Identification method of thin layer of maca powder
By using maca root control medicinal materials and ethanol solution, and combining thin layer chromatography and ultraviolet color development detection, the problem of lack of fast and simple maca powder identification methods in the prior art was solved, and effective qualitative identification of maca powder was achieved.
Patent Information
- Application Number
- CN202311777629.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-12-22
- Publication Date
- 2025-06-03
- Estimated Expiration
- 2043-12-22
AI Technical Summary
There is a lack of a method in the prior art that can quickly, simply and effectively identify maca powder, especially in health food quality control.
The solution was prepared by maca root control medicinal materials and ethanol solution, and the maca powder was inspected by thin-layer chromatography under ultraviolet light 366 nm and sunlight, and the anisaldehyde solution and a specific developer were used to detect the maca powder.
It achieves rapid, simple and effective qualitative identification of maca powder, simple operation, clear and visible spots, moderate Rf value, and good durability.
Smart Images
Figure CN117890525B_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of detection of health foods or food raw materials, and particularly relates to a method for identifying thin layers of maca powder. Background Art
[0002] Maca is a cruciferous plant Lepidium meyenii Walp., with elliptical leaves and a rootstock similar to a small round radish, also known as maca or Inca radish. The hypocotyl is thick, and the hypocotyl fuses with the root to form a rough, inverted pear-shaped tuber. It can be eaten directly, is a natural food with rich nutritional components; is rich in high-unit nutrients, has the function of tonifying and strengthening the body for humans, and has the reputation of "South American ginseng". The shapes and volumes of the roots vary greatly, with triangular, elliptical or rectangular shapes. The hypocotyl of maca may be golden or light yellow, red, purple, blue, black or green.
[0003] Currently, the United States, Japan, Spain, Ecuador, Bolivia, Australia, etc. have successively carried out plantings, but have not formed a scale. China began to introduce and plant in Lijiang, Yunnan in 2002, and has now become the largest maca planting base in the world. With the continuous in-depth scientific research, researchers have found that maca not only has the effect of improving fertility, but also has important effects such as anti-fatigue, relieving menopausal syndrome, anti-prostate hyperplasia, antioxidant, anti-aging and anti-cancer. In 1992, the Food and Agriculture Organization of the United Nations (FAO) recommended maca as a nutritional supplement to the world; in 2001, the US Food and Drug Administration (FDA) approved maca health products to enter the United States. The 13th announcement of the Chinese Ministry of Health approved maca powder as a new resource food. Currently, there are relatively few methods for qualitative identification of maca powder, and exploring and establishing a thin-layer identification method for maca powder to more quickly, simply and effectively control the quality of maca powder in health products and provide more effective data for quantitative analysis has become a research hotspot. Summary of the Invention
[0004] The purpose of the present invention is to overcome one of the above technical defects and provide a method for identifying thin layers of maca powder with mild reaction conditions, without using toxic organic solvents and with a simple process.
[0005] The technical solution of the present invention is: a method for identifying thin layers of maca powder, comprising the following steps:
[0006] (1) Preparation of the reference solution: Weigh 100 mg to 200 mg of the reference medicinal material of maca root, add it to an ethanol solution, reflux in a water bath, vortex to completely precipitate the solution, and filter to obtain the reference solution;
[0007] (2) Preparation of the test solution: Weigh 100 mg - 200 mg of the sample maca powder, add it to an ethanol solution, reflux in a water bath, vortex to completely precipitate the solution, and filter to obtain the test solution;
[0008] (3) Thin-layer chromatography test: Spot the reference solution and the test solution on a silica gel thin-layer plate, develop in the developing agent, take out the thin-layer plate, dry it, and then spray anisaldehyde solution on the thin-layer plate;
[0009] (4) Comparative inspection: Examine the test chromatogram under 366 nm ultraviolet light and sunlight, and compare it with the corresponding position of the reference medicinal material chromatogram. Spots showing the same color indicate the presence of maca powder.
[0010] More specifically, a method for identifying a thin layer of maca powder according to the present invention specifically includes the following steps:
[0011] (11) Preparation of the reference solution: Weigh 100 mg - 200 mg of the reference medicinal material of maca root, add it to 5 ml - 10 ml of an ethanol solution with a concentration of 70%, reflux in a water bath at 50°C - 60°C for 30 - 60 minutes, vortex to completely precipitate the solution, and filter through a 0.4 - 0.5 μm microporous membrane to obtain the reference solution;
[0012] (21) Preparation of the test solution: Weigh 100 mg - 200 mg of the sample maca powder, add it to 5 ml - 10 ml of an ethanol solution with a concentration of 70%, reflux in a water bath at 50°C - 60°C for 30 - 60 minutes, vortex to completely precipitate the solution, and filter through a 0.4 - 0.5 μm microporous membrane to obtain the test solution;
[0013] (31) Thin-layer chromatography (General Rule 0502) test: Spot the reference solution and the test solution on a silica gel G60F254 thin-layer plate, develop in the developing agent (chloroform: acetic acid: methanol: water), take out the thin-layer plate, dry it, and then spray anisaldehyde solution on the thin-layer plate;
[0014] (41) Comparative inspection: Examine the test chromatogram under 366 nm ultraviolet light and sunlight, and compare it with the corresponding position of the reference medicinal material chromatogram. Spots showing the same color indicate the presence of maca powder.
