[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a...[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.展开更多
Objective:To identify Ejiao and its adulterants at the DNA level by using DNA molecula marker.Ejiao(Asini Corii Colla)is a commonly used medicinal material.However,its adulteration is a serious concern.Due to the morp...Objective:To identify Ejiao and its adulterants at the DNA level by using DNA molecula marker.Ejiao(Asini Corii Colla)is a commonly used medicinal material.However,its adulteration is a serious concern.Due to the morphological characteristics of Asini Corii Colla and its adulterants,traditional identification techniques often complex and professional,which is not conducive to the circulation management and safety of the medicinal materials.To improve the distinction between Asini Corii Colla and its adulterants accurately,this study identified and its adulterant samples based on the CytB sequence.Sequence characteristics,Basic Local Alignment Search Tool(BLAST)application,genetic distance,construction of phylogenetic tree showed the CytB sequence to accurately identify Asini Corii Colla from its adulterants.Furthermore,in this study,we designed a specific primer,based on the CytB sequence,and established a PCR detection system for rapid,sensitive,and specific identification of Asini Corii Colla.Compared to DNA barcoding technology,this method has shorter detection time,stronger specificity,and higher sensitivity,which lays the foundation for the rapid identification of Asini Corii Colla.展开更多
文摘[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.
基金supported by Scientific Research Fund Projects of Hubei Food and Drug Administration in 2016-2018 under Grant No.20160212.
文摘Objective:To identify Ejiao and its adulterants at the DNA level by using DNA molecula marker.Ejiao(Asini Corii Colla)is a commonly used medicinal material.However,its adulteration is a serious concern.Due to the morphological characteristics of Asini Corii Colla and its adulterants,traditional identification techniques often complex and professional,which is not conducive to the circulation management and safety of the medicinal materials.To improve the distinction between Asini Corii Colla and its adulterants accurately,this study identified and its adulterant samples based on the CytB sequence.Sequence characteristics,Basic Local Alignment Search Tool(BLAST)application,genetic distance,construction of phylogenetic tree showed the CytB sequence to accurately identify Asini Corii Colla from its adulterants.Furthermore,in this study,we designed a specific primer,based on the CytB sequence,and established a PCR detection system for rapid,sensitive,and specific identification of Asini Corii Colla.Compared to DNA barcoding technology,this method has shorter detection time,stronger specificity,and higher sensitivity,which lays the foundation for the rapid identification of Asini Corii Colla.
