阅读说明：本技术 一种小红参颗粒及标煎液干粉的一板多信息薄层鉴别方法 (One-plate multi-information thin-layer identification method for red ginseng granules and target decoction dry powder ) 是由 牛丽颖 高乐 王相 李宁 王鑫国 于 2021-09-23 设计创作，主要内容包括：本发明涉及一种小红参颗粒及标煎液干粉的一板多信息薄层鉴别方法。其特征在于：用简便、快捷的前处理方法得到供试品与小红参药材溶液,分别点于同一块薄层板上,展开后,在4种不同的检视条件下,检视到小红参颗粒及标煎液干粉的13种信息斑点,归属于约10种不同的化学成分,不同的颜色斑点,代表不同的化学成分,颜色靓丽、易于质量评价,并做到了信息斑点互补,提升了质量检测水平。目前还未查阅到有关小红参颗粒及标煎液干粉快速多信息薄层鉴别报道。具新颖性、创新性和实用性。且方法简便、快捷,效率高、成本低、无环境污染。(The invention relates to a one-plate multi-information thin-layer identification method for rubia yunnanensis granules and target decoction dry powder. The method is characterized in that: a test sample and a rubia yunnanensis medicinal material solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and are unfolded, 13 information spots of rubia yunnanensis particles and standard decoction powder are inspected under 4 different inspection conditions, the 13 information spots belong to about 10 different chemical components, different color spots represent different chemical components, the color is beautiful, the quality evaluation is easy, the information spot complementation is realized, and the quality detection level is improved. At present, the rapid multi-information thin-layer identification report of the rubia yunnanensis granules and the standard decoction dry powder is not found. Has novelty, innovation and practicability. The method is simple, convenient and quick, and has high efficiency, low cost and no environmental pollution.)

1. A one-plate multi-information thin-layer identification method for red ginseng granules and standard decoction dry powder is characterized in that:

taking 0.5g of each of the rubia yunnanensis particles and the standard decoction dry powder, grinding, adding 2ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.5g of radix Rubiae Yunnanensis as reference material, adding 2ml of methanol, and making into reference material solution in the same way; sucking the three solutions with 4-5 mul each, and respectively dropping on the same silica gel GF₂₅₄Developing on a thin layer plate with chloroform-acetone-formic acid as developing agent at volume ratio of 4: 6: 0.5, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the chromatogram of the sample and the corresponding position of the chromatogram of the reference material; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the positions corresponding to those in the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color, inspecting under ultraviolet lamp 365nm, and displaying main fluorescent spots with the same color in the chromatogram of the sample at the corresponding positions of the chromatogram of the reference medicinal material; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the corresponding position of the chromatogram of the test solution and the chromatogram of the reference solution.

2. The thin-layer identification method for radix rubiae yunnanensis granules and standard decoction powder according to claim 1, wherein each 1g of granules is equivalent to 4g of crude drugs; each 1g of standard decoction is equivalent to 4g of crude drug.

3. The method of claim 1, wherein 13 information spots of the powder of the granule and the powder of the standard decoction of the radix rubiae yunnanensis are respectively inspected on the same thin-layer plate under 4 different inspection conditions with the same developing agent and belonging to about 10 chemical components.

Technical Field

The invention relates to a one-plate multi-information thin-layer identification method for rubia yunnanensis granules and target decoction dry powder. Belongs to the field of thin-layer identification of traditional Chinese medicines. Namely, thin-layer chromatography is adopted for identification, 13 information spots of the rubia yunnanensis particles and the standard decoction powder are inspected on the same thin-layer plate under 4 different inspection conditions, and the 13 information spots belong to about 10 chemical components.

Background

The identification method is characterized in that more than ten kinds of information spots are detected on a thin-layer plate under 4 different detection conditions, the method is simple, convenient, rapid and efficient, the sample solution and the contrast medicinal solution do not need to be purified repeatedly, and environmental pollution operation procedures such as evaporation and concentration of toxic solvents are avoided.

Radix Rubiae Yunnanensis is root of Rubia yunnanensis of Rubiaceae, named Yunnan purple ginseng, Xiaohuoxue, and Xiaohongyao. Has effects in promoting blood circulation, stopping bleeding, and dispelling blood stasis. Can be used for treating epistaxis, hematemesis, hematochezia, hematuria, metrorrhagia, menoxenia, amenorrhea, abdominal pain, rheumatic arthralgia, and hepatitis; can be used for treating traumatic injury, furuncle, neurodermatitis, etc.; it can be used singly to decoct and wash feet with radix Rubiae Yunnanensis. Mainly produced in Shaanxi, Henan, Hebei, Shandong provinces, and other provinces, and distributed in China, and belongs to national traditional Chinese medicinal materials. The chemical components of the rubia yunnanensis are complex and various, and mainly comprise anthraquinone and glycosides thereof and arborane triterpenes. In addition, cyclohexapeptides and naphthoquinones are also included. Was recorded in the first part of the Chinese pharmacopoeia 1977, the 1974 edition of the pharmaceutical Standard of Yunnan province, the 1988 edition of the quality Standard of Chinese medicinal materials in Guizhou province, and the 2003 edition of the quality Standard of Chinese medicinal materials and national medicinal materials in Guizhou province. But the standard only stipulates the basic source, properties, microscopic and physicochemical identification of the medicinal materials and other items. The standard technology content is low, and the quality of the medicinal materials and the preparations thereof cannot be scientifically controlled. At present, the research on the chemical components of the national traditional Chinese medicine rubia yunnanensis is reported, but the thin-layer identification is rarely reported, so the standard and systematic research on the quality of the national traditional Chinese medicine and the quality of the granules and the preparation thereof needs to be urgently needed to scientifically control the quality standard.

