Extraction method and application of natural dye from radix saposhnikoviae stalk

Abstract

The invention discloses an extraction method and application of a natural dyestuff from parsnip herb stalks, and relates to the technical field of extraction of natural dyestuff, the method comprises the following steps: pretreating the parsnip herb stalks to prepare parsnip herb stalk fibrillated fine powder; adding the fine powder of the radix saposhnikoviae stalks into a citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value, and adding trichoderma reesei for fermentation to obtain a fermentation mixed solution; uniformly mixing the fermentation mixed solution with water, adding an enzymatic system, carrying out ultrasonic microwave extraction, centrifuging, taking a supernatant, and filtering; circulating the process for three times, and combining filtrate; the enzymatic system contains cellulase, ferriporphyrin and pyridoxal phosphate; adding sodium carbonate into the filtrate obtained in the step S3, and performing heating reaction, solid-phase extraction, suction filtration and freeze drying to obtain the radix saposhnikoviae stalk dye. The extraction process of the radix saposhnikoviae stalk dye is simple, the extraction efficiency is high, the cost is low, and the market prospect is wide. The obtained radix saposhnikoviae stalk extract has the effects of sterilization and the like, is safe and environment-friendly, has good compatibility with the ecological environment, and is biodegradable.

Description

Technical Field

[0001] The invention relates to the technical field of natural dyes, in particular to a natural dye of parsnip stems and an extraction method and application thereof. Background Art

[0002] Natural dyes are derived from plants, animals, minerals, etc. in nature. There is no need to add a large amount of chemical additives during the dyeing process, and no wastewater is generated. The precipitate from the extracted dyes can be used as papermaking raw materials or fertilizers, without causing pollution and achieving "zero emissions". During the dyeing process, its medicinal ingredients will be absorbed by the fabric together with the pigment, so that the dyed fabric has special medicinal health functions for the human body, such as relaxing muscles and tendons, sterilizing, refreshing, anti-ultraviolet rays, etc.

[0003] The Chinese medicine Fangfeng is mostly extracted from the rhizomes of Fangfeng grass to extract active ingredients, which mainly contain coumarin, chromone, volatile oil and polysaccharide and other chemical components. At present, the main medicinal components of the Chinese medicine Fangfeng are Fangfeng root pieces and Fangfeng grass, while the Fangfeng grass stems are mostly discarded as waste. The present invention first analyzes the Fangfeng grass stems (referring to stems, stems, and coarse leaves), which are rich in cellulose, hemicellulose and lignin, which together constitute the main components of the Fangfeng grass stem cell wall. At the same time, the combination between the cellulose and hemicellulose or lignin molecules of the Fangfeng grass stem mainly depends on hydrogen bonds; in addition to hydrogen bonds, there is also a chemical bond between hemicellulose and lignin, resulting in a small amount of carbohydrates in the lignin separated from the Fangfeng grass stem raw material. Cellulose, hemicellulose and lignin are interwoven to form a cell wall, and the degradation of any one type of component is inevitably restricted by other components.

[0004] So far, no one has been found to use parsnip stems as raw materials for extracting natural dyes and using them in dyeing cotton and linen fabrics. Summary of the invention

[0005] The present invention provides a method for extracting a natural dye from a parsnip stem, wherein the parsnip stem is crushed and sterilized, and then fermented and cultured using a homemade Trichoderma reesei (4000-20000UI)-cellulase-coenzyme (porphyrin iron-pyridoxal phosphate) promotion system, and then extracted using an ultrasonic microwave combined extraction instrument, and then cold-dried to obtain a dark yellow powder after solid phase extraction, and the natural dye from the parsnip stem is applied to dyeing silk fabrics. The method is specifically implemented by the following technologies.

[0006] The first aspect of the present invention provides a method for extracting natural dye from parsnip stems, comprising the following steps: S1, pre-treating the stalks of parsnips to prepare fibrillated fine powder of the stalks of parsnips; S2, adding the fine powder of parsnip stems to a citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value, and adding Trichoderma reesei (activity 4000-20000 IU) for fermentation to obtain a fermentation mixture; S3, mixing the fermentation mixture and water in a volume ratio of 1: (25-34), adding an enzymatic system, performing ultrasonic microwave extraction, centrifuging to obtain the supernatant, and filtering; repeating this process 3 times, and combining the filtrate; The enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:(4-12):(6-18); S4. Add sodium carbonate to the filtrate obtained in step S3, heat for reaction, perform solid phase extraction, filter and freeze-dry to obtain the fennel stem dye.

