Thin-layer chromatography detection method for black tea in herba cistanche and herba epimedii black tea
Through the thin-layer chromatography detection method, the existing black tea detection methods are solved, the operation of the existing black tea detection methods, poor separation effect, low accuracy and high cost in Cistanche Epimedium Black Tea, and high accuracy, sensitivity and low cost black tea ingredients detection are achieved.
Patent Information
- Application Number
- CN202510124569.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-26
- Publication Date
- 2025-05-06
AI Technical Summary
When used in Cistanche Epimedium Black Tea, the existing black tea detection methods have problems such as complex operation, poor separation effect, low accuracy and high cost.
A thin layer chromatography detection method is adopted, including preparing test sample solution and reference sample solution, using suitable expanding agent and color developer, and testing through polyamide thin-layer plates to achieve effective separation and high accuracy detection of black tea ingredient tea polyphenols.
It realizes high accuracy and sensitivity detection of black tea ingredients in Cistanche Epimedium Black Tea, which is simple to operate and low cost, improving the accuracy and comprehensiveness of quality control.
Smart Images
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Abstract
Description
Technical Field
[0001] The present application relates to the technical field of food detection, and in particular to a thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea. Background Art
[0002] Cistanche Epimedium Black Tea is a health food with the function of relieving physical fatigue, which is made of Cistanche, jujube, epimedium, Acanthopanax, black tea, dextrin and other raw materials. It adopts Chinese herbal medicine water extraction technology and combines modern technology. This product is a 3.0g / bag tea brewing agent, and its characteristic ingredients are total saponins, tea polyphenols and icariin. Studies have shown that tea contains a variety of anti-fatigue ingredients such as theanine, tea polyphenols, and theophylline, and people can reduce daily fatigue and movement fatigue by drinking tea. Therefore, black tea is one of the important ingredients in Cistanche Epimedium Black Tea, and accurate testing can ensure the quality and efficacy of the product.
[0003] Tea polyphenols is a general term for a class of polyphenol compounds in tea leaves that have strong antioxidant effects. Its content accounts for about 15% to 25% of the dry weight of fresh tea leaves. In recent years, a large number of research results have shown that tea polyphenols have multiple pharmacological effects such as antioxidant, anti-inflammatory, anti-tumor, anti-radiation, and anti-viral effects, and its content has a very important influence on the quality of tea leaves. However, the complex structure of tea polyphenols makes it difficult to accurately measure, which seriously restricts the in-depth research on tea polyphenols.
[0004] At present, there are many methods for detecting black tea, but when they are used to detect the black tea components in Cistanche and Epimedium black tea, there are often problems such as complex operation, poor separation effect, low accuracy and high cost. Summary of the invention
[0005] Based on this, the present application provides a thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea with good separation effect, high detection accuracy, simple operation and low cost.
[0006] The specific technical solutions are as follows:
[0007] A thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea, comprising the following steps:
[0008] Prepare the test solution of Cistanche deserticola and Epimedium black tea, catechin reference solution, epigallocatechin-3-glucopyranoside reference solution and black tea reference solution;
[0009] The test solution of the Cistanche deserticola and Epimedium black tea, the catechin reference solution, the epigallocatechin gallate reference solution and the black tea reference solution are spotted on the same polyamide thin layer plate, developed with a developing agent, taken out, dried and then subjected to color development treatment;
[0010] Wherein, the developing agent is n-butanol, acetone and formic acid in a volume ratio of (1-10):(4-12):(1-6).
[0011] In one embodiment, the developing agent is n-butanol, acetone and formic acid in a volume ratio of (5-10):(4-8):3.
[0012] In one embodiment, the color developing agent used in the color developing process is a ferric chloride ethanol solution with a mass concentration of 0.1% to 5%.
[0013] In one of the embodiments, the color developing agent used in the color developing process is a ferric chloride ethanol solution with a mass concentration of 4% to 5%.
