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.