[0007] In some implementation cases of the present invention, in the above-mentioned method for extracting natural dye from wind grass stems, the specific method of pre-treating the wind grass stems to prepare the wind grass stem fibrillated fine powder can be selected as follows: (1) Pre-treating the stalks (stems, leaves, etc.) of the parsnips to obtain fibrillated fine powder of the stalks, adding the dried stalks to a grinder for crushing and grinding to obtain coarse stalk particles of about 40-80 meshes; and processing the stalks with a planetary ball mill to obtain stalk micro powder (about 200-400 meshes); (2) Take the above-mentioned grass stem powder, add lithium hydroxide, urea and water, homogenize at 8000r / min for 2 minutes, stir and suspend, and place in a low temperature box at -40℃ for 36 hours; (3) After suspension at room temperature, the suspension is placed in a dialysis membrane (MD44-3500, MD25-14000 or MD77-14000) and dialyzed in deionized water for 72 hours until the pH value reaches 6.6-7.4, thereby obtaining a suspension containing fibrillated fine powder of parsnips stems.

[0008] It is easy for a person skilled in the art to know that in order to improve the fermentation efficiency of Trichoderma reesei, Trichoderma reesei may be pre-activated in a suitable environment.

[0009] Optionally, in the method for pretreating parsnips stems, the mass ratio of parsnips stem powder, lithium hydroxide and urea can be (3.5-6):(7-9):(13-17).

[0010] Further optionally, the mass ratio of grass stem powder, lithium hydroxide and urea can be 5:8:15.

[0011] Furthermore, the Trichoderma reesei is Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio is (0.16-0.43):(0.24-0.44):(0.13-0.48).

[0012] Furthermore, in step S2, the amount of Trichoderma reesei added is 0.2-0.45 g / L.

[0013] Furthermore, in step S2, the fermentation temperature is 30-50°C, and the fermentation time is 4-8h.

[0014] Furthermore, in step S3, the total concentration of cellulase, iron porphyrin and pyridoxal phosphate in the enzymatic system is 0.3-0.8 g / L.

[0015] Furthermore, in step S3, the specific method of ultrasonic microwave extraction is: firstly ultrasonic extraction at 33-42°C and 60-180W for 45-60min, and then microwave extraction at 200-600W for 3-15min.

[0016] Furthermore, in step S4, the purpose of adding sodium carbonate is to adjust the pH value of the environment to a pH value of 8.4-9.8.

[0017] Furthermore, in step S4, the amount of sodium carbonate added is 0.2-0.5 g / L.

[0018] Furthermore, in step S4, the heating reaction is carried out by keeping the temperature at 50-70° C. for 60-90 min.

[0019] In some embodiments of the present invention, after the suction filtration in step S4, in order to further improve the purity of the parsnip stem dye, it can be further separated and purified, and then freeze-dried.

[0020] Optionally, the separation and purification method is: (1) Removal of protein The filtrate A was mixed with Sevag reagent (V 氯仿 :V 正丁醇 =(3-4):(1.1-0.8)) are mixed in equal proportions and stirred at room temperature for 30 minutes; After low-temperature centrifugation, the solution was introduced into a separatory funnel, the upper layer solution was collected, and the precipitate was removed by centrifugation. The supernatant was repeatedly centrifuged several times until no precipitate was generated; Add an appropriate volume (for example, 4 times the volume) of anhydrous ethanol to the resulting solution, let it stand for 24 hours, and centrifuge; take the precipitate and wash it with anhydrous ethanol, acetone, and ether respectively, then add distilled water to mix evenly, dialyze it with running water for 72 hours, and vacuum freeze-dry to obtain the preliminarily purified windflower stem dye A.

[0021] (2) Chromatographic separation Weigh the microcrystalline cellulose column material, activate it, wet pack it into a column (70 mm × 500 mm), and balance it with distilled water; Dissolve 0.2 g of parsnip stem dye A in 10 ml of deionized water, pass through a column, and elute with distilled water, NaCl solution (0.05 mol / L), NaCl solution (0.1 mol / L), NaCl solution (0.2 mol / L), NaCl solution (0.4 mol / L) and NaCl solution (0.05 mol / L) in sequence at an elution rate of 1 mL / min. Collect the fractions, concentrate and dialyze, and the resulting product can be subjected to subsequent freeze-drying operations.

[0022] The second aspect of the present invention provides an application of a natural dye made from parsnips stems. The natural dye made from parsnips stems prepared by the above extraction method is used for dyeing cotton, linen or silk fabrics.

[0023] Furthermore, the specific method of dyeing treatment is: P1. Dissolve chitosan in 5% citric acid solution, add non-ionic penetrant, and mix well; soak the fabric in the solution, dry and bake; P2, immersing the fabric treated in step P1 into a diluted solution of a natural dye of parsnip stems for dyeing; P3. The dyed fabric is subjected to post-treatment by a solvent, washed and dried to obtain a finished dyed fabric.