[0014] In one embodiment, the method for preparing the test solution of Cistanche deserticola and Epimedium black tea to be tested comprises the following steps:
[0015] Adding a first solvent to the tested Cistanche deserticola and Epimedium black tea for extraction, evaporating the obtained extract to dryness to prepare a first solid;
[0016] The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid;
[0017] The second solid is added into the third solvent for dissolution to prepare the test solution of the Cistanche deserticola and Epimedium black tea to be tested;
[0018] And / or, the method for preparing a black tea reference solution comprises the following steps:
[0019] A first solvent is added to black tea for extraction, and the obtained extract is evaporated to dryness to prepare a first solid;
[0020] The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid;
[0021] The second solid is added into the third solvent for dissolution to prepare the black tea reference substance solution.
[0022] In one embodiment, the first solvents are each independently an ethanol aqueous solution with a volume concentration of 35% to 45%.
[0023] In one embodiment, the second solvent is each independently water.
[0024] In one embodiment, the third solvent is independently methanol.
[0025] In one embodiment, the step of adding the first solvent for extraction and evaporating the obtained extract to dryness has one or more of the following characteristics:
[0026] (1) The extraction temperature is 75°C to 85°C;
[0027] (2) The number of extractions is 2 to 4 times, and the extraction time for each time is 40 to 50 minutes;
[0028] (3) The evaporation temperature is 55℃~65℃.
[0029] In one embodiment, the step of adding a second solvent, performing ultrasound and centrifugation, and evaporating the obtained extract to dryness has one or more of the following characteristics:
[0030] (1) Ultrasonic conditions include: frequency of 30 kHz to 50 kHz, power of 250 W to 350 W;
[0031] (2) The ultrasound duration is 20 to 40 minutes;
[0032] (3) The centrifugal speed is 3000 r / min to 4000 r / min;
[0033] (4) The centrifugation time is 10 min to 20 min;
[0034] (5) The evaporation temperature is 55℃~65℃.
[0035] The above-mentioned thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea can achieve effective separation of black tea active ingredient tea polyphenols in Cistanche and Epimedium black tea by adopting suitable developing agent and thin layer plate for detection, and then detect it with high accuracy and high sensitivity. The detection operation is simple, the detection time is short, and the cost is low. The corresponding thin layer chromatography spots are clear and without interference, which improves the accuracy and comprehensiveness of quality control of Cistanche and Epimedium black tea, and is conducive to the establishment and overall evaluation of quality standards of Cistanche and Epimedium black tea. BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1 This is a thin layer plate image of Example 1 using n-butanol:acetone:formic acid (5:8:3) as the developing agent and a polyamide plate as the thin layer plate.
[0037] Figure 2 This is a thin layer plate image of Example 2 using n-butanol:acetone:formic acid (1:8:3) as the developing agent and a polyamide plate as the thin layer plate.
[0038] Figure 3 This is a thin layer plate image of Example 3 using n-butanol:acetone:formic acid (10:8:3) as the developing agent and a polyamide plate as the thin layer plate.
[0039] Figure 4 This is a thin layer plate image of Example 4 using n-butanol:acetone:formic acid (5:4:3) as the developing agent and a polyamide plate as the thin layer plate.
[0040] Figure 5 This is a thin layer plate image of Example 5 using n-butanol:acetone:formic acid (5:12:3) as the developing agent and a polyamide plate as the thin layer plate.
[0041] Figure 6 This is a thin layer plate image of Example 6 using n-butanol:acetone:formic acid (5:8:1) as the developing agent and a polyamide plate as the thin layer plate.
[0042] Figure 7 This is a thin layer plate image of Example 7 using n-butanol:acetone:formic acid (5:8:6) as the developing agent and a polyamide plate as the thin layer plate.
[0043] Figure 8 This is a thin layer plate image of comparative example 1 using toluene:acetone:formic acid (9:9:2) as the developing agent and a polyamide plate as the thin layer plate.
