CN108956845B - One-plate four-medicine multi-information thin-layer identification method for three-ingredient decoction freeze-dried powder - Google Patents
One-plate four-medicine multi-information thin-layer identification method for three-ingredient decoction freeze-dried powder Download PDFInfo
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Abstract
The invention relates to a thin-layer identification method for multiple information of one plate of four medicinal materials of three-component decoction freeze-dried powder. The method is characterized in that: obtaining a test sample and a reference medicinal material solution by a simple and quick pretreatment method, and inspecting bright yellow fluorescent spots of the rhubarb on the same thin-layer plate under 4 different inspection conditions by adopting the same test sample solution and the same developing agent; brilliant blue and bluish violet fluorescence main spots of immature bitter orange and notopterygium root; brown speckles of Notopterygium incisum; more than ten traditional Chinese medicine components such as red spots of mangnolia officinalis; the spots were clear and the separation was good. The identification increase rate is 100%, and the identification is completed by one thin-layer plate, only 10ml of sample and reference medicinal material treatment solvent and 15ml of developing agent are needed, the time is 1 hour, the method is simple, convenient, rapid and efficient, and the detection information is more, so that the method is not available in the reported methods at present. Provides an effective quality control method for the quality supervision of the sanhua decoction freeze-dried powder.
Description
Technical Field
The invention relates to a thin-layer identification method for multiple information of one plate of four medicinal materials of three-component decoction freeze-dried powder. The method is to identify 4 medicinal materials on the same thin-layer plate under 4 different inspection conditions, and multiple information refers to information spots of multiple effective components. Belongs to the field of thin-layer chromatography identification of traditional Chinese medicine quality control.
Background
In a traditional Chinese medicine compound preparation, particularly a traditional water-decocted compound preparation, according to a similar compatible extraction principle, a large part of fat-soluble components in the preparation are difficult to extract, more water-soluble components are extracted, the mutual interference is serious, after water extraction, the thin-layer identification of the fat-soluble components is carried out on raw medicinal materials, some of the raw medicinal materials cannot be adopted, the identification characteristic points of the water-soluble components must be searched, and the technical difficulty is limited, so the thin-layer identification increase rate of the water-extracted preparation is low, and the quantitative determination index is few. For example, in 2015 edition of Chinese pharmacopoeia, the decoction of hepatitis B treating granules and thirteen Chinese medicinal herbs are collected, and only thin-layer identification of 4 medicinal herbs is added. The identification rate is low, the identification method is very complicated and complex, 85g or 15g (containing lactose) of a sample is needed for completing 4 items of identification, 386ml of pretreatment solvent is needed only, and the time is 10-12 hours. 4 items identify that 4 sample solutions are prepared and are developed on 4 thin-layer plates for 4 times; the loaded dibutyl particles consist of 4 medicinal materials, and only one thin layer identification is added. The pretreatment needs 15g or 3g (containing lactose) of a sample and 82ml of a treatment solvent for 2 hours; the collected Erbao granules are composed of eleven medicinal materials, and thin-layer identification of 4 medicinal materials is added. And the identification method is very complicated and complicated, and 3 sample solutions are prepared for 4 items of identification and are unfolded on 4 thin-layer plates for 4 times. The total amount of sample is 80g, only 295ml of pretreatment solvent is needed, and the time is 8 hours; with the addition of a developing agent and development time, more organic solvent and time are spent, and the like are not sufficient.
In conclusion, thin-layer identification is basically a variety, and if N items of identification exist, N sample solutions are prepared, N thin-layer plates are unfolded for N times, and a traditional identification mode for identifying N medicinal materials is adopted. In order to eliminate interference, the pretreatment procedure of the sample is complex and troublesome, a large amount of organic reagents are needed for repeated purification treatment, the labor and time are wasted, the reagents are wasted, the environment is polluted, the health is harmed, and the detection period is long. Therefore, the quality standard detection including 6-7 item thin layer identification and two item content measurement is completed, one week of time is generally spent, if the test is repeated, the time is doubled, and the detection speed seriously restricts the modernized production speed of the traditional Chinese medicine. Therefore, the search for a simple and convenient and rapid detection method of the water extraction preparation, the improvement of the detection efficiency and the reduction of the detection cost is a necessary breakthrough in the quality control of the traditional Chinese medicine.
The sanhua decoction is a classic famous prescription published by the State administration of traditional Chinese medicine, and consists of 4 medicinal materials of rhubarb, magnolia officinalis, immature bitter orange and notopterygium root, and the prescription proportion and the preparation process are as follows:
prescription: rhubarb, magnolia bark, immature bitter orange and notopterygium root.
