A method for rapidly removing chlorophyll

Abstract

The present invention belongs to the technical field of extract purification, and particularly relates to a method for rapidly removing chlorophyll, comprising the following steps: (1) preparing a purification column: successively connecting a tetraalkylsilane-bonded silica gel solid-phase extraction column, an octadecylsilane-bonded silica gel solid-phase extraction column, and a chromatographic silica gel solid-phase extraction column in series from top to bottom to obtain a purification column; (2) decolorization: taking the liquid to be decolorized, passing it through the purification column in step (1) to remove chlorophyll, and collecting the effluent to obtain a decolorized extract. The present invention uses a series connection technique to successively connect a tetraalkylsilane-bonded silica gel solid-phase extraction column, an octadecylsilane-bonded silica gel solid-phase extraction column, and a chromatographic silica gel solid-phase extraction column in series from top to bottom, realizing one-step purification, thereby greatly shortening the purification time and significantly improving the pretreatment rate and efficiency.

Description

Technical Field

[0001] The present invention belongs to the technical field of extract purification, and particularly relates to a method for rapidly removing chlorophyll. Background Art

[0002] Currently, in the field of extraction and separation of natural products, chlorophyll often interferes with other target components, thus affecting the purity and content determination of the extract. When the inventor was separating the above-ground parts of common Camellia nitidissima leaves, four-season Camellia nitidissima leaves, and Veronica persica, it was found through chemical reaction color development that the above several plants mainly contain flavonoids, terpenoids, and coumarin components. During TLC analysis, flavonoid components show bright yellow spots under UV254nm and 365nm; terpenoid components show purple-magenta spots under visible light after heating with 10% sulfuric acid-methanol at 110°C; coumarin components show blue spots under visible light after spraying with ferric chloride solution.

[0003] When performing systematic extraction, separation, and purification of the above plants, interference from chlorophyll compounds occurs. Specifically, each part of the medium-polarity fraction of the extract is green-dark green. When analyzed by TLC, a green color band appears from the origin to the front of the TLC plate under visible light, and a red color appears from the origin to the front of the TLC plate under UV365nm. Due to the appearance of dark-colored bands under ultraviolet and visible light, it interferes with the detection of other substances (visible light detection, ultraviolet detection, chemical reaction color development detection). And the conventional removal of chlorophyll compounds requires repeated use of silica gel column chromatography, which will cause irreversible adsorption of other non-chlorophyll substances in the silica gel packing, resulting in losses.

[0004] After using this method for removing chlorophyll, the inventor has separated and purified high-purity flavonoid components such as quercetin, phloretin, and polyhydroxyflavanones from common Camellia nitidissima leaves and four-season Camellia nitidissima leaves; triterpenoid components such as Octyl Oleanolate, α-spinasterol, and Camellenodiol; and sesquiterpenoid components such as loliolide, iso-loliolide, and dethdrovomofoliol. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention proposes a method for rapidly removing chlorophyll.

[0006] The technical solution of the present invention is as follows:

[0007] A method for rapidly removing chlorophyll, comprising the following steps:

[0008] (1) Preparation of purification column: A tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, and a chromatographic silica solid-phase extraction column are connected in series from top to bottom to obtain a purification column;

[0009] (2) Decolorization: Take the liquid to be decolorized, pass it through the purification column in step (1) to remove chlorophyll, and collect the effluent to obtain the decolorized extract.

[0010] Further, in step (1), the ratio of the column lengths of the tetraalkylsilane-bonded silica solid-phase extraction column, the octadecylsilane-bonded silica solid-phase extraction column, and the chromatographic silica solid-phase extraction column is 3:(3 - 5):3.

[0011] Further, in step (1), in the chromatographic silica solid-phase extraction column, the particle size of the silica packing is 300 - 400 mesh.

[0012] Further, in step (2), before loading the column, first rinse the purification column with an organic solvent 1 - 2 times.

[0013] Further, the chlorophyll is one or more of 13 2 -hydroxypheophorbide A ethyl ester, pheelophyll-α, 17c-ethoxyphaeophorbidea.

