Preparation and Identification of Main Ingredients in Effective Parts of Xinjiang Artemisia rupestris / 中国药房

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.

Preparation of impurity C of doxycycline hyclate and its evaluation of in vitro antimicrobial activity and toxicity / 中国药科大学学报

@#This study aimed to isolate and prepare highly purified impurity C from doxycycline hyclate by a preparative HPLC method and to inspect the toxicity and in vitro antimicrobial activity of the impurity C of doxycycline hyclate. The solution of doxycycline hyclate treated with heat produced a solution containing 10% of impurity C which was firstly separated by the Sapphire C18(21. 2 mm×250 mm, 5 μm)column with 0. 1% acetic acid-acetonitrile(83 ∶17)as the mobile phase at 20 mL/min. Secondly, rotary evaporation of the eluted solution at the time of 8. 4 min was performed at 50 °C to remove organic solvent. Then the target product was prepared after freeze drying of evaporated solution adjusting pH to 1. 8 with formic acid. The target product was identified with ultraviolet absorbance(UV), infrared(IR), mass spectrometry(MS)and nuclear magnetic resonance(NMR), and its purity was be determined by HPLC. Meanwhile, cytotoxicity and genotoxicity in the Chinese hamster lung cells, toxicity on the development of zebrafish embryos and in vitro antimicrobial activity were compared among impurity C of doxycycline hyclate, doxycycline, metacycline and β-doxycycline. Results showed that prepared product was confirmed to be the impurity C of doxycycline hyclate. Its purity was 90. 1%, which had been the highest so far. In the cellular toxic tests and genetic toxic tests of Chinese hamster lung cells, impurity C of doxycycline hyclate, doxycycline, metacycline and β-doxycycline were somewhat toxic to Chinese hamster lung cells. Toxicity gradually decreased from doxycycline, impurity C of doxycycline hyclate, β-doxycycline to metacycline from -S9mix test results; toxicity gradually decreased from doxycycline, β-doxycycline, impurity C of doxycycline hyclate to metacycline from +S9mix test results; the aberration rate of all the tested related substances was less than 5%, and no obvious genotoxicity was found. According to test results of the development of zebrafish embryos, impurity C of doxycycline hyclate showed the strongest teratogenicity and lethality. Invitro antimicrobial tests revealed that impurity C of doxycycline hyclate had a weaker antimicrobial activity, and invitro antimicrobial activity potential of the tested compounds followed the order: metacycline, doxycycline, impurity C of doxycycline hyclate, β-doxycycline. Studies on safety and effectiveness indicated that impurity C of doxycycline hyclate belonged to toxic and ineffective impurity and need to be controlled individually in quality standard. A useful suggestion was given to revise the quality standard of doxycycline hyclate and its preparation in the current Pharmacopoeia of the People′s Republic of China.

A New Secoiridoid Glycoside from Swertia cincta / 中草药·英文版

Objective: To study the chemical constituents of Swertia cincta. Methods: Preparative liquid chromatography was employed. The structures of the compounds were elucidated by spectroscopic analysis. Results: Three secoiridoid glycosides were isolated from S. cincta and identified as 8-methoxyl-eustomorusside (1), secoiridoids eustomorusside (2), and eustomoside (3). Conclusion: Compound 1 is a new secoiridoid glycoside. Compounds 2 and 3 are isolated from this plant for the first time. © 2014 Tianjin Press of Chinese Herbal Medicines.

Study of the anti-MRSA activity of Rhizoma coptidis by chemical fingerprinting and broth microdilution methods / 中国天然药物

AIM@#Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogenic bacterium that causes both hospital- and community-acquired infections, and for which single-drug treatments are becoming less efficient. Rhizoma coptidis has been used for more than two thousand years in China to treat diarrhea, fever, and jaundice. In this study, the anti-MRSA activity of Rhizoma coptidis is examined and its effective components sought.@*METHODS@#The mecA and norA genes were determined by PCR amplification and sequencing. Drug susceptibility of Staphylococcus aureus ATCC43300 was performed using the VITEK2 compact system. The chemical fingerprint of Rhizoma coptidis was investigated using HPLC and preparative liquid chromatography, and the anti-MRSA activity was determined using an improved broth microdilution method.@*RESULTS@#The drug susceptibility test revealed that the penicillin-binding protein phenotype of the strain changed in comparison to penicillin-sensitive Staphylococcus aureus. Ten batches of Rhizoma coptidis showed anti-MRSA activity on the norA-negative Staphylococcus aureus strain, as well as the strain that contained a norA gene. The spectrum-effect relationship revealed that the berberine alkaloids were the effective components, within which berberine, coptisine, palmatine, epiberberine, and jatrorrhizine were the major components.@*CONCLUSION@#This study lays a foundation for in vivo studies of Rhizoma coptidis and for the development of multi-component drugs.

Isolation and Purification of Ginkgolide A, B and Bilobalide / 中草药

A simple method for the isolation and purification of ginkgolide A, B and bilobalide(ginkgo terpene trialctone) was developed. A commercially available extract of leaves of Ginkgo biloba L.containing ＞6％ ginkgo terpene trilactone was used as the raw material. After partition in EtOAc, the en-riched extract was separated to give individual terpenes by preparative liquid chromatography. GinkgolideA, B and bilobalide could be isolated with high purity by recrystallizing in MeOH-H2O.