Under the condition of the research background, in order to provide a rapid multi-information thin-layer identification method for national traditional Chinese medicine red ginseng formula particles and standard decoction dry powder and adapt to the convenience of clinical application of patients, firstly, the preparation processes of the red ginseng particles and the standard decoction dry powder are researched; then, thin-layer identification research is carried out on the rubia yunnanensis particles and the standard decoction dry powder. The method for identifying the multiple information thin layers on one plate is simple, convenient and quick, and has no environmental pollution. The preparation process of the red ginseng medicinal materials, the granules and the standard decoction powder is briefly described as follows:

the radix rubiae yunnanensis medicinal material is prepared by removing impurities from collected radix rubiae yunnanensis, cleaning, slicing, and drying in the sun or at a proper temperature according to the processing specification of traditional Chinese medicines to obtain radix rubiae yunnanensis decoction pieces (also called medicinal materials).

The radix rubiae yunnanensis granule is prepared by decocting the radix rubiae yunnanensis decoction pieces with water for 2 times, each time for 1-2 hours, filtering, combining filtrates, concentrating under reduced pressure to an appropriate relative density, spray drying, adding an appropriate amount of dextrin into the dried powder, mixing uniformly, and granulating.

The dry powder of the red ginseng target decoction is prepared by decocting red ginseng decoction pieces with water for 2 times, each time for 1-2 hours, filtering, combining filtrates, concentrating at normal temperature to proper relative density, adding a proper amount of dextrin, stirring uniformly, placing in a 60 ℃ oven, drying, and pulverizing.

Disclosure of Invention

Creates a one-plate multi-information thin-layer identification method for the rubia yunnanensis granules and the target decoction powder. Namely, a test sample and a rubia yunnanensis medicinal material solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and are unfolded, 13 information spots of rubia yunnanensis particles and standard decoction powder are inspected under 4 different inspection conditions, belong to about 10 different chemical components and are in multiple colors and represent multiple chemical components, are bright and beautiful, are easy to judge, and are used for quality evaluation. At present, reports about rapid multi-information thin-layer identification of the rubia yunnanensis particles and the standard decoction dry powder are not found. Has novelty, innovation and practicability. The method is simple, convenient and quick, and has high efficiency, low cost and no environmental pollution.

The technical scheme adopted by the invention for solving the technical problems is as follows:

taking 0.5g of each of the rubia yunnanensis particles and the standard decoction dry powder, grinding, adding 2ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.5g of radix Rubiae Yunnanensis as reference material, adding 2ml of methanol, and making into reference material solution in the same way; sucking the three solutions with 4-5 mul each, and respectively dropping on the same silica gel GF₂₅₄Developing on a thin layer plate with chloroform-acetone-formic acid as developing agent at volume ratio of 4: 6: 0.5, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the chromatogram of the sample and the corresponding position of the chromatogram of the reference material; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the positions corresponding to those in the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color, inspecting under ultraviolet lamp 365nm, and displaying main fluorescent spots with the same color in the chromatogram of the sample at the corresponding positions of the chromatogram of the reference medicinal material; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the corresponding position of the chromatogram of the test solution and the chromatogram of the reference solution.

The principle of the invention is as follows:

according to the chemical structure and properties of each effective component of the traditional Chinese medicine, a test sample and a reference medicinal solution are simply, conveniently and quickly prepared by adopting a proper extraction solvent according to a similar compatible extraction principle. And then proper developing agent is adopted for developing, and various chemical components can be well separated on the thin-layer plate along with the selected developing agent according to different adsorption, desorption, re-adsorption and re-desorption capabilities. By means of effective components with similar polarities, different color spots are displayed on the same thin-layer plate under different inspection conditions, and a thin-layer chromatogram with multiple information is obtained.

The invention has the following innovation points and beneficial effects:

1. creates a rapid thin layer identification method for the rubia yunnanensis granules and the target decoction powder. Namely, a test sample and a rubia yunnanensis medicinal material solution are obtained by a simple and quick pretreatment method, are respectively spotted on the same thin-layer plate, and are unfolded, 13 information spots of rubia yunnanensis particles and standard decoction powder are inspected under 4 different inspection conditions, belong to about 10 different chemical components and are in multiple colors and represent multiple chemical components, are bright and beautiful, are easy to judge, and are used for quality evaluation. At present, the rapid multi-information thin-layer identification report of the rubia yunnanensis granules and the standard decoction dry powder is not found. Has novelty, innovation and practicability. The method is simple, convenient and quick, and has high efficiency, low cost and no environmental pollution.