[0044] Fig. 9 This is the thin layer plate image of comparative example 2 under the conditions of chloroform: acetone: formic acid (10:5:1) as the developing agent and polyamide plate as the thin layer plate.
[0045] Fig.10 This is a thin layer plate image of comparative example 3 using formic acid: ethyl acetate: methanol (4:4:1) as the developing solvent and a polyamide plate as the thin layer plate.
[0046] Fig.11 This is the thin layer plate image of Example 8 using 0.1% ferric chloride ethanol solution as the color developer.
[0047] Fig.12 This is the thin layer plate image of Example 9 using 0.5% ferric chloride ethanol solution as the color developer.
[0048] Fig.13 This is the thin layer plate image of Example 10 using 2% ferric chloride ethanol solution as the color developer.
[0049] Fig.14 This is the thin layer plate image of Example 11 using 3% ferric chloride ethanol solution as the color developer.
[0050] Fig.15 This is the thin layer plate image of Example 12 using 4% ferric chloride ethanol solution as the color developer.
[0051] Fig.16 This is the thin layer plate image of Example 13 using 5% ferric chloride ethanol solution as the color developer.
[0052] Fig.17This is the thin layer plate image of comparative example 4 under the conditions of n-butanol:acetone:formic acid (5:8:3) as the developing solvent and silica gel G thin layer plate (Qingdao Ocean) as the thin layer plate.
[0053] Fig.18 This is the thin layer plate diagram of comparative example 5 with n-butanol:acetone:formic acid (5:8:3) as the developing solvent and silica gel G thin layer plate (Yantai Xinnuo) as the thin layer plate.
[0054] Fig.19 This is the thin layer plate image of comparative example 6 with n-butanol:acetone:formic acid (5:8:3) as the developing solvent and silica gel H thin layer plate (Qingdao Ocean) as the thin layer plate.
[0055] Figures 1 to 19 In the figure, the samples spotted on each thin layer plate are, from left to right, Cistanche deserticola, Epimedium and black tea test solution, black tea reference solution, catechin reference solution, epigallocatechin-3-glucopyranoside reference solution and negative control medicinal material solution.
[0056] Fig. 20 This is a picture of the thin layer plates for testing multiple batches of Cistanche deserticola and Epimedium black tea in Example 14, wherein the samples spotted on each thin layer plate are, from left to right, black tea reference solution, test sample No. 1, test sample No. 2, test sample No. 3, catechin reference solution, epigallocatechin-3-phosphate reference solution, and negative control medicinal material solution. DETAILED DESCRIPTION
[0057] The thin layer chromatography detection method of black tea in the Cistanche and Epimedium black tea of the present application is further described in detail below in conjunction with specific examples. The present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present application more thoroughly understood.
[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.
[0059] The optional scope of the terms "and / or", "or / and", and "and / or" used in this article includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, said any and all combinations include any combination of any two related listed items, any more related listed items, or all related listed items.
[0060] As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.
[0061] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.
[0062] In this application, when it comes to numerical ranges, unless otherwise specified, the above numerical ranges are deemed to be continuous and include the minimum and maximum values of the range, as well as each value between such minimum and maximum values. Further, when a range refers to an integer, each integer between the minimum and maximum values of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges included therein.
[0063] The percentage contents involved in this application, unless otherwise specified, refer to mass percentage for solid-liquid mixing and solid-solid mixing, and refer to volume percentage for liquid-liquid mixing.
[0064] The percentage concentrations mentioned in this application, unless otherwise specified, refer to the final concentration, which refers to the percentage of the added component in the system after the addition of the component.
[0065] The temperature parameters in this application, unless otherwise specified, allow for both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows the temperature to fluctuate within the accuracy range of instrument control.
[0066] The room temperature in the present application generally refers to 4°C to 30°C, preferably 20±5°C.
