CN108956845B - A kind of multi-information thin-layer identification method of Sanhua decoction freeze-dried powder with one plate and four medicinal materials - Google Patents

Abstract

The invention relates to a multi-information thin-layer identification method for one plate and four medicinal materials of Sanhua Tang freeze-dried powder. It is characterized in that: obtaining the test product and the reference medicinal material solution by a simple and fast pretreatment method, using the same test product solution, the same developing agent, on the same thin-layer plate, under 4 different inspection conditions, inspect the rhubarb. There are more than ten kinds of traditional Chinese medicine ingredients such as bright yellow fluorescent spots; bright blue and blue-purple fluorescent main spots of Citrus aurantium and Qianghuo; tan spots of Qianghuo; red spots of Magnolia officinalis; the spots are clear and well separated. The identification and renewal rate is 100%, and it is completed by a thin-layer plate. It only needs 10ml of solvent for sample and reference medicinal materials, 15ml of developing agent, and 1 hour of time. The method is simple, fast, efficient, and has a lot of detection information. It is the method that has been reported so far. None are available. It provides an effective quality control method for the quality supervision of Sanhuatang freeze-dried powder.

Description

A kind of multi-information thin-layer identification method of Sanhua decoction freeze-dried powder with one plate and four medicinal materials

technical field

The invention relates to a multi-information thin-layer identification method for one plate and four medicinal materials of Sanhua Tang freeze-dried powder. It refers to the identification of 4 medicinal materials at the same time under 4 different inspection conditions on the same thin-layer board, and multi-information refers to the information spots of various active ingredients. The invention belongs to the field of thin-layer chromatography identification for quality control of traditional Chinese medicines.

Background technique

In traditional Chinese medicine compound preparations, especially those prepared by traditional decoction, according to the extraction principle of similarity and compatibility, it is difficult to extract a large part of the fat-soluble components in the preparations, while the water-soluble components are extracted more and interfere with each other. Seriously, after water extraction, the original medicinal materials are identified by the thin layer of fat-soluble components, some of which can no longer be used, and the identification feature points of water-soluble components must be found. Due to technical difficulties, the thin layer identification of water-extracted preparations is updated. The rate is low, and there are few quantitative indicators. Taking the 2015 edition of the Chinese Pharmacopoeia as an example, the decocted Yiganning granules contained in the 13 herbs consist of 13 herbs, and only the thin-layer identification of 4 herbs has been added. Not only the identification update rate is low, but the identification method is also very cumbersome and complicated. The completion of the four identifications requires a sample of 85g or 15g (containing lactose), only a pretreatment solvent of 386ml, and a time of 10 to 12 hours. For the 4-item identification, 4 kinds of test solution solutions should be prepared, and four times of unfolding were completed on 4 thin-layer plates; the collected Erding granules were composed of 4 medicinal materials, and only one thin-layer identification was added. The pretreatment requires 15g or 3g of sample (containing lactose), 82ml of solvent, and a time of 2 hours; the Erbao granules contained are composed of eleven medicinal materials, and the thin-layer identification of four medicinal materials has been added. And the identification method is also very cumbersome and complicated. For the 4 identifications, 3 kinds of test solutions should be prepared, and 4 times of expansion are completed on 4 thin-layer plates. A total of 80g of sample, 295ml of pretreatment solvent, and 8 hours of time are required; plus developing agent and developing time, more organic solvent and time are required, etc. There are many similar examples.

To sum up, the thin-layer identification is basically one variety. If there are N identifications, N kinds of test solutions, N thin-layer plates, and N times are prepared to identify the traditional identification mode of N medicinal materials. In order to eliminate interference, the pretreatment procedures of the samples are complicated and cumbersome, and a large number of organic reagents need to be repeatedly purified, which is labor-intensive, time-consuming, reagent-intensive, pollutes the environment, endangers health, and has a long detection cycle. In this way, it generally takes a week to complete a quality standard test containing 6 to 7 thin-layer identification and two content determinations. In case of retest, the time will be doubled, and the detection speed will seriously restrict the modern production speed of traditional Chinese medicine. Therefore, finding a simple and fast detection method for water-extracted preparations, improving detection efficiency, and reducing detection costs is a difficulty that must be overcome in the quality control of traditional Chinese medicine.

Sanhua Decoction is a classic prescription announced by the State Administration of Traditional Chinese Medicine.

Prescription: equal parts of rhubarb, Magnolia, Citrus aurantium and Qianghuo.

