Method for capturing hickory nut byproduct polyphenol through combination of solid-phase extraction instrument sample adsorption and probe type cell crusher treatment
By combining solid-phase extraction sample adsorption with a probe-type cell disruptor, the problem of poor separation and purification of polyphenols from pecan by-products was solved, the total phenol content and antioxidant capacity of the polyphenol extract were improved, and efficient recycling of waste was achieved.
Patent Information
- Application Number
- CN202510918992.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-04
- Publication Date
- 2025-11-07
AI Technical Summary
Existing technologies for separating and purifying polyphenols from pecan byproducts suffer from poor separation efficiency and low purity. In particular, the large molecular weight, complex structure, and active chemical properties of polyphenols lead to unclear objectives during the separation and purification process.
A polyphenol extract from pecan byproducts was prepared by combining solid-phase extraction (SPE) with sample adsorption and a probe-type cell disruptor. The process involved adsorption by the SPE and vibration treatment by the probe-type cell disruptor, followed by crude ethanol extraction, centrifugation, and vacuum filtration.
The method improved the total phenol and total flavonoid content of polyphenol extracts from pecan by-products, enhanced in vitro antioxidant capacity, achieved efficient separation and purification, and recycled waste from pecan processing.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of plant secondary metabolite extraction and separation, and particularly relates to a method for capturing polyphenols in hickory byproduct (hickory seed coat, hickory shell, hickory hull) by using solid phase extraction instrument sample adsorption combined with probe type cell crusher. BACKGROUND
[0002] Hickory is originally from North America and is widely planted in Zhejiang Province of China. In recent years, the market scale of hickory is continuously expanding. The edible part of hickory accounts for only about 25%, and the inedible byproduct (hickory seed coat, hickory shell, hickory hull, etc.) contains phenolic acid, flavonoids and condensed tannins composed of different degrees of polymerization, and many other phenolic substances. These phenolic substances have many biological activities such as antibacterial, anti-inflammatory, antitumor and antioxidant.
[0003] At present, there are some realistic problems in the separation of plant polyphenols, such as poor separation effect and low purity. Domestic and foreign scholars mainly focus on the research on separation and purification methods such as chromatography, membrane separation and metal ion precipitation. However, due to the large molecular weight and complex structure of polyphenols, the chemical properties are relatively active, and there are still problems such as low separation purity and unclear separation target in the process of separation and purification. The solid phase extraction instrument sample adsorption technology is a separation technology that uses the distribution of the extracted substance between the solid phase and the liquid phase combined with the action of the external air pump for sample treatment. The target component can be selectively separated from other compounds that may interfere with the analysis by using the force between the functional groups on the adsorbent and the functional groups of the target compound. The complex matrix effect can be removed. At present, the solid phase extraction instrument sample adsorption technology is mainly used for target cleaning, concentration and fractionation of analytes, and is widely used in the fields of medicine, environment, pharmacy and food. There are many types of polyphenols, and the properties or structures are similar. Therefore, the use of a single method is not enough to achieve the ideal separation and purification effect. Therefore, the combined use of multiple separation technologies will play a greater role. The probe type cell crusher increases the mass transfer by cavitation generated by a high-energy radial focusing vibrator. On the one hand, the vibration treatment process can cause the cell wall of the plant to break, thereby weakening the limitation of the cell structure on the mass transfer; on the other hand, the explosive collapse of the gas bubbles in the liquid-solid extraction produces local pressure, reduces the diffusion boundary layer, and enhances the interaction between the components in the cell and the solvent, thereby improving the extraction efficiency.
[0004] The present application innovatively uses hickory byproduct (hickory seed coat, hickory shell, hickory hull) as raw material to capture polyphenols in hickory byproduct (hickory seed coat, hickory shell, hickory hull) by using solid phase extraction instrument sample adsorption combined with probe type cell crusher, so as to prepare hickory byproduct (hickory seed coat, hickory shell, hickory hull) polyphenol extract with high total phenol content, total flavonoid content and in vitro antioxidant capacity. SUMMARY
[0005] The invention aims to provide a method for capturing polyphenols in pecan byproducts (pecan seed coat, pecan shell, pecan hull) by using a solid-phase extraction instrument sample adsorption combined with a probe-type cell crusher, so as to solve the problems in the prior art.
[0006] To achieve the above-mentioned purpose, the invention adopts the following technical solutions:
[0007] A method for capturing polyphenols in pecan byproducts by using a solid-phase extraction instrument sample adsorption combined with a probe-type cell crusher, comprising the following steps:
[0008] Step 1: raw material pretreatment: after the fresh pecan byproducts are cleaned, they are dried in a vacuum drying oven, crushed, and sieved.
[0009] Step 2: ethanol crude extraction: the pecan byproduct powder obtained in step 1 is mixed with ethanol, and ethanol crude extraction is performed by stirring; the solution after ethanol crude extraction is centrifuged, and the supernatant obtained by centrifugation is filtered under reduced pressure to obtain a crude polyphenol extract of pecan byproducts; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration are subjected to ethanol crude extraction, centrifugation, and reduced pressure filtration again; the crude polyphenol extracts of pecan byproducts are combined; and the crude polyphenol extract of pecan byproducts is concentrated by reduced pressure rotary evaporation.