[0067] In some examples of the present application, a thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea is provided, comprising the following steps:
[0068] Prepare the test solution of Cistanche deserticola and Epimedium black tea, catechin reference solution, epigallocatechin-3-glucopyranoside reference solution and black tea reference solution;
[0069] The test solution of the Cistanche deserticola and Epimedium black tea, the catechin reference solution, the epigallocatechin gallate reference solution and the black tea reference solution are spotted on the same polyamide thin layer plate, developed with a developing agent, taken out, dried and then subjected to color development treatment;
[0070] Wherein, the developing agent is n-butanol, acetone and formic acid in a volume ratio of (1-10):(4-12):(1-6).
[0071] It can be understood that the black tea reference solution mainly contains the active ingredients of black tea, tea polyphenols, including catechins, epigallocatechin gallate, and the like.
[0072] It can be understood that after the color development treatment, it is possible to judge whether the sample contains the active ingredients of black tea and preliminarily determine its content based on the spot positions, colors and other characteristics of the test sample and the reference substance on the thin layer plate.
[0073] Without limitation, the above-mentioned thin layer chromatography detection method further includes the steps of preparing a negative control medicinal material solution and spotting the negative control medicinal material solution on the same polyamide thin layer plate, so as to better reflect the accuracy of the target spot. It can be understood that the "negative control medicinal material" refers to the rest of the samples that are the same as Cistanche deserticola and Epimedium black tea, but do not contain black tea.
[0074] Specifically, the volume ratio of n-butanol, acetone and formic acid in the developing agent includes but is not limited to: 5:8:3, 1:8:3, 10:8:3, 5:4:3, 5:12:3, 5:8:1, 5:8:6 or a range between any two of the foregoing. Further, the developing agent is n-butanol, acetone and formic acid in a volume ratio of (5-10):(4-8):3.
[0075] In some of the examples, the developer used in the color development process is a ferric chloride ethanol solution with a mass concentration of 0.1% to 5%. Specifically, the mass concentration of the ferric chloride ethanol solution includes but is not limited to: 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5% or a range between any two of the foregoing. Further, the developer used in the color development process is a ferric chloride ethanol solution with a mass concentration of 4% to 5%.
[0076] In addition, the accuracy and sensitivity of the detection can be further improved by effectively extracting the active ingredients of the black tea in the Cistanche and Epimedium black tea to be tested.
[0077] In some of the examples, the method for preparing the test solution of the Cistanche deserticola and Epimedium black tea to be tested comprises the following steps:
[0078] Adding a first solvent to the tested Cistanche deserticola and Epimedium black tea for extraction, evaporating the obtained extract to dryness to prepare a first solid;
[0079] The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid;
[0080] The second solid is added into the third solvent for dissolution to prepare the test solution of the Cistanche deserticola and Epimedium black tea to be tested.
[0081] In some of these examples, the method for preparing a black tea reference solution includes the following steps:
[0082] A first solvent is added to black tea for extraction, and the obtained extract is evaporated to dryness to prepare a first solid;
[0083] The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid;
[0084] The second solid is added into the third solvent for dissolution to prepare the black tea reference substance solution.
[0085] Further, in the step of adding a first solvent for extraction and evaporating the obtained extract to dryness:
[0086] In some examples, the first solvent is independently an ethanol aqueous solution with a volume concentration of 35% to 45%. Specifically, the volume concentration of the ethanol aqueous solution includes but is not limited to: 35%, 38%, 40%, 42%, 45% or a range between any two of the foregoing.
[0087] In some examples, the extraction temperature is 75° C. to 85° C. Specifically, the extraction temperature includes, but is not limited to, a range between any two of 75° C., 78° C., 80° C., 82° C., and 85° C.
