CN110596297B - Quality standard detection method for king melon seed medicinal material - Google Patents
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Abstract
The invention discloses a quality standard detection method of a king melon seed medicinal material, which comprises character and thin-layer chromatography identification and water, total ash and extract determination. According to the invention, the quality of the king melon seed medicinal material can be effectively controlled, and the quality and the safety of the medicinal material are improved under the conditions that the properties are revised on the basis of the existing quality standard (2003 edition) of traditional Chinese medicinal materials and national medicinal materials in Guizhou province and the items of thin layer identification, water, total ash content and leaching are added.
Description
Technical Field
The invention relates to the technical field of quality standards of traditional Chinese medicinal materials, in particular to a detection method of the quality standard of a traditional Chinese medicinal material, namely king melon seeds.
Background
The Wang Gua (also named as native melon, Laogua melon (in the picture), Mare melon (hail), and Hali (in the derivation) were recorded in the book, Shizhen (ancient book) is recorded in the book, that is, Mian Wan Gua sprout, with many tendrils and tender branches. Its leaf round like a horseshoe has sharp tip, dark face and light back, and is astringent but not smooth. Six months after the first day, seven months after the first day, five days after the second day, the first day is clustered. The knot is tired, red and yellow in color when cooked, and the skin is coarse and astringent. The king melon seeds are seeds of the king melon of the cucurbitaceae family, namely red hail seeds (materia medica Yanyi), hail seeds (Binghai simplified prescription from Binhu lake), and native melon seeds (materia medica Hui Yan), mature fruits are picked in autumn, split and seeds are taken out. The Wang melon seeds recorded in Fujian medicine Zhi have the functions of clearing heat and cooling blood. It can be used for treating hematemesis. The King melon seeds recorded in compendium for treating regurgitation and vomiting of food are recorded in Chinese materia medica for clearing heat and promoting diuresis, and cooling blood and stopping bleeding. It can be used for treating consumptive lung disease, hematemesis, jaundice, dysentery, and intestinal wind with bloody discharge.
The Wang melon seeds collected in 2003 edition of quality Standard of Chinese herbal and national herbs in Guizhou province have the effects of clearing heat and cooling blood. Can be used for treating consumptive lung disease, hematemesis, jaundice, dysentery, intestinal wind, hemoptysis, and spasm and pain of tendons and bones. But the quality standard is simpler, and is only imperfect under the items of [ character ] and [ microscopic identification ].
Disclosure of Invention
The invention aims to provide a quality standard detection method of king melon seeds, which aims to solve the problems in the prior art.
The technical scheme adopted by the invention for solving the problems is as follows: the characteristics are revised on the basis of the existing quality standard of traditional Chinese medicinal materials and national medicinal materials in Guizhou province (2003 edition), and thin-layer identification, water content, total ash content and leaching items are added.
Wherein, the character detection standard is as follows: the product is rectangular, with a length of 0.8-1.5 cm and a width of 0.5-1.2 cm. The surface is gray brown or gray brown, or is provided with a gray white transparent film, and the film is rough and has fine granular protrusions; a raised wide girdle band is arranged in the middle part, which is commonly called as 'jade girdle and waist winding'; the edge is protruded to form a ridge, both ends are respectively provided with a round blunt part, the top end is provided with a concave hole, and the body is light. The seed coat is hard, 3 chambers are arranged after being broken, no kernel exists in the two end chambers, 1 hole is formed in each chamber, the middle chamber is larger, 1 seed is arranged in the middle chamber, the seed coat is flat triangular or rectangular round, 0.4-1.0 cm in length, 0.2-0.6 cm in width, and a gray-green thin inner seed coat is wrapped at one end of the seed coat. The kernel is peeled into 2 petals, and is yellowish white or milky white, oily, fragrant and light in taste.
