CN1299113C - Inspection for quality control of Longxuejie - Google Patents

Abstract

The invention relates to a method for detecting raw materials of Chinese medicinal materials, in particular to a method for detecting components that can be used for quality control. The quality control detection method of dragon blood jerky described in the invention is a method for simultaneously determining the contents of dragon blood jelly A and dragon blood jelly B by high performance liquid chromatography. The quality control detection method of Dracaena dracaena of the present invention consists of the following steps: 1. Chromatographic conditions and system applicability test; 2. Preparation of reference solution; 3. Preparation of test solution; 4. Determination. In order to comprehensively and accurately control the quality of dragon's blood, the present invention formulates a high-performance liquid chromatography method for simultaneously determining the contents of dragon's blood A and dragon's blood in dragon's blood. After the verification of the analytical method, its precision, reproducibility, recovery rate and stability meet the requirements of the Chinese Pharmacopoeia 2000, an appendix "Guiding Principles for the Validation of Analytical Methods of Quality Standards for Traditional Chinese Medicine".

Description

Quality control detection method of dragon's blood

technical field

The invention relates to a method for detecting raw materials of Chinese medicinal materials, in particular to a method for detecting components that can be used for quality control.

Background technique

Dracaena cochinchinensis (Lour.) Dracaeua cochinchinensis (Lour.) SHChen, a plant of the family Agaveaceae, is a resin extracted and processed from the lipid-containing xylem. It is a commonly used traditional Chinese medicine. It is mainly used for bruises, blood stasis and pain, stagnation of Qi and blood in women, traumatic bleeding, and bedsores that persist for a long time. It has been included in the national drug standard WS3-082(Z-016)-99(Z). The currently implemented standard quality control only measures the content of dracaena B (C ₁₈ H ₂₀ O ₅ ). It is found through research that the main components of dracaena japonicus are flavonoids, and it is difficult to accurately control the amount of only one type of flavonoid in the medicinal material. reflect product quality.

Contents of the invention

The purpose of the present invention is to provide a comprehensive and accurate quality control detection method for dragon's blood to control the quality of dragon's blood.

The quality control detection method of dragon's blood in the invention is a method for simultaneously measuring the contents of dragon's blood A and dragon's blood in dragon's blood by high-performance liquid chromatography.

In order to comprehensively and accurately control the quality of dragon's blood, the applicant has conducted research on flavonoids in dragon's blood, and proposed to increase the content of dragon's blood A (C ₁₇ H ₁₈ O ₄ ), which is high in content and easy to quantify. , the results showed that the content of dracaine A in Dracaena dracaena produced in Yunnan was 5 to 6 times higher than that produced in Guangxi, and the content of dracaena in Dracaena dracaena from different origins was quite different. ₁₇ H ₁₈ O ₄ ) and dragon's blood B (C ₁₈ H ₂₀ O ₅ ) belong to the flavonoid chalcone compounds, and are the main active ingredients of dragon's blood. In order to ensure that the product quality is controllable, safe and effective, the applicant has researched and developed a high-performance liquid chromatography method for the simultaneous determination of the contents of dracaine A and dracaine B in dragon's blood. After the verification of the analytical method, its precision, reproducibility, recovery rate and stability meet the requirements of the Chinese Pharmacopoeia 2000, an appendix "Guiding Principles for the Validation of Analytical Methods of Quality Standards for Traditional Chinese Medicine".

Dragon's blood quality control detection method of the present invention is made up of the following steps:

1. Chromatographic conditions and system suitability test

Octadecylsilane bonded silica gel is used as filler; glacial acetic acid solution 1→100-acetonitrile 63±1∶37±1 is used as mobile phase, detection wavelength is 275±2nm, column temperature: 30±1℃, number of theoretical plates According to the calculation of dragon blood A and B peaks, it should not be less than 5000, and the separation degree of dragon blood A and B peaks should be greater than 2.0;

Two, the preparation of reference substance solution

Weigh an appropriate amount of ascarin A and ascarin B reference substances dried under reduced pressure by a desiccant for 24 hours, add methanol to dissolve and make a mixed solution containing 0.18 mg of ascarin A and 0.04 mg of ascarin B per 1 ml, instant;

Three, the preparation of test solution

Take about 0.25g of the powder of this product, accurately weigh it into a 25ml measuring bottle, add about 20ml of methanol, sonicate for 10 minutes to dissolve the dragon's blood, let it cool to room temperature, dilute to the mark with methanol, shake well, 0.45μm micropore Filtrate through a filter membrane, discard the initial filtrate, and obtain;

4. Determination

Precisely draw 10 μl of the reference substance solution and 5 μl of the test solution, inject it into the liquid chromatograph, measure it, and obtain it.