[0014] Further, the liquid to be decolorized includes an ethanol extract, a dichloromethane extract, or an ethyl acetate extract of plants.

[0015] Further, the preparation method of the liquid to be decolorized includes the following steps: Take Camellia nitidissima leaves and soak them successively in ethanol with a volume concentration of 95%, 80%, and 50%, collect and combine all the extracts, and obtain the total extract by low-temperature vacuum distillation; The total extract is extracted with petroleum ether and ethyl acetate successively, and concentrated under reduced pressure to obtain a petroleum ether extract and an ethyl acetate extract respectively; Take the ethyl acetate extract, use silica gel column chromatography, and perform gradient elution with a mixed solution of petroleum ether and ethyl acetate with different volume ratios as the eluent to obtain different fractions, and take the dark green fraction as the liquid to be decolorized.

[0016] Further, in the mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 40:1 - 0:1.

[0017] Further, the preparation method of the liquid to be decolorized comprises the following steps: soaking the golden camellia leaves in ethanol with a volume concentration of 95%, 80%, and 50% in sequence, collecting and combining all the extracts, and obtaining the total extract by low-temperature vacuum distillation; extracting the total extract with petroleum ether, dichloromethane, and ethyl acetate in sequence to obtain petroleum ether extract, dichloromethane extract, and ethyl acetate extract respectively; taking the dichloromethane extract, using silica gel column chromatography, and performing gradient elution with a mixed solution of petroleum ether, dichloromethane, and methanol with different volume ratios as the eluent to obtain different fractions, and taking the dark green fraction as the liquid to be decolorized.

[0018] Further, in the mixed solution of petroleum ether, dichloromethane, and methanol, the volume ratio of petroleum ether, dichloromethane, and methanol is 20:1:0 - 0:10:1.

[0019] The beneficial effects of the present invention are as follows:

[0020] (1) The present invention uses a tandem technology to connect a tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, and a chromatographic silica solid-phase extraction column in series from top to bottom, realizing one-step purification, thus greatly shortening the purification time and significantly improving the pretreatment rate and efficiency.

[0021] (2) The present invention has a high specific removal rate for chlorophyll substances, overcoming the loss of non-chlorophyll substances caused by traditional methods such as activated carbon adsorption of chlorophyll. Description of the Drawings

[0022] Figure 1 It is the HPLC chromatogram of the chlorophyll in the liquid to be decolorized in Example 2;

[0023] Figure 2 It is the HPLC chromatogram of the decolorized extract in Example 2, where a is 13 2 -hydroxypheophorbide A ethyl ester, b is pheelophyll-α, and c is 17c-ethoxyphaeophorbidea;

[0024] Figure 3 It is the TLC thin-layer chromatogram before and after removing chlorophyll under UV365nm, where I is the TLC thin-layer chromatogram before removing chlorophyll and II is the TLC thin-layer chromatogram after removing chlorophyll;

[0025] Figure 4 It is the TLC thin-layer chromatogram before and after removing chlorophyll under visible light, where III is the TLC thin-layer chromatogram before removing chlorophyll and IV is the TLC thin-layer chromatogram after removing chlorophyll;

[0026] Figure 5It is the HPLC chromatogram of the chlorophyll in the liquid to be decolorized in Example 4, where d is chlorophylla;

[0027] Figure 6 It is the HPLC chromatogram of Effluent A in Example 4, where d is chlorophylla;

[0028] Figure 7 It is the HPLC chromatogram of Effluent B in Example 4, where d is chlorophylla;

[0029] Figure 8 It is the HPLC chromatogram of Effluent C in Example 4, where d is chlorophylla;

[0030] Figure 9 It is the HPLC chromatogram of Effluent D in Example 4, where d is chlorophylla. Detailed implementation mode

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0032] Example 1

[0033] Preparation of purification column:

[0034] (1) Prepare a tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, and a chromatographic silica solid-phase extraction column respectively.