Wherein, the thin-layer chromatography identification steps are as follows: collecting powder of the product 1.0g, adding petroleum ether (60)
10ml at 90 deg.C, ultrasonic treating for 20 min, filtering, adding 10ml methanol into the residue, ultrasonic treating for 20 min, filtering, evaporating, and dissolving the residue with 1ml methanol to obtain sample solution. Taking 1.0g of semen Trichosanthis Cucumeroidis reference medicinal material, and making reference medicinal solution in the same way. Performing thin layer chromatography (0502 of the four parts of the design reside in the Chinese pharmacopoeia 2015), sucking 10 μ l of each of the two solutions, dropping on the same silica gel G plate, developing with chloroform-methanol-formic acid (20:1:0.01) as developing agentTaking out, air drying, and inspecting under ultraviolet lamp (365nm) to show fluorescent spots of the same color in the chromatogram of the sample at the position corresponding to the chromatogram of the reference medicinal material; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and inspecting under ultraviolet lamp (365 nm). In the chromatogram of the test solution, fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution.
Further, the method for measuring the moisture comprises the following steps: the water content is measured according to the second method of 0832 water content measurement in accordance with the general rules of the four departments of the edition of Chinese pharmacopoeia 2015, which is not more than 8.0 percent.
Further, the method for determining the total ash content comprises the following steps: the total ash content is measured according to a total ash content measuring method of 2302 of the four-part general rule of the edition of Chinese pharmacopoeia 2015, which is not more than 4.0 percent.
Further, the extract determination method comprises the following steps: the alcohol-soluble extract of semen Benincasae is measured by hot dipping method under 2201 item of general rules of four parts of the edition of Chinese pharmacopoeia 2015 with ethanol as solvent, and should not be less than 10.0%.
The invention has the beneficial effects that:
according to the invention, the quality of the king melon seed medicinal material can be effectively controlled, and the quality and the safety of the medicinal material are improved under the conditions that the properties are revised on the basis of the existing quality standard (2003 edition) of traditional Chinese medicinal materials and national medicinal materials in Guizhou province and the items of thin layer identification, water, total ash content and leaching are added.
Drawings
FIG. 1 is a drawing of Wang melon seed
FIG. 2 is a thin-layer chromatography identification system applicability test of king melon seeds; 1-3, testing a sample; 4. reference medicinal materials; (A: inspection under 365nm with an ultraviolet lamp; B: inspection under 365nm with an ultraviolet lamp after spraying 10% ethanol sulfate);
FIG. 3 different brands of thin layer chromatography plates; 1-3, testing a sample; 4. reference medicinal materials; (A is Qingdao ocean silica gel G plate, B is Yangtai Jiangyou yellow sea HSG, C is Qingdao Dingkang silica gel G plate as dot);
FIG. 4 thin layer chromatography plates at different temperatures; 1-3, testing a sample; 4. reference medicinal materials; (plate A at 10 deg.C, plate B at 25 deg.C, and plate C at 35 deg.C);
FIG. 5 relative humidity 32%, 58%, 72%; 1-3, testing a sample; 4. reference medicinal materials; (RH for plate A32%, RH for plate B58%, RH for plate C72%);
FIG. 6 shows TLC chart for stability examination of test solution; 1. the same day; 2. standing for 1 day; 3. standing for 2 days; 4. standing for 3 days; 5. reference medicinal materials;
FIG. 7 thin-layer chromatography identification of different batches of king melon seeds; 1-4, different batches of the king melon seed medicinal material (detailed information is shown in table 1); 5. and (5) comparison medicinal materials.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.
The embodiment of the invention comprises the following steps:
the original plant and the sample were identified as seeds of Trichosanthes cucumerides (Ser.) Maxim. of Cucurbitaceae plant by the pharmaceutical plant of the institute of medicine of Guizhou medical university and by Zhang Xue professor of Physciaenopsis in the department of pharmacy. The certificate specimen is stored in a key laboratory of pharmaceutical preparations of Guizhou province of Guizhou medical university.