[0010] Step 3: sample adsorption: the solid-phase extraction instrument is adsorbed under the action of an external air pump; methanol is used to activate the solid-phase extraction instrument, and then HCl is added to remove the methanol; the crude polyphenol extract of pecan byproducts obtained in step 2 is added to the activated solid-phase extraction instrument for adsorption; then HCl is added, and after it passes through, ethyl acetate is added to remove the unadsorbed impurities; finally, methanol is added to elute the adsorbed polyphenols, and the polyphenol extract of pecan byproducts is collected.
[0011] Step 4: probe-type cell crusher treatment: the polyphenol extract of pecan byproducts collected in step 3 is subjected to probe-type cell crusher treatment; the polyphenol extract of pecan byproducts treated by the probe-type cell crusher is concentrated by reduced pressure rotary evaporation to obtain a polyphenol extract of pecan byproducts.
[0012] Further, in step 1, the pecan byproducts include pecan seed coat, pecan shell, and pecan hull.
[0013] Further, in step 1, the fresh pecan byproducts are cleaned, dried in a vacuum drying oven at 35-45℃ for 40-50h, crushed, and sieved through a 40-80 mesh sieve.
[0014] Further, in step two, the hickory by-product powder obtained in step one is mixed with 40-70 v / v% ethanol at a solid-liquid ratio of 1 g:20-40 mL, and then placed in an electronic constant-temperature oil bath at 35-45℃ and stirred at 500-800 rpm for 1-2 h for ethanol crude extraction; the solution after ethanol crude extraction is centrifuged at 4-15℃ and 6000-8000 rpm for 15-30 min, and the supernatant obtained by centrifugation is filtered under reduced pressure at -0.05 --0.098 MPa to obtain a hickory by-product crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced-pressure filtration are subjected to 1-3 times of ethanol crude extraction, centrifugation and reduced-pressure filtration; and the hickory by-product crude polyphenol extracts obtained in 2-4 times are combined.
[0015] Further, in step two, the hickory by-product powder obtained in step one is mixed with 40-70 v / v% ethanol at a solid-liquid ratio of 1 g:20-40 mL, and then placed in an electronic constant-temperature oil bath at 35-45℃ and stirred at 500-800 rpm for 1-2 h for ethanol crude extraction; the solution after ethanol crude extraction is centrifuged at 4-15℃ and 6000-8000 rpm for 15-30 min, and the supernatant obtained by centrifugation is filtered under reduced pressure at -0.05 --0.098 MPa to obtain a hickory by-product crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced-pressure filtration are subjected to 1-3 times of ethanol crude extraction, centrifugation and reduced-pressure filtration; and the hickory by-product crude polyphenol extracts obtained in 2-4 times are combined.
[0016] Further, in step three, the solid-phase extraction instrument is adsorbed under the action of an external air pump, the air flow rate of the external air pump is 0.5-10 L / min, the working pressure is 0.1-0.5 MPa, the adsorption rate of the solid-phase extraction instrument is 1-5 mL / min, and the adsorption temperature is 20-30℃.
[0017] Further, in step three, the solid-phase extraction instrument is adsorbed under the action of an external air pump, the air flow rate of the external air pump is 0.5-10 L / min, the working pressure is 0.1-0.5 MPa, the adsorption rate of the solid-phase extraction instrument is 1-5 mL / min, and the adsorption temperature is 20-30℃.
[0018] Further, in step four, the hickory by-product polyphenol extract collected in step three is treated by a probe-type cell disrupter, the probe diameter is 25 mm, the excitation voltage is 80-120 V, the vibration frequency is 20-25 kHz, the vibration amplitude is 10-70%, and the excitation time is 5-30 min, wherein, on 1 s off 1 s, and the treatment is carried out under ice bath.
[0019] Further, in step four, the vibration amplitude is 40%, and the excitation time is 10 min.
[0020] Further, in step four, the hickory by-product polyphenol extract liquid treated by the probe type cell crusher is concentrated to 10%-20% of the original volume under the condition of 30-42 DEG C and-0.05--0.098 MPa by rotary evaporation under reduced pressure to obtain the hickory by-product polyphenol extract.
[0021] The beneficial effects of the present application are as follows:
[0022] The present application uses hickory by-products (hickory seed coat, hickory shell, hickory hull) as raw materials, and prepares hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract through sample adsorption by a solid phase extraction instrument and treatment by a probe type cell crusher with high energy delivery efficiency, and optimizes the preparation process conditions, so that the prepared hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract has high total phenol content, total flavonoid content and in vitro antioxidant capacity, and the preparation operation is simple and the purification efficiency is high. The present application recycles the waste (hickory seed coat, hickory shell, hickory hull) in the hickory processing process, provides a reference for evaluating the overall value of hickory, and has great significance for promoting the transformation of hickory by-products into high-value products. DETAILED DESCRIPTION
[0023] The present application will be further described below in conjunction with specific examples, but the present application is not limited by the following examples.
[0024] The hickory seed coat, hickory shell and hickory hull involved in the following examples are provided by Lin'an Chenghe Agricultural Product Processing Factory.
[0025] The solid phase extraction instrument involved in the following examples is purchased from Hunan Bikeman Biotechnology Co., Ltd., the solid phase extraction column filler is C18 (octadecyl), the filler gram weight is 500 mg, and the column tube capacity is 6 mL; the probe type cell crusher is purchased from Ningbo Xinzhi Biological Technology Co., Ltd., and the model is JY99-IIDN.
