ABSTRACT
OBJECTIVE:To establish the method for the content determination of L-Hyp and collagen in gelatinous medicinal material,and to compare the contents of two components in reference medicinal material and commercially available medicinal material.METHODS:Pre-column derivatization was adopted for pretreatment.The content of L-Hyp was determined by HPLC.The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1 mol/L sodium acetate buffer (pH 6.5,7 ∶ 93,V/V)-acetonitrile-water (4 ∶ 1,V/V) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 254 urn,and column temperature was set at 43 ℃.The sample size was 20 μL.The content of collagen was calculated by using convert coefficient.RESULTS:The linear range of L-Hyp were 2.5-40 μg/mL(r=0.999 9).LOQ was 0.20 μg/mL,and LOD was 0.05 t g/mL.RSDs of precision,stability and reproducibility tests were all lower than 4.0％.The recoveries were 96.03％-102.07％ (RSD =2.20％,n =9).There was difference in the contents of L-Hyp and collagen among 28 batches of commercially available medicinal material.The contents of two components in 13 batches of commercially available Colla Corii Asini were relative close to reference medicinal material;5 batches of Colla Carapacis et Plastri Testudinis and 7 batches of Cervi Comus Colla were much higher than those of reference medicinal material.CONCLUSIONS:The method is accurate,reliable and suitable for the content determination of L-Hyp and collagen in gelatinous medicinal material.
ABSTRACT
OBJECTIVE:To establish the method for the content determination of L-Hyp and collagen in gelatinous medicinal material,and to compare the contents of two components in reference medicinal material and commercially available medicinal material.METHODS:Pre-column derivatization was adopted for pretreatment.The content of L-Hyp was determined by HPLC.The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1 mol/L sodium acetate buffer (pH 6.5,7 ∶ 93,V/V)-acetonitrile-water (4 ∶ 1,V/V) (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 254 urn,and column temperature was set at 43 ℃.The sample size was 20 μL.The content of collagen was calculated by using convert coefficient.RESULTS:The linear range of L-Hyp were 2.5-40 μg/mL(r=0.999 9).LOQ was 0.20 μg/mL,and LOD was 0.05 t g/mL.RSDs of precision,stability and reproducibility tests were all lower than 4.0％.The recoveries were 96.03％-102.07％ (RSD =2.20％,n =9).There was difference in the contents of L-Hyp and collagen among 28 batches of commercially available medicinal material.The contents of two components in 13 batches of commercially available Colla Corii Asini were relative close to reference medicinal material;5 batches of Colla Carapacis et Plastri Testudinis and 7 batches of Cervi Comus Colla were much higher than those of reference medicinal material.CONCLUSIONS:The method is accurate,reliable and suitable for the content determination of L-Hyp and collagen in gelatinous medicinal material.
ABSTRACT
OBJECTIVE:To investigate the effects of Astragalus polysaccharides on heart function and myocardial fibrosis in spontaneously hypertensive rats (SHR) and corresponding mechanism. METHODS:50 SHR were randomly divided into a model group,a captopril tablets group (positive drug,30 mg/kg) and the groups of high,medium and low-dose (100,50,25 mg/kg) Astragalus polysaccharides,with 10 SHR in each group. Another 10 Wistar Kyoto rats were included into the normal group. The rats in the drug administration groups were given corresponding drugs ip,while those in the normal group and the model group were given isometric distilled water ip,once a day,for 12 consecutive weeks. The left ventricular systolic pressure (LVSP),left ventricular end diastolic pressure (LVEDP),left ventricular pressure rise rate (+dp/dtmax) and left ventricular pressure decline rate (-dp/dtmax) of the rats were recorded. The heart mass index (HMI) and left ventricular mass index (LVMI) thereof were deter-mined. The level of hydroxyproline and the mRNA and protein expression of transforming growth factor-β1(TGF-β1)and peroxi-some proliferator-activated receptor-γ(PPAR-γ)in the cardiac muscle tissue thereof were detected. RESULTS:Compared to the nor-mal group,the rats in the model group had lower LVSP,+dp/dtmax and-dp/dtmax,higher LVEDP,HMI and LVMI,as well as high-er levels of hydroxyproline and the mRNA and protein expression of TGF-β1 and lower level of the mRNA and protein expression of PPAR-γ in the cardiac muscle tissue (P0.05) in all the above-mentioned indexes was shown between the model group and the group of low-dose Astragalus polysaccha-rides,except that the-dp/dtmax of the latter was significantly higher and the level of the mRNA expression of TGF-β1 was obvious-ly lower (P<0.05). CONCLUSIONS:Astragalus polysaccharide can improve heart function and myocardial fibrosis in SHR by a mechanism which may be related to downregulating the expression of TGF-β1 and upregulating the expression of PPAR-γin the car-diac muscle tissue.