[0035] Among them, the preparation method of the tetraalkylsilane-bonded silica solid-phase extraction column is as follows: Take a solid-phase extraction empty column, press a sieve plate at the bottom of the column tube, fill 2-5 g of tetraalkylsilane-bonded silica filler, compact it, and press a sieve plate above the filler. The diameter of the sieve plate is the same as the inner diameter of the solid-phase extraction column, and the thickness is 3 mm;

[0036] The preparation method of the octadecylsilane-bonded silica solid-phase extraction column is as follows: Take a solid-phase extraction empty column, press a sieve plate at the bottom of the column tube, fill 2 g of octadecylsilane-bonded silica filler, compact it, and press a sieve plate above the filler. The diameter of the sieve plate is the same as the inner diameter of the solid-phase extraction column, and the thickness is 3-5 mm;

[0037] The preparation method of the chromatographic silica gel solid-phase extraction column is as follows: Take an empty solid-phase extraction column, press a sieve plate at the bottom of the column tube, fill it with 300-400 mesh chromatographic silica gel filler dried at 105°C for 30 minutes, compact it, and press a sieve plate above the filler. The diameter of the sieve plate is the same as the inner diameter of the solid-phase extraction column, and the thickness is 3 mm;

[0038] (2) Install a quark adapter valve at the lower opening of the tetraalkylsilane-bonded silica gel solid-phase extraction column and connect it to the upper opening of the octadecylsilane-bonded silica gel solid-phase extraction column; then install a quark adapter valve at the lower opening of the octadecylsilane-bonded silica gel solid-phase extraction column and connect it to the upper opening of the chromatographic silica gel solid-phase extraction column to obtain the purification column.

[0039] The inner diameter of the above-mentioned empty solid-phase extraction column is 0.8-1.5 cm.

[0040] Example 2

[0041] (1) Prepare a purification column according to Example 1, where the column lengths of the tetraalkylsilane-bonded silica gel solid-phase extraction column, octadecylsilane-bonded silica gel solid-phase extraction column, and chromatographic silica gel solid-phase extraction column are all 3 mm. The inner diameter of the empty solid-phase extraction column is selected to be 0.8 cm.

[0042] (2) Soak 5 kg of ordinary golden camellia leaves from Fangchenggang in 20 kg of ethanol with a volume concentration of 95%, 80%, and 50% in sequence. Each soaking time is 7 days. Collect and combine all the extracts, and obtain the total extract by low-temperature vacuum distillation; extract the total extract with petroleum ether, dichloromethane, and ethyl acetate 3 times respectively to obtain petroleum ether extract, dichloromethane extract, and ethyl acetate extract. Take the dichloromethane extract and use silica gel column chromatography. Use a mixed solution of petroleum ether and ethyl acetate with different volume ratios as the eluent for gradient elution to obtain different fractions. Take the dark green fraction as the liquid to be decolorized. Among them, in the mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 40:1, 30:1, 20:1, 10:1, 0:1; Take the liquid to be decolorized through the purification column in step (1) (before loading the column, first rinse the purification column with methanol once), collect the effluent, and obtain the decolorized extract.

[0043] (3) Calculation of the chlorophyll removal rate in the liquid to be decolorized:

[0044] The determination of the chlorophyll peak area in the liquid to be decolorized is as follows:

[0045] Take the fraction to be decolorized, dissolve it in acetonitrile, and prepare a 2.0 mg / mL test solution for HPLC analysis. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, Ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 80% acetonitrile - 0.1% phosphoric acid - 19.9% water. The HPLC chromatogram is as Figure 1As shown, where t R = 12.545 min corresponds to chlorophyll compounds: 13 2 -hydroxypheophorbide A ethyl ester (a), with a corresponding peak area of 778223, t R = 18.666 min corresponds to chlorophyll compounds: pheelophyll-α (b), with a corresponding peak area of 381393, t R = 19.366 min corresponds to chlorophyll compounds: 17c-ethoxyphaeophorbide a (c), with a corresponding peak area of 388355.