4 batches of the king melon seeds are all from Guizhou. Wherein Guizhou Bijie (batch No. 20180912) is used as a reference drug.
Table 1 sample source table
[ PROPERTIES ] the properties of a plurality of batches of king melon seeds are observed and revised:
the product is rectangular, and has a length of about 0.9-1.2 cm and a width of 0.2-0.5 cm. The surface is gray brown or gray brown, or is provided with a gray white transparent film, and the film is rough and has fine granular protrusions; a raised wide girdle band is arranged in the middle part, which is commonly called as 'jade girdle and waist winding'; the edge is protruded to form a ridge, both ends are respectively provided with a round blunt part, the top end is provided with a concave hole, and the body is light. The seed coat is hard, 3 chambers are arranged after the seed coat is broken, no kernel exists in the two end chambers, 1 hole is formed in each chamber, the middle chamber is larger, 1 seed is arranged in the middle chamber, the seed coat is flat triangular or rectangular round, 0.5-0.7 cm in length, 0.3-0.5 cm in width, and a gray-green thin inner seed coat is coated at one end of the seed coat. The kernel is peeled into 2 petals, and is yellowish white or milky white, oily, fragrant and light in taste. The semen Trichosanthis Cucumeroidis shown in figure 1.
[ IDENTIFICATION ] thin-layer chromatography for identification
1. Chromatographic conditions and System suitability test
Through investigating the influence thin-layer identification factors such as different extraction methods, extraction solvents, developing agents and the like of the king melon seeds, the identification method of the king melon seeds by thin-layer chromatography is established, namely: collecting powder of the product 1.0g, adding petroleum ether (60)
10ml at 90 deg.C, ultrasonic treating for 20 min, filtering, adding 10ml methanol into the residue, ultrasonic treating for 20 min, filtering, evaporating, and dissolving the residue with 1ml methanol to obtain sample solution. Taking 1.0g of semen Trichosanthis Cucumeroidis reference medicinal material, and making reference medicinal solution in the same way. Performing thin layer chromatography (0502 of the four ministry of the national pharmacopoeia 2015), sucking 10 μ l of each of the two solutions, respectively dropping on the same silica gel G plate, developing with chloroform-methanol-formic acid (20:1:0.01) as developing agent, taking out, air drying, and viewing under ultraviolet lamp (365nm) to show fluorescent spots of the same color in the chromatogram of the test sample at the position corresponding to the chromatogram of the control medicinal material; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and inspecting under ultraviolet lamp (365 nm). The test chromatogram shows the same color of fluorescent spot at the corresponding position of the control chromatogram, FIG. 2.
2. Durability test for identifying king melon seeds by thin-layer chromatography
2.1 silica gel G thin-layer chromatography plates of different brands
According to the proposed chromatographic conditions, a Qingdao ocean silica gel G plate, a Yangtai Jiangyou yellow sea silica gel HSG plate and a Qingdao Dingkang silica gel G plate are respectively used for spotting, developing and visually inspecting. As shown in fig. 3.
The results show that: the three brands of silica gel G plates can know that the separation degrees of Qingdao ocean and Qingdao Dingkang silica gel G plates are basically the same, which shows that the durability is better, the Yangtze river friend yellow sea silica gel HSG plate is a high-efficiency plate, the separated spots are more than those of other two common silica gel G plates, and the common silica gel G plate is selected from the economical and applicable angle.
2.2 investigation of different development temperatures
According to the proposed chromatographic conditions, the Qingdao marine silica gel G plate is used for spotting, developing and developing, the Qingdao marine silica gel G plate is respectively developed under the conditions of 10 ℃, 25 ℃, 35 ℃ and 58% of relative humidity, and the influence of different temperatures on the development is examined, as shown in figure 4.
The results show that: the bands of the chromatogram map at different temperatures are clear, and the difference of the Rf values is not obvious, which indicates that the medicinal material is not greatly influenced by the temperature.