The preparation method comprises the following steps: water such as water is big like lima bean, three or two water is taken every time, three liters of water is decocted to one liter and one half, concentrated and freeze-dried to obtain sanhua decoction freeze-dried powder (the term explains that the water such as water is used for chopping medicinal materials in a right-side prescription to the water according to the ancient writing format from right to left and from top to bottom).
In order to ensure the quality of the preparation, thin-layer identification research is carried out on the sanhua decoction freeze-dried powder.
Disclosure of Invention
The invention relates to a traditional identification mode for identifying N medicinal materials by preparing N sample solutions and N thin-layer plates and expanding the solutions for N times aiming at the current variety for identifying N medicinal materials. Through the research on different combinations and the parameters such as proportion, color developing agent, thin layer carrier, detection conditions and the like of the developing agent, a new mode of thin layer identification with one test and multiple evaluations is created, wherein the new mode adopts the same sample solution and the same developing agent to simultaneously identify 4 medicinal materials and more than ten kinds of effective components of traditional Chinese medicines on the same thin layer plate under 4 different inspection conditions.
The technical scheme adopted by the invention for solving the technical problems is as follows:
taking 0.5g of sanhua decoction freeze-dried powder, adding 2ml of methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.1g of control medicinal material powder of cortex Magnolia officinalis, radix et rhizoma Rhei, Notopterygii rhizoma and fructus Aurantii Immaturus respectively, adding 2ml of methanol, respectively, ultrasonic treating for 20 min, and taking supernatant as respective control medicinal material solution; respectively dropping 4-6 μ l of the reference medicinal material solution and 5-8 μ l of the sample solution on the same silica gel GF254Spreading on a thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 10: 5: 0.2, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain bright yellow main fluorescence spot (FIG. 1) in the sample chromatogram at the position corresponding to the chromatogram of the radix et rhizoma Rhei reference material; displaying the same bright blue fluorescence main spot on the position corresponding to the color spectrum of the immature bitter orange control drug (figure 1); spraying 10% ethanol sulfate solution, heating at 105 deg.C until main spot of Notopterygii rhizoma is clearly developed, inspecting under ultraviolet lamp 365nm, and subjecting to sample chromatographyDisplaying the same color fluorescence main spots on the corresponding positions of the reference drug chromatogram (figure 2); then placing in a darkroom, observing through light, and displaying the same color of main spot in the chromatogram of the sample at the position corresponding to the chromatogram of the Notopterygii rhizoma control medicinal material (FIG. 3); spraying 5% vanillin sulfuric acid solution-ethanol mixed solution at volume ratio of 1: 8, heating at 105 deg.C until color development of spot is clear, placing in dark room, and observing with lamp light to obtain red main spot (figure 4) in the chromatogram of the test sample at the position corresponding to the chromatogram of cortex Magnolia officinalis control material.
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. By adopting the developing agents with different components, different proportions and different polarities for development, various chemical components can be well separated on respective thin-layer plates along with different developing agents according to different adsorption, desorption, re-adsorption and re-desorption capacities. And by means of effective components with similar polarities, the effective components are overlapped on the same thin-layer plate under different inspection conditions, but are not interfered with each other on different layers through different color development means, different spot colors are displayed, and the thin-layer chromatogram with multiple information is obtained. The thin layer desire of simplicity, convenience, rapidness, low cost and high efficiency is realized.
The invention has the following innovation points and beneficial effects:
1. the SANHUATANG lyophilized powder is developed with cyclohexane-ethyl acetate-formic acid at volume ratio of 10: 5: 0.2, and then inspected under ultraviolet lamp 365nm to obtain bright yellow fluorescent main spot of radix et rhizoma Rhei; a bright blue fluorescent main spot of immature bitter orange; spraying 10% ethanol sulfate solution, heating at 105 deg.C until the main spot of Notopterygii rhizoma is clearly developed, and detecting new main fluorescent spot of fructus Aurantii Immaturus and Notopterygii rhizoma after fluorescence increasing under ultraviolet lamp 365 nm; then placing in a dark room to inspect the main brown spots of the notopterygium root by lamp light; finally spraying 5% vanillin sulfuric acid solution-ethanol mixed solution with volume ratio of 1: 8, and inspecting red main spot of cortex Magnolia officinalis by light.
