HPLC分析大蒜多糖中的单糖

Determination of Monosaccharide Compositions and Contents in Polysaccharide of Garlic by HPLC

关键词:  大蒜多糖  衍生化  1-苯基-3-甲基-5-吡唑啉酮  反相高效液相色谱法

Key words:  polysaccharide of garlic  derivation  PMP  RP-HPLC

摘要:

目的建立一种反相高效液相色谱法(RP-HPLC)分析大蒜多糖中的单糖组成及摩尔比。方法大蒜多糖样品经2 mol·L-1硫酸降解为单糖后，经1-苯基-3-甲基-5-吡唑啉酮( PMP)衍生化，采用Shimadzu VP-ODS柱(150 mm×4.6 mm，5 μm)分离，以乙腈-醋酸铵缓冲溶液(取醋酸铵7.708 g加水溶解并稀释至1 000 mL，用醋酸调pH 5.5 (22∶78)为流动相，流速为0.8 mL·min-1，在245 nm处检测单糖的衍生化产物。结果大蒜多糖中甘露糖(Man)、鼠李糖(Rha)、葡萄糖醛酸(GlcUA)半乳糖醛酸(GalUA)、葡萄糖(Glc)、半乳糖(Gal)、阿拉伯糖(Arab)7种单糖的摩尔比为4.31∶4.23∶4.19∶9.13∶27.4∶4.99∶3.81。结论本方法灵敏度高，结果准确可靠，可用于大蒜多糖的单糖组成测定及质量控制。

Abstract:

OBJECTIVE To establish a RP-HPLC method for the determination of the monosaccharide compositions and contents in polysaccharide of garlic. METHODS The garlic polysaccharide were derivatization with 1-phenyl-3-methyl-5- pyrazolone (PMP) after hydrolyzed into monosaccharides with 2 mol·L-1 H2SO4, monosaccharide derivatives were separated on Shimadzu VP-ODS volum (150 mm×4.6 mm, 5 μm) with acetonitril-Acetate buffer adjusted to pH 5.5 with Acetate (22∶78) as mobile phase and UV detection at 245 nm. RESULTS The content of monosaccharide compositions were measured in polysaccharide of garlic, molar ratio of mannose, rhamnose, glucuronide, galacturonicacid, glucose, galactose, arabinose was 4.31∶4.23∶4.19∶9.13∶27.4∶4.99∶3.81. CONCLUSION The methods is simple, sensitive, accurate and reliable. The method can be applied for the analysis of quality control and monosaccharide composition and molar ratio in polysaccharide of garlic.