Determination of fatty acids in bio-samples based on the pre-column fluorescence derivatization with 1,3,5,7-tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene by high performance liquid chromatography

• Published in: Journal of Chromatography A Vol. 1291; pp. 84 - 91
• Main Authors: Wang, Fei-Hua, Xiong, Xu-Jie, Guo, Xiao-Feng, Wang, Hong, Zhang, Hua-Shan
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 24.05.2013; Elsevier
• Subjects: 1,3,5,7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene; Analytical chemistry; Animals; Applied sciences; Biological and medical sciences; Boron Compounds - chemistry; Chemical industry and chemicals; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Derivatization; detection limit; Essential oils, perfumes; Exact sciences and technology; Fat industries; Fatty acids; Fatty Acids - analysis; Fatty Acids - blood; Fatty Acids - chemistry; fluorescence; Fluorescence detection; Fluorescent Dyes - chemistry; fluorescent labeling; Food industries; Fundamental and applied biological sciences. Psychology; High performance liquid chromatography; Humans; Indenes - chemistry; Investigative techniques, diagnostic techniques (general aspects); Linear Models; Liver - chemistry; Medical sciences; Mice; Miscellaneous. Technology; Other chromatographic methods; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Plant Oils - chemistry; rapid methods; Reproducibility of Results; Spectrometry, Fluorescence - methods; Washing products. Cosmetics and toiletries. Perfumes; 1,3,5,7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene; Derivatization; Fatty acids; High performance liquid chromatography; Fluorescence detection; Biological fluid; Chemical analysis; Camellia; Liver; HPLC chromatography; Vegetable oil; Blood; Tissue; Amine borane; Corn oil; Dicotyledones; Angiospermae; Clinical biology; Chemical composition; Essential oil; Human; Fluorescence detector; Healthy subject; Precolumn; Rodentia; Patient; Malignant tumor; Edible oil; 1,3,5.7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene; Vertebrata; Reversed phase chromatography; Mammalia; Mouse; Fluorescent labelling; Animal; Spermatophyta; Blood serum; Cancer; Theaceae
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2013.03.062

•We have designed and synthesized TMBB-EDAN as a new fluorescent labeling reagent for fatty acids (FAs).•TMBB-EDAN has high fluorescence quantum yields (Φ=0.70) and high reactivity with FAs.•An HPLC-fluorescence method was developed for the detection of twelve FAs by using TMBB-EDAN.•The derivatization of TMBB-EDAN with FAs could be completed at room temperature in 30min.•The detection limits for FAs were in the 0.2–0.4nM range. 1,3,5,7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene (TMBB-EDAN) has been designed and synthesized as a highly fluorescent labeling reagent for carboxylic acids. By using TMBB-EDAN, a sensitive and rapid method based on high performance liquid chromatography-fluorescence detection for the determination of twelve fatty acids (FAs) in bio-samples has been developed. Under optimized conditions, these FAs were tagged with TMBB-EDAN in the presence of 1-ethyl-3-(3-dimethyla-minopropyl) carbodiamide at 20°C for 30min and then the baseline separation was achieved on a C18 column with a linear gradient elution in 26min. With fluorescence detection at λex/λem=490nm/510nm, the linear ranges of FAs were from 3.0 to 300nM and the detection limits with a signal-to-noise ratio of 3 were in the 0.2–0.4nM range. The proposed method offers advantages of milder derivatization condition and much better sensitivity for the determination of FAs, when compared to the reported fluorescence derivatization-based methods.