A thin layer chromatography method for rapid identification of Chlorella and Spirulina

Abstract

The present invention relates to a thin layer chromatography analysis method, and in particular to a thin layer chromatography analysis method for quickly identifying Chlorella and Spirulina. The present invention solves the technical problem that the existing detection method for identifying Chlorella and Spirulina must be judged by microscopic appearance and shape, and when the products are processed into powders, capsules and other different solid forms, or when other products are used to replace Chlorella and Spirulina, the identification cannot be performed. The thin layer chromatography analysis method of the present invention comprises the following steps: 1) preparing a standard control solution and a sample solution to be tested; 2) performing thin layer spotting; 3) preparing a developing agent; 4) developing; 5) identifying. The thin layer chromatography identification method of the present invention has multiple advantages such as rapidity and high efficiency, simple operation, low cost, intuitive and reliable detection results, and can quickly and efficiently identify and distinguish Chlorella and Spirulina.

Description

Technical Field

[0001] The invention relates to a thin layer chromatography analysis method, in particular to a thin layer chromatography analysis method for quickly identifying chlorella and spirulina. Background Art

[0002] Chlorella is a common single-cell green algae of the genus Chlorella in the Chlorophyta. It is a spherical single-cell freshwater algae with a diameter of 3 to 8 microns. It appeared more than 2 billion years ago and is one of the earliest life on Earth. It is widely distributed all over the world. Chlorella is an efficient photosynthetic plant that grows and reproduces by photosynthesis and autotrophy. It mostly lives in shallow water areas, and there are also marine species. Under natural conditions, there are few individuals of Chlorella, but it can be reproduced in large quantities through artificial cultivation. The cells of Chlorella contain abundant nutrients such as proteins, vitamins, minerals, dietary fiber, nucleic acids and chlorophyll that are indispensable for maintaining and promoting human health. In particular, the biologically active substances it contains, such as glycoproteins, polysaccharides, and up to 13% of nucleic acids, have the functions of enhancing human immunity, preventing virus proliferation, inhibiting cancer cell proliferation, inhibiting blood sugar rise, lowering serum cholesterol levels, eliminating toxins and quickly repairing body damage. In addition, the CG (Chlorella Growth Factor) rich in Chlorella can quickly restore the damage caused by the body, and can be used as a food flavor improver, and is widely used in the fields of food and fermentation.

[0003] Spirulina, which is shaped like a clockwork and is blue-green in color, is also called blue-green algae. It belongs to the phylum Cyanobacteria, class Cyanobacteria, family Oscillatoriaceae, and genus Spirulina. It has a life history of 3.5 billion years and is an ancient low-level prokaryotic single-cell or multi-cell aquatic plant. Spirulina is the most nutritious and comprehensive organism in nature and is a very good natural food. Spirulina is rich in high-quality protein, γ-linolenic acid fatty acids, carotenoids, vitamins, and a variety of trace elements such as iron, iodine, selenium, zinc, etc. It has biological activities such as antioxidant, antiviral, antibacterial, immunomodulatory and anti-cancer.

[0004] Nowadays, with the rapid development of economy, people's quality of life is constantly improving. In addition to daily food consumption, people are paying more and more attention to the health needs of themselves and their families, and are also paying more and more attention to nutritional balance and health care. Algae not only has a comprehensive nutritional balance, but also has the characteristics of pure natural ingredients. It is known as a "micro-nutrient bank". Therefore, algae has always been a hot product in the field of health and immunity in nature. Among the many algae we know, chlorella and spirulina are supplements with higher nutrition.