Preparation method: You cut as big as Madou, take three taels per serving, three liters of water, fry to one and a half liters, concentrate, freeze-dry, get Sanhua Tang freeze-dried powder (noun explanation: You cut as big as Madou, refers to pressing In the ancient right-to-left, top-down writing format, the medicinal materials in the prescription on the right side are cut into pieces to the size of hemp beans).

In order to ensure the quality of the preparation, a thin-layer identification study was carried out on the freeze-dried powder of Sanhua Decoction.

SUMMARY OF THE INVENTION

The present invention is aimed at the current one variety, and if there are N items of identification, N kinds of test solution solutions, N pieces of thin-layer plates, and N times are unfolded to identify the traditional identification mode of N medicinal materials. By studying the parameters of different combinations of developing agents and their ratios, color developing agents, thin-layer carriers, and detection conditions, four different inspections were created using the same test solution, the same developing agent, and the same thin-layer plate. Under the condition of simultaneous identification of 4 medicinal materials and more than ten kinds of active ingredients of traditional Chinese medicine, a new mode of TLC identification with one measurement and multiple evaluations.

The scheme adopted by the present invention to solve its technical problem is:

Take 0.5 g of Sanhua Tang freeze-dried powder, add 2 ml of methanol, ultrasonically treat it for 10 minutes, centrifuge, and use the supernatant as the test solution; also take 0.1 g of Magnolia officinalis, rhubarb, Qianghuo and Citrus aurantium reference medicinal powders, respectively add Methanol 2ml, ultrasonically treated for 20 minutes, take the supernatant as the respective reference medicinal material solution; pipette 4-6 μl of the above-mentioned control medicinal material solution and 5-8 μl of the test solution, respectively, and place them on the same silica gel GF ₂₅₄ thin-layer plate, respectively, by volume Cyclohexane-ethyl acetate-formic acid with a ratio of 10:5:0.2 was used as a developing agent, developed, taken out, dried with hot air, and inspected under an ultraviolet lamp at 365 nm. At the position, the same bright yellow fluorescent main spot was displayed (Figure 1); at the position corresponding to the chromatogram of the control medicinal material of Citrus aurantium, the same bright blue fluorescent main spot was displayed (Figure 1); sprayed with 10% sulfuric acid ethanol solution, Heated at 105°C until the main spot of Qianghuo was clearly colored, and inspected under a UV lamp at 365 nm. In the chromatogram of the test product, the main spots of the same color were displayed in the chromatogram corresponding to the chromatogram of Citrus aurantium and Qianghuo control material respectively (Figure 2). ; Put it in a dark room to check through the light, and in the chromatogram of the test sample, the main spots of the same color appear on the position corresponding to the chromatogram of the Qiang living control medicinal material (Figure 3); then spray with 5% vanilla in a volume ratio of 1:8 Aldehyde sulfuric acid solution-ethanol mixed solution, heated at 105 ℃ until the spot color is clear, placed in a dark room, inspected through light, in the chromatogram of the test sample, at the position corresponding to the chromatogram of Magnolia officinalis, the main red color was the same. spots (Figure 4).

The principle of the present invention is as follows:

According to the chemical structure and properties of the active ingredients of traditional Chinese medicine, following the extraction principle of similarity and compatibility, and using suitable extraction solvents, the test sample and the reference medicinal material solution were prepared simply and quickly. Using developing agents with different components, different proportions, and different polarities to carry out development, various chemical components will vary in their respective good separation on the thin layer plate. Then, with the help of various active ingredients with similar polarities, on the same thin-layer board, under different inspection conditions, although overlapping, through different color rendering methods, they can be displayed at different levels without interfering with each other. Individually different spot colors to obtain informative thin-layer chromatograms. Realize the desire for simple, fast, low-cost, and high-efficiency thin layers.

The innovations and beneficial effects of the present invention are as follows:

1. The lyophilized powder of Sanhua Decoction uses cyclohexane-ethyl acetate-formic acid with a volume ratio of 10:5:0.2 as a developing agent, and after development, firstly inspected under an ultraviolet lamp at 365 nm, the bright yellow fluorescent main spot of rhubarb; The bright blue fluorescent main spots of Citrus aurantium; sprayed with 10% sulfuric acid ethanol solution, heated at 105 ℃ until the color of the main spots of Qianghuo was clear, and checked the new fluorescent main spots of Citrus aurantium and Qianghuo after the increase of fluorescence under the UV lamp 365nm; In a dark room, check the main spots of tan and tan with light; finally, spray with a 1:8 volume ratio of 5% vanillin sulfuric acid solution-ethanol mixture solution, check the red main spots of Magnolia officinalis through light.

