ABSTRACT
Objective To develop an HPLC method for simultaneous determination of jasminoidin, baicalin, berberine hydro-chloride and emodin in Zhizi Jinhua(ZZJH) Pills. Methods The analysis was performed on a Zorbax Eclipse XDB-C18(4.6 mm×150 mm, 5 μm) with gradient elution by using the mobile phase of acetonitrile-0.2% phosphoric acid solution. The column temperature was maintained at room temperature and the flow rate was 1.0 ml/min. The detection wavelengths were set at 240 nm and 275 nm, respectively. Results The calibration curves were linear within the range of 0.056~0.3360 μg (r=0.9998) for gardenoside, 0.208~0.624μg (r=0.9990) for baicalin, 0.116~0.696μg (r=0.9994) for berberine hydrochloride, and 0.040~0.240 μg (r=0.9993) for emodin, respectively. The recoveries of gardenoside, baicalin, berberine hydrochloride and emodin were 91.18 %(RSD=1.60%)、91.55%(RSD=0.75%)、91.19%(RSD=1.08%) and 92.29%(RSD=1.98%), respectively. Conclusion The method is convenient, fast, sensitive and reproducible, with good precision, specificity and accuracy. So it can be used for simultaneous determination of jasminoidin, baicalin, berberine hydrochloride and emodin in ZZJH Pills.
ABSTRACT
Objective:To investigate the effects of GnRH agonist on the expressions of FSHR proteins in pituitary and ovaries in ewes,also to analyse the bioinformatics characteristics of ovine pituitary FSHR.Methods: Forty-two prepubertal ewes (Ovis aries) were randomly assigned to six groups (n=7).The ewes in experimental group EG-Ⅰ,EG-Ⅱand EG-Ⅲwere subcutaneously injected with 200μg,300μg and 400 μg alarelin antigens twice (at day 0 and 14),respectively.Ewes in EG-Ⅳand EG-Ⅴwere injected sub-cutaneously with 200μg and 300μg alarelin antigen four times (at day 0,7,14 and 21),respectively.Ewes in the control group (CG) were subcutaneously injected with 2.0 ml solvent twice (at day 0 and 14).Fluorescence quantitative RT-PCR was used to detect gene expression of FSHR in pituitary glands.The expression of FSHR protein in the ovaries was detected using Western blot.Results:The 2-ΔΔCt values of GnRHR,FSHR and LHR mRNAs in the pituitary gland of EG were lower than that in control group (CG).The 2-ΔΔCt values in EG-Ⅳ( P<0.05 ) and EG-Ⅴ( P<0.05 ) were lower than those in EG-Ⅰand EG-Ⅱ.Expressions of FSHR proteins in EG ovaries increased.Levels of FSHR proteins in EG-Ⅳand EG-Ⅴ( P<0.05 ) were higher than CG.The length of sheep FSHR sequence of nucleotides were 1 091 bp,the homology of FSHR sequences was 100% with that reported in NCBI.The theory isoelectric point , half-life,unstable index,fat index,hydrophobic average were respectively 1.2 h,5.05,47.75,0.836 and 30.61.Conclusion:Alarelin active immunization can inhibit the expression of FSHR mRNA in the pituitary gland ,but increase FSHR expression in ovarian of ewes.FSHR is an unstable hydrophobic protein.