ABSTRACT
OBJECTIVE:To establish a method for simultaneous determination of 6 components in Niuhuang ninggong tablets.METHODS:HPLC method was adopted.The determination was performed on TC-C18 column with mobile phase consisted of methanol-0.05％ phosphate acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 280 nm,and the column temperature was 25 ℃.The sample size was 10 μL.RESULTS:The linear ranges of glycyrrhizin,berberine hydrochloride,baicalin,baicalin,emodin and chrysophanol were 3.2-320 μg/mL(r=0.999 9),8.8-880 μg/mL(r=0.999 8),5.6-560 μg/mL (r=0.999 5),2.0-200 μg/mL(r=0.999 9),4.4-440 μg/mL (r=0.999 9),2.0-200 μg/mL(r=0.999 7),respectively.RSDs of precision,stability and reproducibility tests were all lower than 6.0％.The recoveries were 96.34％-97.25％ (RSD=0.33％,n=6),96.12％-98.06％ (RSD=0.82％,n=6),96.36％-99.09％ (RSD=1.02％,n=6),95.84％-97.32％ (RSD=0.65％,n=6),95.83％-97.92％(RSD=0.88％,n=6),98.60％-99.65％(RSD=0.42％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for content determination of 6 components in Niuhuang ninggong tablets.
ABSTRACT
OBJECTIVE:To establish a method for simultaneous determination of 6 components in Niuhuang ninggong tablets.METHODS:HPLC method was adopted.The determination was performed on TC-C18 column with mobile phase consisted of methanol-0.05％ phosphate acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 280 nm,and the column temperature was 25 ℃.The sample size was 10 μL.RESULTS:The linear ranges of glycyrrhizin,berberine hydrochloride,baicalin,baicalin,emodin and chrysophanol were 3.2-320 μg/mL(r=0.999 9),8.8-880 μg/mL(r=0.999 8),5.6-560 μg/mL (r=0.999 5),2.0-200 μg/mL(r=0.999 9),4.4-440 μg/mL (r=0.999 9),2.0-200 μg/mL(r=0.999 7),respectively.RSDs of precision,stability and reproducibility tests were all lower than 6.0％.The recoveries were 96.34％-97.25％ (RSD=0.33％,n=6),96.12％-98.06％ (RSD=0.82％,n=6),96.36％-99.09％ (RSD=1.02％,n=6),95.84％-97.32％ (RSD=0.65％,n=6),95.83％-97.92％(RSD=0.88％,n=6),98.60％-99.65％(RSD=0.42％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for content determination of 6 components in Niuhuang ninggong tablets.
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Objective: To establish a method for the simultaneous determination of paeoniflorin, hesperidine, baicalin and chry-sophanol in Xiaoshi Lidan capsules by dual wavelength HPLC. Methods:An Inertsil ODS-3 C18 column (150 mm × 4. 6 mm,5 μm) was used. The mobile phase was acetonitrile-0. 2% phosphoric acid with gradient elution. The flow rate was 1. 0 ml·min-1 , the de-tection wavelength was 230nm for paeoniflorin, hesperidine and baicalin, and 280nm for hrysophanol, and the column temperature was 40℃. Results:The linear range of paeoniflorin, hesperidine,baicalin and chrysophanol was 0. 030-0. 592 μg(r=0. 999 9), 0. 063-1. 264 μg(r=1. 000 0), 0. 205-4. 094 μg(r=1. 000 0) and 0. 008 2-0. 164 μg(r=1. 000 0), respectively. The average recovery of the four components was 99.49% (RSD =0.85%), 99.74%(RSD =0.71%), 99.75%(RSD =0.47%) and 99.08%(RSD =1. 28%), respectively. Conclusion:The method is simple and accurate, which can be used for the content determination of paeoniflor-in,hesperidine,baicalin and chrysophanol in Xiaoshi Lidan capsules.
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OBJECTIVE:To establish a method for simultaneous determination of geniposide,berberine hydrochloride and ba-icalin in Annao pill. METHODS:HPLC was performed on the column of Inertsil ODS-3 C18 with mobile phase of acetonitrile-0.2% phosphoric acid solution (gradient elution) at a flow rate of 1.0 ml/min,the column temperature was 30 ℃,the detection wavelength was 230 nm,and the injection volume was 10 μl. RESULTS:The linear range was 0. 041 0-0.820 0 μg for geniposide (r=0.999 9),0.037 6-0.752 0μg for berberine hydrochloride(r=0.999 9)and 0.095 2-1.904 0μg for baicalin(r=0.999 9),respec-tively;recoveries were 97.31%-100.41%(RSD=1.10%,n=6),98.32%-101.76%(RSD=1.20%,n=6) and 99.16%-101.30%(RSD=0.76%,n=6),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the quality control of Annao pill.
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AIM: To establish the method of fingerprinting analysis onZedoary turmeric oil by GC. Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them. Based on the cluster analysis, all the Zedoary turmeric oils could be divided into two categories, self-made and market purchasing oil, the latter oils purchased from Liaoning province and Jiangsu province varied considerably. Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the marketpurchasing Zedoary turmeric oil, quality control requires serious consideration.
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Objective To reconstitute a transactivation system in yeast (yeast model) for screening the pesticides acting on ecdysone metabolism route and eventually influencing the process of ecdysis. Methods The fragment of 5 times repeated EcRE from Drosophila melanogaster was synthesized and the HSP27 promoter from D. melanogaster genome was amplified with PCR. The two sequences were connected and followed by a reporting gene——green fluorescence protein(GFP) gene. The EcRE-HSP27 promoter-GFP fragment was inserted into the expression plasmid pPIC3.5 and integrated into the yeast chromosome to construct yeast A. EcR and USP coding sequences of Aedes albopictus were synthesized, and these two fragments were inserted into Pichia pastoris expression plasmid pGAPZ as two respective reading frames. The two reading frames were integrated into Pichia pastoris chromosome in another recombinant site(pGAPZ and pPIC3.5k share different recombinant sites while being integrated into Pichia pastoris yeast chromosome). EcR and USP were constituted and expressed in the yeast. This recombinant yeast was called yeast B. The model yeast was thus constructed. A known ecdysone agonist-tebufenozide was used to test the yeast model. The effect of tebufenozide on the model yeast was observed under fluorescent microscope. Semi-quantitative RT-PCR was used to test the transcrip-tion level of GFP in the tebufenozide affected yeast and the control. Results In the model yeast, the intracellular expressed EcR and USP constituted EcR/USP heterodimer interacting with EcRE, the expression of GFP was activated, and green fluorescence was observed in model yeast under fluorescent microscope. Tebufenozide affected model yeast showed less fluorescence in comparison to the control model yeast, indicating that the transcription of GFP was suppressed by tubufenozide. Yeast housekeeping gene Actin-1 was used as inner control, semi-quantitative RT-PCR was operated and the result was scanned. The ratio of the brightness of GFP to Actin-1 was calculated automatically, and that of tubufenozide added yeast and the control yeast was 0.614 and 1.134 respectively. This result showed a low transcription level of GFP in tebufenozide affected model yeast, comparing to that of the control. Conclusion The ecdysone-related transacting system in yeast has been constructed, and the model yeast can be used to screen the ecdysone agonists which can act on the ecdysone metabolic route.
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AIM: To establish the method of fingerprinting analysis on Zedoary turmeric oil by GC.Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them.Based on the cluster analysis,all the Zedoary turmeric oils could be divided into two categories,self-made and market purchasing oil,the latter oils purchased from Liaoning province and Jiangsu province varied considerably.Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the market purchasing Zedoary turmeric oil,quality control requires serious consideration.