[0005] At present, the main detection method for identifying chlorella and spirulina is microscopic identification, which is to distinguish and judge by observing the morphology of chlorella and spirulina under a microscope. Chlorella appears as a round ball under a microscope, and spirulina appears as a spiral ribbon. The above method must be judged by microscopic appearance. When the product is processed into powder, capsules and other different solid forms, or when other products are used to replace chlorella and spirulina, it is impossible to identify, which will have an adverse impact on the market sales of chlorella and spirulina products. Summary of the invention

[0006] The purpose of the present invention is to solve the existing detection method for identifying chlorella and spirulina, which must be judged by microscopic appearance shape. When the product is processed into other different solid forms such as powder, capsule, or other products are used to replace chlorella and spirulina, it is impossible to identify, which in turn has an adverse effect on the market sales of chlorella and spirulina products. A thin layer chromatography analysis method for quickly identifying chlorella and spirulina is provided. The thin layer chromatography analysis method for quickly identifying chlorella and spirulina provided by the present invention is to distinguish the two plants of chlorella and spirulina by the difference in chemical components contained in them, so that the quality can be more effectively checked, and a reliable basis can be provided for the health of users. At the same time, the thin layer chromatography analysis method for identifying chlorella and spirulina of the present invention has the advantages of low cost, strong specificity, intuitive detection results, and simplicity and efficiency.

[0007] The technical solution of the present invention is:

[0008] A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina, which is special in that it comprises the following steps:

[0009] 1) Prepare standard control solution and sample solution to be tested

[0010] 1.1) taking a chlorella raw material and a spirulina raw material, placing them in sealable test containers respectively, adding 5-10 times the mass of the chlorella raw material and the spirulina raw material and 50%-95% ethanol respectively, and performing ultrasonic extraction for 30-60 minutes, cooling to room temperature, and then filtering with a microporous filter membrane, collecting the filtrate as the standard control solution of chlorella and spirulina respectively;

[0011] 1.2) Take the sample to be identified, place it in a sealable test container, add 5 to 10 times the mass of the sample, and 50% to 95% ethanol, and perform ultrasonic extraction for 30 to 60 minutes, let it cool to room temperature, and then filter it with a microporous membrane, and collect the filtrate as the sample solution to be tested;

[0012] 2) Perform thin layer spotting

[0013] Place the standard control solutions of Chlorella and Spirulina prepared in step 1.1) and the sample solution to be tested prepared in step 1.2) on the same silica gel G thin layer plate at different positions at the same height to form a spot origin;

[0014] 3) Preparation of developing agent

[0015] 3.1) Ethyl acetate, formic acid, acetic acid and water are uniformly mixed in a volume ratio of 10-20:0.5-1.5:0.5-1.5:1.5-2.5 to obtain a developing agent;

[0016] 3.2) Place the developing agent in the developing cylinder, seal the developing cylinder, and pre-saturate for 15 to 20 minutes;

[0017] 4) Expand

[0018] 4.1) Open the development cylinder after pre-saturation in step 3), and quickly put the silica gel G thin layer plate spotted in step 2) into the development cylinder, so that the silica gel G thin layer plate is tilted against the development cylinder;

[0019] 4.2) When the front edge of the developing agent moves up to 6 cm to 10 cm from the sample origin, take out the silica gel G thin layer plate;

[0020] 5) Identification

[0021] 5.1) After the reagent on the silica gel G thin layer plate developed in step 4.2) is dried, the silica gel G thin layer plate is placed under 365 nm ultraviolet light for inspection to obtain an inspection image;

[0022] 5.2) Compare the spots formed at the sampling origin of the sample solution to be tested on the inspection diagram with the spots formed at the sampling origin of the standard control solutions of Chlorella and Spirulina;

[0023] If the spot of the sample liquid to be tested is consistent with the spot of the standard control solution of Chlorella, then the sample to be tested is Chlorella; if the spot of the sample liquid to be tested is consistent with the spot of the standard control solution of Spirulina, then the sample to be tested is Spirulina; if the spot of the sample to be tested is inconsistent with the spot of the standard control solution of Chlorella and is also inconsistent with the spot of the standard control solution of Spirulina, then the sample to be tested is other samples.