[0026] Example 1
[0027] Step one, raw material pretreatment: after the fresh hickory by-products (hickory seed coat, hickory shell and hickory hull, which are tested respectively) are cleaned, they are dried in a vacuum drying oven at 40 DEG C for 48 h, crushed and sieved through a 60 mesh sieve;
[0028] Step two, ethanol crude extraction: the hickory by-products (hickory seed coat, hickory shell, hickory hull) powder obtained from step one and ethanol (70 v / v%) were poured into a beaker (1 g: 20 mL of liquid), which was placed in an electronic constant temperature oil bath at 40℃, and ethanol crude extraction was carried out using an electric stirrer (600 rpm) for 1 h; the solution after ethanol crude extraction was centrifuged at 4℃, 6000 rpm for 20 min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to ethanol crude extraction, centrifugation and reduced pressure filtration for a second time; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts obtained in the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098 MPa.
[0029] Step three, sample adsorption: the solid phase extraction instrument was activated by an external air pump at a flow rate of 5 L / min and a working pressure of 0.2 MPa, and the adsorption rate of the solid phase extraction instrument was 1 mL / min at room temperature; 12 mL of methanol was first used to activate the solid phase extraction instrument, and then 18 mL of 0.01 v / v% HCl (0.01 v / v% HCl contains 0.01 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid is 36 wt% to 38 wt% concentrated hydrochloric acid; the same below) was added to remove methanol; 30 mL of the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract obtained in step two was then added to the activated solid phase extraction instrument for adsorption; then 30 mL of 0.01 v / v% HCl was added, and after it passed through, 18 mL of ethyl acetate was added to remove unadsorbed impurities; finally, 24 mL of methanol was added to elute the adsorbed polyphenols, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract was collected;
[0030] Step four, probe-type cell disruptor treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three was treated by a probe-type cell disruptor, with a probe diameter of 25 mm, an excitation voltage of 80 V, a vibration frequency of 22 kHz, a vibration amplitude of 20%, an excitation time of 10 min (on for 1 s and off for 1 s), and treatment in an ice bath; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by the probe-type cell disruptor was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098 MPa, and a hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract was obtained.
[0031] Example 2
[0032] Step one, raw material pretreatment: the fresh pecan by-products (pecan seed coat, pecan shell, pecan hull, pecan seed coat, pecan shell, pecan hull were tested respectively) were cleaned and dried in a vacuum drying oven at 40℃ for 48h, then crushed and sieved through a 60 mesh screen;
[0033] Step two, ethanol crude extraction: the pecan by-product (pecan seed coat, pecan shell, pecan hull) powder obtained in step one and ethanol (70v / v%) were poured into a beaker (1g:20mL of solid-liquid ratio), and placed in a 40℃ electronic constant temperature oil bath, using an electric mixer (600rpm) to stir for 1h for ethanol crude extraction; the ethanol crude extraction solution was centrifuged at 4℃, 6000rpm for 20min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098MPa) to obtain the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull); the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to 2 more times of ethanol crude extraction, centrifugation and reduced pressure filtration; the crude polyphenol extracts of the pecan by-product (pecan seed coat, pecan shell, pecan hull) obtained in the three times were combined; the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull) was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098MPa;
[0034] Step three, sample adsorption: the solid phase extraction instrument was activated by an external air pump at a flow rate of 5L / min and a working pressure of 0.2MPa, and the adsorption rate of the solid phase extraction instrument was 1mL / min at room temperature; 12mL of methanol was used to activate the solid phase extraction instrument, and then 18mL of 0.01v / v% HC1 (0.01v / v% HC1 means 0.01mL of concentrated hydrochloric acid in 100mL, and the concentrated hydrochloric acid is 36wt%-38wt% concentrated hydrochloric acid; the same below) was added to remove methanol; 30mL of the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull) obtained in step two was added to the activated solid phase extraction instrument for adsorption; then 30mL of 0.01v / v% HC1 was added, and after it passed through, 18mL of ethyl acetate was added to remove the unadsorbed impurities; finally, 24mL of methanol was added to elute the adsorbed polyphenols, and the pecan polyphenol extract was collected;
[0035] Step four, probe type cell crusher treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three was treated by probe type cell crusher, the probe diameter was 25 mm, the excitation voltage was 80 V, the vibration frequency was 22 kHz, the vibration amplitude was 20%, the excitation time was 20 min (on 1 s off 1 s), and the treatment was carried out under ice bath; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by probe type cell crusher was concentrated to 15% of the original volume under the conditions of 40℃ and -0.098 MPa reduced pressure rotary evaporation, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract was obtained.