[0015] Furthermore, the reference medicinal material of maca root (source: ALKEMIST LABS, batch number: 949004); maca powder sample (source: Ntural); the preparation processes of the reference solution and the test solution in steps (1) and (2) are simple. Add a 70% ethanol solution, reflux in a water bath at 50°C - 60°C for 30 - 60 minutes. Vortex to completely precipitate the solution, and filter through a 0.4 - 0.5 μm microporous membrane.
[0016] Further, in step (3), the developing agent is chloroform: acetic acid: methanol: water, and the volume ratio of chloroform: acetic acid: methanol: water is 5-8: 2-5: 0.5-2: 0.5-2.
[0017] Further, in step (3), spray with anisaldehyde solution. Take 0.45-0.55 ml of anisaldehyde, 10 ml of glacial acetic acid, 85 ml of methanol and 5 ml of sulfuric acid, and mix them evenly.
[0018] Preferably, in step (3), air dry and then dry at 100°C - 105°C for 5 - 10 min.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] 1. The present invention uses the reference medicinal material of maca root as a control, chloroform: acetic acid: methanol: water as the developing agent, sprays anisaldehyde solution, air dries, and then examines under ultraviolet light at 366 nm and white light. This method is simple to operate, targeted, the spots are clearly visible, the Rf value is moderate, and it has good durability. Description of the Drawings
[0021] Figure 1 and Figure 2 are the chromatograms of Example 1 of the present invention examined under ultraviolet light at 366 nm and sunlight;
[0022] Figure 3 and Figure 4 are the chromatograms of Comparative Example 1 of the present invention examined under ultraviolet light at 366 nm and sunlight. Detailed Embodiments
[0023] In order to make the objectives, technical solutions and advantages of the present invention clearer, the following embodiments are used to further elaborate on the present invention in detail, so that those skilled in the art can fully understand the technical content of the present invention. It should be understood that the following embodiments are used to further illustrate the present invention and should not be construed as limiting the protection scope of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the above content of the present invention fall within the protection scope of the present invention. The specific process parameters and the like in the following examples are also only an example within a suitable range, that is, those skilled in the art can make selections within a suitable range through the description herein, rather than being limited to the specific values in the following examples.
[0024] The method for identifying the thin layer of maca powder of the present invention specifically includes the following steps:
[0025] S1 Reference solution: Weigh 100 mg to 200 mg of the reference medicinal material of maca root (source: ALKEMISTLABS, batch number: 949004), add 5 ml to 10 ml of 70% ethanol solution, place in a water bath at 50 °C to 60 °C and reflux for 30 to 60 minutes. Vortex to completely precipitate the solution, and filter through a 0.4 - 0.5 μm microporous membrane.
[0026] S2 Test solution: Weigh 100 mg to 200 mg of the sample (source: Ntural), add 5 ml to 10 ml of 70% ethanol solution, place in a water bath at 50 °C to 60 °C and reflux for 30 to 60 minutes. Vortex to completely precipitate the solution, and filter through a 0.4 - 0.5 μm microporous membrane.
[0027] S3 Thin layer chromatography identification: Conduct the test according to the thin layer chromatography method (General Principles 0502). Spot the reference solution and the test solution on a silica gel G60F254 thin layer plate, develop in the developing solvent (chloroform: acetic acid: methanol: water), take out, air dry, spray with anisaldehyde solution (take 0.45 ml to 0.55 ml of anisaldehyde, 10 ml of glacial acetic acid, 85 ml of methanol and 5 ml of sulfuric acid), mix well. Then air dry in the air and dry at 100 °C - 105 °C for 5 - 10 min.
[0028] S4 Visual inspection for identification: Place under ultraviolet light at 366 nm and sunlight for visual inspection. In the chromatogram of the test sample, at the position corresponding to the chromatogram of the reference medicinal material, spots showing the same color indicate the presence of maca powder.
[0029] In the method of the present invention, when visually inspecting the chromatogram of the test sample under ultraviolet light at 366 nm and sunlight, at the position corresponding to the chromatogram of the reference medicinal material, spots showing the same color appear. It does not require complex treatment methods, reagents, instruments or equipment, can quickly identify maca powder, and provides a convenient and practical way for the qualitative identification of maca powder.