2. The technique of developing color first with 10% sulfuric acid ethanol solution, and then with 5% vanillin sulfuric acid solution-ethanol mixed solution with a volume ratio of 1:8, is the first report. Its innovation and practicability lie in the use of the color development sequence, specificity and sensitivity of the color developer, that is, in the case of color development in 10% sulfuric acid ethanol solution, Magnolia officinalis does not show red spots, but only the main spots of Qiang live. ; The predicament that the Qianghuo reference medicinal material was directly sprayed with vanillin and presented multiple information spots, while the test solution presented few spots, could not be described.

3. The main bright yellow fluorescent spots of rhubarb directly inspected under the ultraviolet light of 365nm on the thin-layer plate; the main bright blue fluorescent spots of Citrus aurantium, spray 10% sulfuric acid ethanol solution, after heating, these original fluorescent spots disappear, Only the new fluorescent spots after increased fluorescence are displayed. These spots are not the same component as those directly inspected under the UV lamp at 365nm, which greatly improves the detection index and is more conducive to quality supervision.

4. Compared with the traditional thin-layer identification exemplified under the background technology, the identification renewal rate is 100%, and it is completed by a thin-layer plate, requiring only 0.5g of the sample, 0.1g of each of the four reference medicinal materials, 10ml of processing solvent, and Dosage 15ml, time 1 hour. The method is simple, fast, efficient, and has a lot of detection information, which are not available in the reported methods so far. It provides an effective quality control method for the quality supervision of Sanhuatang freeze-dried powder. It fully explained the necessity of detection technology innovation and the significant beneficial effects it brought.

Description of drawings

Fig. 1 is the TLC image of rhubarb and citrus aurantium examined under the UV lamp 365nm of the lyophilized powder of Sanhua decoction.

Figure 2 is the TLC image of the fluorescent spots of Qiang living and Citrus aurantium examined under a UV lamp at 365 nm after the 10% sulfuric acid ethanol solution of the lyophilized powder of Sanhua Tang was developed.

Figure 3 shows the TLC image of the main spot of Qianghuo in a dark room after color development of 10% sulfuric acid ethanol solution of the freeze-dried powder of Sanhua decoction.

Figure 4 is a TLC image of Magnolia officinalis spots viewed through light after the freeze-dried powder of Sanhua Tang was sprayed with a 5% vanillin sulfuric acid solution-ethanol mixed solution with a volume ratio of 1:8 for color development.

Figures 1, 2, 3, and 4 are the TLC chromatograms of the same thin-layer plate under different inspection conditions, among which, 1. Magnolia officinalis blank 2. Magnolia officinalis reference material 3. Rhubarb blank 4. Rhubarb reference medicinal material 5.6.7 Sanhua Tang freeze-dried powder sample solution 8. Citrus aurantium reference medicinal material 9 Citrus aurantium blank 10. Qianghuo reference medicinal material 11. Qianghuo blank

The specific embodiments of the present invention are as follows:

Take 0.5 g of Sanhua Tang freeze-dried powder, add 2 ml of methanol, ultrasonically treat it for 10 minutes, centrifuge, and use the supernatant as the test solution; also take 0.1 g of Magnolia officinalis, rhubarb, Qianghuo and Citrus aurantium reference medicinal powders, respectively add Methanol 2ml, ultrasonically treated for 20 minutes, take the supernatant as the respective reference medicinal material solution; pipette 4-6 μl of the above-mentioned control medicinal material solution and 5-8 μl of the test solution, respectively, and place them on the same silica gel GF ₂₅₄ thin-layer plate, respectively, by volume Cyclohexane-ethyl acetate-formic acid with a ratio of 10:5:0.2 was used as a developing agent, developed, taken out, dried with hot air, and inspected under an ultraviolet lamp at 365 nm. In the position, the same bright yellow fluorescent main spot was displayed; in the corresponding position on the chromatogram of the citrus reference medicinal material, the same bright blue fluorescent main spot was displayed; sprayed with 10% sulfuric acid ethanol solution, heated at 105 ℃ until the main spot of Qianghuo was displayed. The color is clear and inspected under UV light at 365 nm. In the chromatogram of the test product, the main fluorescent spots of the same color are displayed at the positions corresponding to the chromatograms of the reference medicinal materials of Citrus aurantium and Qianghuo; In the product chromatogram, the main spots of the same color appear at the corresponding positions of the chromatogram of the Qianghuo reference medicinal materials; then spray with a 5% vanillin sulfuric acid solution-ethanol mixed solution with a volume ratio of 1:8, and heat at 105 °C until the spots develop color. Clear, placed in a dark room, inspected through light, in the chromatogram of the test product, the same red main spot was displayed on the corresponding position of the chromatogram of the reference medicinal material of Magnolia officinalis.

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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