[0036] Example 3
[0037] Step one, raw material pretreatment: the fresh hickory by-product (hickory seed coat, hickory shell, hickory hull, hickory seed coat, hickory shell, hickory hull were tested respectively) was cleaned and dried in a vacuum drying oven at 40℃ for 48h, then crushed and sieved through a 60 mesh sieve;
[0038] Step two, ethanol crude extraction: the hickory by-product (hickory seed coat, hickory shell, hickory hull) powder obtained in step one and ethanol (70v / v%) were poured into a beaker (solid-liquid ratio 1g:20mL), and the mixture was placed in a 40℃ electronic constant temperature oil bath, and an electric stirrer (600rpm) was used for stirring for 1h for ethanol crude extraction; the solution after ethanol crude extraction was centrifuged at 4℃ and 6000rpm for 20min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to 2 more times of ethanol crude extraction, centrifugation and reduced pressure filtration; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts of the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under the conditions of 40℃ and -0.098 MPa reduced pressure rotary evaporation;
[0039] Step three, sample adsorption: the solid phase extraction instrument is adsorbed under the action of an external air pump, the air flow rate of the external air pump is 5 L / min, the working pressure is 0.2 MPa, the adsorption rate of the solid phase extraction instrument is 1 mL / min, and the adsorption temperature is room temperature; 12 mL of methanol is used to activate the solid phase extraction instrument, then 18 mL of 0.01 v / v% HCl (0.01 v / v% HCl refers to 0.01 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid is 36 wt%-38 wt% concentrated hydrochloric acid; the same below) is added to remove methanol; 30 mL of the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract obtained in step two is added to the activated solid phase extraction instrument for adsorption; then 30 mL of 0.01 v / v% HCl is added, and after it passes through, 18 mL of ethyl acetate is added to remove unadsorbed impurities; finally, 24 mL of methanol is added to elute the adsorbed polyphenols, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is collected;
[0040] Step four, probe-type cell crusher treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three is treated by a probe-type cell crusher, the probe diameter is 25 mm, the excitation voltage is 80 V, the vibration frequency is 22 kHz, the vibration amplitude is 40%, the excitation time is 10 min (on for 1 s and off for 1 s), and the treatment is carried out in an ice bath; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by the probe-type cell crusher is concentrated to 15% of the original volume under reduced pressure at 40°C and -0.098 MPa, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is obtained.
[0041] Example 4
[0042] Step one, raw material pretreatment: the fresh hickory by-product (hickory seed coat, hickory shell, hickory hull, and the hickory seed coat, hickory shell, and hickory hull are tested respectively) is cleaned and dried in a vacuum drying oven at 40°C for 48 h, crushed, and sieved through a 60-mesh sieve;
[0043] Step two, ethanol crude extraction: the hickory by-products (hickory seed coat, hickory shell, hickory hull) powder obtained from step one and ethanol (70 v / v%) were poured into a beaker (1 g: 20 mL of liquid), which was placed in an electronic constant temperature oil bath at 40℃, and ethanol crude extraction was carried out using an electric stirrer (600 rpm) for 1 h; the solution after ethanol crude extraction was centrifuged at 4℃, 6000 rpm for 20 min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to ethanol crude extraction, centrifugation and reduced pressure filtration for a second time; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts obtained in the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098 MPa.
[0044] Step three, sample adsorption: the solid phase extraction instrument was activated by an external air pump at a flow rate of 5 L / min and a working pressure of 0.2 MPa, and the adsorption rate of the solid phase extraction instrument was 1 mL / min at room temperature; 12 mL of methanol was first used to activate the solid phase extraction instrument, and then 18 mL of 0.01 v / v% HCl (0.01 v / v% HCl contains 0.01 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid is 36 wt% to 38 wt% concentrated hydrochloric acid; the same below) was added to remove methanol; 30 mL of the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract obtained in step two was then added to the activated solid phase extraction instrument for adsorption; then 30 mL of 0.01 v / v% HCl was added, and after it passed through, 18 mL of ethyl acetate was added to remove unadsorbed impurities; finally, 24 mL of methanol was added to elute the adsorbed polyphenols, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract was collected;
[0045] Step four, probe-type cell disruptor treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three was treated by a probe-type cell disruptor, with a probe diameter of 25 mm, an excitation voltage of 80 V, a vibration frequency of 22 kHz, a vibration amplitude of 40%, and an excitation time of 20 min (1 s on and 1 s off) under ice bath treatment; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by the probe-type cell disruptor was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098 MPa to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract.
[0046] Example 5
[0047] Step one, raw material pretreatment: the fresh pecan by-products (pecan seed coat, pecan shell, pecan hull, pecan seed coat, pecan shell, pecan hull were tested respectively) were cleaned and dried in a vacuum drying oven at 40℃ for 48h, then crushed and sieved through a 60 mesh screen;
[0048] Step two, ethanol crude extraction: the pecan by-product (pecan seed coat, pecan shell, pecan hull) powder obtained in step one and ethanol (70v / v%) were poured into a beaker (1g:20mL of solid-liquid ratio), and placed in a 40℃ electronic constant temperature oil bath, using an electric mixer (600rpm) to stir for 1h for ethanol crude extraction; the ethanol crude extraction solution was centrifuged at 4℃, 6000rpm for 20min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098MPa) to obtain the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull); the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to 2 more times of ethanol crude extraction, centrifugation and reduced pressure filtration; the crude polyphenol extracts of the pecan by-product (pecan seed coat, pecan shell, pecan hull) obtained in the three times were combined; the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull) was concentrated to 15% of the original volume under reduced pressure at 40℃ and -0.098MPa;
[0049] Step three, sample adsorption: the solid phase extraction instrument was activated by an external air pump at a flow rate of 5L / min and a working pressure of 0.2MPa, and the adsorption rate of the solid phase extraction instrument was 1mL / min at room temperature; 12mL of methanol was used to activate the solid phase extraction instrument, and then 18mL of 0.01v / v% HC1 (0.01v / v% HC1 means 0.01mL of concentrated hydrochloric acid in 100mL, and the concentrated hydrochloric acid is 36wt%-38wt% concentrated hydrochloric acid; the same below) was added to remove methanol; 30mL of the crude polyphenol extract of the pecan by-product (pecan seed coat, pecan shell, pecan hull) obtained in step two was added to the activated solid phase extraction instrument for adsorption; then 30mL of 0.01v / v% HC1 was added, and after it passed through, 18mL of ethyl acetate was added to remove the unadsorbed impurities; finally, 24mL of methanol was added to elute the adsorbed polyphenols, and the pecan polyphenol extract was collected;
[0050] Step four, probe type cell crusher treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three was treated by probe type cell crusher, the probe diameter was 25 mm, the excitation voltage was 80 V, the vibration frequency was 22 kHz, the vibration amplitude was 60%, the excitation time was 10 min (on 1 s off 1 s), and the treatment was carried out under ice bath; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by probe type cell crusher was concentrated to 15% of the original volume under the conditions of 40°C and -0.098 MPa reduced pressure rotary evaporation, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract was obtained.
