Determination of trace elements in Bazhen pills an

Determination of paeoniflorin in Bazhen pills and Bazhen Yimu pills by high performance capillary electrophoresis Abstract: Objective: to establish a high performance capillary electrophoresis method for the determination of paeoniflorin in Bazhen pills and Bazhen Yimu pills. Method: the running buffer is 50mmol/L borax buffer (ph0.6); The operating voltage is 20kV; The detection wavelength is 230nm. Results: the linear range of paeoniflorin was o.0625 ~ 1.0mg/ml, r=o.9995; The recovery rate of samples was 98.19% - 98.45%; The intra day and intra day precision were 1.61% and 3.15% respectively: the content of paeoniflorin in Bazhen pill and Bazhen Yimu pill was determined, and the results of the former were consistent with those of HPLC. Conclusion: this analytical method is simple, rapid and economical; Accurate

key words: high performance capillary electrophoresis; Paeoniflorin; Bazhen pill; Bazhen Yimu pill

determination of paeoniflorin in Bazhen piers and Bazhen Yim u piers by h igh performance capability electrophoresis (HPCE)

abstract:objective:a HPCE method for determination of paeoniflorin in Bazhen piers and Bazhen Yimu piers was developed.methods:m emergencies were carried out in a buffer solution containing 50MPa mol/L borax solution (pH 10.6), At deteenon wavclength 230nm with 20kV run, you can Ning voltage applied to a 50cm only according to simple installation steps × 5O "m capability.results:the linear range of dacohifierinwasinthe rangeof0.0.0mg/ml (r-0.9995), rsdwithin dayswas1.61%, RSD between dayswas 3.15%. The rcc() vcrv of pae() niflorin was 98．19％ ～ 98．45％ ．The results of detection for Bazhen pills and Bazhen Yimu pills were highly correlated tO those obtained by HPLC．CONCLUSION ：The method iS simple，rapid，economical and accurate．

KEY WoRDS：HPCE； paeoniflorin； Bazhen pill； Bazhen Yimu pill

Bazhen Yimu nine are both varieties included in the Chinese Pharmacopoeia 2000 edition. The content determination of Bazhen pill adopts high performance liquid chromatography (HPLC) to take paeoniflorin contained in white peony as the quantitative chemical index. At present, there is no content determination method of paeoniflorin in Bazhen Yimu pill in China. There are foreign literature reports. If there is no clear and profound understanding of this work, further measures should be taken to standardize it, It is impossible to do this good thing that benefits the country and the people

1instruments and reagents

bio focus 3000 capillary electrophoresis apparatus (bio rad company of the United States); Fused quartz capillary column (uncoated) 501 μ m × 50cm (the effective separation K is 45.5cm, Hebei Yongnian optical fiber factory); Microporous membrane (0.45 μ m) And filter unit (millipore, USA); Milli qplus (USA) paeoniflorin reference substance (China Institute for the control of pharmaceutical and biological products); Medicinal materials (provided by Fengyuan traditional Chinese medicine room); The methanol of Bazhen pills (Shuimi pills) and Bazhen Yimu pills (Shuimi pills) (commercially available products) is chromatographically pure, and the other reagents are analytically pure

2 methods and results

2.1 determination conditions: running buffer: 50mmol/L borax buffer, pH 10.6 with sodium hydroxide; Operating voltage: 20kV, polarity from positive to negative; Detection wavelength: 230nm; Column temperature: 20 ℃; Pressure injection is adopted, and the pressure injection constant is 10psi × S both running buffer and sample solution have passed 0.45 μ M microporous filter membrane after filtration, use capillary flushing method: wash with ultra pure water for 3min each time, then wash with 0.1mo1/L sodium hydroxide for 3Rain, and then wash with ultra pure water for 3min, and then operate slowly. It is expected that the sales of material science and technology in 2014 will show an increase in the number of units (under the coordination of foreign exchange and investment) for 3min. Between each determination, flush lmin