Methodological investigation of the present invention:

Linear relationship investigation:

Accurately draw 1, 2, 5, 10, 15, and 20 ul of the reference solution respectively, and measure the peak area according to the above-mentioned chromatographic conditions. The sample volume was regressed by the peak area integral value, and a good linear relationship was shown in the range of 0.1840 μg---3.680 μg for ascarin A and 0.0419 μg---0.8380 μg for ascarin B. The regression equation of dragon blood A is: Y=3411.92X+34.40, r=0.99989; the regression equation of dragon blood B is: Y=2694.85X+14.79, r=0.99960; where Y is the peak area, X is the injection volume, r is the correlation coefficient.

Table 1 Linear relationship inspection results Injection volume (μl) Dragon blood A Dragon blood B Injection amount (μg) Peak area Injection amount (μg) Peak area 125101520 0.18400.36800.92001.8402.7603.680 634.91269.53179.96468.99433.612527.0 0.04190.08380.20950.41900.62850.8380 119.0235.2588.11195.91690.12259.7

Precision test:

Take the same reference substance solution of dragon blood A and dragon blood B, according to the above-mentioned chromatographic conditions, measure continuously 6 times (injection 10 μ l), the measured peak area integral value is shown in Table 2.

Table 2 precision test results frequency Dragon blood A peak area Dragon blood B peak area 123456 mean RSD(%) 6389.56351.96363.36364.96361.06361.16365.30.20 1116.01112.11115.81118.01119.31116.91116.40.22

Stability test:

Take the same test solution, according to the above chromatographic conditions, inject 10 μl of sample, and measure at different times, the results are shown in Table 3.

Table 3 Stability test results time Dragon blood A Dragon blood B 05hr25hr45hr96hr166hr average RSD 10525.510510.610645.610684.210544.010738.410608.050.89% 1620.31627.81644.81652.21663.21678.21647.751.31%

The results showed that the sample was relatively stable within 166hr (about 7 days).

Reproducibility test:

Take about 0.25 g of dragon's blood powder (passed through No. 3 sieve, small sample), weigh 6 parts in total, and test according to the content determination method. The results are shown in Table 4.

Table 4 Reproducibility test results serial number Sampling amount (g) Dragon blood A (%) Dragon blood B (%) 123456 0.25820.24570.24640.26550.24880.2310 2.882.892.852.922.932.84 0.580.580.580.590.590.59 Average content 2.885 0.585 RSD 1.2 0.94

The results showed good reproducibility.

Sample recovery test:

Take about 0.12g of dragon’s blood jerky powder (pilot test sample, 010101; containing 2.80% asdracaine A and 0.59% asdracaine B) whose content has been determined, weigh 6 parts in total, weigh them accurately, and put them in 25ml volume In the bottle, add 10, 15, 20ml of dragon blood A and B reference solution (each 1ml contains dragon blood A 0.1840mg/ml, dragon blood B 0.0419mg/ml), add methanol to about 20ml, Sonicate for 10 minutes, let it cool to room temperature, add methanol to dilute to the mark, shake well, filter through a 0.45 μm microporous membrane, take the subsequent filtrate, measure the content and calculate the recovery rate, the results are shown in Table 5 and Table 6.

Table 5 Dragon blood A sample addition recovery rate test result serial number Sampling volume Sample dragon blood A content Sample volume Measured total Recovery rate 123456 0.1166g0.1138g0.1216g0.1131g0.1208g0.1291g 3.26mg3.19mg3.40mg3.17mg3.38mg3.61mg 1.84mg1.84mg2.76mg2.76mg3.68mg3.68mg 5.06mg4.95mg6.04mg5.93mg6.94mg7.18mg 97.8% 95.7% 95.7% 100.0% 96.7% 97.0%

Table 6 Dragon's blood B addition recovery rate test result serial number Sampling volume Sample dragon blood B content Sample volume Measured total Recovery rate 123456 0.1166g0.1138g0.1216g0.1131g0.1208g0.1291g 0.69mg0.67mg0.72mg0.67mg0.71mg0.76mg 0.419mg0.419mg0.629mg0.629mg0.838mg0.838mg 1.09mg1.08mg1.32mg1.30mg1.53mg1.59mg 95.5% 97.9% 95.4% 100.2% 97.9% 99.0%

The results show that the sample recovery rate of this product meets the requirements.