[0030] Example 1:
[0031] 1.0 Preparation of solutions:
[0032] (1) Reference solution: Take 100 mg of the reference medicinal material of maca root (source: ALKEMISTLABS, batch number: 949004), add 5 ml of 70% ethanol solution, place in a water bath at 55 °C and reflux for 30 minutes, vortex to completely precipitate the solution, and filter through a 0.45 μm microporous membrane.
[0033] (2) Test solution: Take 100 mg of maca powder (source: Ntural), add 5 ml of 70% ethanol solution, place in a water bath at 55 °C and reflux for 30 minutes, vortex to completely precipitate the solution, and filter through a 0.45 μm microporous membrane.
[0034] 1.1 Thin layer chromatography identification:
[0035] Absorb 5 μl each of the reference substance solution and the test solution, and spot them separately on the same silica gel G60F254 thin layer plate (source: Merck, TLC Silicagel 60F254, 10×20 cm). Use chloroform: acetic acid: methanol: water (6:3:1:1, V / V / V / V) as the developing solvent, develop, take out, air dry, spray anisaldehyde solution, air dry again, heat at 100 - 105 °C for 5 minutes, and examine under ultraviolet light at 366 nm and daylight. The results are shown in the appendix Figure 1 、 2 。
[0036] Example 2:
[0037] Preparation of 2.0 solution:
[0038] (1) Reference substance solution: Take 200 mg of the reference crude drug of Maca root (source: ALKEMIST LABS, batch number: 949004), add 10 ml of 70% ethanol solution, reflux in a water bath at 50 °C for 50 minutes, vortex to completely precipitate the solution, and filter through a 0.5 μm microporous membrane.
[0039] (2) Test solution: Take 200 mg of Maca powder (source: Ntural), add 10 ml of 70% ethanol solution, reflux in a water bath at 50 °C for 50 minutes, vortex to completely precipitate the solution, and filter through a 0.5 μm microporous membrane.
[0040] 2.1 TLC identification:
[0041] Absorb 5 μl each of the reference substance solution and the test solution, and spot them separately on the same silica gel G60F254 thin layer plate (source: Merck, TLC Silicagel 60F254, 10×20 cm). Use chloroform: acetic acid: methanol: water (5:4:1.5:1.5, V / V / V / V) as the developing solvent, develop, take out, air dry, spray anisaldehyde solution, air dry again, heat at 100 - 105 °C for 5 minutes, and examine under ultraviolet light at 366 nm and daylight. The results are similar to Figure 1 、 2 and both show the presence of Maca powder.
[0042] Example 3:
[0043] Preparation of 3.0 solution:
[0044] (1) Reference substance solution: Take 150 mg of the reference crude drug of Maca root (source: ALKEMIST LABS, batch number: 949004), add 7 ml of 70% ethanol solution, reflux in a water bath at 60 °C for 40 minutes, vortex to completely precipitate the solution, and filter through a 0.4 μm microporous membrane.
[0045] (2) Test solution: Take 150 mg of maca powder (source: Ntural), add 7 ml of 70% ethanol solution, place it in a water bath at 55 °C and reflux for 45 minutes, vortex to completely precipitate the solution, and filter with a 0.4 μm microporous filter membrane.
[0046] 3.1 Thin layer chromatography identification:
[0047] Absorb 5 μl each of the reference solution and the test solution, and spot them on the same silica gel G60F254 thin layer plate (source: Merck, TLCSilicagel60F254, 10×20 cm) respectively. Use chloroform: acetic acid: methanol: water (8:4:1.8:1.6, V / V / V / V) as the developing agent, develop, take out, dry in air, spray anisaldehyde solution, dry in air, heat at 100 - 105 °C for 5 minutes, and examine under ultraviolet light at 366 nm and daylight. The results are similar to Figure 1 、 2 , and all show the presence of maca powder.
[0048] Control example 1:
[0049] Preparation of D1.0 solution:
[0050] (1) Reference solution: Take 0.2 mg of linoleamide (source: Beijing Zhongkejianyu Biotechnology Co., Ltd., batch number: Z - 326 - 004, purity ≥98%), add 1 ml of n - hexane to make a reference solution.
[0051] (2) Test solution: Take 2 g of maca powder (source: Ntural), add 20 ml of n - hexane, heat under reflux in a water bath for 30 minutes, cool, filter, and use the filtrate as the test solution.