[0024] Furthermore, in step P2, the concentration of the natural dye of parsnip stems in the dilution liquid is 5 g / L, the pH value of the dilution liquid is 3-7; the dyeing temperature is 60-75° C., and the dyeing time is 30-60 min.

[0025] Furthermore, in step P3, the post-mordant treatment method is: immersing the dyed fabric in a mordant solution at a bath ratio of 1:(30-50) for mordanting, the mordant temperature is 60-80°C, and the mordant time is 30-45min; potassium aluminum sulfate is used as a mordant in the mordant solution, and the concentration is 5-15g / L.

[0026] Compared with the prior art, the invention is beneficial in that the extraction process of the sesame stem dye is simple, the extraction efficiency is high, the cost is low, and the market prospect is broad. The obtained sesame stem extract has bactericidal effects, is safe and environmentally friendly, has good compatibility with the ecological environment, and is biodegradable. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the infrared spectrum analysis diagram of the parsnip stem extract obtained in Example.

[0028] Figure 2 This is the H-NMR spectrum of the natural dye from parsnip stems extracted in Example 1. DETAILED DESCRIPTION

[0029] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] In some implementation cases of the present invention, a method for extracting natural dye from parsnip stems is provided, comprising the following steps: S1, pre-treating the stalks of parsnips to prepare fibrillated fine powder of the stalks of parsnips; S2, adding the fine powder of parsnips stems to a citric acid-disodium hydrogen phosphate buffer solution, and adding Trichoderma reesei for fermentation to obtain a fermentation mixture; S3, mixing the fermentation mixture and water in a volume ratio of 1: (25-34), adding an enzymatic system, performing ultrasonic microwave extraction, centrifuging to obtain the supernatant, and filtering; repeating this process 3 times, and combining the filtrate; The enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:(4-12):(6-18); S4. Add sodium carbonate to the filtrate obtained in step S3, heat for reaction, perform solid phase extraction, filter and freeze-dry to obtain the burdock dye.

[0031] In some embodiments of the present invention, in the above-mentioned method for extracting natural dye from wind grass stems, the specific method of pre-treating the wind grass stems to prepare the wind grass stem fibrillated fine powder can be selected as follows: (1) Pre-treating the stalks (stems, leaves, etc.) of the parsnips to obtain fibrillated fine powder of the stalks, adding the dried stalks to a grinder for crushing and grinding to obtain coarse stalk particles of about 40-80 meshes; and processing the stalks with a planetary ball mill to obtain stalk micro powder (about 200-400 meshes); (2) Take the above-mentioned grass stem powder, add lithium hydroxide, urea and water, homogenize at 8000r / min for 2 minutes, stir and suspend, and place in a low temperature box at -40℃ for 36 hours; (3) After suspension at room temperature, the suspension was placed in a dialysis membrane (MD44-3500, MD25-14000, MD77-14000), and dialyzed in deionized water for 72 hours until the pH value reached 6.6-7.4, thereby obtaining a suspension containing fibrillated fine powder of parsnips stems.

[0032] It is easy for a person skilled in the art to know that in order to further improve the fermentation efficiency of Trichoderma reesei, Trichoderma reesei may be pre-activated in a suitable environment.

[0033] Optionally, in the method for pretreating parsnips stems, the mass ratio of parsnips stem powder, lithium hydroxide and urea can be (3.5-6):(7-9):(13-17).

[0034] Further optionally, the mass ratio of grass stem powder, lithium hydroxide and urea can be 5:8:15.

[0035] In some embodiments of the present invention, the Trichoderma reesei is Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of these is (0.28-0.42):(0.24-0.44):(0.16-0.48).

[0036] Further specifically, the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 can be 0.28:0.36:0.4, 0.28:0.44:0.38, 0.35:0.36:0.29, 0.35:0.44:0.21, 0.43:0.36:0.21, 0.43:0.44:0.13, 0.32:0.24:0.48 or 0.16:0.36:0.48.

[0037] Furthermore, in step S2, the amount of Trichoderma reesei added is 0.2-0.45 g / L.

[0038] Furthermore, the addition amount of Trichoderma reesei can be 0.2 g / L, 0.25 g / L, 0.3 g / L, 0.35 g / L, 0.4 g / L or 0.45 g / L.

[0039] Furthermore, in step S2, the fermentation temperature is 30-50°C, and the fermentation time is 4-8h.

[0040] Furthermore, the fermentation temperature can be 30, 35, 40, 45 or 50°C, and the fermentation time can be 4-8h.

[0041] In some embodiments of the present invention, in step S3, the mass ratio of cellulase, iron porphyrin and pyridoxal phosphate is 100:(4-12):(6-18).

[0042] Furthermore, the mass ratio of cellulase, iron porphyrin and pyridoxal phosphate can be selected as: 100:4:6, 100:4:18, 100:12:6 or 100:12:18.

