RESUMEN
OBJECTIVE：To establish a method for the simultaneous determination of euphol and euphorbol in Euphorbium resinifera，and to optimize the extraction technology. METHODS ：HPLC method was adopted. The determination was performed on Agilent ZORBAX Eclipse Plus C 8 column with mobile phase consisted of acetonitrile-water （90∶10，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and column temperature was 30 ℃. The sample size was 10 μL. Using the contents of euphol and euphorbol ，yield of the extract as evaluation index ，comprehensive score was conducted. The extraction technology was optimized by L （9 34）orthogonal tests ，with ethanol volume fraction ，extraction time and solvent dosage as factors. RESULTS：The linear ranges of euphol and euphorbol were 0.030 4-1.216 mg/mL（r＝0.999 6）and 0.01-0.4 mg/mL（r＝0.999 9）， respectively. RSDs of precision ，stability（24 h）and producibility tests were all lower than 2％. Average recoveries were 100.46％ （RSD＝1.03％，n＝6）and 99.36％（RSD＝0.91％，n＝6）. The optimized technology was extracting with 40 mL 95％ ethanol for 1 h. After 3 times of validation tests showed that average content of euphol was 94.43 mg/g（RSD＝0.92％，n＝3），and that of euphorbol was 25.42 mg/g（RSD＝0.98％，n＝3）；average yield of the extract was 51.42％（RSD＝1.95％，n＝3），and average comprehensive score was 98.87（RSD＝0.92％，n＝3）. CONCLUSIONS ：Established method is simple ，accurate and reproducible ， which can be used for the quality control of E. resinifera . The optimized extraction technology is simple and stable.
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OBJECTIVE:To compare the differences between artificially cultivated and wild Xinjiang Artemisia rupestris,and screen the different components. METHODS:HPLC-MS was adopted to establish the fingerprints of artificially cultivated and wild Xinjiang A. rupestris from different origin and harvest time. Principal component analysis was conducted by Marker ViewTM soft-ware and SIMCA-P 11.5 software,the characteristics of principal components were analyzed,difference variable was screened, and different components of artificially cultivated and wild varieties were obtained. RESULTS:Fingerprints of 22 batches of A. rup-estris(12 batches of wild varieties,10 batches of artificially cultivated varieties)were established. According to the principal com-ponent analysis,artificially cultivated and wild varieties were well grouped,with obvious differences;the principal components of artificially cultivated varieties with different harvest time showed certain difference,mainly before and after flowering,concentrat-ing in to-be flowering and full flowering periods. Wild varieties from different origins had obvious regional difference,showing cer-tain differences in composition and content. 268 variables were found in matrix of positive ion mode and 155 in negative ion mode. 28 groups of variables were extracted by difference variable,and 19 variables were determined. CONCLUSIONS:Artificially culti-vated and wild varieties have obvious difference in principal component,mainly in flowering period and picking places. It can pro-vide theoretical basis for the standardized cultivation and origin protection of Xinjiang A. rupestris.
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OBJECTIVE:To establish a method for rapid identification and efficient preparation of main ingredients in effective parts of Xinjiang Artemisia rupestris,and provide reference for researching the ethnic medicines. METHODS:LC-HRMS/MS was conducted for the preliminary study of main ingredients in effective parts of A. rupestris. HPLC,UV and MS were used to compare and analyze parts of the compounds and its reference substances,their names were determined. Column separation and preparative liquid chromatography were used for the undetermined compounds to receive monomer rapidly,and the structures were identified. RESULTS:5 compounds were separated from the effective parts,2 of which were identified as artemetin and casticin. A monomer-ic compound was obtained (yield was 0.35 mg/g,the purity was 98.5%),which was confirmed to be 6-demethoxy-4′-O-me-thoxy-capillarisin-7-O-β-D-glucoside by the structure. CONCLUSIONS:The method has achieved rapid separation,identification and preparation of target ingredients,which can be used for the fundamental research of ethnic medicine complex materials.
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OBJECTIVE:To study the effect of peiminine on increasing the chemosensitivity of 5 kinds of cancer cells. METH-ODS:Using human esophageal cancer Eca-109 cell,human breast cancer MCF-7 cell,human small cell lung cancer A549 cell,hu-man hepatoma HepG2 cell and human cervical cancer HeLa cell as objects,MTT colorimetric method was used to detect the growth inhibition rate of above-mentioned 5 kinds of cancer cells after treated by peiminine with maximal non-toxic mass concentra-tion(20 μg/mL)and adriamycin with different gradient mass concentrations(0.026-2.1,0.026-2.1,0.125-2.0,0.125-2.0,0.0625-0.10μg/mL)for 72 h. The half inhibitory concentration(IC50)was calculated. Crystal violet staining method was adopted to observe the proliferation of above-mentioned cancer cells after treated by peiminine with maximal non-toxic mass concentration and adriamycin with low mass concentrations(0.02,0.005,0.04,0.02,0.01 μg/mL)for 7 d. Solvent control,single use of peiminine and adriam-ycin control were conducted. RESULTS:Compared with single use of adriamycin,the combination use of peiminine and adriamy-cin can improve the growth inhibition rate of 5 kinds of cancer cells to certain degree,most of the differences were statistically sig-nificant (P<0.05 or P<0.01);and IC50 was obviously decreased,with statistical significances (P<0.05 or P<0.01). Compared with solvent control,single use of peiminine or adriamycin had no obvious effects on the proliferation of above-mentioned cancer cells in 7 d,and the combination use of peiminine and adriamycin can obviously inhibit the proliferation of above-mentioned cancer cells in 7 d. CONCLUSIONS:Peiminine can enhance the sensitivity of above-mentioned-mentioned 5 kinds of cancer cells to cer-tain degree,showing certain chemosensitivity increasing effect.