[0052] D1.1 Thin layer chromatography identification:
[0053] Absorb 5 μl each of the reference solution and the test solution, and spot them on the same silica gel G thin layer plate with sodium carboxymethyl cellulose as the binder (source: Qingdao Ocean). Use petroleum ether: acetone: ammonia water (30:5:1, V / V / V) as the developing agent, place it in the developing agent test solution for pre - saturation for 10 min and then develop, take out, dry in air, spray with 0.5% sulfuric acid ethanol test solution, heat at 105 °C until the spots are clearly visible, and examine under ultraviolet light at 366 nm and daylight. The results are shown in Appendix Figure 3 、 4 . No spots appear for the linoleamide reference substance, and the method reproducibility is poor.
[0054] Control example 2:
[0055] Preparation of D2.0 solution:
[0056] (1) Reference solution: Take 0.4 mg of linoleamide (source: Beijing Zhongke Jianyu Biotechnology Co., Ltd., batch number: Z-326-004, purity ≥98%) and add 1 ml of n-hexane as the reference solution.
[0057] (2) Test solution: Take 4 g of maca powder (source: Ntural), add 20 ml of n-hexane, heat under reflux in a water bath for 30 min, cool, filter, and use the filtrate as the test solution.
[0058] D2.1 Thin layer chromatography identification:
[0059] 5 μl of reference solution and test solution were taken and spotted on the same silica gel G thin layer plate with sodium hydroxymethyl cellulose as adhesive (source: Qingdao Ocean), and petroleum ether: acetone: ammonia water (50:10:5, V / V / V) was used as the developing agent. The developing agent solution was pre-saturated for 10 minutes and then developed. The plate was taken out, dried, sprayed with 0.5% sulfuric acid ethanol solution, heated at 105℃ until the spots were clearly colored, and then placed under ultraviolet light 366nm and sunlight for inspection. The results are shown in the attached Figure 3 , 4 Similarly, no spots were observed for the linoleamide control substances, indicating that the method had poor reproducibility.
[0060] The present invention is illustrated by the above embodiments, the purpose of which is to enable people familiar with the art to understand the content of the present invention and implement it accordingly, and it cannot be used to limit the scope of protection of the present invention. However, it should be understood that the present invention is not limited to the specific examples and implementation schemes described here. The purpose of including these specific examples and implementation schemes here is to help those skilled in the art to practice the present invention. Any technician in the field can easily make further improvements and perfections without departing from the spirit and scope of the present invention. Therefore, the present invention is only limited by the content and scope of the claims of the present invention, and it is intended to cover all alternatives and equivalent schemes included in the spirit and scope of the present invention defined by the appendix claims. All equivalent changes or modifications made according to the spirit and essence of the present invention should be included in the scope of protection of the present invention.
Claims
1. A method for identifying a thin layer of maca powder, characterized in that, it comprises the following steps: (1) Preparation of the reference solution: Weigh 100 mg to 200 mg of the reference medicinal material of maca root, add it to an ethanol solution, reflux in a water bath, vortex to completely precipitate the solution, and filter to obtain the reference solution; (2) Preparation of the test solution: Weigh 100 mg to 200 mg of the sample maca powder, add it to an ethanol solution, reflux in a water bath, vortex to completely precipitate the solution, and filter to obtain the test solution; (3) Thin layer chromatography test: Spot the reference solution and the test solution on a silica gel thin layer plate, and develop in the developing agent. After taking out the thin layer plate and drying it in the air, then spray anisaldehyde solution on the thin layer plate; The developing agent is a mixture of chloroform: acetic acid: methanol: water; The volume ratio of chloroform: acetic acid: methanol: water is (5 - 8):(2 - 5):(0.5 - 2):(0.5 - 2); The anisaldehyde solution is obtained by mixing 0.45 ml to 0.55 ml of anisaldehyde, 10 ml of glacial acetic acid, 85 ml of methanol and 5 ml of sulfuric acid evenly; (4) Comparative inspection: Examine the test sample chromatogram under 366 nm ultraviolet light and sunlight, and then compare it with the corresponding position of the reference medicinal material chromatogram. Spots showing the same color indicate the presence of maca powder.
2. The identification method according to claim 1, characterized in that: In steps (1) and (2), the amount of the ethanol solution added is 5 ml to 10 ml with a concentration of 70%.
3. The identification method according to claim 1 or 2, characterized in that: In steps (1) and (2), the filtration is carried out using a 0.4 μm to 0.5 μm microporous filter membrane.
4. The identification method according to claim 1 or 2, characterized in that: The water bath reflux conditions in steps (1) and (2) are to reflux in a 50°C to 60°C water bath for 30 to 60 minutes.
5. The identification method according to claim 1, characterized in that: The thin layer chromatography method in step (3) adopts the chromatography method in General Principles 0502 of the Chinese Pharmacopoeia.
6. The identification method according to claim 1 or 5, characterized in that: The silica gel thin layer plate in step (3) is G60F254.
7. The identification method according to claim 6, characterized in that: The thin layer plate in step (3) is dried in the air and dried at 100°C to 105°C for 5 to 10 minutes.