[0043] In some implementation cases of the present invention, in step S3, the total concentration of cellulase, iron porphyrin and pyridoxal phosphate in the enzymatic system is 0.3-0.8 g / L.

[0044] Further, the total concentration of cellulase, iron porphyrin and pyridoxal phosphate can be selected to be 0.3 g / L, 0.4 g / L, 0.5 g / L, 0.6 g / L, 0.7 g / L or 0.8 g / L.

[0045] In some implementation cases of the present invention, in step S3, the specific method of ultrasonic microwave extraction is: first 33-42°C, 60-180W ultrasonic extraction for 45-60min, and then 200-600W microwave extraction for 3-15min.

[0046] Furthermore, the temperature of ultrasonic extraction can be selected to be 33°C, 35°C, 37°C, 40°C or 42°C.

[0047] Furthermore, the power of ultrasonic extraction can be selected as 60W, 70W, 80W, 90W, 100W, 110W, 120W, 130W, 140W, 150W, 160W, 170W or 180W.

[0048] Furthermore, the ultrasonic extraction time can be selected to be 45 min, 50 min, 55 min or 60 min.

[0049] Furthermore, the power of microwave extraction can be selected as 200W, 250W, 300W, 350W, 400W, 450W, 500W, 550W or 600W.

[0050] Furthermore, the time of microwave extraction can be selected as 3 min, 4 min, 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, 11 min, 12 min, 13 min, 14 or 15 min.

[0051] In some implementation cases of the present invention, in step S4, the purpose of adding sodium carbonate is to adjust the pH value of the environment to a pH value of 8.4-9.8.

[0052] Furthermore, in step S4, the amount of sodium carbonate added is 0.2-0.5 g / L.

[0053] Furthermore, the amount of sodium carbonate added is 0.2g / L, 0.25g / L, 0.3g / L, 0.35g / L, 0.4g / L, 0.45g / L or 0.5g / L.

[0054] In some implementation cases of the present invention, in step S4, the heating reaction is carried out by keeping the temperature at 50-70° C. for 60-90 min.

[0055] In some embodiments of the present invention, after the suction filtration in step S4, in order to further improve the purity of the parsnip stem dye, it can be further separated and purified, and then freeze-dried.

[0056] Optionally, the separation and purification method is: (1) Removal of protein The filtrate A was mixed with Sevag reagent (V 氯仿 :V 正丁醇 =(3-4):(1.1-0.8)) in equal proportions and stirred at room temperature for 30 minutes; After low-temperature centrifugation, the solution was introduced into a separatory funnel, the upper layer solution was collected, and the precipitate was removed by centrifugation. The supernatant was repeatedly centrifuged several times until no precipitate was generated; Add an appropriate volume (for example, 4 times the volume) of anhydrous ethanol to the resulting solution, let it stand for 24 hours, and centrifuge; take the precipitate and wash it with anhydrous ethanol, acetone, and ether respectively, then add distilled water to mix evenly, dialyze it with running water for 72 hours, and vacuum freeze-dry to obtain the preliminarily purified windflower stem dye A.

[0057] (2) Chromatographic separation Weigh the microcrystalline cellulose column material, activate it, wet pack it into a column (70 mm × 500 mm), and balance it with distilled water; Dissolve 0.2 g of parsnip stem dye A in 10 ml of deionized water, pass through a column, and elute with distilled water, NaCl solution (0.05 mol / L), NaCl solution (0.1 mol / L), NaCl solution (0.2 mol / L), NaCl solution (0.4 mol / L) and NaCl solution (0.05 mol / L) in sequence at an elution rate of 1 mL / min. Collect the fractions, concentrate and dialyze, and the resulting product can be subjected to subsequent freeze-drying operations.

[0058] Example 1 The method for extracting the natural dye from the stalks of parsnip used in this embodiment comprises the following steps: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, adding 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value to 6.4-6.8, adding Trichoderma reesei (4000-20000IU) for fermentation (fermentation temperature 40°C, fermentation time 5h), and obtaining a fermentation mixture, wherein the concentration of Trichoderma reesei is 0.4g / L; the Trichoderma reesei is composed of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of the three is 0.16:0.36:0.48; S3, mixing the fermentation mixture and water in a volume ratio of 1:25, adding an enzymatic system, wherein the enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:4:6, and the total concentration of the three is 0.5 g / L; Ultrasonic microwave extraction (first 40°C, 100W ultrasonic extraction for 60min, then 500W microwave extraction for 10min), centrifugal separation, supernatant, filtration; cycle this process 3 times, and combine the filtrate; S4. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 60° C. for 30 min, perform 12-phase solid phase extraction, and filter the solution through a 0.22 μm microporous filter membrane at room temperature. Freeze-dry the filtered liquid to obtain a parsnip stem dye.

[0059] It was determined that the extraction amount of the parsnip stem dye obtained in this example was 61.72 mg / kg.

