1.Optimization of Extraction Conditions for Four Major Effective Components in Wudang Ⅱ Flos lonicerae Caulis by Orthogonal Design and Content Determination
Cong LI ; Peng LI ; Fang ZHENG ; Huimin LIU ; Xuesong ZHU ; Changhu KE ; Xueru DING ; Yang ZHAO
China Pharmacist 2016;19(9):1766-1769
Objective:To study the content determination method for the effective components in WudangⅡFlos lonicerae Caulis to lay foundation for the quality evaluation. Methods: An ultrasonic method was used. The effects of extraction solvent, ultrasonic time, ultrasonic power and ratio of solid to liquid on the contents of rutin and mignonette nucleoside were studied, and the extraction conditions were optimized by a 4-factor and 3-level orthogonal experiment. The chromatographic conditions were as follows:a Phenome-nex Luna-C18(250 mm ×4.60 mm, 5 μm) column was adopted for chlorogenic acid, and a Fortis Xi Phenyl column (250 mm × 4. 6 mm, 5 μm) was used for rutin, loganin and luteoloside;the mobile phase was acetonitrile (B)-0. 4% phosphoric acid (C) solu-tion (15 ∶85) for chlorogenic acid and loganin, and acetonitrile (B) -0. 5% glacial acetic acid aqueous solutjion (D) with gradient e-lution for rutin and luteoloside;the column temperature was 30℃, and the detection wavelength was 327,237,354 and 348 nm, re-spectively. Results:The optimum extraction conditions for rutin and luteoloside from WudangⅡFlos lonicerae Caulis were as follows:the extraction solvent was 60% ethanol, the solid-liquid ratio was 1 ∶30, the ultrasonic power and the ultrasonic time were 350 W and 50 min for rutin, and 250W and 60min for luteoloside. The content of chlorogenic acid, loganin, rutin and luteoloside was 10. 27, 6. 33, 0. 401 and 0. 450 mg·g-1 in the samples, respectively. Conclusion:The method is simple and convenient, accurate and re-producible, which can be used to control the quality of WudangⅡFlos lonicerae Caulis and provide reference for the further develop-ment.
2.Determination of thiabendazole in honeysuckle flower by LC-MS/MS
Jun ZHU ; Bing SHANG ; Xueru DING ; Lin XIONG ; Qinhua CHEN
International Journal of Traditional Chinese Medicine 2018;40(3):258-261
Objective To establish a rapid and sensitive liquid chromatograph-mass spectrometry (LC-MS/MS) method for the determination of thiabendazole in honeysuckle flowers. Methods The acetic ether was selected as extraction solvent. The mass spectrometer analysis was conducted in the positive ionization electrospray mode using SIM. The transitions m/z 202→175 was used to quantify thiabendazole. Results The satisfactory linearity was obtained in the range of 0.1×10-5-2×10-5mg for thiabendazole (r=0.999 5), and the limit of detection (LOD) of 10.0 ng/ml and the mean recovery of 93.70% were obtained by this LC-MS/MS method. Conclusions The method of LC-MS/MS is sensitive, simple and accurate, and it proved to be suitable for the determination of thiabendazole in Honeysuckle flowers.
3.Effects of Atractylenolide Ⅰ,Atractylenolide Ⅱ and Atractylenolide Ⅲ on Cytokines Expression of Inflam-matory Macrophages
Qinhua CHEN ; Fei YU ; Hongmei WANG ; Xueru DING ; Jun ZHU ; Yushuang LIU ; Lun ZHANG
China Pharmacist 2017;20(12):2112-2116
Objective:To study the effect of atractylodes lactideⅠ,Ⅱand Ⅲ on the expression of cytokines by inducing M1 type macrophage model. Methods:Apoptotic macrophages were induced by sodium thioglycolate, and then the rat peritoneal inflammatory macrophages were purified by a differential adherence method. The expression of macrophage marker antigen (ED1) was identified by flow cytometry macrophages. Tumor necrosis factor ( TNF-α) , proinflammatory cytokines ( IL-1β) and interleukin-6 ( IL-6 ) were measured by ELISA in vitro. The levels of expression of arginase 1, inducible nitric oxide synthase (iNOS), inflammatory cytokines (IL-1β) chemokine (CCL22) and transforming growth factor (TGF-β) in inflammatory macrophages were determined by RT-PCR. Results:As for the purification of cultured rat inflammatory macrophage expressing ED1, atractylenolideⅠand Ⅲ could reduce the ex-pression levels of iNOS and IL-1βand increase the expression levels of arginase1 CCL22 and TGF-β. The release of inflammatory fac-tor IL-1β decreased, and the release levels of TGF-β and chemokines CCL22 were promoted(P<0. 05). Atropine lactoneⅠ and Ⅲ had significant inhibitory effects on TNF-α, IL-1β and IL-6 in macrophages(P<0. 05). Conclusion: Atractylodes lactone Ⅰand Ⅲ with anti-inflammatory activity can promote the expression of cytokines in inflammatory macrophages, while the change induced by at-ractylaxanthin Ⅱ is not obvious.