[0046] The determination of the peak area of chlorophyll in the decolorized extract is as follows:

[0047] Take the decolorized extract, concentrate it at low temperature to prepare a test solution of 2.0 mg / mL for HPLC analysis. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, UV detector, detection wavelength: 210 nm, chromatographic mobile phase: 80% acetonitrile - 0.1% phosphoric acid - 19.9% water. The HPLC chromatogram is as Figure 2 shown, where t R = 12.668 min corresponds to chlorophyll compounds: 13 2 -hydroxypheophorbide A ethyl ester (a), with a corresponding peak area of 2647, t R = 18.4646 min corresponds to chlorophyll compounds: pheelophyll-α (b), with a corresponding peak area of 154725, t R = 19.366 min corresponds to chlorophyll compounds: 17c-ethoxyphaeophorbide a (c), with a corresponding peak area of 220256.

[0048] According to Figure 1 and Figure 2 The removal rates of the liquid to be decolorized calculated from the peak areas are shown in Table 1.

[0049] Table 1

[0050]

[0051] Conclusion: After decolorization by the above method, the corresponding peak areas of the three chlorophyll compounds decreased significantly. Especially for 13 2 -hydroxypheophorbide A ethyl ester (a), the removal effect reached 99.65%, which also indicates that the purification column of the present invention has a good adsorption effect on chlorophyll compounds.

[0052] Example 3

[0053] (1) Prepare the purification column according to Example 1, wherein the column lengths of the tetraalkylsilane-bonded silica solid-phase extraction column, octadecylsilane-bonded silica solid-phase extraction column, and chromatography silica solid-phase extraction column are 3 mm, 5 mm, and 3 mm respectively. The inner diameter of the solid-phase extraction empty column is selected to be 1.5 cm.

[0054] (2) Soak 10 kg of Jasminum sambac Ait. leaves in 50 kg of ethanol with volume concentrations of 95%, 80%, and 50% successively, with each soaking time being 7 days. Collect and combine all the extracts, and obtain the total extract by low-temperature vacuum distillation; extract the total extract with 10 kg of petroleum ether and ethyl acetate three times each, and concentrate under reduced pressure to obtain petroleum ether extract and ethyl acetate extract respectively. Take the ethyl acetate extract, and use silica gel column chromatography. Use a mixed solution of petroleum ether, dichloromethane, and methanol with different volume ratios as the eluent for gradient elution to obtain different fractions. Take the dark green fraction as the liquid to be decolorized. Among them, in the mixed solution of petroleum ether, dichloromethane, and methanol, the volume ratio of petroleum ether, dichloromethane, and methanol is 20:1:0, 10:5:1, 0:10:1; pass the liquid to be decolorized through the purification column in step (1) (before loading the column, first rinse the purification column with methanol twice), and collect the effluent to obtain the decolorized extract.

[0055] (3) Comparison of the effects before and after dechlorophyllization of the liquid to be decolorized:

[0056] Use TLC thin-layer chromatography to compare the effects before and after dechlorophyllization. Among them, the thin-layer chromatography plate is a silica gel G plate, and the developing agent is: chloroform:toluene:methanol = 10:5:1.

[0057] The TLC thin-layer chromatography chromatograms before and after dechlorophyllization under UV365 nm are as Figure 3 shown. The TLC thin-layer chromatography chromatograms before and after dechlorophyllization under visible light are as Figure 4 shown.

[0058] Before and after dechlorophyllization, before dechlorophyllization, other component components shown under UV365 nm and visible light are interfered by chlorophyll-like substances. After thin-layer chromatography, the thin-layer plate is covered with a chlorophyll band from the top to the origin, interfering with the identification of other substances;

[0059] After dechlorophyllization, as shown under UV365 nm and visible light, most of the chlorophyll-like color bands are removed, and the spots on the thin-layer plate are clearly visible.

[0060] It shows that the purification column of the present invention has a good adsorption effect on chlorophyll-like compounds, and can preferably remove the interference caused by chlorophyll-like components to the detection and separation of other substances during the phytochemical separation and purification process.