[0051] Example 6
[0052] Step one, raw material pretreatment: the fresh hickory by-product (hickory seed coat, hickory shell, hickory hull, hickory seed coat, hickory shell, hickory hull were tested respectively) was cleaned and dried in a vacuum drying oven at 40°C for 48 h, crushed, and sieved through a 60 mesh sieve;
[0053] Step two, ethanol crude extraction: the hickory by-product (hickory seed coat, hickory shell, hickory hull) powder obtained in step one and ethanol (70 v / v%) were poured into a beaker (solid-liquid ratio 1 g:20 mL), and the mixture was placed in a 40°C electronic constant temperature oil bath, and an electric stirrer (600 rpm) was used for stirring for 1 h for ethanol crude extraction; the solution after ethanol crude extraction was centrifuged at 4°C and 6000 rpm for 20 min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to 2 more times of ethanol crude extraction, centrifugation and reduced pressure filtration; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts of the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under the conditions of 40°C and -0.098 MPa reduced pressure rotary evaporation;
[0054] Step three, sample adsorption: the solid phase extraction instrument is adsorbed under the action of an external air pump, the air flow rate of the external air pump is 5 L / min, the working pressure is 0.2 MPa, the adsorption rate of the solid phase extraction instrument is 1 mL / min, and the adsorption temperature is room temperature; 12 mL of methanol is used to activate the solid phase extraction instrument, then 18 mL of 0.01 v / v% HCl (0.01 v / v% HCl refers to 0.01 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid is 36 wt%-38 wt% concentrated hydrochloric acid; the same below) is added to remove methanol; 30 mL of the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract obtained in step two is added to the activated solid phase extraction instrument for adsorption; then 30 mL of 0.01 v / v% HCl is added, and after it passes through, 18 mL of ethyl acetate is added to remove unadsorbed impurities; finally, 24 mL of methanol is added to elute the adsorbed polyphenols, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is collected;
[0055] Step four, probe-type cell crusher treatment: the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract collected in step three is treated by a probe-type cell crusher, the probe diameter is 25 mm, the excitation voltage is 80 V, the vibration frequency is 22 kHz, the vibration amplitude is 60%, the excitation time is 20 min (on for 1 s and off for 1 s), and the treatment is carried out in an ice bath; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract treated by the probe-type cell crusher is concentrated to 15% of the original volume under reduced pressure at 40°C and -0.098 MPa, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is obtained.
[0056] Comparative example 1
[0057] Step one, raw material pretreatment: the fresh hickory by-product (hickory seed coat, hickory shell, hickory hull, and the hickory seed coat, hickory shell, and hickory hull are tested respectively) is cleaned and dried in a vacuum drying oven at 40°C for 48 h, crushed, and sieved through a 60-mesh sieve;
[0058] Step two, ethanol crude extraction: the hickory by-products (hickory seed coat, hickory shell, hickory hull) powder obtained in step one and ethanol (70 v / v%) were poured into a beaker (1 g: 20 mL of solid-liquid ratio), which was placed in an electronic constant-temperature oil bath at 40°C, and ethanol crude extraction was performed using an electric stirrer (600 rpm) for 1 h; the solution after ethanol crude extraction was centrifuged at 4°C and 6000 rpm for 20 min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to ethanol crude extraction, centrifugation and reduced pressure filtration for a second time; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts obtained in the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under reduced pressure at 40°C and -0.098 MPa to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract.
[0059] Comparative example 2
[0060] Step one, raw material pretreatment: fresh hickory by-products (hickory seed coat, hickory shell, hickory hull, hickory seed coat, hickory shell and hickory hull were tested respectively) were cleaned and dried in a vacuum drying oven at 40°C for 48 h, crushed and sieved through a 60-mesh sieve;
[0061] Step two, ethanol crude extraction: the hickory by-products (hickory seed coat, hickory shell, hickory hull) powder obtained in step one and ethanol (70 v / v%) were poured into a beaker (1 g: 20 mL of solid-liquid ratio), which was placed in an electronic constant-temperature oil bath at 40°C, and ethanol crude extraction was performed using an electric stirrer (600 rpm) for 1 h; the solution after ethanol crude extraction was centrifuged at 4°C and 6000 rpm for 20 min, and the supernatant obtained by centrifugation was filtered under reduced pressure (-0.098 MPa) to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration were subjected to ethanol crude extraction, centrifugation and reduced pressure filtration for a second time; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extracts obtained in the three times were combined; the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract was concentrated to 15% of the original volume under reduced pressure at 40°C and -0.098 MPa to obtain a hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract.
