阅读说明：本技术 一种瓜蒌配方颗粒基源鉴定的方法 (Method for identifying base source of snakegourd fruit formula particles ) 是由 付静 张志强 高扬 姜筱璇 于 2019-11-29 设计创作，主要内容包括：本发明涉及中药配方颗粒检测技术领域,公开了一种瓜蒌配方颗粒基源鉴定的方法,所述方法以甲醇-0.1％甲酸水溶液为流动相,采用梯度洗脱并结合紫外荧光检测器检测的液相色谱分析得到瓜蒌配方颗粒的特征图谱,然后以5号色谱峰的香草酸色谱峰为参照峰,计算3号色谱峰与4号色谱峰相对峰面积的比值,根据两者的比值即可判定所述瓜蒌配方颗粒的基源,操作简便、高效,弥补了瓜蒌配方颗粒基源鉴定技术领域的空缺,该方法可作为栝楼配方颗粒或双边栝楼配方颗粒的专属鉴别方法。(The invention relates to the technical field of traditional Chinese medicine formula particle detection, and discloses a trichosanthes kirilowii Maxim formula particle base source identification method, wherein a methanol-0.1% formic acid aqueous solution is used as a mobile phase, a characteristic spectrum of trichosanthes kirilowii Maxim formula particles is obtained by adopting gradient elution and liquid chromatography analysis combined with ultraviolet fluorescence detector detection, then a vanillic acid chromatographic peak of a No. 5 chromatographic peak is used as a reference peak, the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated, the base source of the trichosanthes kirilowii Maxim formula particles can be judged according to the ratio of the two, the operation is simple and convenient, the efficiency is high, the vacancy in the technical field of trichosanthes kirilowii Maxim formula particle base source identification is compensated, and the method can be used as.)

1. A method for identifying a base source of trichosanthes kirilowii maxim formula particles is characterized by comprising the following steps:

analyzing the test sample solution of the snakegourd fruit formula particles by adopting a liquid chromatography, wherein in the obtained characteristic spectrum, the No. 5 chromatographic peak is the characteristic peak of vanillic acid, and the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated by taking the characteristic peak of vanillic acid as a reference peak.

2. The method of claim 1, wherein the chromatographic conditions of the liquid chromatography are:

octadecylsilane chemically bonded silica is used as a filling column, methanol is used as a phase A, and 0.1% formic acid aqueous solution is used as a phase B to form a mobile phase; adopting a gradient elution mode, wherein the elution process comprises the following steps: 0-2.5min, phase A: 2% and B phase: 98 percent; 2.5-4min, phase A: 2-15%, phase B: 98-85%; 4-4.5min, phase A: 15% and phase B: 85 percent; 4.5-7.5min, phase A: 15-20% of phase B and 85-80% of phase B; 7.5-8.5min, phase A: 20-35% of phase B and 80-65% of phase B; 8.5-10min, 35-2% of phase A and phase B: 65-98 percent; 10-12min, phase A: 2% and 98% of phase B; the detection wavelength was 250 nm.

3. The method according to claim 2, wherein the chromatographic conditions are a flow rate of 0.35mL/min, a column temperature of 35 ℃ and a sample size of 1 μ L.

4. The method of any one of claims 1-3, wherein the sample solution is prepared by: taking the snakegourd fruit formula particles, grinding, weighing 0.2g, adding 10mL of water, carrying out ultrasonic extraction for 25-35 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely the test solution of the snakegourd fruit formula particles.

5. The method as claimed in claim 4, wherein the preparation method of the trichosanthes formula granule comprises the following steps: adding water 13 times the mass of the fructus trichosanthis decoction pieces, boiling and extracting for 1 hour, filtering, adding water 11 times the mass of the filter residue into the filter residue, boiling and extracting for 1 hour, filtering, combining the secondary filtrates, and concentrating under reduced pressure at 70 ℃.

6. The method of claim 5, wherein the method of preparing the trichosanthes formula further comprises: concentrating under reduced pressure until the density of the feed liquid is 1.30-1.40 g/cm at 55 DEG C³And drying at 80 ℃ to obtain the trichosanthes kirilowii maxim formula particles.

7. The method of claim 5, wherein the method of preparing the trichosanthes formula further comprises: concentrating under reduced pressure until the density of the feed liquid at 60 ℃ is 1.10-1.15 g/cm³Adding auxiliary materials accounting for 20% of the mass of the snakegourd fruit decoction pieces, and drying at 170-180 ℃ to obtain the snakegourd fruit formula particles.