[0060] Example 2 The method for extracting the natural dye from the stalks of parsnip used in this embodiment comprises the following steps: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, adding 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value to 6.4-6.8, adding Trichoderma reesei (4000-20000IU) for fermentation (fermentation temperature 30°C, fermentation time 8h), and obtaining a fermentation mixture, wherein the concentration of Trichoderma reesei is 0.2g / L; the Trichoderma reesei is composed of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of the three is 0.28:0.44:0.38; S3, mixing the fermentation mixture and water in a volume ratio of 1:25, adding an enzymatic system, wherein the enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:4:18, and the total concentration of the three is 0.8 g / L; Ultrasonic microwave extraction (first 33°C, 180W ultrasonic extraction for 45min, then 200W microwave extraction for 15min), centrifugal separation, take the supernatant, filter; cycle this process 3 times, and combine the filtrate; S4. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 50°C for 90 minutes, perform 12-phase solid phase extraction, and filter the mixture through a 0.22 μm microporous filter membrane at room temperature. Freeze-dry the filtered liquid to obtain a parsnip stem dye.

[0061] It was determined that the extraction amount of the parsnip stem dye obtained in this example was 54.21 mg / kg.

[0062] Example 3 The method for extracting the natural dye from the stalks of parsnip used in this embodiment comprises the following steps: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, adding 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value to 6.4-6.8, adding Trichoderma reesei (4000-20000IU) for fermentation (fermentation temperature 50°C, fermentation time 4h), and obtaining a fermentation mixture, wherein the concentration of Trichoderma reesei is 0.45g / L; the Trichoderma reesei is composed of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of the three is 0.35:0.36:0.29; S3, mixing the fermentation mixture and water in a volume ratio of 1:25, and adding an enzymatic system, wherein the enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:12:18, and the total concentration of the three is 0.3 g / L; Ultrasonic microwave extraction (first 33°C, 180W ultrasonic extraction for 45min, then 200W microwave extraction for 15min), centrifugal separation, take the supernatant, filter; cycle this process 3 times, and combine the filtrate; S4. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 50°C for 90 minutes, perform 12-phase solid phase extraction, and filter the mixture through a 0.22 μm microporous filter membrane at room temperature. Freeze-dry the filtered liquid to obtain a parsnip stem dye.

[0063] The extraction amount of the burdock stem dye obtained in this example was measured to be 78.65 mg / kg.

[0064] Example 4 The method for extracting the natural dye from the stalks of parsnip used in this embodiment comprises the following steps: S1, pre-treating the stalks of parsnips to prepare fibrillated fine powder of the stalks of parsnips; S1-1, pre-treating the stalks (stems, leaves, etc.) of the parsnips to prepare fibrillated fine powder of the stalks, adding the dried stalks to a grinder for crushing and grinding to prepare coarse stalk particles of about 40-80 meshes; and preparing stalk micro powder (about 200-400 meshes) after treatment with a planetary ball mill; S1-2, take the above-mentioned grass stem powder, add lithium hydroxide, urea and water, homogenize at 8000r / min for 2min, stir and suspend, and place in a low temperature box at -40℃ for 36h; S1-3. After suspension at room temperature, the suspension was placed in a dialysis membrane (MD44-3500, MD25-14000, MD77-14000), and placed in deionized water for running water dialysis for 72 hours until the pH value was 6.6-7.4, to obtain a suspension containing fibrillated fine powder of parsnips stems.

[0065] S2, adding 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value to 6.4-6.8, adding Trichoderma reesei (4000-20000IU) for fermentation (fermentation temperature 50°C, fermentation time 4h), and obtaining a fermentation mixture, wherein the concentration of Trichoderma reesei is 0.45g / L; the Trichoderma reesei is composed of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of the three is 0.43:0.44:0.13; S3, mixing the fermentation mixture and water in a volume ratio of 1:25, and adding an enzymatic system, wherein the enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:12:18, and the total concentration of the three is 0.3 g / L; Ultrasonic microwave extraction (first 33°C, 180W ultrasonic extraction for 45min, then 200W microwave extraction for 15min), centrifugal separation, take the supernatant, filter; cycle this process 3 times, and combine the filtrate; S4. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 50°C for 90 minutes, perform 12-phase solid phase extraction, and filter the solution through a 0.22-μm microporous filter membrane at room temperature. Separate and purify the filtered liquid, and freeze-dry it to obtain the fennel stem dye.

