedical Applications 619(1): 1-8

2-(4-Hydrazinocarbonylphenyl)-4,5-diphenylimidazole as a versatile fluorescent derivatization reagent for the high-performance liquid chromatographic analysis of free fatty acids

A new fluorescent derivatization reagent with a lophine skeleton, 2-(4-hydrazinocarbonylphenyl)-4,5-diphenylimidazole (HCPI), has been synthesized and applied to the assay of saturated free fatty acids. The HCPI derivatives with five representative saturated fatty acids [caproic acid (C6), lauric acid (C12), palmitic acid (C16), stearic acid (C18) and arachidic acid (C20)] were synthesized to examine their fluorescence properties. The fluorescence spectra (lambda ex ca. 335 nm, lambda em ca. 455 nm) of the derivatives were almost identical in methanol, n-heptane and acetonitrile. For the high-performance liquid chromatographic assay, fatty acids were derivatized with HCPI using a condensing agent, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and pyridine at room temperature. The HCPI derivatives were separated on a C18 column with a gradient elution of methanol-water, and the eluates were monitored at 455 nm using an excitation wavelength of 335 nm. By this method, seven kinds of free fatty acid (C6, C10, C12, C14, C16, C18 and C20) could be determined using margaric acid (C17) as an internal standard. Calibration curves were linear over the range 0.2-400 pmol per 20-microliters injection (r = 0.994-1.000). Relative standard deviations of the peak-height ratio for five replicate measurements of fatty acids (100 pmol per injection) were 0.7-2.6%. Detection limits were 7-57 fmol at a signal-to-noise ratio of 3. The method could be successfully applied to the determination of four saturated fatty acids (C12, C14, C16 and C18) in normal human serum.

Accession: 002549858

PMID: 8245146

DOI: 10.1016/0378-4347(93)80440-f

Download PDF Full Text: 2-(4-Hydrazinocarbonylphenyl)-4,5-diphenylimidazole as a versatile fluorescent derivatization reagent for the high-performance liquid chromatographic analysis of free fatty acids



Related references

5,6-dimethoxy-2-(4'-hydrazinocarbonylphenyl)benzothiazole as a highly sensitive and stable fluorescence derivatization reagent for carboxylic acids in high performance liquid chromatography. Journal of Liquid Chromatography 18(15): 2991-3006, 1995

Selective derivatization and high performance liquid chromatographic analysis of free fatty acids in lipid extracts. Cereal Chemistry 60(6): 411-413, 1983

Pre-column derivatization of free bile acids for high-performance liquid chromatographic and gas chromatographic-mass spectrometric analysis. Journal of Chromatography 562(1-2): 697-712, 1991

6-oxy-(acetyl piperazine) fluorescein as a new fluorescent labeling reagent for free fatty acids in serum using high-performance liquid chromatography. Journal of Chromatography. A 1169(1-2): 77-85, 2007

High-performance liquid chromatographic analysis of serum short-chain fatty acids by direct derivatization. Journal of Chromatography 421(1): 33-41, 1987

Simple derivatization method for sensitive determination of fatty acids with fluorescence detection by high-performance liquid chromatography using 9-(2-hydroxyethyl)-carbazole as derivatization reagent. Analytica Chimica Acta 436(1): 163-172, 2001

An improved derivatization method for sensitive determination of fatty acids by high-performance liquid chromatography using 9-(2-hydroxylethyl)-carbazole as derivatization reagent with fluorescence detection. Chromatographia 54(5/6): 316-322, 2001

High-performance liquid chromatographic analysis of serum long-chain fatty acids by direct derivatization method. Journal of Chromatography 416(2): 237-245, 1987

High-performance liquid chromatographic analysis of free amino acids in fruit juices using derivatization with 9-fluorenylmethyl-chloroformate. Journal of Chromatographic Science 40(1): 14-18, January, 2002

Automated high-performance liquid chromatographic determination of plasma free fatty acids using on-line derivatization with 9-bromomethylacridine based on micellar phase-transfer catalysis. Journal of Chromatography 500: 443-452, 1990