2.3 investigation of different deployment humidities
According to the proposed chromatographic conditions, a Qingdao marine silica gel G plate is used for spotting, developing and developing, the samples are respectively developed at the relative humidity of 32%, 58% and 72% (relative humidity is adjusted by sulfuric acid-water), and the development is examined at the temperature of 25 ℃, and the influence of different humidities on the development is examined, as shown in figure 5.
The results show that: although the bands of the chromatogram map under different humidity are clear, the Rf value has certain difference, but the medicinal materials can still be well identified, which shows that the humidity has better durability.
2.4 examination of the stability of the test solutions
Preparing test solution by the proposed method before 3 days, 2 days, 1 day and the same day, respectively dropping on the same silica gel G plate, developing, and developing as shown in FIG. 6.
The results show that: and (3) the chromatograms of the test solution which is placed for 1-3 days after being unfolded are basically consistent, and the test solution is good in stability in 3 days.
3. Determination of samples
The results of the 4 batches were measured according to the proposed thin layer chromatography identification method and are shown in FIG. 7.
[ WATER ] the water content of 4 batches of king melon seeds is 6.04-7.12%, the average value is 6.72%, but not more than 8.0%, measured according to the second method of 0832 water content measurement method in accordance with the general rules of the four ministry of the government (2015) of the pharmacopoeia of China.
[ Total Ash ] the total ash content was measured by the method of Total Ash according to the general rule 2302 of the four ministry of the pharmacopoeia of China 2015 edition, and the total ash content in 4 batches of the king melon seeds was 3.11% -3.34%, the average value was 3.27%, but not more than 4.0%.
[ EXTRACT ] the extract determination method is according to hot dipping method under 2201 item of general rules of the four departments of the 2015 pharmacopoeia, ethanol is used as solvent to determine alcohol-soluble extract of king melon seeds, wherein the alcohol-soluble extract of 4 batches of king melon seeds is 11.74% -13.16%, the average value is 12.51%, and is not less than 10.0%.
TABLE 2 measurement results of water, ash and extract of Wang melon seeds
The revised standard is as follows
[ PRODUCT ] the product is dry mature seed of Trichosanthes cucumeroides (Ser.) of Cucurbitaceae. Picking in autumn when the fruit is ripe and turns red, taking out the seeds, cleaning and drying.
[ PROPERTIES ] the product is rectangular, with a length of 0.8-1.5 cm and a width of 0.5-1.2 cm. The surface is gray brown or gray brown, or is provided with a gray white transparent film, and the film is rough and has fine granular protrusions; a raised wide girdle band is arranged in the middle part, which is commonly called as 'jade girdle and waist winding'; the edge is protruded to form a ridge, both ends are respectively provided with a round blunt part, the top end is provided with a concave hole, and the body is light. The seed coat is hard, 3 chambers are arranged after the seed coat is broken, no kernel exists in the two end chambers, 1 hole is formed in each chamber, the middle chamber is larger, 1 seed is arranged in the middle chamber, the seed coat is flat triangular or rectangular round, 0.4-1.0 cm in length, 0.2-0.6 cm in width, and a gray-green thin inner seed coat is wrapped at one end of the seed coat. The kernel is peeled into 2 petals, and is yellowish white or milky white, oily, fragrant and light in taste.