[0062] Step three, sample adsorption: the solid phase extraction instrument is adsorbed under the action of an external air pump, the air flow rate of the external air pump is 5 L / min, the working pressure is 0.2 MPa, the adsorption rate of the solid phase extraction instrument is 1 mL / min, and the adsorption temperature is room temperature; 12 mL of methanol is used to activate the solid phase extraction instrument, then 18 mL of 0.01 v / v% HCl (0.01 v / v% HCl refers to 100 mL containing 0.01 mL of concentrated hydrochloric acid, and the concentrated hydrochloric acid is 36 wt%-38 wt% concentrated hydrochloric acid; the same below) is added to remove methanol; then 30 mL of the hickory by-product (hickory seed coat, hickory shell, hickory hull) crude polyphenol extract obtained in step two is added to the activated solid phase extraction instrument for adsorption; then 30 mL of 0.01 v / v% HCl is added, and after it passes through, 18 mL of ethyl acetate is added to remove unadsorbed impurities; finally, 24 mL of methanol is added to elute the adsorbed polyphenols, and the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is collected; the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is concentrated to 15% of the original volume under reduced pressure at 40°C and -0.098 MPa to obtain the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract.
[0063] The total phenol content, total flavonoid content, total antioxidant capacity (FRAP method), DPPH radical scavenging capacity, and ABTS radical scavenging capacity of the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract obtained in the above examples and comparative examples are determined.
[0064] The total phenol content determination method is as follows:
[0065] The total phenol content is determined by the Folin-Ciocalteu method. First, a 1 mg / mL gallic acid standard stock solution is prepared, and it is sequentially diluted to 100 μg / mL, 80 μg / mL, 60 μg / mL, 40 μg / mL, 20 μg / mL, and 10 μg / mL to prepare standard curve working solutions. Then, 0.5 mL of the gallic acid working solution and 0.5 mL of 50-fold diluted sample solution (i.e., the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract of each example and comparative example diluted with distilled water (1:50) (v / v)) are removed into a colorimetric tube, 2.5 mL of 10% (v / v) Folin phenol solution is added, mixed for 2 min, then 2 mL of 7.5% (w / v) Na2CO3 solution is added, and the reaction is carried out at 45°C for 20 min in the dark. Then, the absorbance value is determined at 760 nm using a UV-visible spectrophotometer. The gallic acid standard curve equation is y=10.339x-0.0199 (R2=0.9999), and the total phenol content of the sample is calculated according to the standard curve equation. 2= 0.9999), the total phenol content of the hickory by-products (hickory seed coat, hickory shell, hickory hull) of each example and the comparative example was calculated according to the gallic acid standard curve, and the total phenol content was expressed as gallic acid equivalent per gram of dry weight of sample (mg GAE / g DW).
[0066] The total flavonoid content determination method was as follows:
[0067] First, a rutin standard stock solution of 1 mg / mL was prepared and sequentially diluted to 100 μg / mL, 80 μg / mL, 60 μg / mL, 40 μg / mL, 20 μg / mL, and 10 μg / mL to prepare a standard curve working solution. Subsequently, 0.5 mL of the rutin working solution and 0.5 mL of a 50-fold diluted sample solution (i.e., the hickory by-products (hickory seed coat, hickory shell, hickory hull) polyphenol extract of each example and the comparative example diluted with distilled water (1:50 (v / v)) were removed into a cuvette, 0.3 mL of a 5% (w / v) NaNO3 solution and 0.3 mL of a 10% (w / v) AlCl3 solution were sequentially added, and light shielding reaction was performed for 6 min, respectively. Subsequently, 4 mL of a 4% (w / v) NaOH solution was added, and the volume was made up to 10 mL with methanol, and light shielding was performed for 15 min. Finally, the absorbance was determined at 510 nm using a UV-visible spectrophotometer. The rutin standard curve equation was y = 9.7069x + 0.0124 (R 2 = 0.999), the total flavonoid content of the hickory by-products (hickory seed coat, hickory shell, hickory hull) of each example and the comparative example was calculated according to the rutin standard curve, and the total flavonoid content was expressed as rutin equivalent per gram of dry weight of sample (mg RE / g DW).
[0068] The total antioxidant capacity (FRAP method) determination method was as follows:
[0069] First, TPTZ (10 mM), FeCl3 (20 mM), and acetate buffer (0.3 M, pH 3.6) were mixed at a volume ratio of 1:1:10 to react for 5 min at 37°C to obtain a FRAP working solution. 190 μL of the FRAP working solution and 10 μL of a 50-fold diluted sample solution (i.e., the hickory by-products (hickory seed coat, hickory shell, hickory hull) polyphenol extract of each example and the comparative example diluted with distilled water (1:50 (v / v)) were sequentially added to a 96-well plate, and after reaction at 37°C for 10 min, the absorbance of the above sample was determined at 593 nm by an enzyme marker. A FeSO4·7H2O solution was used to construct a standard curve. The total antioxidant capacity (FRAP method) was expressed as mmol Fe 2+ equivalent per gram of dry weight (mmol Fe 2+ / g DW).