8. The method according to any one of claims 1 to 7, comprising the steps of:

preparation of reference solutions

Taking 1g of a trichosanthes kirilowii maxim reference medicinal material, adding 10mL of water, carrying out ultrasonic extraction for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely a trichosanthes kirilowii maxim reference medicinal material reference solution;

taking 1g of bilateral trichosanthes kirilowii Maxim reference medicinal material, adding 10mL of water, carrying out ultrasonic extraction for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain filtrate, namely bilateral trichosanthes kirilowii Maxim reference medicinal material solution;

adding methanol into vanillic acid reference substance, and dissolving completely to obtain solution containing vanillic acid 20 μ g per 1mL as reference substance solution;

preparation of test solution

Taking the snakegourd fruit formula particles, grinding, weighing 0.2g, adding 10mL of water, ultrasonically extracting for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely a test solution of the snakegourd fruit formula particles;

liquid chromatography assay

And respectively sucking the reference substance solution and the test sample solution, injecting the reference substance solution and the test sample solution into a high performance liquid chromatograph for analysis, comparing a plurality of maps, and selecting a common characteristic peak, wherein the No. 5 chromatographic peak is a characteristic peak of vanillic acid, the characteristic peak of vanillic acid is taken as a reference peak, and the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated.

Technical Field

The invention relates to the technical field of detection of traditional Chinese medicine formula particles, and particularly relates to a method for identifying a base source of a trichosanthes kirilowii maxim formula particle.

Background

Fructus Trichosanthis is dry mature fruit of Trichosanthes kirilowii Maxim or Trichosanthes rosthornii Harms of Trichosanthes rosthornii of Trichosanthes of Cucurbitaceae, and its pericarp, seed and root are respectively treated with pericarpium Trichosanthis, semen Trichosanthis and Trichosanthis radix. According to the examination, Gua Lou is recorded in Shen nong Ben Cao Jing, listed as the middle-grade product, and is a commonly used Chinese medicine for clearing heat and removing toxicity. The original plant of snakegourd fruit used in the past generation is vine, tendril and single leaf, 3-5 cracks or does not crack, the fruit is round or long, the tip is provided with a stylobate or does not exist, the fruit is yellow after ripening, the shape description accords with the main characteristics of the cucurbitaceae plant, the snakegourd fruit is taken as the main source of the snakegourd fruit serving as the medicinal material in the distribution area of China by combining the cucurbitaceae plant of the snakegourd fruit, but the snakegourd fruit serving as the medicinal material is also used by fruits of the same family plants such as bilateral snakegourd fruit and the king melon fruit. The original plant of fructus Trichosanthis specified in pharmacopoeia of people's republic of China in 2015 is fructus Trichosanthis or bilateral fructus Trichosanthis of Trichosanthes of Cucurbitaceae.

Disclosure of Invention

Therefore, the technical problem to be solved by the invention is to overcome the defect that the trichosanthes kirilowii maxim formula particle and the bilateral trichosanthes kirilowii maxim formula particle cannot be distinguished in the prior art, so that the method for identifying the base source of the trichosanthes kirilowii maxim formula particle is provided.

In order to solve the technical problems, the invention provides a method for identifying the base source of the snakegourd fruit formula particles, which comprises the following steps:

analyzing the test sample solution of the snakegourd fruit formula particles by adopting a liquid chromatography, wherein in the obtained characteristic spectrum, the No. 5 chromatographic peak is the characteristic peak of vanillic acid, and the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated by taking the characteristic peak of vanillic acid as a reference peak.

Further, the chromatographic conditions of the liquid chromatography are as follows:

octadecylsilane chemically bonded silica is used as a filling column, methanol is used as a phase A, and 0.1% formic acid aqueous solution is used as a phase B to form a mobile phase; adopting a gradient elution mode, wherein the elution process comprises the following steps: 0-2.5min, phase A: 2% and B phase: 98 percent; 2.5-4min, phase A: 2-15%, phase B: 98-85%; 4-4.5min, phase A: 15% and phase B: 85 percent; 4.5-7.5min, phase A: 15-20% of phase B and 85-80% of phase B; 7.5-8.5min, phase A: 20-35% of phase B and 80-65% of phase B; 8.5-10min, 35-2% of phase A and phase B: 65-98 percent; 10-12min, phase A: 2% and 98% of phase B; the detection wavelength was 250 nm.