[0066] The separation and purification method is: S4-1. Protein removal The filtrate A was mixed with Sevag reagent (V 氯仿 :V 正丁醇 =(3-4):(1.1-0.8)) in equal proportions and stirred at room temperature for 30 minutes; After low-temperature centrifugation, the solution was introduced into a separatory funnel, the upper layer solution was collected, and the precipitate was removed by centrifugation. The supernatant was repeatedly centrifuged several times until no precipitate was generated; Add an appropriate volume (for example, 4 times the volume) of anhydrous ethanol to the resulting solution, let it stand for 24 hours, and centrifuge; take the precipitate and wash it with anhydrous ethanol, acetone, and ether respectively, then add distilled water to mix evenly, dialyze it with running water for 72 hours, and vacuum freeze-dry to obtain the preliminarily purified windflower stem dye A.

[0067] S4-2, Chromatographic Separation Weigh the microcrystalline cellulose column material, activate it, wet pack it into a column (70 mm × 500 mm), and balance it with distilled water; 0.2 g of parsnip stem dye A was dissolved and dispersed in 10 ml of deionized water, passed through a column, and eluted with distilled water, NaCl solution (0.05 mol / L), NaCl solution (0.1 mol / L), NaCl solution (0.2 mol / L), NaCl solution (0.4 mol / L) and NaCl solution (0.05 mol / L) in sequence at an elution rate of 1 mL / min. The fractions were collected, concentrated and dialyzed. The obtained product is freeze-dried to obtain the parsnip stem dye.

[0068] According to the measurement, the extraction amount of the parsnip stem dye obtained in this example is 82.33 mg / kg.

[0069] Comparative Example 1 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 1, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.2:0.55:0.25.

[0070] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 42.61 mg / kg.

[0071] Comparative Example 2 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 2, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.6:0.2:0.2.

[0072] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 53.42 mg / kg.

[0073] Comparative Example 3 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 3, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.15:0.15:0.7.

[0074] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 47.87 mg / kg.

[0075] Comparative Example 4 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 4, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.55:0.15:0.3.

[0076] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 51.47 mg / kg.

[0077] Comparative Example 5 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 4, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.22:0.52:0.26.

[0078] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 46.86 mg / kg.

[0079] Comparative Example 6 The method for extracting the natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 1, except that the mass ratio of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932 is 0.54:0.16:0.3.

[0080] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 50.97 mg / kg.

[0081] Comparative Example 7 The method for extracting the natural dye from the stalks of parsnips provided in this comparative example is basically the same as that in Example 1, except that the mass ratio of cellulase, iron porphyrin and pyridoxal phosphate is 100:2:20.

[0082] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 55.42 mg / kg.

[0083] Comparative Example 8 The method for extracting the natural dye from the stalks of parsnips provided in this comparative example is basically the same as that in Example 1, except that the mass ratio of cellulase, iron porphyrin and pyridoxal phosphate is 100:14:4.

[0084] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 58.91 mg / kg.

[0085] Comparative Example 9 The method for extracting natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 1, except that in step S3, the enzymatic system contains only cellulase and iron porphyrin, and the mass ratio of the two is 100:10.

[0086] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 49.24 mg / kg.

[0087] Comparative Example 10 The method for extracting natural dye from parsnip stems provided in this comparative example is basically the same as that in Example 1, except that in step S3, the enzymatic system contains only cellulase and pyridoxal phosphate, and the mass ratio of the two is 100:15.

[0088] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 47.23 mg / kg.

[0089] Comparative Example 11 Compared with Example 1, the method for extracting the natural dye from the stalk of parsnip provided in this comparative example does not involve the fermentation treatment with Trichoderma reesei, nor the enzymatic system decomposition. The specific method is as follows: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, add 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjust the pH value to 6.4-6.8, perform ultrasonic microwave extraction (first 33°C, 180W ultrasonic extraction for 45min, then 200W microwave extraction for 15min), centrifuge to obtain the supernatant, filter; repeat this process 3 times, and combine the filtrate; S3. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 50° C. for 90 min, perform 12-phase solid phase extraction, and filter the mixture through a 0.22 μm microporous filter membrane at room temperature. Freeze-dry the filtered liquid to obtain a parsnip stem dye.

[0090] It was determined that the extraction amount of the burdock stem dye obtained in this comparative example was 17.76 mg / kg.

[0091] Comparative Example 12 The method for extracting the natural dye from the stalk of parsnip provided in this comparative example, compared with Example 1, only performs the fermentation treatment with Trichoderma reesei, and does not perform the enzymatic system decomposition. The specific method is as follows: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, adding 5g of fine powder of parsnip stem to 50g of citric acid-disodium hydrogen phosphate buffer solution, adjusting the pH value to 6.4-6.8, adding Trichoderma reesei (4000-20000IU) for fermentation (fermentation temperature 40°C, fermentation time 5h), and obtaining a fermentation mixture; the concentration of Trichoderma reesei is 0.4g / L, and the Trichoderma reesei is composed of Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio of the three is 0.16:0.36:0.48; S3, mixing the fermentation mixture and water at a volume ratio of 1:25, performing ultrasonic microwave extraction (first 40°C, 100W ultrasonic extraction for 60min, then 500W microwave extraction for 10min), centrifuging, taking the supernatant, filtering; repeating this process 3 times, and combining the filtrate; S4. Add sodium carbonate (concentration 0.5 g / L) to the filtrate obtained in step S3, heat the reaction at 50°C for 90 minutes, perform 12-phase solid phase extraction, and filter the mixture through a 0.22 μm microporous filter membrane at room temperature. Freeze-dry the filtered liquid to obtain a parsnip stem dye.

