The Preparation Of Pouzolzia Zeylanica (L.) Benn. Var. Microphylla (Wedd.) W.T.Wang.Ointment

Pouzolzia Zeylanica (L.) Benn. Var. Microphylla (Wedd.)W.T.Wang belongs to the plant family of Urticaceae and Genus is Pouzolzia. In China, it’s mainly distributed throughout Jiangxi, Fujian, Taiwan, Guangdong, Guangxi And Yunan, and has been used as a folk medicine for the treatment of skin and soft tissue infections, including skin abscess, gangrenous ulcers, sores, boils, dysentery, syphilis, and gonorrhea. Phytochemical studies have indicated that total flavonoids extracted from the aerial parts of Pouzolzia zeylanica (L.) Benn. var. microphylla (Wedd.)W. T.Wang (FPZ) were the major constituents. In order to develop a new drug on skin abscess infected with bacteria by using FPZ as the raw material, we have done a series of study. Firstly, we optimize enrichment and purification technology of FPZ. Secondly, we establish a sensitive and specific LC-MS fingerprint for the quality controlling of FPZ.Thirdly, we determined the prescription of ointment matrix and the preparation process. Furthermore, FPZ was used as the main drug of ointment and the S. aureus-induced skin abscess in mice, S. aureus-induced skin ulcer in mice and carrageenan-induced hind paw oedema mode were adopted to investigate the pharmacological effects of Pouzolzia Zeylanica (L.) Benn. Var. Microphylla (Wedd.)W. T.Wang ointment. Finally, the quality standard of the ointment were studied. The mainly results are presented as follow.1. It was proved that AB-8 resin could effectively enrich FPZ, optimum purification process was as following:the concentration of sample solution 0.3g/ml, pH of 5, the amount of sample solution of 5 BV, flow rate for sampling of 3 BV/h, removed impurities by 2 BV of water and then eluted total flavonoids with 2 BV of 60% ethanol at flow rate of 2BV/h; purity of total flavonoids increased from 28.81% in sample solution to 68.18% in eluant.2. The flavonoids composition of Pouzolzia zeylanica (L.) Benn.Var. microphylla (Wedd.) W.T.Wang was determined on an ACQUITY UPLC BEH SHIELD RP18 chromatographic column with acetonitrile(A)-0.2% formic acid(B) water as the mobile phase in gradient elution (0～22 min) 15%～19%A, (22～26 min) 19%～24%A, (26～32min)24%A, (32～40 min) 24%～38%A; The flow rate is 0.3mL·min－1 and the injection volume is 0.2μL. The detection wavelength was set at 330 nm. The method is simple, accurate and repeatable. It can be used for research and quality control of the effective components in Pouzolzia zeylanica (L.) Benn. Var. microphylla (Wedd.) W.T.Wang. Quercetin-3,7-O-α-L-dirhamnopyranoside and kaempferol-3,7-O-α-L-dirhamno pyranoside were the major constituents of FPZ.3. The best prescription was present as follow:Stearic acid 1g, Albolene 1g, Setanol 2g, Liquid paraffin 1.5g, Glycerin monostearate 0.8g, Span-600.4g were used as oil phase and Propylene glycol 2g, Azone 1g, Triethanolamine1g, Tween-80 1g, water 19.1g were used as water phase. The oil phrase and the water phrase were heal to 85℃ separately then slowing pouring the oil phrase into the water phrase mixing them evenly in 5 mins to get the matrix. The FPZ extract was added at 2.5%, 5%and 10%(w/w) as main drug. The three kinds of ointment samples in certain ration was use to evaluating its pharmacological effects.4. The healing activity of three kinds of Pouzolzia zeylanica (L.) Benn. Var. microphylla(W edd.)W. T.Wang ointment were investigated on S. aureus-induced skin abscess in mice and S. aureus-induced skin ulcer in mice. In addition, the topical anti-inflammatory activity of these ointment was studied on carrageenan-induced hind paw oedema in mice. By comparing the vehicle group, The ratio of wound area was decreased in ointment-treated mice. This observation strongly suggests the essential role of the ointment in S. aureus infection of skin abscess and wound healing. Compared with vehicle group, the swelling in high dose group was decreased at the time of 180min,270min and 360min, the difference had statistical significance(P<0.05).5. The study of the quality standard of Pouzolzia zeylanica (L.) Benn.Var.microphylla(W edd.)W.T.Wang ointment:The ointment has no stimulate to skin. The amount of quercetin-3,7-O-α-L-dirhamnopyranoside and kaempferol-3,7-O-α-L-dirhamnopyranoside of the ointment no less than 1000μg·g-1 and 110μg·g-1, Separatly.