[0061] Example 4

[0062] (1) Prepare a tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, a chromatography silica solid-phase extraction column and a purification column respectively according to Example 1.

[0063] (2) Determination of the peak area of chlorophyll before decolorization:

[0064] Take 500.7 mg of chlorophyll a reference substance (Shanghai Aladdin Biochemical Technology Co., Ltd., LOT#H2209387), add it to a mixed solution of 48.5 mL of acetonitrile + 1.5 mL of ethyl acetate to dissolve, and filter through a 0.45 μm microporous filter membrane to obtain a chlorophyll reference substance solution.

[0065] Take 20 μL of the above reference substance solution for HPLC analysis. The HPLC chromatogram is as Figure 5 shown. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 95% acetonitrile - 0.1% phosphoric acid - 4.9% water. Among them, t R = 7.178 min corresponds to the chlorophyll compound: chlorophyll a. The corresponding peak area is 5991095.

[0066] (3) Comparison of the removal effects of different chromatographic columns on chlorophyll compounds:

[0067] Take the above chlorophyll reference substance solution under the condition of not being in series, and separately use a tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, a chromatography silica solid-phase extraction column, and compare the removal effects of the above three solid-phase extraction columns in series on chlorophyll compounds.

[0068] A. Take 10 mL of the above reference substance solution and pass it through a tetraalkylsilane-bonded silica solid-phase extraction column (inner diameter 1.5 cm, packing thickness 9 mm) to obtain an effluent A. Take 20 μL of the effluent A for HPLC analysis. The HPLC chromatogram is as Figure 6 shown. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 95% acetonitrile - 0.1% phosphoric acid - 4.9% water. Among them, t R = 7.400 min corresponds to the chlorophyll compound: chlorophyll a. The corresponding peak area is 4080539.

[0069] B. Take 10 mL of the above reference solution and pass it through an octadecylsilyl-bonded silica solid-phase extraction column (inner diameter 1.5 cm, packing thickness 9 mm) to obtain the effluent B. Take 20 μL of the effluent B for HPLC analysis, and the HPLC chromatogram is as shown in Figure 7 Figure [0000181]. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 95% acetonitrile - 0.1% phosphoric acid - 4.9% water. Among them, t R = 7.264 min corresponds to the chlorophyll compound: chlorophylla. The corresponding peak area is 2,891,857.

[0070] C. Take 10 mL of the above reference solution and pass it through a chromatographic silica gel solid-phase extraction column (inner diameter 1.5 cm, packing thickness 9 mm) to obtain the effluent C. Take 20 μL of the effluent C for HPLC analysis, and the HPLC chromatogram is as shown in Figure 8 Figure [0000185]. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 95% acetonitrile - 0.1% phosphoric acid - 4.9% water. Among them, t R = 7.404 min corresponds to the chlorophyll compound: chlorophylla. The corresponding peak area is 2,926,957.

[0071] D. Take 10 mL of the above reference solution and pass it through the purification column of the present invention (wherein the inner diameters of the tetraalkylsilyl-bonded silica solid-phase extraction column, octadecylsilyl-bonded silica solid-phase extraction column, and chromatographic silica gel solid-phase extraction column are all 1.5 cm, and the packing thicknesses are all 3 mm), collect the effluent D, take 20 μL of the effluent D for HPLC analysis, and the HPLC chromatogram is as shown in Figure 9 Figure [0000189]. Among them, the chromatographic column: Yuexu XB-C18, 4.6×250 mm, 5 μm, ultraviolet detector, detection wavelength: 210 nm, chromatographic mobile phase: 95% acetonitrile - 0.1% phosphoric acid - 4.9% water. Among them, t R = 7.279 min corresponds to the chlorophyll compound: chlorophylla. The corresponding peak area is 750,757.

[0072] According to Figures 5 - 9 the peak areas, the decolorization rates of the liquid to be decolorized are calculated as shown in Table 2.