[0088] In some examples, the number of extractions is 2 to 4 times, and the extraction time for each time is 40 to 50 minutes. Specifically, the number of extractions includes but is not limited to: 2 times, 3 times, 4 times, or a range between any two of the foregoing; the extraction time for each time includes but is not limited to: 40 minutes, 43 minutes, 45 minutes, 47 minutes, 50 minutes, or a range between any two of the foregoing.
[0089] In some examples, the evaporation temperature is 55° C. to 65° C. Specifically, the evaporation temperature includes but is not limited to: 55° C., 58° C., 60° C., 62° C., 65° C., any range between any two of the foregoing.
[0090] Further, in the step of adding a second solvent, performing ultrasound and centrifugation, and evaporating the obtained extract to dryness:
[0091] In some examples, the second solvent is each independently water.
[0092] In some examples, the ultrasonic conditions include: a frequency of 30kHZ to 50kHZ, and a power of 250W to 350W. Specifically, the frequency includes but is not limited to: 30kHZ, 35kHZ, 40kHZ, 45kHZ, 50kHZ or a range between any two of the foregoing; the power includes but is not limited to: 250W, 280W, 300W, 320W, 350W or a range between any two of the foregoing.
[0093] In some examples, the ultrasound time is 20 min to 40 min. Specifically, the ultrasound time includes but is not limited to: 20 min, 25 min, 28 min, 30 min, 32 min, 35 min, 40 min or a range between any two of the foregoing.
[0094] In some examples, the centrifugal speed is 3000 r / min to 4000 r / min. Specifically, the centrifugal speed includes but is not limited to: 3000 r / min, 3300 r / min, 3500 r / min, 3700 r / min, 4000 r / min or a range between any two of the foregoing.
[0095] In some examples, the centrifugation time is 10 min to 20 min. Specifically, the centrifugation time includes but is not limited to: 10 min, 13 min, 15 min, 17 min, 20 min or a range between any two of the foregoing.
[0096] In some examples, the evaporation temperature is 55° C. to 65° C. Specifically, the evaporation temperature includes, but is not limited to, 55° C., 58° C., 60° C., 62° C., 65° C., or a range between any two of the foregoing.
[0097] Further, in the step of adding a third solvent for dissolving:
[0098] In some examples, the third solvent is each independently methanol.
[0099] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.
[0100] The raw materials and reagents involved in the following specific embodiments can be obtained from the market, or those skilled in the art can prepare them according to known means. The Cistanche Epimedium Black Tea used in the embodiment is from Xinjiang Huachun Pharmaceutical Co., Ltd., with the following proportions: Cistanche Tubulosa 1410g, jujube 990g, epimedium 990g, Acanthopanax Senticosus 990g, black tea 2010g, dextrin 99g, and this product is a 3.0g / bag bagged tea brewing agent.
[0101] Example 1
[0102] This embodiment is a thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea.
[0103] (1) Preparation of reference solution
[0104] Take catechin and epigallocatechin ester reference substances and prepare solutions containing 1 mg per 1 mL with methanol, respectively, as catechin and epigallocatechin ester reference substance solutions.
[0105] (2) Black tea reference solution
[0106] Take 5 g of black tea leaves, add 50 mL of 40% ethanol, heat and reflux at 80°C for extraction 3 times, 45 min each time, filter, evaporate the filtrate at 60°C, add 50 mL of water to the residue to dissolve it, ultrasonicate for 30 min, centrifuge at 3500 r / min for 15 min, evaporate the liquid at 60°C, add 1 mL of methanol to dissolve it, and use it as the black tea reference solution.
[0107] (3) Test solution
[0108] Take 5g of Cistanche deserticola and Epimedium black tea, add 50mL of 40% ethanol, heat and reflux at 80℃ for extraction 3 times, 45min each time, filter, evaporate the filtrate at 60℃, add 50mL of water to the residue to dissolve, ultrasonicate for 30min, centrifuge at 3500r / min for 15min, evaporate the liquid at 60℃, add 1mL of methanol to dissolve, and use it as the test solution.