2. Firstly, 10% sulfuric acid ethanol solution is used for color development, and then 5% vanillin sulfuric acid solution-ethanol mixed solution with the volume ratio of 1: 8 is used for color development, which is reported for the first time. The innovativeness and the practicability are that the color development sequence, the specificity and the sensitivity of a color developing agent are utilized, namely, under the condition of developing color by a 10% sulfuric acid ethanol solution, the mangnolia officinalis does not show red spots and only shows main spots of notopterygium; the dilemma that the notopterygium root is directly sprayed with vanillin as a reference medicinal material to present a plurality of information spots, and a test solution presents few spots which cannot be described is eliminated.
3. The bright yellow fluorescent main spot of the rhubarb is directly detected by the thin-layer plate under the 365nm ultraviolet lamp; the main bright blue fluorescent spots of the immature bitter orange are sprayed with 10% sulfuric acid ethanol solution, after heating, the original fluorescent spots disappear, only new fluorescent spots after fluorescence enhancement are presented, and the spots and direct inspection under 365nm ultraviolet light are not of the same component, so that the detection index is greatly improved, and the quality supervision is facilitated.
4. Compared with the traditional thin-layer identification exemplified in the background technology, the identification increase rate is 100%, and the identification is completed by one thin-layer plate, and only 0.5g of sample, 0.1g of each of four control medicinal materials, 10ml of treatment solvent, 15ml of developing solvent and 1 hour are needed. The method is simple, convenient, fast and efficient, has a large amount of detection information, and is not available in the reported methods at present. Provides an effective quality control method for the quality supervision of the sanhua decoction freeze-dried powder. The necessity of detection technology innovation and the brought significant beneficial effects are fully illustrated.
Drawings
FIG. 1 is T L C picture of radix et rhizoma Rhei and fructus Aurantii Immaturus inspected by SANHUANG decoction lyophilized powder under 365nm ultraviolet lamp.
FIG. 2 is a T L C diagram of fluorescence-enhanced spots of Notopterygii rhizoma and Aurantii Immaturus, which is observed under an ultraviolet lamp 365nm after the SANHUATANG lyophilized powder is developed with 10% sulphuric acid ethanol solution.
FIG. 3 is a diagram of Notopterygium incisum main spot T L C inspected in a darkroom through lamplight after the sanhua decoction lyophilized powder is developed with 10% sulfuric acid ethanol solution.
FIG. 4 is a T L C diagram of Magnolia officinalis spots inspected by light after the lyophilized powder of the three-ingredient decoction is sprayed with 5% vanillin-sulfuric acid solution-ethanol mixed solution at a volume ratio of 1: 8 for color development.
FIG. 1, 2, 3, 4 are thin layer chromatograms of the same thin layer plate under different inspection conditions, wherein 1, Magnolia officinalis blank 2, Magnolia officinalis contrast medicinal material 3, Rheum officinale blank 4, Rheum officinale contrast medicinal material 5.6.7 sanhua decoction lyophilized powder sample solution 8, Citrus aurantium contrast medicinal material 9, Citrus aurantium blank 10, Notopterygium incisum contrast medicinal material 11, Notopterygium incisum blank
The specific implementation mode of the invention is as follows:
taking 0.5g of sanhua decoction freeze-dried powder, adding 2ml of methanol, carrying out ultrasonic treatment for 10 minutes, centrifuging, and taking supernate as a test solution; taking 0.1g of control medicinal material powder of cortex Magnolia officinalis, radix et rhizoma Rhei, Notopterygii rhizoma and fructus Aurantii Immaturus respectively, adding 2ml of methanol, respectively, ultrasonic treating for 20 min, and taking supernatant as respective control medicinal material solution; respectively dropping 4-6 μ l of the reference medicinal material solution and 5-8 μ l of the sample solution on the same silica gel GF254Spreading on a thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 10: 5: 0.2, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain bright yellow main fluorescence spots in the chromatogram of the sample at the positions corresponding to those of the chromatogram of the radix et rhizoma Rhei reference material; displaying the same bright blue fluorescence main spot on the position corresponding to the color spectrum of the immature bitter orange control medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the main spot of Notopterygii rhizoma develops color clearly, and inspecting under ultraviolet lamp 365nm to show the same color fluorescence main spot in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal materials of fructus Aurantii Immaturus and Notopterygii rhizoma; then placing in a darkroom, and observing through lamplight, wherein main spots with the same color appear in the chromatogram of the sample at the positions corresponding to the chromatogram of the notopterygium root control medicinal material; spraying 5% vanillin sulfuric acid solution-ethanol mixed solution at volume ratio of 1: 8, heating at 105 deg.C until color development of spot is clear, placing in dark room, and observing with lamp light to show the same red main spot in the chromatogram of the sample at the position corresponding to the chromatogram of cortex Magnolia officinalis control material.
