Key words: capillary zone electrophoresis; ferulic acid; Angelica <br> high performance capillary electrophoresis (high performance capillary electrophoresis, HPCE) is a novel separation and analysis technology developed rapidly over the past decade, with higher education, fast, low-power, sample The amount is small and so on. Capillary zone electr0phoresls (CZE) is a separation mode of HPCE. It is separated according to the difference in electrophoretic mobility of charged components under an applied electric field, and has been widely used in the analysis of traditional Chinese medicine. Angelica is one of the commonly used traditional Chinese medicines. Ferulic acid is the main active ingredient in inhibiting platelet aggregation. Therefore, it is determined by thin layer scanning method and high performance liquid phase method as one of the indexes for evaluating the quality of Angelica sinensis and its preparation. We used the CZE method to determine the content of ferulic acid in Angelica and achieved satisfactory results.
1 Materials and methods
L_l Instruments and reagents Waters Capillary Ion Analyzer. The hollow fused silica capillary was 75 μm × 100 cm. The standard reference substance for ferulic acid and diclofenac is provided by the Chinese pharmaceutical biological product Orange. Angelica is a commercial item. Borax and hexanol are of analytical grade.
1.2 Electrophoresis condition buffer is 20mmol/L borax solution, operating voltage is 25kV, temperature is 25°C, siphon injection, time is 10s, UV detection on column, wavelength 214nm.
1.3 Sample determination Precisely weighed the appropriate amount of Angelica fine powder, placed in a 25ml volumetric flask, add 70% ethanol, and ultrasonically shake for 45min. The internal standard diclofenac was added to make a concentration of 40 μg/ml, and the supernatant was centrifuged for measurement.
1.4 Sample recovery rate The precision of the Angelica fine powder was accurately weighed and placed in a 25 ml volumetric flask. Add the appropriate amount of ferulic acid reference substance, follow the steps under the sample determination and calculate the recovery rate.
2 Results and discussion
2.1 The choice of each part of electrophoresis Zui's large absorption wavelength of ferulic acid is about 313nm, but due to the limitations of the instrument conditions, this experiment uses 214nm for detection, and still achieves good results. In this experiment, the sand buffer was used for separation. By investigating the effect of sand concentration on the separation, it was determined that 20mmol/L borax solution was used as a buffer for dans. Under these conditions, the other components of Angelica and ferulic acid were The selected internal standard produced a good separation.
2.2 Standard curve Separate ferulic acid reference substance 10, 2O, 40, 60, 80μg in a 1ml volumetric flask, add 40μg of internal standard, and make up to volume with 70% ethanol. Separating and measuring the electrophoresis conditions, and recording the ratio of the peak area of ​​ferulic acid to the internal standard Yn. The regression equation of Y and concentration X in this concentration range is: Y = 30.12X - 0.020 76 (r = 0.9992).
2.3 Precision The intra- and inter-day precision experiments were carried out on ferulic acid at high, medium and low concentrations. The results showed that the method was of good rationality.
2.4 Sample recovery rate The recovery rate of ferulic acid was 97.25%, and the RSD was 1.94% (n=5).
2.5 Sample preparation Using the established method, three batches of commercially available angelica were measured five times each. The results showed that the content of ferulic acid in Angelica was significantly different.
This is related to the difference in the origin of the angelica or the harvest season, which also indicates the necessity of quality control of the crude drug Angelica.
3 Conclusions This study established a good linear relationship and reproducibility of CZE separation and determination of ferulic acid in the traditional Chinese medicine Angelica. The method is accurate and simple, and provides a new methodological method for the quality control of Angelica. It can be foreseen that CZE The characteristics of the law will make it have an extremely attractive development prospect in the analysis of traditional Chinese medicine.
references
1 Liu Zhisong, Fang Yulun, Application of High Performance Capillary Electrophoresis in Pharmaceutical Analysis[J].Chinese Journal of Chromatography,1996,14(5):364
1 Materials and methods
L_l Instruments and reagents Waters Capillary Ion Analyzer. The hollow fused silica capillary was 75 μm × 100 cm. The standard reference substance for ferulic acid and diclofenac is provided by the Chinese pharmaceutical biological product Orange. Angelica is a commercial item. Borax and hexanol are of analytical grade.
1.2 Electrophoresis condition buffer is 20mmol/L borax solution, operating voltage is 25kV, temperature is 25°C, siphon injection, time is 10s, UV detection on column, wavelength 214nm.
1.3 Sample determination Precisely weighed the appropriate amount of Angelica fine powder, placed in a 25ml volumetric flask, add 70% ethanol, and ultrasonically shake for 45min. The internal standard diclofenac was added to make a concentration of 40 μg/ml, and the supernatant was centrifuged for measurement.
1.4 Sample recovery rate The precision of the Angelica fine powder was accurately weighed and placed in a 25 ml volumetric flask. Add the appropriate amount of ferulic acid reference substance, follow the steps under the sample determination and calculate the recovery rate.
2 Results and discussion
2.1 The choice of each part of electrophoresis Zui's large absorption wavelength of ferulic acid is about 313nm, but due to the limitations of the instrument conditions, this experiment uses 214nm for detection, and still achieves good results. In this experiment, the sand buffer was used for separation. By investigating the effect of sand concentration on the separation, it was determined that 20mmol/L borax solution was used as a buffer for dans. Under these conditions, the other components of Angelica and ferulic acid were The selected internal standard produced a good separation.
2.2 Standard curve Separate ferulic acid reference substance 10, 2O, 40, 60, 80μg in a 1ml volumetric flask, add 40μg of internal standard, and make up to volume with 70% ethanol. Separating and measuring the electrophoresis conditions, and recording the ratio of the peak area of ​​ferulic acid to the internal standard Yn. The regression equation of Y and concentration X in this concentration range is: Y = 30.12X - 0.020 76 (r = 0.9992).
2.3 Precision The intra- and inter-day precision experiments were carried out on ferulic acid at high, medium and low concentrations. The results showed that the method was of good rationality.
2.4 Sample recovery rate The recovery rate of ferulic acid was 97.25%, and the RSD was 1.94% (n=5).
2.5 Sample preparation Using the established method, three batches of commercially available angelica were measured five times each. The results showed that the content of ferulic acid in Angelica was significantly different.
This is related to the difference in the origin of the angelica or the harvest season, which also indicates the necessity of quality control of the crude drug Angelica.
3 Conclusions This study established a good linear relationship and reproducibility of CZE separation and determination of ferulic acid in the traditional Chinese medicine Angelica. The method is accurate and simple, and provides a new methodological method for the quality control of Angelica. It can be foreseen that CZE The characteristics of the law will make it have an extremely attractive development prospect in the analysis of traditional Chinese medicine.
references
1 Liu Zhisong, Fang Yulun, Application of High Performance Capillary Electrophoresis in Pharmaceutical Analysis[J].Chinese Journal of Chromatography,1996,14(5):364
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