[0073] Table 2

[0074]

[0075] From the data of the peak area changes before and after passing through different solid-phase extraction columns above, it can be seen that the purification column of the present invention has a better effect on removing chlorophyll a than the single tetraalkylsilane-bonded silica gel solid-phase extraction column, octadecylsilane-bonded silica gel solid-phase extraction column, and chromatographic silica gel solid-phase extraction column.

[0076] The above description is a detailed description of the preferred feasible embodiments of the invention, but the embodiments are not intended to limit the scope of the patent application of the present invention. Any equivalent changes or modifications made under the technical spirit disclosed by the present invention shall fall within the scope of the patent covered by the present invention.

Claims

1. A method for rapidly removing chlorophyll, characterized in that, It includes the following steps: (1) Prepare a purification column: Connect a tetraalkylsilane-bonded silica solid-phase extraction column, an octadecylsilane-bonded silica solid-phase extraction column, and a chromatographic silica solid-phase extraction column in series from top to bottom to obtain a purification column; (2) Decolorize: Take the liquid to be decolorized, pass it through the purification column in step (1) to remove chlorophyll, and collect the effluent to obtain a decolorized extract; the liquid to be decolorized includes an ethanol extract, a dichloromethane extract, or an ethyl acetate extract of plants.

2. The method for rapidly removing chlorophyll according to claim 1, characterized in that, In step (1), the ratio of the column lengths of the tetraalkylsilane-bonded silica solid-phase extraction column, the octadecylsilane-bonded silica solid-phase extraction column, and the chromatographic silica solid-phase extraction column is 3:(3 - 5):

3.

3. The method for rapidly removing chlorophyll according to claim 1, characterized in that, In step (1), in the chromatographic silica solid-phase extraction column, the particle size of the silica packing is 300 - 400 mesh.

4. The method for rapidly removing chlorophyll according to claim 1, characterized in that: In step (2), before loading the column, first rinse the purification column with an organic solvent 1 - 2 times.

5. The method for rapidly removing chlorophyll according to claim 1, characterized in that: The chlorophyll is one or more of 13 2 -hydroxypheophorbide A ethyl ester, pheelophyll-α, and 17c-ethoxyphaeophorbide a.

6. The method for quickly removing chlorophyll according to claim 1, characterized in that: The preparation method of the liquid to be decolorized includes the following steps: Soak Camellia nitidissima leaves in ethanol with a volume concentration of 95%, 80%, and 50% in sequence, collect and combine all the extracts, and obtain a total extract by low-temperature vacuum distillation; extract the total extract with petroleum ether and ethyl acetate in sequence, and concentrate under reduced pressure to obtain a petroleum ether extract and an ethyl acetate extract respectively; take the ethyl acetate extract, use silica gel column chromatography, and perform gradient elution with a mixed solution of petroleum ether and ethyl acetate with different volume ratios as the eluent to obtain different fractions, and take the dark green fraction as the liquid to be decolorized.

7. The method for rapidly removing chlorophyll according to claim 6, characterized in that: In the mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 40:1 - 0:

1.

8. The method for rapidly removing chlorophyll according to claim 1, characterized in that: The preparation method of the liquid to be decolorized includes the following steps: Soak Camellia nitidissima leaves in ethanol with a volume concentration of 95%, 80%, and 50% in sequence, collect and combine all the extracts, and obtain a total extract by low-temperature vacuum distillation; extract the total extract with petroleum ether, dichloromethane, and ethyl acetate in sequence to obtain a petroleum ether extract, a dichloromethane extract, and an ethyl acetate extract respectively; take the dichloromethane extract, use silica gel column chromatography, and perform gradient elution with a mixed solution of petroleum ether, dichloromethane, and methanol with different volume ratios as the eluent to obtain different fractions, and take the dark green fraction as the liquid to be decolorized.

9. The method for rapidly removing chlorophyll according to claim 8, characterized in that: In the mixed solution of petroleum ether, dichloromethane, and methanol, the volume ratio of petroleum ether, dichloromethane, and methanol is 20:1:0 - 0:10:1.

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