[0024] Furthermore, in step 5.1), after inspection under 365nm ultraviolet light, a sulfuric acid ethanol color developer with a mass concentration of 5% to 10% is sprayed on the silica gel G thin layer plate for color development, and then heated at 100-110°C for 3 to 5 minutes. After cooling to room temperature, the silica gel G thin layer plate is inspected under 365nm ultraviolet light to obtain an inspection diagram.

[0025] Furthermore, in step 5.1), the mass concentration of the sulfuric acid ethanol color developer is 10%, and the heating temperature is 105°C.

[0026] Furthermore, in step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.0 to 1.5 cm.

[0027] Furthermore, in step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 15:1:1:2.

[0028] Furthermore, in step 3.2), the developing cylinder adopts a double-slot developing cylinder, the size of the double-slot developing cylinder is 10cm×10cm, 20cm×10cm or 10cm×20cm, and the dosage of the developing agent in the double slots is kept consistent, and pre-saturation is performed for 15 minutes.

[0029] Furthermore, step 4.2) is specifically as follows: when the front edge of the developing agent moves upward to a distance of 8 cm to 10 cm from the sample spot origin, the silica gel G thin layer plate is taken out.

[0030] Furthermore, in step 1.1), ethanol having a mass concentration of 70% and an amount of 6 times the mass of the chlorella raw material and the spirulina raw material is added respectively, and ultrasonic extraction is performed for 60 minutes;

[0031] In step 1.2), ethanol with a mass concentration of 70% and an amount of 6 times the mass of the sample to be tested is added, and ultrasonic extraction is performed for 60 minutes.

[0032] Furthermore, in step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.0 cm;

[0033] Step 4.2) Specifically, when the front edge of the developing agent moves upward to 8 cm from the sample spot origin, the silica gel G thin layer plate is taken out;

[0034] In step 3.2), the size of the double-slot expansion cylinder is 10 cm×10 cm.

[0035] The beneficial effects of the present invention are:

[0036] 1. The present invention adopts thin layer chromatography to identify and distinguish Chlorella and Spirulina, which has the advantages of simple operation, rapidity and high efficiency, good separation effect, fast identification, high sensitivity, low cost and intuitive results. It solves the problem of distinguishing Chlorella and Spirulina well and can control the quality at the same time.

[0037] 2. The present invention uses ethyl acetate, formic acid, acetic acid and water in a volume ratio of 10-20: 0.5-1.5: 0.5-1.5: 1.5-2.5 to prepare a developing agent. The obtained spot separation effect is good, the characteristic spots are prominent, and there is no spot tailing and spot diffusion phenomenon. The result is intuitive and obvious and easy to judge.

[0038] 3. The present invention adopts silica gel G thin layer plate to perform non-color inspection and color inspection respectively, making the results more valuable for reference.

[0039] 4. The present invention adopts 5% to 10% ethanol sulfuric acid as a color developer. The color developer is simple to prepare. After color development, the spots are richer and clearer, and the specificity is strong.

[0040] 5. The present invention uses an appropriate amount of ethanol to directly perform ultrasonic treatment on the raw material and filter it, and uses the filtrate as the detection solution. The sample pretreatment is simple, low in cost, and easy to operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 This is an inspection diagram of the G thin layer plate under 365nm light in an embodiment of the present invention;

[0042] Figure 2 This is an inspection diagram of the G thin layer plate after color development under 365nm light in an embodiment of the present invention. DETAILED DESCRIPTION

[0043] The present invention is described in detail below through specific embodiments.

[0044] The present invention adopts a thin layer chromatography analysis method to perform chromatographic analysis on the raw materials of chlorella and spirulina and the sample to be tested, and then compares the spots of the sample to be tested with the spots of chlorella and spirulina, respectively, so as to identify and distinguish whether the sample to be tested is chlorella or spirulina, or neither chlorella nor spirulina. The raw materials of chlorella and spirulina in the present invention are standard raw materials that have been tested and confirmed, and the sample to be tested is commercially available.

