Analysis Of The Chemical Composition Of Daozhen Lonicera Flos And Preliminary Study Of The Immunological Activity Of Polysaccharides

Objective To provide the basis for further study of Lonicerae Flos in Daozhen, Zunyi,two analytical methods, HPLC and UV-VIS, were employed to determinate the contents ofmain constituents. Meanwhile, fingerprint of Lonicerae Flos was established by HPLC–DAD and preliminary immunology screening test was carried out, so as to find activepolysaccharides in Lonicerae Flos.Methods The contents of main ingredients in Lonicerae Flos were determinted byUV-VIS and HPLC, and the system adaptability and methodology were investigated at thesame time. HPLC fingerprint was established using10different batches of Lonicerae Flos,and the chromatogram similarity was calculated. The polyccharides extraction wasoptimized by U15(54) uniform design, impact factors, extract time, temerature, solid-liquidratio and extract times were selected. An immunocompromised mouse model induced byCTX were employed, the control group, model group and polysaccharides low, mediumand high dose groups were designed. The organ index of spleen and thymus was calculated,the phagocytosis of mouse peritoneal macrophages, HC50, and spleen lymphocyteproliferation index were determinated.Results The average contents of total flavonoids, chlorogenic acid composition and totalsaponins of Lonicerae Flos determinated by UV-VIS were22.09%,9.45%,29.62%(g/g),respectively. And the linear range was25.85~51.70μg,42.32~105.80μg, and150.50~365.50μg, respectively. While the rutin, chlorogenic acid and macranthoides saponin Bdeterminated by HPLC were1,12%,2.48%, and7.77%, respectively. The linear range was0.1625~4.880μg,2.175~6.960μg, and0.966~7.728μg, respectively. The similarity ofHPLC fingerprint was above0.95. The optimized extraction was100oC, solid-liquid ratiowas1∶30(g/v),3times, and each time was2h. Compared with control group, Thephagocytic capacity of macrophages reduces significantly (P <0.01) in model group. The high-dose group of Lonicerae Flos polysaccharides compared with model group, thephagocytic acivity of macrophages elevated significantly (P<0.01). Compared with controlgroup, the organ index decreased significantly (P<0.5) in model group, compared withmodel group, the organ index increased significantly (P<0.01) in the polysaccharidemedium group. The HC50in model group was significantly lower than control group(P<0.05), compared with model group, the HC50were significantly higher (P<0.01) inpolysaccharide low and medium-dose group. Spleen lymphocyte proliferation index showsthat different concentration of Lonicerae Flos polysaccharide can promote the proliferationof spleen lymphocytes, and the proliferation index was1.521when the polysaccharideconcentration was500μg/mL.Conclusion The Lonicerae Flos in Daozhen was in conformity with the requirements ofChinese Pharmacopoeia (2010). There were13common peaks in10batches HPLCfingerprint. The established standard HPLC fingerprint can provided quality control ofLonicerae Flos in Daozhen. The extraction process optimized by uniform design was stableand simple. The Lonicerae Flos polysaccharides may enhance mouse macrophagephagocytosis, humoral immunity, and immune cells in vitro.