[0109] (4) Test solution
[0110] To prepare the negative control medicinal material of Cistanche deserticola and Epimedium black tea, the four medicinal materials of Cistanche tubulosa, jujube, Epimedium and Acanthopanax senticosus were mixed and crushed according to the compound ratio, and the negative control medicinal material solution was prepared in the same way as the test sample solution.
[0111] (5) Chromatographic conditions
[0112] According to the thin layer chromatography method (Appendix VI B of Part I of the 2010 edition of the Chinese Pharmacopoeia), 2 μL of each of the above solutions were taken and spotted on the same polyamide thin layer plate, and the plate was developed with n-butanol:acetone:formic acid (5:8:3) as the developing solvent. The plate was taken out, dried, and sprayed with 1% ferric chloride ethanol solution for color development.
[0113] The thin layer chromatography detection methods of Examples 2 to 7 and Comparative Examples 1 to 3 are the same as those of Example 1, the main difference being that different developing agents are used. The specific developing agents are shown in Table 1 below. The detection results of Examples 1 to 7 and Comparative Examples 1 to 3 are shown in Table 1 below. Figures 1 to 10 As shown:
[0114] Table 1
[0115]
[0116] Combination Figures 1 to 10It can be seen that the developing agents of Examples 1 to 7 can effectively separate the black tea polyphenol components catechins and epigallocatechin gallate in Cistanche Epimedium Black Tea, and the spots are clear, the impurity spot separation is high, the interference is small, and the detection results are accurate, among which Examples 1, 3, and 4 are the best. Comparative Examples 1 to 3 change the type of developing agent, and there are problems such as overlap with adjacent spots, unclear spots, and excessive development, which makes it difficult to detect black tea in Cistanche Epimedium Black Tea.
[0117] The thin layer chromatography detection method of Examples 8 to 13 is the same as that of Example 1, the main difference being that the mass concentration of the color developer used is different, as shown in Table 2 below. The detection results of Examples 8 to 13 are as follows: Figures 11 to 16 As shown:
[0118] Table 2
[0119] Color developer Attached photos Example 1 1.0% Ferric chloride ethanol solution Figure 1 Example 8 0.1% ferric chloride ethanol solution Fig.11 Example 9 0.5% ferric chloride ethanol solution Fig.12 Example 10 2.0% Ferric chloride ethanol solution Fig.13 Embodiment 11 3.0% Ferric chloride ethanol solution Fig.14 Example 12 4.0% Ferric chloride ethanol solution Fig.15 Embodiment 13 5.0% Ferric chloride ethanol solution Fig.16
[0120] Combination Figure 1 and 11 It can be seen from Examples 1 to 16 that the color development of characteristic spots can be well achieved within the mass concentration range of 0.1% to 5%, among which Examples 12 and 13 are better.
[0121] The thin layer chromatography detection method of Comparative Examples 4 to 6 is the same as that of Example 1, the main difference being that different thin layer plates are used, as shown in Table 3 below. The detection results of Comparative Examples 4 to 6 are as follows: Figures 17 to 19 As shown:
[0122] Table 3
[0123] Experiment No. Thin layer plate Attached photos Comparative Example 4 Silica G Thin Layer Plate (Qingdao Ocean) Fig.17 Comparative Example 5 Silica G Thin Layer Plate (Yantai Xinnuo) Fig.18 Comparative Example 6 Silica gel H thin layer plate (Qingdao Ocean) Fig.19
[0124] Combination Figure 1 and 17 It can be seen from Figure 19 that the use of polyamide plates can make each spot clearer, with less interference from miscellaneous peaks and accurate detection results.
[0125] The thin layer chromatography detection method of Example 14 is the same as that of Example 1, and three different batches of Cistanche Epimedium black tea are respectively taken to prepare Sample No. 1, Sample No. 2 and Sample No. 3. It can be seen that the active ingredient spots in different batches of Cistanche Epimedium black tea samples have good specificity and high clarity, and can achieve accurate detection of black tea in Cistanche Epimedium black tea.
