A method for efficiently extracting and separating chemical components from peanut shells

Abstract

The present invention discloses a method for efficiently extracting and separating chemical components from peanut shells. The method comprises the following steps: crushing raw peanut shells, weighing the powder, adding 95% ethanol and soaking for 3 hours, heating and refluxing for extraction for 3 times, each time for 40 minutes, filtering the extract, concentrating under reduced pressure, and steaming until there is no alcohol taste. The extract is extracted twice with an equal amount of petroleum ether, discarding the petroleum ether phase, and then extracting three times with an equal amount of ethyl acetate. The extracts are combined and evaporated to dryness under reduced pressure to obtain a crude extract of raw peanut shells. The method is characterized in that the crude extract of raw peanut shells is separated using a solvent in a head-to-tail elution mode of high-speed countercurrent chromatography. The present invention can effectively separate the crude extract of raw peanut shells.

Description

Technical Field

[0001] The invention relates to a method for efficiently extracting and separating chemical components in peanut shells. Background Art

[0002] Peanut shells, the outer shells of peanut pods, are an abundant and inexpensive agricultural byproduct of the peanut plant. While a small amount is used as fuel and roughage, or made into medium-density fiberboard, adhesives, plastic fillers, and other applications, the vast majority of these agricultural byproducts are incinerated or discarded without proper utilization. In addition to crude protein, carbohydrates, and ash, peanut shells also contain functional ingredients such as polyphenols, luteolin, carotene, and isosaponins, which are of great significance for the development of high-value-added products.

[0003] Peanut shells are also a good source of natural antioxidants and have attracted widespread attention from nutrition and chemistry researchers. Rich in antioxidants and other potentially health-promoting compounds, peanut shells can be used in food applications, and numerous physical and chemical techniques have been employed to extract antioxidants from them. Therefore, the rational utilization of peanut shell waste not only helps prevent environmental problems but also plays an important role in the efficient use of natural resources. However, the chemical composition of peanut shells is complex, making extraction difficult. Currently, no methods exist for extracting chemical components from peanut shells, let alone separating the different chemical components within them. Summary of the Invention

[0004] The invention provides a method for efficiently extracting and separating chemical components in peanut shells, and solves the technical problem of extracting and separating different chemical components in peanut shells.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A method for efficiently extracting and separating chemical components from peanut shells comprises the following steps: crushing raw peanut shells, weighing the powder, soaking it in 95% ethanol for 3 hours, heating and refluxing for 3 times, each time for 40 minutes, filtering the extract, concentrating under reduced pressure, and distilling until the alcohol taste is gone to obtain a concentrate, extracting it twice with an equal amount of petroleum ether to the concentrate, discarding the petroleum ether phase, and then extracting it three times with an equal amount of ethyl acetate to the concentrate, combining the extracts, and distilling it to dryness under reduced pressure to obtain a crude raw peanut shell extract. The crude raw peanut shell extract is separated using a solvent using a high-speed countercurrent chromatography in a head-to-tail elution mode.

[0007] The solvent is a mixture of n-hexane, ethyl acetate, methanol and water;

[0008] The chemical formulas of the peanut shell extracts are

[0009] (1)

[0011] (2)

[0013]

[0014] (3).

[0015] The solvent is a mixture of n-hexane, ethyl acetate, methanol and water, wherein the volume ratio of n-hexane, ethyl acetate, methanol and water is 3:7:5:5.

[0016] The solvent is carried out according to the following steps:

[0017] Weigh n-hexane, ethyl acetate, methanol and water into a separatory funnel, shake thoroughly, let stand for 30 min, and separate the layers into upper and lower phases.

[0018] The high-speed countercurrent chromatography head-to-tail elution mode is

[0019] Use a pipette to draw up the upper and lower phases of the solvent system;

[0020] Take the crude extract of raw peanut shells and add 5 mL of the upper phase and 5 mL of the lower phase mixed solution, and dissolve it by ultrasonication to serve as the sample solution;

[0021] The upper phase is the stationary phase and the lower phase is the mobile phase;

[0022] Open the cold hydrazine, maintain the temperature at 25°C, pump the stationary phase into the chromatographic column at a flow rate of 30 mL / min. After ensuring that it is completely filled, adjust the speed to 800 rpm in the forward direction and start equilibration of the mobile phase at a flow rate of 2 mL / min until fluid dynamics equilibrium is reached. Manually inject the sample solution into the chromatographic column through the injection valve;

