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A quality control (QC) strategy for quantitative and qualitative analysis of “common peaks” in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD-MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C18 column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the “common peaks”. For quantification of “common peaks”, the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of “common peaks”, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC-MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.
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Objective To explore associations between SG13S114A/T and SG13S32A/C polymorphisms of ALOX5AP gene and the genetic susceptibility of ischemic cerebrovascular diseases (ICVD) in Henan Han population.Methods Two hundred and forty-six ICVD patients and 245 healthy controls were recruited from Han population in Henan province. Polymorphisms of SG13S114A/T and SG13S32A/C in ALOX5AP gene were genotyped in these samples by SnaPshot minisequencing method.Each genotype frequency and allele frequency were statistically analyzed and compared between ICVD group and control group using SPSS16.0 software.Haplotype and linkage disequilibrium were analyzed by SHEsis software.Results The SG13S114 AA genotype frequency ( 18.7％ ) and A allele frequency (41.3％) in ICVD group were significantly higher than those in control group (9.0％ and 32.7％,respectively; P =0.002 and P =0.005 ).It was also found that in male ICVD group and in younger ICVD group ( ＜50 years old),the SG13S114 AA genotype frequencies (22.1％ and 22.0％,respectively) and A allele frequencies (42.1％ and 42.7％,respectively) were significantly higher than those in male control group and younger control group (SG13S114 AA genotype:9.0％ and 8.9％ ; P =0.010 and P =0.006,respectively) ;A allele frequencies,34.0％ and 32.0％ ; P =0.048 and P =0.020,respectively.Finally,the prevalence of A-A haplotype in ICVD group was significantly higher than that in control group(30.4％ vs 23.5％,OR =1.419,95％ CI 1.068-1.885,P =0.015).T-C haplotype frequency of ICVD group was significantly lower than that in control group (22.0％ vs 28.8％,OR =0.698,95％ CI 0.523-0.932,P =0.014 ).Conclusions The A allele in SG13S114 loci of ALOX5AP may be a genetic risk factor for ICVD in Han population in Henan province.The association is predominant in ICVD patients of male and younger than 50 years old.Maybe A-A haplotype increases the risk of ICVD and T-C haplotype and has a protective effect against ICVD in Henan Han population.
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OBJECTIVE: To establish a method for the content determination of domiphen by potassium chromate indicator method.METHODS: The contents of domiphen were determined based on the theory that bromide ion in domiphon could react with AgNO3 to produce silver bromide precipitation.The method was compared with the sodium tetraphenylborate method issued in China Pharmacopeica(2005 edition).RESULTS: The RSD of contents was 0.18%,and the average recovery was 100.2% in the potassium chromate indicator method.There was no significant difference between the results of two determination methods by t-test.CONCLUSION: The potassium chromate indicator method is simple,fast and accurate,which can be used for the content determination of domiphen.
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Objective To establish and optimize AFLP reaction system used in providing necessary technique basis for genetic diversity analysis in Asarum sieboldii.Methods Leaves of A.sieboldii were used as experimental materials to analyze various essential elements of the whole process,such as quality of extracted DNA,time of restriction digest and ligation,concentration of Mg2+,primers and Taq DNA polymerase during PCR amplification etc.,so that the most optimal AFLP reaction system could be built up. Results The optimal AFLP reaction system of A.sieboldii has been constructed: in the genomic DNA extraction,mercaptoethanol being utilized and samples being incubated about 30 min at 65 ℃;the genomic DNA being digested by Trul Ⅰ and Pst Ⅰ for 3 h respectively;in PCR amplification,the final concentration of Mg2+ being 1.5 mmol/L and the volume of Taq DNA polymerase being 0.2 ?L.Conclusion The present reaction system for A.sieboldii is able to gain the favorable results of AFLP analysis and can be used in the genetic diversity research of the species.