[0126] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0127] The above-described embodiments only express several implementation methods of the present application, which is convenient for understanding the technical solutions of the present application in detail, but cannot be understood as limiting the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present application, which all belong to the scope of protection of the present application. It should be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the scope of protection of the claims attached to the present application. Therefore, the scope of protection of the patent of the present application shall be based on the contents of the attached claims, and the description can be used to interpret the contents of the claims.
Claims
1. A thin layer chromatography detection method for black tea in Cistanche and Epimedium black tea, characterized in that: The steps include: Prepare the test solution of Cistanche deserticola and Epimedium black tea, catechin reference solution, epigallocatechin-3-glucopyranoside reference solution and black tea reference solution; The test solution of the Cistanche deserticola and Epimedium black tea, the catechin reference solution, the epigallocatechin gallate reference solution and the black tea reference solution are spotted on the same polyamide thin layer plate, developed with a developing agent, taken out, dried and then subjected to color development treatment; Wherein, the developing agent is n-butanol, acetone and formic acid in a volume ratio of (1-10):(4-12):(1-6).
2. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 1, characterized in that: The developing agent is n-butanol, acetone and formic acid in a volume ratio of (5-10):(4-8):
3.
3. The thin layer chromatography detection method of black tea in the Cistanche and Epimedium black tea according to claim 1, characterized in that: The color developing agent used in the color developing treatment is a ferric chloride ethanol solution with a mass concentration of 0.1% to 5%.
4. The thin layer chromatography detection method of black tea in the Cistanche and Epimedium black tea according to claim 3, characterized in that: The color developing agent used in the color developing treatment is a ferric chloride ethanol solution with a mass concentration of 4% to 5%.
5. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to any one of claims 1 to 4, characterized in that: The method for preparing the test solution of the Cistanche deserticola and Epimedium black tea to be tested comprises the following steps: Adding a first solvent to the tested Cistanche deserticola and Epimedium black tea for extraction, evaporating the obtained extract to dryness to prepare a first solid; The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid; The second solid is added into the third solvent for dissolution to prepare the test solution of the Cistanche deserticola and Epimedium black tea to be tested; And / or, the method for preparing a black tea reference solution comprises the following steps: A first solvent is added to black tea for extraction, and the obtained extract is evaporated to dryness to prepare a first solid; The first solid is added to a second solvent for ultrasonic and centrifugation, and the obtained extract is evaporated to dryness to prepare a second solid; The second solid is added into the third solvent for dissolution to prepare the black tea reference substance solution.
6. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 5, characterized in that: The first solvents are each independently an ethanol aqueous solution with a volume concentration of 35% to 45%.
7. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 5, characterized in that: The second solvent is each independently water.
8. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 5, characterized in that: The third solvent is each independently methanol.
9. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 5, characterized in that: The step of adding the first solvent for extraction and evaporating the obtained extract to dryness has one or more of the following characteristics: (1) The extraction temperature is 75°C to 85°C; (2) The number of extractions is 2 to 4 times, and the extraction time for each time is 40 to 50 minutes; (3) The evaporation temperature is 55℃~65℃.
10. The thin layer chromatography detection method of black tea in Cistanche and Epimedium black tea according to claim 5, characterized in that: The steps of adding a second solvent, performing ultrasound and centrifugation, and evaporating the obtained extract to dryness have one or more of the following characteristics: (1) Ultrasonic conditions include: frequency of 30 kHz to 50 kHz, power of 250 W to 350 W; (2) The ultrasound duration is 20 to 40 minutes; (3) The centrifugal speed is 3000 r / min to 4000 r / min; (4) The centrifugation time is 10 min to 20 min; (5) The evaporation temperature is 55℃~65℃.