Determination of hesperidin in Weili capsules by t

2022-08-15
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Objective: to establish a method for the determination of hesperidin in Weili capsules

method: determine by high performance liquid chromatography. Ywg-c18 analytical column (4.6 mm) is selected × 250 mm,10 μ m) Mobile phase: acetonitrile water methanol phosphoric acid (20 ∶ 79.6 ∶ 0.4 ∶ 0.01), detection wavelength: 283 nm, flow rate: 1.5 n-feedback is current 1, column temperature: room temperature

results: this method is simple, sensitive and accurate. Linear range: 0.2 ~ 1.5 μ g(r=0.9993)。 Average recovery: 99.39%, rsd=3.5%

conclusion: the established quantitative method can be used for the quality control standard of Weili capsule

Weili capsule is composed of Atractylodes macrocephala, Fructus aurantii, bupleurum, hawthorn and other medicinal flavors. This prescription is a compound preparation added on the basis of Zhangzhongjing's famous prescription Fructus aurantii pill, which has the effects of strengthening the spleen and eliminating PI, regulating qi and Zhong. It is clinically used for epigastric fullness, vomiting, nausea, stupidity, pantothenic acid caused by gastrointestinal motility disorders, which belong to spleen deficiency and qi stagnation, stomach loss and decline. In order to effectively control the product quality and ensure the safety and effectiveness of clinical medication, reference reports [1 ~ 3 fish can swim under the sieve]. This paper studies and establishes a method for the determination of hesperidin in Fructus aurantii Immaturus in the prescription by high performance liquid chromatography. Now it is reported as follows:

1 instrument and test drug

high performance liquid chromatography Beckman 338, 427 data processor; 406 UV detector; Hesperidin was purchased from China Institute for the control of pharmaceutical and biological products; Radix Atractylodis Macrocephalae, Fructus aurantii Immaturus, Radix Bupleuri, Fructus crataegi and other medicinal materials have been identified by Professor Guo Yimin of Shanxi staff medical college; All reagents used are analytically pure

2 chromatographic conditions

ywg-c18 analytical column (4.6 mm × 250 mm,10 μ m) , mobile phase: acetonitrile water methanol phosphoric acid (20 ∶ 79.6 ∶ 0.4 ∶ 0.01); Detection wavelength: 283 nm [3]; Flow rate 1.5 n-1; Column temperature: room temperature

3 experimental method

3.1 selective test according to the prescription proportion and production preparation method, prepare the negative sample without Fructus aurantii Immaturus, prepare the negative solution according to the sample solution preparation method under the sample determination, and inject 10 samples μ 50. Take another hesperidin reference solution and inject 10 samples μ L。 The chromatograms of the negative solution and the reference solution are shown in Figure 1. The peaks of the reference solution and the negative solution do not overlap

3.2 linear relationship investigation accurately prepare methanol solution containing 0.1 mg hesperidin per ml, and accurately suck 2.0, 4.0, 8.0, 12.0, 15.0 respectively μ L inject samples in sequence, measure the peak area according to the above chromatographic conditions, and take the integral value of the peak area as the ordinate, and the injection volume of the reference substance( μ g) Draw the standard curve for the abscissa, and the regression equation is:

y=173755.4x-1724.8 r=0.9993

there is a good linear relationship between the peak area of hesperidin and the injection volume, and the linear range is 0.2 ~ 1.5 μ G

3.3 precision test take the reference solution under the sample determination item and repeat the injection for 5 times, 10 times each time μ 50. Determine the peak area, rsd=1.90%

3.4 stability test take the same test sample and inject 10 samples at 0, 3, 6 and 12 hours respectively μ 50. Record the peak area, rsd=0.23%

3.5 repeatability test take the same batch of samples with batch number 960528 and determine it for 6 times according to the sample determination method. Calculate the orange peel. Rest assured to know the glycoside content of our company Jinan new era gold assay instrument Co., Ltd., rsd=2.6%

3.6 sample recovery test take 5 parts of the content of this product (batch No. 960528) with known content, accurately weigh, add an appropriate amount of the control solution each, volatilize it dry, measure it according to the sample determination method, and calculate the recovery rate respectively. The average recovery rate is 99.39%, RSD is 3.5%

4 sample determination

4.1 preparation of sample solution take about 0.5g of Weili capsule content, accurately weigh it, put it into a 500 ml volumetric flask, add methanol to the near scale, ultrasonic treatment for 40 minutes, cool it, add methanol to the scale, filter it, and take the continuous filtrate as the test solution

4.2 the preparation of the reference solution has a linear relationship

4.3 determination method accurately suck 10% of the reference solution and 10% of the sample solution respectively μ 50. Inject it into the high-performance liquid chromatograph and determine it according to the above chromatographic conditions. See Fig. 1 for the chromatogram. Calculate the content of hesperidin in the samples by the external standard two-point method. Each batch of samples is measured three times, and the results are shown in Table 1

Table 1 hesperidin content (%)

batch number content RSD/%

831.2

. 671.3

. 281.5

5 discussion

5.1 the extraction rate of hesperidin by ultrasonic extraction at different times (10 ~ 80 min) was compared, which showed that hesperidin could be extracted completely after 40 min treatment

5.2 under the chromatographic conditions determined in this paper, the hesperidin peak can achieve baseline separation from other component peaks, and the number of theoretical plates is greater than 1500

5.3 this method is simple, sensitive and accurate, which provides a basis for the quality control of the preparation

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