[0023] Set the UV detector to continuously monitor the absorbance at 280 nm and turn on the recorder to record the chromatographic data graph;

[0024] The eluate was continuously collected using an automated collector. Each peak fraction was collected into a reagent bottle according to the chromatogram. After eluting the desired peak, the speed and elution were stopped, the instrument was shut down, and the components in the column were collected in a graduated cylinder using a vacuum pump. The collected fractions were concentrated under reduced pressure. After a single-step separation, HPLC analysis revealed three main peaks: Peak 1, Peak 2, and Peak 3, all with purities exceeding 98%. The target compound was enriched using a continuous injection method, with three consecutive injections of 100 mg each time, to obtain a sufficient sample quality for further structural identification.

[0025] The invention has the following beneficial technical effects:

[0026] Different chemical components in peanut shells are extracted and separated. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 HPLC chromatogram of chemical components in peanut shells;

[0028] Figure 2 High-speed countercurrent chromatogram of chemical components in peanut shells; DETAILED DESCRIPTION

[0029] The present invention is further described below with reference to specific examples.

[0030] Example 1

[0031] Crush raw peanut shells, weigh 700 g of powder, add 4000 mL of 95% ethanol and soak for 3 hours. Heat to 90°C and reflux for extraction three times, each time for 40 minutes. Filter the extract, concentrate under reduced pressure, and evaporate until there is no alcohol smell to obtain a concentrate. Extract twice with an equal amount of petroleum ether to the concentrate, discard the petroleum ether phase, and then extract three times with an equal amount of ethyl acetate to the concentrate. Combine the extracts.

[101] , and evaporated to dryness under reduced pressure to obtain 20.02 g of crude peanut shell extract.

[0032] The solvent is a composition of n-hexane, ethyl acetate, methanol and water in a volume ratio of 3:7:5:5;

[0033] The solvent is carried out according to the following steps:

[0034] Weigh n-hexane, ethyl acetate, methanol and water into a separatory funnel, shake thoroughly, let stand for 30 min, and separate the layers into upper and lower phases.

[0035] The high-speed countercurrent chromatography head-to-tail elution mode is

[0036] Use a pipette to draw up the upper and lower phases of the solvent system;

[0037] Take the crude extract of raw peanut shells and add 5 mL of the upper phase and 5 mL of the lower phase mixed solution, and dissolve it by ultrasonication to serve as the sample solution;

[0038] The upper phase is the stationary phase and the lower phase is the mobile phase;

[0039] Open the cold hydrazine, maintain the temperature at 25°C, pump the stationary phase into the chromatographic column at a flow rate of 30 mL / min. After ensuring that it is completely filled, adjust the speed to 800 rpm in the forward direction and start equilibration of the mobile phase at a flow rate of 2 mL / min until fluid dynamics equilibrium is reached. Manually inject the sample solution into the chromatographic column through the injection valve;

[0040] Set the UV detector to continuously monitor the absorbance at 280 nm and turn on the recorder to record the chromatographic data graph;

[0041] The eluate was continuously collected using an automatic collector, and each peak fraction was collected into a reagent bottle according to the chromatogram. After the desired peak was eluted, the speed and elution were stopped, the instrument was shut down, and the components in the chromatographic column were collected in a measuring cylinder using a vacuum pump. The collected fractions were concentrated under reduced pressure. After a single-step separation, three main peaks were obtained by HPLC analysis: Peak 1, Peak 2, and Peak 3, all with purities exceeding 98%. The target compounds, Compound 1, Compound 2, and Compound 3, were enriched using a continuous injection method. Three consecutive injections, each with a 100 mg injection volume, were injected to obtain sufficient sample quality for further structural identification.

[0042] Detection was performed using high performance liquid chromatography:

[0043] The test conditions are:

[0044] Column: Phenomenex C 18 column (250×4.6mm, 5 μm); mobile phase: A-0.1% formic acid in water, B-acetonitrile; gradient elution conditions: 0-30 min, 80-74% A, 20-26% B; 30-35 min, 74-70% A, 26-30% B; 35-35.5 min, 70-80% A, 30-20% B; 35.5-42 min, 80% A, 20% B; flow rate: 1 mL / min; column temperature: 25°C; detection wavelength: 280 nm; injection volume: 10 μL. HPLC chromatogram: see Figure 1 .