[0070] The DPPH radical scavenging capacity determination method is as follows:
[0071] Test group: First, the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract of each example and comparative example was prepared into sample solutions of different concentrations (10, 20, 40, 80, 160 μg / mL) with methanol, 2 mL of each was mixed with DPPH methanol solution (0.1 mM) in equal volume, and reacted for 45 min in the dark. DPPH methanol solution plus equal volume of methanol was used as a blank control group, and sample solution plus equal volume of methanol (without DPPH) was used as a reagent blank group. After the reaction, the absorbance was measured at 517 nm, and the DPPH radical scavenging rate was calculated according to the following formula:
[0072]
[0073] Wherein, A0 is the absorbance of the blank control group at 517 nm, A1 is the absorbance of the reagent blank group at 517 nm, and A2 is the absorbance of the test group at 517 nm. The DPPH radical scavenging capacity of the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract is expressed as IC value (the concentration of the hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract when the DPPH radical scavenging rate is 50%, μg / mL).
[0074] The ABTS radical scavenging capacity determination method is as follows:
[0075] Test group: First, 7.0 mM ABTS solution and 2.45 mM potassium persulfate solution were mixed in equal volume, and reacted for 12 h in the dark to generate a stable ABTS radical solution. The ABTS radical solution was diluted with methanol to an initial absorbance of 0.70±0.02 at 734 nm. The hickory by-product (hickory seed coat, hickory shell, and hickory hull) polyphenol extract of each example and comparative example was prepared into sample solutions of different concentrations (10, 20, 40, 80, 160 μg / mL) with methanol, 0.2 mL of each was mixed with 2.0 mL of diluted ABTS radical solution, and mixed thoroughly, and then reacted for 6 min in the dark. 2.0 mL of diluted ABTS radical solution was mixed with 0.2 mL of methanol as a blank control group, and 0.2 mL of sample solution was mixed with 2.0 mL of methanol (without ABTS) as a reagent blank group. After the reaction, the absorbance was measured at 734 nm, and the scavenging rate was calculated according to the following formula:
[0076]
[0077] A0= A1- A2, wherein A0is the absorbance value of the blank control group at 734 nm, A1is the absorbance value of the reagent blank group at 734 nm, and A2is the absorbance value of the test group at 734 nm. The ABTS free radical scavenging capacity of the polyphenol extract of the hickory by-product (hickory seed coat, hickory shell, hickory hull) is expressed as an IC value (the concentration of the polyphenol extract of the hickory by-product (hickory seed coat, hickory shell, hickory hull) when the ABTS free radical scavenging rate is 50%, μg / mL).
[0078] Table 1 Comparison of effects of hickory seed coat examples and comparative examples
[0079]
[0080]
[0081] Table 2 Comparison of effects of hickory shell examples and comparative examples
[0082]
[0083] Table 3 Comparison of effects of hickory hull examples and comparative examples
[0084]
[0085] As shown in the above tables, it is found by comparing examples 1-6 that the polyphenols captured from hickory seed coat treated by the solid phase extraction instrument sample adsorption combined with the probe type cell crusher have the best total phenol content, total flavonoid content, total antioxidant capacity, DPPH free radical scavenging capacity and ABTS free radical scavenging capacity, which are 405.8 ± 10.3 mg GAE / g DW, 46.40 ± 0.57 mg RE / g DW, 76.10 ± 1.22 mmol Fe 2+ / g DW, 1.64 ± 0.13 μg / mL (IC50), and 4.87 ± 0.12 μg / mL (IC50), respectively. The physicochemical indicators of the polyphenols captured from hickory by-products (hickory seed coat, hickory shell, hickory hull) treated only by 70 v / v% ethanol extraction without solid phase extraction instrument sample adsorption combined with probe type cell crusher treatment are not good. In addition, it is found by comparing comparative example 2 with each example that the solid phase extraction instrument sample adsorption and the probe type cell crusher treatment have a synergistic effect, and the combination of the two can effectively improve the total phenol content, total flavonoid content and in vitro antioxidant capacity of the polyphenols captured from hickory by-products (hickory seed coat, hickory shell, hickory hull), wherein the vibration amplitude is 40% and the excitation time is 10 min, which is the best condition.
[0086] In summary, the present application takes hickory by-products (hickory seed coat, hickory shell, hickory hull) as raw materials, and prepares hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract through sample adsorption of a solid-phase extraction instrument and treatment of a probe type cell crusher with high energy delivery efficiency, and the preparation process conditions are optimized, the prepared hickory by-product (hickory seed coat, hickory shell, hickory hull) polyphenol extract has high total phenol content, total flavonoid content and in-vitro antioxidant capacity, and the preparation operation is simple, and the purification efficiency is high. The present application recycles waste in the hickory processing process, provides a reference for evaluating the overall value of hickory, and has great significance for promoting the transformation of hickory by-products into high-value products.
[0087] Although the present application has been disclosed with the preferred embodiments as above, it is not intended to limit the present application, and any person skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present application, therefore the protection scope of the present application should be defined by the claims.
Claims
1. A method for processing hickory by-product polyphenols captured by a solid phase extraction instrument sample adsorption combined probe type cell disruptor, characterized by, Comprising the following steps: Step one: raw material pretreatment: after washing the fresh pecan by-products clean, drying in a vacuum drying oven, crushing, sieving; Step two: ethanol crude extraction: mixing the pecan by-product powder obtained in step one with ethanol, stirring for ethanol crude extraction; Centrifuging the solution after ethanol crude extraction, obtaining the supernatant by centrifugation, and obtaining the crude polyphenol extract of pecan by-products by reduced pressure filtration; re-extracting the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration with ethanol, centrifuging and reducing pressure filtration; combining the crude polyphenol extracts of pecan by-products; reducing pressure rotary evaporation to concentrate the crude polyphenol extract of pecan by-products; Step three: sample adsorption: the solid phase extraction instrument is adsorbed under the action of the external air pump; first activate the solid phase extraction instrument with methanol, then add HCl to remove methanol; Then add the crude polyphenol extract of pecan by-products obtained in step two to the activated solid phase extraction instrument for adsorption; then add HCl, and after it passes through, add ethyl acetate to remove unabsorbed impurities; finally, add methanol to elute the absorbed polyphenols, and collect the polyphenol extract of pecan by-products; Step four: probe type cell crusher treatment: the polyphenol extract of pecan by-products collected in step three is treated by a probe type cell crusher; The polyphenol extract of pecan by-products treated by the probe type cell crusher is concentrated by reduced pressure rotary evaporation to obtain the polyphenol extract of pecan by-products.
