Rapid quantitative method for tea polysaccharide monosaccharide composition

Abstract

The invention discloses a rapid quantitative method for tea polysaccharide monosaccharide composition. The steps includes: (1) hydrolyzing tea polysaccharide with trifluoroacetic acid; (2) dissolving the hydrolyzed tea polysaccharid sample with ultra pure water to obtain a hydrolyzed polysaccharide solution, and adding trehalose to serve as the internal standard substance; (3) conducting derivatization on the internal standard containing hydrolyzed polysaccharide solution with 1-phenyl-3-methyl-5-pyrazolone (PMP); (4) at the same time of hydrolyzed polysaccharide derivatization, conducting derivatization on standard monosaccharide; and (5) carrying out high performance liquid chromatographic analysis, thus obtaining the content of each monosaccharide. The high performance liquid detection system conditions employed by the method include: octadecyl silane bonded silica gel is a chromatographic column of filler, the mobile phase is a buffer solution mixed according to certain ratio and an organic agent, the flow speed is 1.0ml / min, and the detection wavelength of an ultraviolet detector is 250nm. The method is simple, fast, convenient and, stable, achieves separation and analysis of monosaccharide within 20min, and improves the accuracy and precision of tea polysaccharide monosaccharide detection.

Description

Technical field [0001] The invention belongs to the field of tea chemistry, and more specifically relates to a rapid quantitative method of tea polysaccharide monosaccharide composition. Background technique [0002] Tea polysaccharide is an important active substance in tea, which has many functions such as immunity, lowering blood sugar, anti-radiation, anti-oxidation, and lowering blood fat. [0003] Tea polysaccharide is an acid glycoprotein whose activity is closely related to molecular weight and monosaccharide composition. Detecting the monosaccharide composition of tea polysaccharides is very helpful for studying its biological activity. The detection methods of monosaccharide composition in polysaccharides mainly include chromogenic method, capillary electrophoresis, gas chromatography, thin layer chromatography, liquid chromatography, etc. Su Bingxia (Separation and Content Determination of Polysaccharides in Camellia Sinensis, 2012, Journal of Tropical Crops, Vol.33(3)...

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Problems solved by technology

Su Bingxia (Isolation and Content Determination of Polysaccharides in Camellia Kucha, 2012, Journal of Tropical Crops, Vol.33(3): 567-571), Fu Boqiang (Determination of Polysaccharide Content in Tea, 2001, Food Science, 22(11): 69 The non-patent literature of people such as -73) shows that the chromogenic method can only measure the content of total sugar in the tea polysaccharide, and cannot quantitatively analyze the monosaccharide composition of the tea polysaccharide; Application, 1999, Chromatography, Vol.17(4):346-252), Zhang Lizhi (research progress of monosaccharide composition analysis methods, 2013, progress in microbiology and immunology, Vol.41(1)77-81) et al. Non-patent literature shows that the polysaccharide composition is separated by capillary electrophoresis, and the separation degree of the polysaccharide is poor; ), Chen Haixia (Studies on Primary Structure Characterization, Space Conformation and Biological Activity of Highly Active Tea Polysaccharides, 2002, Huazhong Agricultural University) and others showed that in gas chromatography, monosaccharides are not easy to volatilize, leading to derivatization The process is complicated and unstable, isomers are easily produced after derivatization, and the content of uronic acid cannot be detected; Yan Jun (TLC rapid analysis of monosaccharide composition of polysaccharides, 2006, Food Science, Vol.27(12): 603-607) The non-patent literature of et al. shows that the sensitivity of thin chromatography is relatively low and the analysis is inaccurate; The literature shows that ordinary liquid chromatography analysis uses sugar column or amino column, which is expensive and has high maintenance costs, and it takes a long time to equilibrate

Deng Jing (Application of High Performance Liquid Chromatography and Chemometrics in the Quality Control of White Tea and Its Polysaccharides, 2014, Nanchang University), Guo Wei (Pre-column Derivative HPLC Method to Analyze the Monosaccharide Composition of Pu-erh Tea Polysaccharides, Agricultural Science and Technology, 2013, Vol.14(4):556-558) et al. used pre-column derivatization HPLC method to analyze the monosaccharide composition of white tea and Pu'er tea polysaccharides, but the analysis time was more than 40 minutes, and the separation effect of some monosaccharides needs improvement

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