[0045] The chemical components in peanut shells were determined by HPLC, and the spectrum is shown in Table 2.

[0046] Compound structure identification and analysis

[0047] Compound 1: white powder, ESI-MS m / z 178.77 [M+H] + , molecular formula is C9H6O4; the NMR data in Table 1 and the literature reports

[102] The results were basically consistent, and compound 1 was identified as 5,7-dihydroxychromone, with the structural formula

[0048]

[0049] Compound 1 1 H-NMR (400 MHz) and 13 C-NMR (100 MHz) data, see Table 1

[0050] Table 1

[0051]

[0052] Compound 2: white powder, ESI-MS m / z 288.99 [M+H] + , molecular formula is C 15 H 12 O6; The NMR data in Table 2 are basically consistent with those reported in the literature, and compound 2 is identified as eriodictyol, and its structural formula is

[0053]

[0054] Table 2 Compound 2 1 H-NMR (400 MHz) and 13 C-NMR (100 MHz) data

[0055] Table 2

[0056]

[0057] Compound 3: yellow powder, ESI-MS m / z 287.00 [M+H] + , the molecular formula is C 15 H 10 O6; Table 3 NMR data and literature reports ] The results were basically consistent, and compound 3 was identified as luteolin, whose structural formula is

[0058]

[0059] Table 3 Compound 3 1 H-NMR (400 MHz) and 13 C-NMR (100 MHz) data

[0060] Table 3

[0061]

Claims

1. A method for efficiently extracting and separating chemical components from peanut shells, comprising crushing raw peanut shells, weighing the powder, adding 95% ethanol and soaking for 3 hours, heating and refluxing for 3 times, each time for 40 minutes, filtering the extract, concentrating under reduced pressure, and steaming until there is no alcohol smell to obtain a concentrate, extracting twice with an equal amount of petroleum ether to the concentrate, discarding the petroleum ether phase, and then extracting three times with an equal amount of ethyl acetate to the concentrate, combining the extracts, and evaporating under reduced pressure to dryness to obtain a crude extract of raw peanut shells, characterized in that: High-speed countercurrent chromatography in head-to-tail elution mode was used to separate the crude peanut shell extract using solvent; The solvent is a mixture of n-hexane, ethyl acetate, methanol and water; The chemical formulas of the peanut shell extracts are The solvent is a mixture of n-hexane, ethyl acetate, methanol and water, wherein the volume ratio of n-hexane, ethyl acetate, methanol and water is 3:7:5:5; the high-speed countercurrent chromatography head-to-tail elution mode is Use a pipette to draw up the upper and lower phases of the solvent system; Take the crude extract of raw peanut shells and add 5 mL of the upper phase and 5 mL of the lower phase mixed solution, and dissolve it by ultrasonication to serve as the sample solution; The upper phase is the stationary phase and the lower phase is the mobile phase; Open the cold hydrazine, maintain the temperature at 25 ° C, pump the stationary phase into the chromatographic column at a flow rate of 30 mL / min. After ensuring that it is completely filled, adjust the speed to 800 rpm in the forward direction and start equilibration of the mobile phase at a flow rate of 2 mL / min until fluid dynamics equilibrium is reached. Manually inject the sample solution into the chromatographic column through the injection valve; Set the UV detector to continuously monitor the absorbance at 280 nm and turn on the recorder to record the chromatographic data graph; The eluate was continuously collected by an automatic collector, and each peak fraction was collected into a reagent bottle according to the chromatogram. After the desired peak was eluted, the rotation speed and elution were stopped, the instrument was turned off, and the components in the chromatographic column were collected in a measuring cylinder using a vacuum pump. The collected fractions were concentrated under reduced pressure. After one-step separation, three main peaks were obtained by HPLC determination, namely Peak 1, Peak 2 and Peak 3, with a purity of more than 98%. The target compound was enriched by a continuous injection method, and three consecutive injections were made, with an injection volume of 100 mg each time, to obtain a sample of sufficient quality for further structural identification.

2. The method for efficiently extracting and separating chemical components from peanut shells according to claim 1, wherein: The solvent is carried out according to the following steps: Weigh n-hexane, ethyl acetate, methanol and water into a separatory funnel, shake thoroughly, let stand for 30 minutes, and separate into upper and lower phases.

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