[0092] It was determined that the extraction amount of the parsnip stem dye obtained in this example was 33.54 mg / kg.

[0093] Comparative Example 13 The method for extracting the natural dye from the stalk of parsnip provided in this comparative example, compared with Example 1, does not perform the Trichoderma reesei fermentation treatment, but only performs the enzymatic system decomposition. The specific method is as follows: S1. Pre-treating the stalks of parsnips (washing, air-drying, hot-drying, 400-600 mesh freeze-grinding, sterilizing at 124°C for 30 min) to prepare fibrillated fine powder of the stalks of parsnips; S2, 5g of fine powder of parsnip stems was mixed with citric acid-disodium hydrogen phosphate buffer solution at a ratio of 1:25, the pH value was adjusted to 6.4-6.8, and an enzymatic system was added, wherein the enzymatic system contained cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three was 100:4:6, and the total concentration of the three was 0.5g / L; Ultrasonic microwave extraction (first 40°C, 100W ultrasonic extraction for 60min, then 500W microwave extraction for 10min), centrifugal separation, supernatant, filtration; cycle this process 3 times, and combine the filtrate; S3. Add sodium carbonate (concentration 0.5 g / L) to the obtained filtrate, heat the reaction at 50°C for 90 min, and after 12-phase solid phase extraction, use a 0.22-μm microporous filter membrane to filter at room temperature, and freeze-dry the filtered liquid to obtain the wind grass stem dye.

[0094] It was determined that the extraction amount of the parsnip stem dye obtained in this example was 37.35 mg / kg.

[0095] Test Example 1: Characterization and Analysis of Parsnip Stem Dye The parsnip stem dye prepared in Example 1 was characterized and analyzed. Its infrared spectrum analysis chart and H 1 -NMR spectra are as follows Figure 1 , 2 shown.

[0096] By comparing with the literature, the water-soluble components identified in the stems of the herb are mainly chromone components such as cimifugin and hamaudol, and the structures are shown below.

[0097]

[0098] There are also some water-soluble coumarins such as nodakenetin, scopoletin and deltoin, the structures of which are shown below.

[0099]

[0100] The prepared parsnip stem dye was subjected to infrared spectroscopy analysis. Figure 1 FTIR ν cm -1 (KBr) 3417.52 (OH), 2952.83, 2933.41 (CH2), 2856.44 (CH3), 1646.40 (C=C), 1464.36 (CH2), 1371.47 (CH3), 1157.96, 1081.64, (C-O), 1019.38, 992.46, 800.16 (=CH), 674.291 and 599.57, which are basically consistent with the structures of chelidonol and coumarin in parsnip stems.

[0101] like Figure 2 The NMR spectrum of the natural dye from parsnip stems is shown. The dye is yellow-brown needle-shaped crystals with a molecular formula of C 16 H 18 O6, molecular weight 306. 1 HNMR(CDCl3)δ 7.35 (1H, d, J=9.6Hz, H-3), 7.26 (1H, s, H-8), 6.98 (1H, s, H-5), 5.06 (1H, t, J =9.0Hz, H-2'), 4.08 (2H, s, H-3'), 2.84 (3H, s, -OCH3), 2.60 (2.79 (2H, br.s, C 4’ -OH and C2-CH2OH), 1.29 (3H, s, 4'-CH3). 1Comprehensive analysis of HNMR data suggested that the compound was scopoletin.

[0102] Test Example 2: Evaluation of the dyeing effect of the natural dye of parsnip stems prepared in Example 1 on pure cotton woven fabrics Pure cotton woven fabrics are selected for dyeing, specifically: (1) Dissolve 12 g / L chitosan (85% deacetylation degree) in 5% citric acid solution, add non-ionic penetrant JFC at a concentration of 1 g / L, soak the pure cotton fabric in the chitosan solution, and treat it by padding. The specific treatment process is two dips and two pads, drying at 100 °C, and baking at 125 °C for 3 min.

[0103] (2) adding the dried parsnip stem natural dye into deionized water to dilute it to a concentration of 5 g / L to obtain a uniform dilution solution, adjusting the pH of the dilution solution to 3-7, and then immersing the pre-treated cotton and linen woven fabrics into the dilution solution to dye the fabrics at 70°C for 60 min; (3) After dyeing, the fabric is treated with potassium aluminum sulfate, washed and dried to obtain the dyed fabric.