2. The method for processing the captured polyphenol of hickory by-product according to claim 1, wherein the solid phase extraction instrument sample adsorption combined probe type cell crusher is characterized in that, In step one, the pecan by-products include pecan seed coat, pecan shell, and pecan hull.
3. The method for processing the by-product polyphenols of Carya cathayensis according to claim 1 or 2, wherein the solid phase extraction instrument sample adsorption combined probe type cell crusher is characterized in that, In step one, the fresh pecan by-products are washed clean and dried in a vacuum drying oven at 35-45℃ for 40-50h, crushed, and sieved through a 40-80 mesh sieve.
4. The method for processing the by-product polyphenol of Carya cathayensis according to claim 1 or 2, wherein the solid phase extraction instrument sample adsorption combined probe type cell crusher is used. In step two, the pecan by-product powder obtained in step one is mixed with 40-70v / v% ethanol at a solid-liquid ratio of 1g:20-40mL, and then placed in a 35-45℃ electronic constant temperature oil bath, stirred at 500-800rpm for 1-2h for ethanol crude extraction; the solution after ethanol crude extraction is centrifuged at 4-15℃ and 6000-8000rpm for 15-30min, and the supernatant obtained by centrifugation is filtered by reduced pressure to obtain the crude polyphenol extract of pecan by-products; the precipitate obtained by centrifugation and the filter residue obtained by reduced pressure filtration are re-extracted with ethanol, centrifuged and reduced pressure filtered for 1-3 times; the crude polyphenol extracts of pecan by-products of 2-4 times are combined. 5. The method of claim 4, wherein the solid phase extraction instrument sample adsorption combined probe type cell disruptor processing captures pecan by-product polyphenols, characterized by, In step two, the crude polyphenol extract of pecan by-products is concentrated by reduced pressure rotary evaporation to 10%-20% of the original volume at 30-42℃ and -0.05--0.098MPa.
6. The method for processing the captured polyphenol of hickory by-product according to claim 1 or 2, wherein the method is characterized by, In step three, the solid phase extraction instrument is adsorbed under the action of the external air pump, the airflow rate of the external air pump is 0.5-10L / min, the working pressure is 0.1-0.5MPa, the adsorption rate of the solid phase extraction instrument is 1-5mL / min, and the adsorption temperature is 20-30℃.
7. The method of claim 6, wherein the solid phase extraction instrument sample adsorption combined probe type cell disruptor processing captures hickory by-product polyphenols, characterized by, In step three, the solid phase extraction instrument was activated with 10-15 mL of methanol, and then 15-35 mL of 0.01-0.1 v / v% HCl was added, wherein the 0.01-0.1 v / v% HCl contained 0.01-0.1 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid was 36 wt% to 38 wt% concentrated hydrochloric acid, to remove the methanol; then 20-40 mL of the crude polyphenol extract of hickory by-products obtained in step two was added to the activated solid phase extraction instrument for adsorption; then 15-35 mL of 0.01-0.1 v / v% HCl was added, wherein the 0.01-0.1 v / v% HCl contained 0.01-0.1 mL of concentrated hydrochloric acid in 100 mL, and the concentrated hydrochloric acid was 36 wt% to 38 wt% concentrated hydrochloric acid, and after the 0.01-0.1 v / v% HCl passed through, 10-20 mL of ethyl acetate was added to remove the unabsorbed impurities; finally, 10-30 mL of methanol was added to elute the adsorbed polyphenols, and the hickory by-product polyphenol extract was collected.
8. The method for processing the captured polyphenol of hickory by-product according to claim 1 or 2, wherein the sample adsorption combined probe type cell crusher is a solid phase extraction instrument. In step four, the hickory by-product polyphenol extract collected in step three was treated with a probe-type cell disrupter, the probe diameter was 25 mm, the excitation voltage was 80-120 V, the vibration frequency was 20-25 kHz, the vibration amplitude was 10-70%, and the excitation time was 5-30 min, wherein the machine was turned on for 1 s and then turned off for 1 s, and the treatment was performed under ice bath.
9. The method of claim 8, wherein the solid phase extraction instrument sample adsorption combined probe type cell disruptor processing captures hickory by-product polyphenols, characterized by, In step four, the vibration amplitude was 40%, and the excitation time was 10 min.
10. The method of claim 8, wherein the solid phase extraction instrument sample adsorption combined with a probe type cell disruptor processing captured pecan by-product polyphenols, characterized in that, In step four, the hickory by-product polyphenol extract treated with the probe-type cell disrupter was concentrated to 10%-20% of the original volume under reduced pressure at 30-42 °C and -0.05--0.098 MPa, to obtain a hickory by-product polyphenol extract.