[0045] Example 1

[0046] The present invention discloses a thin layer chromatography analysis method for quickly identifying Chlorella and Spirulina, comprising the following steps:

[0047] 1) Prepare standard control solution and sample solution to be tested

[0048] 1.1) Take a chlorella raw material and a spirulina raw material, place them in a stoppered conical flask, add 5 to 10 times the mass of the chlorella raw material and the spirulina raw material, and perform ultrasonic extraction for 30 to 60 minutes. In this embodiment, 70% ethanol is added, and the amount of ethanol is 6 times the mass of the chlorella raw material or the spirulina raw material, and ultrasonic extraction is performed for 60 minutes; let it cool to room temperature, and then filter it with a microporous filter membrane, collect the filtrate as the standard control solution of chlorella and spirulina, respectively; the standard control solutions of the chlorella raw material and the spirulina raw material are numbered as No. 1 and No. 5, respectively.

[0049] 1.2) Take 6 samples to be identified, place them in stoppered conical bottles, add 5 to 10 times the mass of the samples, and ethanol with a mass concentration of 50% to 95%, and perform ultrasonic extraction for 30 to 60 minutes. In this embodiment, ethanol with a mass concentration of 70% is added, and the amount of ethanol is 6 times the mass of the Chlorella raw material or the Spirulina raw material, and ultrasonic extraction is performed for 60 minutes; let it reach room temperature, and then filter it with a microporous filter membrane, and collect the filtrate as the sample liquid to be tested; the sample liquids of the 6 samples to be identified and distinguished are numbered 2, 3, 4, 6, 7, and 8 respectively.

[0050] 2) Perform thin layer spotting

[0051] The standard control solutions of Chlorella and Spirulina prepared in step 1.1) and the sample solution to be tested prepared in step 1.2) are sequentially spotted at different positions at the same height on the same silica gel G thin layer plate according to the numbers to form 8 spot origins; and the distance between the spot origin position and the lower edge of the silica gel G thin layer plate is 1.0 to 1.5 cm. In this embodiment, the distance between the spot origin position and the lower edge of the silica gel G thin layer plate is 1.0 cm.

[0052] 3) Preparation of developing agent

[0053] 3.1) Ethyl acetate, formic acid, acetic acid and water are uniformly mixed in a volume ratio of 10-20:0.5-1.5:0.5-1.5:1.5-2.5 to prepare 19 mL of a developing solvent. In this embodiment, the volume ratio of ethyl acetate, formic acid, acetic acid and water is 15:1:1:2, and ethyl acetate, formic acid and acetic acid are all pure products.

[0054] 3.2) Place the developing agent in a developing cylinder, seal the developing cylinder, and pre-saturate for 15 to 20 minutes; in this embodiment, the developing cylinder adopts a double-slot developing cylinder, the size of the double-slot developing cylinder is 10 cm×10 cm, and the dosage of the developing agent in the double slots is kept consistent, and the pre-saturation time is 15 minutes.

[0055] 4) Expand

[0056] 4.1) Open the pre-saturated developing cylinder in step 3) and quickly place the silica gel G thin layer plate spotted in step 2) into the developing cylinder, so that the silica gel G thin layer plate is tilted against the developing cylinder and the developing agent in the developing cylinder cannot cover the spotting origin.

[0057] 4.2) When the front edge of the developing agent moves upward to 8 cm from the sample spot origin, the silica gel G thin layer plate is taken out. In other embodiments, the distance from the front edge of the developing agent to the sample spot origin may be 6 cm to 10 cm.

[0058] 5) Identification

[0059] 5.1) Dry the reagent on the silica gel G thin layer plate developed in step 4.2) with a hair dryer, leave it for 10 minutes, and inspect it under 365nm ultraviolet light to obtain an inspection image.