2.2 with running buffer. Preparation and determination of reference solution: accurately weigh 10.0mg of paeoniflorin dried in phosphorus pentoxide vacuum dryer for 36h, and fix the volume with methanol to form a reference stock solution with a concentration of 1.0mg/ml. Take stock solution 500 μ 1. Put it into a LML volumetric flask, add running buffer to the scale, inject samples and determine

2.3 preparation and determination of test solution: Bazhen pills and Bazhen Yimu pills are prepared according to the preparation method of test solution under "determination of Bazhen pills" in Chinese Pharmacopoeia on characteristics of pressure burst tester (2000 EDITION), diluted to an appropriate concentration with running buffer, injected and determined

negative control solution: according to the prescription proportion and production preparation method, prepare blank samples of Bazhen l pill and Bazhen Yimu pill without white peony, prepare negative control solution according to the preparation method of test solution, inject samples and determine

2.4 preparation of standard curve: take paeoniflorin stock solution 62.5, 125, 250, 500 and 1000 respectively μ l. Fix the volume with ultra pure water to LML, inject samples in sequence, and then regress with the ratio of drug concentration (c) to peak area migration time (x). The regression equation is: c=-1.2 blonde technology is called the world's most complete new material enterprise 765 × 100+2．7579 × 106X，r=0．9995； The linear range is: 0.0625 ~ 1.0mg/ml

2.5 precision test: take the reference solution with the concentration of 0.25mg/ml, repeat the injection for 5 times in a day and a day, measure the ratio of peak area to migration time, and calculate the precision. The RSD in a day is 1.61%; The daily RSD is 3.15%

2.6. Sample recovery test: add appropriate paeoniflorin reference solution to the sample solution of Bazhen pill and Bazhen Yimu pill with known concentration respectively for sample injection analysis, and determine the content. The recovery results are shown in table 12.7. Sample content determination: determine the content of Bazhen pill and Bazhen Yimu pill samples with HPCE method, and determine the content according to the HPLC method under "Bazhen pill" in Chinese Pharmacopoeia (2000 EDITION), Results table 2

3 discussion

the buffer in the pH range was investigated in the experiment. The results showed that when the pH was greater than 9.0, paeoniflorin could be separated, and the peak shape was better; When the pH is less than 9.0, the paeoniflorin peak deformation is bad and the migration time is prolonged, so the optimal pH is finally determined to be 10.6. Under this condition, paeoniflorin can be well separated and the peak shape is good, and the migration time is moderate. The author investigated the borax concentration in the range of 25 ~ 100mmol/L, and found that with the increase of concentration, the resolution increases, but at the same time, the increase of current also causes the baseline instability, taking into account the relationship between the two, Finally, the concentration of borax buffer was determined to be 50mmol/L

from the analysis results of this experiment, it can be seen that the HPCE method is accurate and effective for the quantitative determination of paeoniflorin in Bazhen pills and Bazhen Yimu pills, and the HPCE method of Bazhen pills is basically consistent with the HPLC method. However, compared with HPLC method, HPCE method has the characteristics of high efficiency, rapidity, small injection volume, less solvent consumption and strong anti pollution ability. Chinese Pharmacopoeia (2000 EDITION) (Volume I) takes HPLC method as the quantitative method of paeoniflorin in Bazhen pills, requiring the number of theoretical plates not less than 2000. In this experiment, the number of theoretical plates of HPCE method can reach more than 100000, and the separation efficiency is much higher than HPLC method. According to the results of methodological investigation, this method has high precision, good stability and reproducibility

the results of this experiment undoubtedly provide a new way for the quantitative determination of paeoniflorin in Bazhen pills and Bazhen Yimu pills. It can also provide reference for the quality control of similar Chinese patent medicines

References:

[1] Pharmacopoeia Committee of the people's Republic of China. Chinese Pharmacopoeia (Volume 1) [S]. 2000 edition. Beijing: Chemical Industry Press, 2000:341.

[2] Honda s, Suzuki K, Kataoka m, El a1.analysisof the corncomponents of Paeonia radio by capillary zone electrophoresis [J j-j chromalogr, 1990515:653