Separation and Purification of Costunolide and Dehydrocostus Lactone in Aucklandiae Radix by Supercritical Fluid Chromatography / 中国实验方剂学杂志

Objective:To establish a supercritical fluid chromatography(SFC) method for separating and purifying costunolide and dehydrocostus lactone in Aucklandiae Radix. Method:With supercritical carbon dioxide as the mobile phase,the effect of six factors, such as type of chromatographic columns,modifiers and modifiers ratio, flow rate of mobile phase,pressure and temperature, on the separation process of supercritical fluid chromatography were explored. The target components were separated and prepared by semi-preparative supercritical fluid chromatography. High performance liquid chromatography and nuclear magnetic resonance were used to analyze the components and study the thermodynamic regularity of the chromatographic process. Result:C18 column (10 mm×250 mm,5 μm) was adopted, with supercritical fluid dioxide as the mobile phase,the ratio of methanol was 0.13%,the flow rate was 12 mL·min-1,column pressure was 13 MPa,column temperature was 318℃, and detection wavelength was 225 nm. The sample was injected for 20 times,crude extract was 4 mg,and each target component was collected according to the chromatogram. Its purity was determined to be more than 99%by HPLC,and its structure was determined as costunolide and dehydrocostus lactone by NMR. Under this condition,the SFC separation process was normal-phase chromatography. Conclusion:The method can be used to prepare effective components of Aucklandiae Radix with a high purity and low solvent residue.

Math1 gene therapy for kanamycin and furosemide-induced deaf guinea pigs / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the morphology and function changes of cochlear hair cells before and after math1 gene injection into the cochlea of deaf guinea pigs which were induced by kanamycin and furosemide. To explore the feasibility of Math1 gene for medicine-induced deafness therapy.</p><p><b>METHODS</b>Kanamycin (500 mg/kg) and furosemide (50 mg/kg) were given to the healthy adult guinea pigs intramuscularly and intravenously to establish the deafness model. The guinea pigs whose auditory brainstem response (ABR) threshold > 95 dB SPL were randomly divided into five groups. Blank control group (without any treatment, n = 3), operation control group (right ear scala tympani operation, n = 3), artificial perilymph group (right ear scala tympani injection artificial perilymph, n = 3), virus vector group [right ear scala tympani injection adenovirus which carrying enhanced green fluorescent protein (EGFP) gene (Ad. EGFP) , n = 4], Math1 gene therapy group [right ear scala tympani injection adenovirus which carrying Math1 and EGFP gene (Ad. Math1-EGFP), n = 6]. Each animal received ABR test before and after injection. The cochlear tissue was observed by scanning electronic microscopy.</p><p><b>RESULTS</b>The ABR thresholds of tone burst( 4, 8, 16, 20 kHz ) were not statistically significant in different groups (P > 0.05). The number of hair cells increased in some of severe deaf guinea pigs after the injection of Ad. Math1-EGFP gene. However, there was no obvious difference with morphology and numbers of cochlea hair cells in other groups.</p><p><b>CONCLUSIONS</b>The injection of Math1 gene to cochlea can regenerate or repair the hair cells of medicine-induced deaf guinea pigs, but there was no improvement on the hearing loss.</p>

Separation and identification of alkaloids in Stephania tetrandra by capillary electrophoresis electrospray-MS/MS / 药学学报

<p><b>AIM</b>To establish a method for separation and identification of alkaloids in Stephania tetrandra S. Moore methanol extracts by using non-aqueous capillary electrophoresis interfaced with electrospray ionization ion trap mass spectrometry.</p><p><b>METHODS</b>The molecular ions or adducts of alkaloids and fragments of specific parent ions were used for the identification. An uncoated capillary (86 cm x 75 microm ID, on-line UV detection occurred at 21 cm from the inlet of the capillary) was used. Ammonium acetate (50 mmol x L(-1)) containing 4% HAc in methanol was used as the running buffer; separation voltage was 25 kV. A coaxial sheath flow interface was used as the CE-MS interface; the electrospray voltage was 4.5 kV; the temperature of aluminium capillary was 170 degrees C; 60% isopropanol-39% water-1% HAc was used as the sheath liquid with the flow rate of 5 microL x min(-1); the collision energy of MS-MS was set at 30% and the least ion counts was 1 x 10(5).</p><p><b>RESULTS AND CONCLUSION</b>The alkaloids in Stephania tetrandra S. Moore methanol extracts were separated and identified by CE-ESI-MS/MS. The proposed method is of high accuracy and can be used for the investigation of traditional Chinese medicine.</p>