Further, the flow rate in the chromatographic conditions is 0.35mL/min, the column temperature is 35 ℃, and the sample injection amount is 1 μ L.

Further, the preparation method of the test solution comprises the following steps: taking the snakegourd fruit formula particles, grinding, weighing 0.2g, adding 10mL of water, carrying out ultrasonic extraction for 25-35 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely the test solution of the snakegourd fruit formula particles.

Further, the preparation method of the trichosanthes formula particle comprises the following steps: adding water 13 times the mass of the fructus trichosanthis decoction pieces, boiling and extracting for 1 hour, filtering, adding water 11 times the mass of the filter residue into the filter residue, boiling and extracting for 1 hour, filtering, combining the secondary filtrates, and concentrating under reduced pressure at 70 ℃.

Further, the preparation method of the trichosanthes formula particle also comprises the following steps: concentrating under reduced pressure until the density of the feed liquid is 1.30-1.40 g/cm at 55 DEG C³Drying at 80 deg.C to obtainAnd (3) preparing the snakegourd fruit formula particles.

Further, the preparation method of the trichosanthes formula particle also comprises the following steps: concentrating under reduced pressure until the density of the feed liquid at 60 ℃ is 1.10-1.15 g/cm³Adding auxiliary materials accounting for 20% of the mass of the snakegourd fruit decoction pieces, and drying at 170-180 ℃ to obtain the snakegourd fruit formula particles.

Furthermore, the trichosanthes decoction pieces are prepared from trichosanthes medicinal materials.

Further, the identification method comprises the following steps:

preparation of reference solutions

Taking 1g of a trichosanthes kirilowii maxim reference medicinal material, adding 10mL of water, carrying out ultrasonic extraction for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely a trichosanthes kirilowii maxim reference medicinal material reference solution;

taking 1g of bilateral trichosanthes kirilowii Maxim reference medicinal material, adding 10mL of water, carrying out ultrasonic extraction for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain filtrate, namely bilateral trichosanthes kirilowii Maxim reference medicinal material solution;

adding methanol into vanillic acid reference substance, and dissolving completely to obtain solution containing vanillic acid 20 μ g per 1mL as reference substance solution;

preparation of test solution

Taking the snakegourd fruit formula particles, grinding, weighing 0.2g, adding 10mL of water, ultrasonically extracting for 30 minutes, cooling to room temperature, shaking up, and filtering to obtain a filtrate, namely a test solution of the snakegourd fruit formula particles;

liquid chromatography assay

And respectively sucking the reference substance solution and the test sample solution, injecting the reference substance solution and the test sample solution into a high performance liquid chromatograph for analysis, comparing a plurality of maps, and selecting a common characteristic peak, wherein the No. 5 chromatographic peak is a characteristic peak of vanillic acid, the characteristic peak of vanillic acid is taken as a reference peak, and the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated.

The technical scheme of the invention has the following advantages:

1. the method for identifying the base source of the snakegourd fruit formula particles provided by the invention is characterized in that methanol-0.1% formic acid aqueous solution is used as a mobile phase, a characteristic map of the snakegourd fruit formula particles is obtained by adopting gradient elution and liquid chromatography analysis combined with ultraviolet fluorescence detector detection, then the ratio of the relative peak areas of the No. 3 chromatographic peak and the No. 4 chromatographic peak is calculated by taking the vanillic acid characteristic peak of the No. 5 chromatographic peak as a reference peak, and the base source of the snakegourd fruit formula particles can be judged according to the ratio of the No. 3 chromatographic peak to the No. 4 chromatographic peak.

2. According to the method for identifying the base source of the trichosanthes kirilowii maxim formula particles, the reference substance solution and the test solution of the reference medicinal material are extracted by a water extraction method, the preparation method is the same, and the reference substance chromatogram and the test solution of the reference medicinal material obtained by the method are more accurate in comparison and analysis and have stronger specificity.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a characteristic map of a reference solution of Trichosanthes kirilowii Maxim and Trichosanthes bilateral Maxim in example 1 of the present invention (wherein, peak No. 5 is vanillic acid S peak);

fig. 2 is a characteristic map of the trichosanthes kirilowii maxim formula particle in example 1 of the present invention (wherein, the peak No. 5 is a vanillic acid S peak).

Detailed Description

The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.