Claims (2)
1. A thin-layer identification method for one-plate four-medicine multi-information of three-ingredient decoction freeze-dried powder is characterized by comprising the following steps:
taking 0.5g of SANHUATANG lyophilized powder, adding 2ml of methanol, and performing ultrasonic treatment 10Centrifuging the mixture, and taking the supernatant as a test solution; taking 0.1g of control medicinal material powder of cortex Magnolia officinalis, radix et rhizoma Rhei, Notopterygii rhizoma and fructus Aurantii Immaturus respectively, adding 2ml of methanol, respectively, ultrasonic treating for 20 min, and taking supernatant as respective control medicinal material solution; respectively dropping 4-6 μ l of the reference medicinal material solution and 5-8 μ l of the sample solution on the same silica gel GF254Spreading on a thin layer plate with cyclohexane-ethyl acetate-formic acid as developing agent at volume ratio of 10: 5: 0.2, taking out, drying with hot air, and inspecting under ultraviolet lamp 365nm to obtain bright yellow main fluorescence spots in the chromatogram of the sample at the positions corresponding to those of the chromatogram of the radix et rhizoma Rhei reference material; displaying the same bright blue fluorescence main spot on the position corresponding to the color spectrum of the immature bitter orange control medicinal material; spraying 10% sulphuric acid ethanol solution, heating at 105 deg.C until the main spot of Notopterygii rhizoma develops color clearly, and inspecting under ultraviolet lamp 365nm to show the same color fluorescence main spot in the chromatogram of the sample at the position corresponding to the chromatogram of the control medicinal materials of fructus Aurantii Immaturus and Notopterygii rhizoma; then placing in a darkroom, and observing through lamplight, wherein main spots with the same color appear in the chromatogram of the sample at the positions corresponding to the chromatogram of the notopterygium root control medicinal material; spraying 5% vanillin sulfuric acid solution-ethanol mixed solution at volume ratio of 1: 8, heating at 105 deg.C until color development of spot is clear, placing in dark room, and observing with lamp light to show the same red main spot in the chromatogram of the sample at the position corresponding to the chromatogram of cortex Magnolia officinalis control material.
2. The thin-layer identification method for three-ingredient decoction freeze-dried powder, one plate and four medicinal materials with multiple information according to claim 1, wherein each 1g of the freeze-dried powder is equivalent to 4.5-5.3 g of original medicinal materials.
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| CN111024834B (en) * | 2019-11-27 | 2022-12-27 | 浙江金大康动物保健品有限公司 | Gradient full-information thin-layer identification method for notopterygium root medicinal material |
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| CN111735901B (en) * | 2020-07-09 | 2021-08-10 | 四川大学 | Gradient full-information thin-layer identification method for multi-source rhubarb medicinal material |
| CN112051353B (en) * | 2020-09-23 | 2022-10-11 | 浙江金大康动物保健品有限公司 | Gradient full-information thin-layer identification method for radix peucedani medicinal material |
| CN117871766B (en) * | 2023-12-31 | 2024-08-06 | 湖南易能生物医药有限公司 | Thin-layer identification method for medicinal materials notopterygium root and rheum officinale in traditional Chinese medicine composition containing rheum officinale and application of thin-layer identification method |
| CN118191129A (en) * | 2024-03-04 | 2024-06-14 | 湖南易能生物医药有限公司 | Fingerprint construction method of traditional Chinese medicine composition containing rheum officinale and application of fingerprint construction method |
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Inventor after: Niu Liying Inventor after: Wang Xinguo Inventor after: Wang Xiang Inventor after: Gao Le Inventor before: An Lina Inventor before: Niu Liying Inventor before: Wang Xiang Inventor before: Gao Le Inventor before: Han Guiru Inventor before: Wang Xinguo |
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Effective date of registration: 20220317 Address after: 675099 Zhuangdian pharmaceutical Park, Chuxiong high tech Zone, Chuxiong Yi Autonomous Prefecture, Yunnan Province Patentee after: YUNNAN SHENWEI SHIPURUI PHARMACEUTICAL Co.,Ltd. Patentee after: SHINEWAY PHARMACEUTICAL Group Ltd. Patentee after: HEBEI University OF CHINESE MEDICINE Address before: 050200 No.3 Xingyuan Road, economic development zone, Luquan District, Shijiazhuang City, Hebei Province Patentee before: HEBEI University OF CHINESE MEDICINE |