2. Trichloromethane-acetone-formic acid with the volume ratio of 4: 6: 0.5 is used as a developing agent, after the rubia yunnanensis particles and the standard decoction dry powder are developed, the red ginseng particles and the standard decoction dry powder are placed under an ultraviolet lamp 365nm for inspection, 4 fluorescent spots are inspected on the medicinal materials and the standard decoction dry powder, 1 bright blue, one bright blue with yellowish and 2 red spots are inspected, 1 yellow spot is also formed on the particles except for the 4 fluorescent spots, but the amount of the 2 red fluorescent spots is reduced; the reason for this is that the main fluorescent spots are consistent (fig. 1) due to the difference in composition caused by low-temperature drying and high-temperature instant spray drying; inspecting under 254nm ultraviolet lamp to obtain clear brown spots and 2 dark brown spots (FIG. 2) on the medicinal materials, granules and the dried powder of the decoction; spraying 10% ethanol sulfate solution for color development, and inspecting under 365nm ultraviolet lamp to obtain 3 fluorescent spots of 1 bright blue and 2 bright blue green (figure 3); the thin layer plate was placed in a dark room, and the dry powder of the herbs, granules, and target decoction showed 3 red spots by light inspection, of which 1 was higher (fig. 4). The same thin-layer plate is inspected for 13 information spots under 4 inspection conditions, and the information spots are analyzed by comparison according to the size, content and Rf value of the spots and are attributed to about 10 different chemical components.

3. The innovation point of the invention is that the polarity of the developing agent is proper, so that the chemical components of the rubia yunnanensis are uniformly distributed in the whole range of the thin-layer plate, the Rf values of the several main spots with the highest content are all 0.3-0.6, and the brown spots, the colored red spots and the fluorescent main spots are similar in Rf value, but are not the same chemical components in comparison of the sizes and the shapes of the spots, the brown spots under 254nm and the colored red spots seem to be the same components, and the fluorescent spots before and after color development seem not to be the same components, so that the component spots under different conditions are complementary. On the premise of not increasing any workload and detection cost, the detection index is greatly improved, and supervision and inspection are facilitated.

4. From the analysis of the composition and proportion of the developing agent, about 10 components detected by the identification belong to medium-polarity components.

5. The identification method can obtain the sample and the medicinal material solution only by respectively 0.5g of the rubia yunnanensis medicinal material powder, the granule and the standard decoction powder and 6ml of methanol for 15 minutes, and can perform multi-index evaluation on the sample quality by adding 12ml of the developing agent and 30 minutes of developing time, thereby spending 1 hour totally, being simple, convenient and quick and having very high practical value.

Drawings

FIG. 1 is a thin-layer TLC chart of the granule and the target decoction powder of Rubia yunnanensis DC under 365nm UV light.

FIG. 2 is a thin-layer TLC chart of the granule of Rubia yunnanensis and the dry powder of the standard decoction under 254nm UV lamp.

FIG. 3 is a TLC image of thin layer observed under 365nm UV light after developing the granule of Rubia yunnanensis and the dry powder of the standard decoction with 10% ethanol sulfate solution.

FIG. 4 is a thin-layer TLC chart of light inspection in a dark room after color development of the Rubia yunnanensis granules and the target decoction powder with 10% sulfuric acid ethanol solution.

FIGS. 1, 2, 3 and 4 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1 and 2 are radix Rubiae Yunnanensis standard decoction powder; 3. 4, radix rubiae Yunnanensis is taken as a medicinal material; 5.6. is granule of radix Rubiae Yunnanensis;

detailed description of the invention

Taking 0.5g of each of the rubia yunnanensis particles and the standard decoction dry powder, grinding, adding 2ml of methanol respectively, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; get another0.5g of radix Rubiae Yunnanensis reference material, adding 2ml of methanol, and making into reference material solution by the same method; sucking the three solutions with 4-5 mul each, and respectively dropping on the same silica gel GF₂₅₄Developing on a thin layer plate with chloroform-acetone-formic acid as developing agent at volume ratio of 4: 6: 0.5, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain main fluorescent spots with the same color in the chromatogram of the sample and the corresponding position of the chromatogram of the reference material; inspecting under 254nm ultraviolet lamp to show main spots of the same color in the chromatogram of the sample at the positions corresponding to those in the chromatogram of the reference medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the spots develop color, inspecting under ultraviolet lamp 365nm, and displaying main fluorescent spots with the same color in the chromatogram of the sample at the corresponding positions of the chromatogram of the reference medicinal material; inspecting in dark room through lamplight, wherein the main spot with the same color appears at the corresponding position of the chromatogram of the test solution and the chromatogram of the reference solution.