[0060] After inspection under 365nm ultraviolet light, the silica gel G thin layer plate is sprayed with a sulfuric acid ethanol color developer with a mass concentration of 5% to 10% for color development. In this embodiment, the mass concentration of the sulfuric acid ethanol color developer is 10%; then it is placed on an electric heating plate and heated at 100-110°C for 3 to 5 minutes. In this embodiment, the heating temperature is 105°C; after it is allowed to cool to room temperature, the silica gel G thin layer plate is inspected again under 365nm ultraviolet light to obtain an inspection diagram.

[0061] 5.2) The spots formed by the spotting origins of the sample solutions to be tested (No. 2, 3, 4, 6, 7 and 8) on the inspection diagram are compared and analyzed with the spots formed by the spotting origins of the standard control solutions of Chlorella (No. 1) and Spirulina (No. 5). The analysis principle is: if the spots of the sample solution to be tested are consistent with the spots of the standard control solution of Chlorella, the sample to be tested is Chlorella; if the spots of the sample solution to be tested are consistent with the spots of the standard control solution of Spirulina, the sample to be tested is Spirulina; if the spots of the sample solution to be tested are inconsistent with the spots of the standard control solution of Chlorella and the spots of the standard control solution of Spirulina, the sample to be tested is other samples.

[0062] The two inspection images obtained in the above embodiment are respectively as follows: Figure 1 and Figure 2 As shown in the figure, the horizontal axis represents the sample number, and the vertical axis represents the Rf value of the thin layer chromatography, among which No. 1 and No. 5 are known Chlorella and Spirulina, respectively, and No. 2-4 and No. 6-8 are unknown samples to be tested.

[0063] Figure 1 This is the inspection diagram obtained under 365nm ultraviolet light, in which the spots of the samples to be tested No. 2, 3, and 4 are consistent with the spot of No. 1 in quantity and Rf position, and the spots of No. 6, 7, and 8 are consistent with the spot of No. 5 in quantity and Rf. It can be judged that the samples to be tested No. 2, 3, and 4 are Chlorella, and the samples to be tested No. 6, 7, and 8 are Spirulina. Figure 1 Under the non-color inspection method, the spots between Chlorella and Spirulina are obviously different and easy to identify. The distinguishing features between Chlorella and Spirulina are mainly that Chlorella has yellow and red spots between Rf values ​​of 0.1-0.2, and red spots at Rf values ​​of 0.9; Spirulina has blue spots between Rf values ​​of 0.15-0.2, and green spots at Rf values ​​of 0.02 and 0.52.

[0064] In order to ensure the accuracy and reliability of the results, a color development inspection was also performed in this embodiment to obtain Figure 2View of the Figure 2 It can be seen that the spots of the samples to be tested No. 2, 3, and 4 are consistent with the spots of No. 1 in quantity and Rf position, and the spots of No. 6, 7, and 8 are consistent with the spots of No. 5 in quantity and Rf. It can be judged that the samples to be tested No. 2, 3, and 4 are Chlorella, and the samples to be tested No. 6, 7, and 8 are Spirulina. Under the color development inspection method, the difference in spots between Chlorella and Spirulina is also very obvious and easy to identify. In the inspection diagram after color development with 10% sulfuric acid ethanol color developer, the distinguishing features of Chlorella and Spirulina are that Chlorella has blue spots at Rf values ​​of 0.1-0.15 and red spots at Rf values ​​of 0.9; Spirulina has blue spots at Rf values ​​of 0.02 and green spots at Rf values ​​of 0.52.

[0065] Example 2

[0066] The method steps of this embodiment are the same as those of Embodiment 1, except that:

[0067] In step 1.1), the amount of ethanol added is 5 times the mass of the Chlorella raw material or the Spirulina raw material, the ethanol concentration is 95%, and the ultrasonic extraction time is 30 minutes.

[0068] In step 1.2), the amount of ethanol added is 5 times the mass of the sample to be tested, the ethanol concentration is 95%, and the ultrasonic extraction time is 30 minutes.

[0069] In step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.5 cm.