[ IDENTIFICATION ] this product is reddish brown in color. The epidermal cells of the seed coat are 1 column of grid cells. Some epidermal cells contain fine calcium oxalate crystals. The pericarp sclerenchyma cells exist in a plurality of sheets, are large, thick, yellowish-brown, polygonal, rectangular or irregular, have extension, truncated ends, diameters of about 81 microns, lengths of about 200 microns, wall thicknesses of 5-11 microns, are lignified, and have round or oblong holes and obvious pits. A plurality of stone cells are grouped or scattered singly, and have a square-like shape, a round-like shape, a conical shape or a rectangular-like shape, some stone cells have a few branches, the diameter is 25-68 mu m, the length is about 132 mu m, pore channels are obvious, yellow brown substances are filled in cavities, and some stone cells contain calcium oxalate square crystals; the stellate stone cells are irregular and have a plurality of branched protrusions, the diameter of the cells is 11-27 μm, the cells with thicker walls are colorless or light brown, and the cells with thicker walls are brown. The calcium oxalate cubic crystal is square, biconical or polygonal, the diameter is 3-11 μm, and the length is 17 μm. The cotyledon cells are filled with round or polygonal aleurone particles and contain fat oil drops and lipid substances.
(2) Collecting powder of the product 1.0g, adding petroleum ether (60)
10ml at 90 deg.C, ultrasonic treating for 20 min, filtering, adding 10ml methanol into the residue, ultrasonic treating for 20 min, filtering, evaporating, and dissolving the residue with 1ml methanol to obtain sample solution. Taking 1.0g of semen Trichosanthis Cucumeroidis reference medicinal material, and making reference medicinal solution in the same way. Performing thin layer chromatography (0502 of the four ministry of the national pharmacopoeia 2015), sucking 10 μ l of each of the two solutions, respectively dropping on the same silica gel G plate, developing with chloroform-methanol-formic acid (20:1:0.01) as developing agent, taking out, air drying, and inspecting under ultraviolet lamp (365nm) to show fluorescent spots of the same color in the chromatogram of the test sample at the position corresponding to the chromatogram of the control medicinal material; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and inspecting under ultraviolet lamp (365 nm). In the chromatogram of the test solution, fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution.
[ EXAMINATION ] the water content should not exceed 8.0% (0832 second method in the fourth division of the design reside in the pharmacopoeia of China 2015).
The total ash content is not more than 4.0% (the general rule 2302 in the four departments of the 2015 edition in China pharmacopoeia).
[ EXTRACT ] is measured by hot dipping method under alcohol soluble extract measuring method (2201 in the four ministry of the pharmacopoeia of China 2015 edition), and should not be less than 10.0%.
[ CHARANTIANJING ] is sour, bitter and neutral in nature. It enters lung, stomach and large intestine meridians.
[ FUNCTIONS ] can clear away heat and cool blood. Can be used for treating consumptive lung disease, hematemesis, jaundice, dysentery, intestinal wind, hemoptysis, and spasm and pain of tendons and bones.
[ dosage ] 3-9 g.
[ STORAGE ] standing in a cool and dry place.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes in the embodiments and/or modifications of the invention can be made, and equivalents and modifications of some features of the invention can be made without departing from the spirit and scope of the invention.
Claims (2)
1. A quality standard detection method of a king melon seed medicinal material is characterized by comprising the following steps: the method comprises the steps of character identification, thin-layer chromatography identification, water content determination, total ash content determination and extract determination; the thin-layer chromatography identification steps are as follows: taking the powder, adding petroleum ether, ultrasonic treating, filtering, adding methanol into residue, ultrasonic treating, filtering, evaporating to dryness, and dissolving the residue with methanol to obtain sample solution; taking another reference material of semen Trichosanthis Cucumeroidis, and making reference material solution by the same method; performing thin-layer chromatography test, dropping the above two solutions on the same silica gel G plate respectively, developing with chloroform-methanol-formic acid at a ratio of 20:1:0.01 as developing agent, taking out, air drying, and inspecting under ultraviolet lamp to show fluorescent spots of the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the control solution; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the spots are clearly developed, and inspecting under ultraviolet lamp; in the chromatogram of the test solution, fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution.
2. The quality standard detection method of the king melon seed medicinal material as claimed in claim 1, which is characterized in that: the extract is measured by hot dipping method, ethanol is used as solvent, and the alcohol-soluble extract of semen Cucurbitae should not be less than 10.0%.
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