Description of drawings

Figure 1 is a standard curve diagram of dragon blood A.

Figure 2 is a standard curve diagram of dragon blood B.

Detailed ways

Example:

Chromatographic conditions and system suitability test Octadecylsilane bonded silica gel was used as filler; glacial acetic acid solution (1→100)-acetonitrile (63:37) was used as mobile phase, and the detection wavelength was 275nm. Column temperature: 30°C. The number of theoretical plates should not be less than 5,000 based on the calculation of the peaks A and B of dragon blood, and the separation degree between peak A and peak B of dragon blood should be greater than 2.0.

Preparation of Reference Substance Solution Accurately weigh the appropriate amount of reference substances of dragon blood A and dragon blood B which have been dried under reduced pressure by phosphorus pentoxide for 24 hours, add methanol to dissolve and make each 1ml containing dragon blood A 0.18mg, containing dragon blood A mixed solution of 0.04 mg of element B is obtained.

Preparation of the test solution Take about 0.25g of the powder of this product (passed through a No. 3 sieve), accurately weigh it into a 25ml measuring bottle, add about 20ml of methanol, and sonicate for 10 minutes to dissolve the dragon’s blood, let it cool to room temperature, and use Dilute with methanol to the mark, shake well, filter through a 0.45 μm microporous membrane, discard the initial filtrate, and obtain the product.

Determination method Precisely draw 10 μl of the reference substance solution and 5 μl of the test solution, inject it into the liquid chromatograph, measure it, and obtain it.

Claims (2)

1, a kind of dragon's blood quality control detection method, is the method for simultaneously measuring dragon's blood A and dragon's blood B content in dragon's blood by high performance liquid chromatography, is characterized in that being made up of following steps: 1. Chromatographic conditions and system suitability test Use octadecylsilane bonded silica gel as filler; 1% glacial acetic acid aqueous solution-acetonitrile 63±1:37±1 as mobile phase, detection wavelength is 275±2nm, column temperature: 30±1°C, the number of theoretical plates should be no less than 5000 based on the calculation of the peaks of draconin A and B, and the separation degree of the peaks of dracaine A and B should be greater than 2.0; 2. Preparation of reference substance solution Weigh appropriate amount of reference substances of ascarin A and ascarin B that have been dried under reduced pressure in a desiccant for 24 hours, add methanol to dissolve and make 0.18 mg of ascarin A and 0.18 mg of ascarinin per 1 ml The mixed solution of B0.04mg is ready; 3. Preparation of the test solution Take 0.25g of the powder of this product, accurately weigh it into a 25ml measuring bottle, add 20ml of methanol, and sonicate for 10 minutes to dissolve the dragon’s blood, let it cool to room temperature, dilute to the mark with methanol, shake Evenly, filter through a 0.45 μm microporous membrane, discard the initial filtrate, and obtain; 4. Determination Precisely draw 10 μl of the reference substance solution and 5 μl of the test solution, inject it into the liquid chromatograph, measure it, and get it. 2. The method for detecting the quality of dragon's blood according to claim 1, characterized in that it consists of the following steps: 1. Chromatographic conditions and system suitability test Use octadecylsilane bonded silica gel as filler; 1% glacial acetic acid aqueous solution-acetonitrile 63:37 as mobile phase, detection wavelength is 275nm, column temperature: 30°C, number of theoretical plates According to the calculation of dragon blood A and B peaks, it should not be less than 5000, and the separation degree of dragon blood A and B peaks should be greater than 2.0; 2. Preparation of reference substance solution Accurately weigh the appropriate amount of reference substances of ascarin A and ascarin B that have been dried under reduced pressure by phosphorus pentoxide for 24 hours, add methanol to dissolve and make each 1ml containing ascarin A 0.18mg, containing The mixed solution of dragon blood B 0.04mg is ready; 3. Preparation of the test solution Take 0.25g of the powder of this product passed through the No. 3 sieve, accurately weigh it into a 25ml measuring bottle, add 20ml of methanol, and sonicate for 10 minutes to dissolve the dragon's blood, let it cool to room temperature, and dissolve it with methanol Dilute to the mark, shake well, filter through a 0.45 μm microporous membrane, discard the initial filtrate, and obtain; 4. Determination method Precisely draw 10 μl of the reference substance solution and 5 μl of the test solution, inject it into the liquid chromatograph, measure it, and get it.

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