[0070] In step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 10:0.8:0.8:1.8;

[0071] In step 3.2), the size of the double-slot development cylinder is 20 cm×10 cm, and the pre-saturation time is 20 minutes.

[0072] In step 4.2), the front edge of the developing agent moves upward to a distance of 6 cm from the sample spot origin.

[0073] In step 5.1), the mass concentration of the sulfuric acid ethanol color developer is 5%, and the heating temperature is 100°C.

[0074] The inspection diagram obtained is basically consistent with that of Example 1.

[0075] Example 3

[0076] The method steps of this embodiment are the same as those of Embodiment 1, except that:

[0077] In step 1.1), the amount of ethanol added is 10 times the mass of the Chlorella raw material or the Spirulina raw material, the ethanol concentration is 50%, and the ultrasonic extraction time is 40 minutes.

[0078] In step 1.2), the amount of ethanol added is 10 times the mass of the sample to be tested, the ethanol concentration is 50%, and the ultrasonic extraction time is 40 minutes.

[0079] In step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.3 cm.

[0080] In step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 20:1.2:1.2:2.2.

[0081] In step 3.2), the size of the double-slot development cylinder is 10 cm×20 cm, and the pre-saturation time is 17 minutes.

[0082] In step 4.2), the front edge of the developing agent moves upward to a distance of 10 cm from the sample spot origin.

[0083] In step 5.1), the mass concentration of the sulfuric acid ethanol color developer is 7%, and the heating temperature is 110°C.

[0084] The inspection diagram obtained is basically consistent with that of Example 1.

[0085] Example 4

[0086] The method steps of this embodiment are the same as those of Embodiment 1, except that:

[0087] In step 1.1), the amount of ethanol added is 7 times the mass of the Chlorella raw material or the Spirulina raw material, the ethanol concentration is 80%, and the ultrasonic extraction time is 50 minutes.

[0088] In step 1.2), the amount of ethanol added is 7 times the mass of the sample to be tested, the ethanol concentration is 80%, and the ultrasonic extraction time is 50 minutes.

[0089] In step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.2 cm.

[0090] In step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 10:0.5:0.5:1.5.

[0091] In step 5.1), the mass concentration of the sulfuric acid ethanol color developer is 8%, and the heating temperature is 107°C.

[0092] The inspection diagram obtained is basically consistent with that of Example 1.

[0093] Example 5

[0094] The method steps of this embodiment are the same as those of Embodiment 1, except that:

[0095] In step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 20:1.5:1.5:2.5.

[0096] Step 5.1) is specifically as follows: after drying the reagent on the silica gel G thin layer plate developed in step 4.2) with a hair dryer, spray the silica gel G thin layer plate with 10% sulfuric acid ethanol color developer for color development, and then place it on an electric heating plate and heat it at 105° C. for 3 to 5 minutes; after cooling to room temperature, inspect the silica gel G thin layer plate under 365nm ultraviolet light to obtain an inspection diagram.

[0097] The inspection diagram obtained is similar to that of Example 1. Figure 2 Basically the same.