[0104] The post-mordant treatment process is: bath ratio 1:50, mordant dosage 10g / L, mordant temperature 80℃, and mordant time 45min.

[0105] The dyed pure cotton fabric was subjected to dyeing colorimetry test after dyeing, and the test results are shown in Table 1 below.

[0106] Table 1 Colorimetric test results of dyeing

[0107] From Table 1 above, it can be seen that the dye dilution ratio has a certain influence on the color characteristic value of the extracted parsnip stem dye on cotton and linen woven fabrics. With the increase of the dilution ratio, the L lightness value slowly increases, the color concentration weakens, the b value and the K / S value gradually decrease, and the color saturation changes to a certain extent. The color performance is mainly a transition from yellow-brown to light yellow; after the dilution ratio is 1:50, the color characteristic value decreases significantly. It can be seen that in the absence of a mordant, the extracted parsnip stem dye is dyed on cotton and linen woven fabrics. With the increase of the dye dilution ratio, the apparent color yield of the cotton and linen fabric gradually decreases. After the dye dilution ratio is 1:50, the color yield of the fabric decreases and the fabric is light yellow.

[0108] The above specific embodiments describe the implementation of the present invention in detail, but the present invention is not limited to the specific details in the above embodiments. Within the scope of the claims and technical concept of the present invention, the technical solution of the present invention can be modified and changed in many simple ways, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. A method for extracting natural dye from parsnip stems, characterized in that: The following steps are involved: S1, pre-treating the stalks of parsnips to prepare fibrillated fine powder of the stalks of parsnips; S2, adding the fine powder of parsnips stems to a citric acid-disodium hydrogen phosphate buffer solution, and adding Trichoderma reesei for fermentation to obtain a fermentation mixture; S3, mixing the fermentation mixture and water in a volume ratio of 1: (25-34), adding an enzymatic system, performing ultrasonic microwave extraction, centrifuging to obtain the supernatant, and filtering; repeating this process 3 times, and combining the filtrate; The enzymatic system contains cellulase, iron porphyrin and pyridoxal phosphate, and the mass ratio of the three is 100:(4-12):(6-18); S4. Add sodium carbonate to the filtrate obtained in step S3, heat for reaction, perform solid phase extraction, filter and freeze-dry to obtain the burdock dye.

2. The method for extracting natural dye from parsnip stems according to claim 1, characterized in that: The Trichoderma reesei are Trichoderma reesei CICC 2626, Trichoderma reesei 41494 and Trichoderma reesei CICC40932, and the mass ratio is (0.16-0.43):(0.24-0.44):(0.13-0.48).

3. The method for extracting natural dye from parsnip stems according to claim 1, characterized in that: In step S2, the added amount of Trichoderma reesei is 0.2-0.45 g / L.

4. The method for extracting natural dye from parsnip stems according to claim 1, characterized in that: In step S2, the fermentation temperature is 30-50°C and the fermentation time is 4-8h.

5. The method for extracting natural dye from parsnip stems according to claim 1, characterized in that: In step S3, the total concentration of cellulase, iron porphyrin and pyridoxal phosphate in the enzymatic system is 0.3-0.8 g / L.

6. The method for extracting natural dye from parsnip stems according to claim 1, characterized in that: In step S3, the specific method of ultrasonic microwave extraction is: firstly ultrasonic extraction at 33-42°C and 60-180W for 45-60min, and then microwave extraction at 200-600W for 3-15min.

7. An application of a natural dye made from parsnip stems, characterized in that: The natural dye of parsnip stems is prepared by the extraction method according to any one of claims 1 to 6 and is used for dyeing cotton, linen or silk fabrics.

8. The use according to claim 7, characterized in that: The specific method of dyeing treatment is: P1. Dissolve chitosan in 5% citric acid solution, add non-ionic penetrant, and mix well; soak the fabric in the solution, dry and bake; P2, immersing the fabric treated in step P1 into a diluted solution of a natural dye of parsnip stems for dyeing; P3. The dyed fabric is subjected to post-treatment by a solvent, washed and dried to obtain a finished dyed fabric.

9. The use according to claim 8, characterized in that: In step P2, the concentration of the natural dye of parsnip stems in the diluent is 5 g / L, the pH value of the diluent is 3-7, the dyeing temperature is 60-75° C., and the dyeing time is 30-60 min.

10. The use according to claim 8, characterized in that: In step P3, the post-mordant treatment method is: immersing the dyed fabric in a mordant solution at a bath ratio of 1:(30-50) for mordanting, the mordant temperature is 60-80° C., and the mordant time is 30-45 min; potassium aluminum sulfate is used as a mordant in the mordant solution, and the concentration is 5-15 g / L.

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