Claims

1. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina, characterized in that: The following steps are involved: 1) Prepare standard control solution and sample solution to be tested 1.1) taking a chlorella raw material and a spirulina raw material, placing them in sealable test containers respectively, adding 5-10 times the mass of the chlorella raw material and the spirulina raw material and 50%-95% ethanol respectively, and performing ultrasonic extraction for 30-60 minutes, cooling to room temperature, and then filtering with a microporous filter membrane, collecting the filtrate as the standard control solution of chlorella and spirulina respectively; 1.2) Take the sample to be identified, place it in a sealable test container, add 5 to 10 times the mass of the sample, and 50% to 95% ethanol, and perform ultrasonic extraction for 30 to 60 minutes, let it cool to room temperature, and then filter it with a microporous membrane, and collect the filtrate as the sample solution to be tested; 2) Perform thin layer spotting Place the standard control solutions of Chlorella and Spirulina prepared in step 1.1) and the sample solution to be tested prepared in step 1.2) on the same silica gel G thin layer plate at different positions at the same height to form a spot origin; 3) Preparation of developing agent 3.1) Ethyl acetate, formic acid, acetic acid and water are mixed at a ratio of 10 to 20: The mixture is uniformly mixed in a volume ratio of 0.5-1.5:0.5-1.5:1.5-2.5 to obtain a developing agent; 3.2) Place the developing agent in the developing cylinder, seal the developing cylinder, and pre-saturate for 15 to 20 minutes; 4) Expand 4.1) Open the development cylinder after pre-saturation in step 3), and quickly put the silica gel G thin layer plate spotted in step 2) into the development cylinder, so that the silica gel G thin layer plate is tilted against the development cylinder; 4.2) When the front edge of the developing agent moves up to 6 cm to 10 cm from the sample point, take out the silica gel G thin layer plate; 5) Identification 5.1) After the reagent on the silica gel G thin layer plate developed in step 4.2) is dried, the silica gel G thin layer plate is placed under 365 nm ultraviolet light for inspection to obtain an inspection image; 5.2) Compare the spots formed at the sampling origin of the sample solution to be tested on the inspection diagram with the spots formed at the sampling origin of the standard control solutions of Chlorella and Spirulina; If the spot of the sample liquid to be tested is consistent with the spot of the standard control solution of Chlorella, then the sample to be tested is Chlorella; if the spot of the sample liquid to be tested is consistent with the spot of the standard control solution of Spirulina, then the sample to be tested is Spirulina; if the spot of the sample to be tested is inconsistent with the spot of the standard control solution of Chlorella and is also inconsistent with the spot of the standard control solution of Spirulina, then the sample to be tested is other samples.

2. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 1, characterized in that: In step 5.1), after inspection under 365nm ultraviolet light, spray 5% to 10% sulfuric acid ethanol color developer on the silica gel G thin layer plate for color development, then heat at 100-110°C for 3 to 5 minutes, let it cool to room temperature, and then inspect the silica gel G thin layer plate under 365nm ultraviolet light to obtain an inspection diagram.

3. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 2, characterized in that: In step 5.1), the mass concentration of the sulfuric acid ethanol color developer is 10%, and the heating temperature is 105°C.

4. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 3, characterized in that: In step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.0 to 1.5 cm.

5. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 4, characterized in that: In step 3.1), the volume ratio of ethyl acetate, formic acid, acetic acid and water is 15:1:1:

2.

6. The thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 5, characterized in that: In step 3.2), the developing cylinder adopts a double-slot developing cylinder, the size of which is 10cm×10cm, 20cm×10cm or 10cm×20cm, and the dosage of the developing agent in the double slots is kept consistent, and pre-saturation is performed for 15 minutes.

7. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 6, characterized in that: Step 4.2) is specifically as follows: when the front edge of the developing agent moves upward to a distance of 8 cm to 10 cm from the sample spot origin, the silica gel G thin layer plate is taken out.

8. The thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 7, characterized in that: In step 1.1), ethanol with a mass concentration of 70% and an amount of 6 times the mass of the Chlorella raw material and Spirulina raw material is added respectively, and ultrasonic extraction is performed for 60 minutes; In step 1.2), ethanol with a mass concentration of 70% and an amount of 6 times the mass of the sample to be tested is added, and ultrasonic extraction is performed for 60 minutes.

9. A thin layer chromatography analysis method for rapid identification of Chlorella and Spirulina according to claim 8, characterized in that: In step 2), the distance between the sample spot origin and the lower edge of the silica gel G thin layer plate is 1.0 cm; Step 4.2) Specifically, when the front edge of the developing agent moves upward to 8 cm from the sample spot origin, the silica gel G thin layer plate is taken out; In step 3.2), the size of the double-slot expansion cylinder is 10 cm×10 cm.

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