CN110501410B - Electrochemical method for rapidly detecting total amide content in pericarpium zanthoxyli - Google Patents
Electrochemical method for rapidly detecting total amide content in pericarpium zanthoxyli Download PDFInfo
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Abstract
The invention belongs to the field of food detection, and relates to an electrochemical method for rapidly detecting the content of total amide in pepper peel; pulverizing dried pericarpium Zanthoxyli, sieving, extracting the dry powder with anhydrous ethanol, washing residue, diluting, and diluting to desired volume to obtain Zanthoxylamide extractive solution; mixing the extracting solution with an electrolyte solution to prepare a Zanthoxylum bungeanum amide detection working solution, wherein the electrolyte solution is a phosphoric acid buffer solution containing ethanol; measuring a differential pulse voltammetry curve of a detection working solution in the process of adding a standard substance of hydroxy-alpha-sanshool for multiple times, drawing a working curve graph of a multiple standard addition method, calculating to obtain the total amide content in the detection solution, and converting to obtain the total amide content in a pepper peel sample, namely the equivalent value of the hydroxy-alpha-sanshool; the invention solves the problem that the existing method for quantitatively detecting the total amide in the pepper can not realize the combination of rapidness, low cost and accuracy, and the detection method can be used for rapidly, conveniently and accurately detecting the total amide content in the pepper peel.
Description
Technical Field
The invention belongs to the field of food detection, and relates to an electrochemical method for rapidly detecting the content of total amide in pepper.
Background
The Chinese prickly ash is one of the traditional eight major seasonings in China, and is deeply favored by the nation because of the unique numb taste. The main component causing the numb taste of the zanthoxylum is chain polyunsaturated fatty acid amide, which is generally called zanthoxylum amide, wherein Hydroxyl-alpha-sanshool (Hydroxyl-alpha-sanshool, alpha-SOH), Hydroxyl-beta-sanshool (Hydroxyl-alpha-sanshool, alpha-SOH), Hydroxyl-gamma-sanshool (Hydroxyl-alpha-sanshool, alpha-SOH) and Hydroxyl-epsilon-sanshool (Hydroxyl-alpha-sanshool, alpha-SOH) are the most representative amide substances and account for more than 98 percent of the total amide content.
The content of the Zanthoxylum bungeanum amide is an important index for measuring the quality of the Zanthoxylum bungeanum and also an important basis for grading the quality of the Zanthoxylum bungeanum. The existing instrument detection method capable of accurately measuring the content of the zanthoxylum bungeanum amide is High Performance Liquid Chromatography (HPLC) or high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS); patent document 201711217038.4 discloses a method for detecting the content of zanthoxylum amide in zanthoxylum bungeanum maxim samples by using HPLC equipment, but the limitations of expensive high performance liquid chromatography equipment, complex operation procedures, high detection environment requirements and no portable use prevent the large-scale application of the method in the industry.
The prior document discloses a method for rapidly detecting the content of numb-taste substances of zanthoxylum, which comprises the following steps: extracting total amide extract; the amide is converted into ammonium nitrogen; and (4) measuring the content of ammonium nitrogen. The method belongs to a titration method for indirectly measuring the Zanthoxylum bungeanum amide, wherein the total time consumption of the pretreatment processes of total amide extraction, amide conversion into ammonium nitrogen and the like is 2-6 hours; and the titration operation procedure is complicated, and the titration end point is easy to be misjudged. At present, no method is disclosed for detecting the content of the Zanthoxylum bungeanum amide quickly, conveniently, accurately and at low cost.
Disclosure of Invention
The invention aims to solve the technical problems of complex operation, high cost and the like of the existing method for detecting the content of the zanthoxylum amide in the zanthoxylum bungeanum maxim sample, and provides the electrochemical method which is simple, convenient and quick to operate, low in detection cost and capable of realizing quick, accurate and portable detection of the content of the total zanthoxylum amide in the zanthoxylum bungeanum maxim raw material.
The technical scheme for solving the technical problems comprises the following steps: an electrochemical method for rapidly detecting the content of total amide in pepper comprises the following specific steps:
(1) preparing a zanthoxylum peel sample;
removing wilting or browning dried pericarpium Zanthoxyli, pulverizing, sieving to obtain fructus Zanthoxyli powder, and refrigerating at-20 deg.C;
(2) detecting the preparation of mother liquor;
weighing the pepper powder obtained in the step (1), adding absolute ethyl alcohol A with the mass of W, shaking up, and performing ultrasonic extraction at normal temperature to obtain a crude extract; centrifuging the crude extract, and collecting supernatant; collecting filter residue, adding absolute ethyl alcohol B, shaking, centrifuging again, and collecting supernatant; repeating the steps, mixing the supernatant liquid collected for multiple times to obtain a detection mother liquid, recording the total volume V, and refrigerating for later use; the whole step is operated under the condition of keeping out of the sun;
(3) differential pulse voltammetry analysis;
dripping a certain volume of hydroxy-alpha-sanshool standard solution (alpha-SOH) into a sample detection solution according to the principle of a multiple standard addition method, and then detecting a differential pulse voltammetry curve of a pepper amide substance in a mixed solution by using a three-electrode system;
preparing an electrolytic detection solution, wherein the electrolytic detection solution is a mixed solution of absolute ethyl alcohol and a PBS buffer solution; absorbing the detection mother liquor prepared in the step (2), and recording the volume as V1Diluting with electrolytic detection solution to obtain sample detection solution A, and recording volume as V2Detecting a differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system;
absorbing the detection mother solution for the second time, wherein the volume is V1, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding an alpha-SOH standard solution, and marking the added volume as V3 to obtain a mixed solution, and detecting the differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system;
absorbing the detection mother solution with the volume of V1 for the third time, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding 2 times of the alpha-SOH standard solution with the volume of V3, and detecting the differential pulse voltammetry curve of the Zanthoxylum piperitum amide substance in the mixed solution by using a three-electrode system;
absorbing the detection mother solution with the volume of V1 for the fourth time, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding 3 times of the alpha-SOH standard solution with the volume of V3, and detecting the differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system; repeating the operation by analogy;
finally, taking the mass concentration of the alpha-SOH standard solution as an abscissa, taking the corresponding peak current as an ordinate to perform linear regression, taking the corresponding differential pulse voltammetry peak current as an ordinate to perform linear regression curve, and taking the absolute value of the intersection point of the linear regression curve and the abscissa as the total amide content in the sample detection solution, and marking the absolute value as m;
(4) calculating the total amide content;
and (4) according to the total amide content in the sample detection solution obtained in the step (3), further converting the total amide content in the detection mother liquor according to the following formula, namely the content of alpha-SOH equivalent:
in this formula, M: the total amide content in the pepper sample is mu g/g; m: detecting the total amide content (calculated according to a working curve of a multi-time standard addition method) in the solution by using a sample, wherein the content is mu g of alpha-SOH equivalent/mL; v1: the volume of the detection mother liquor used when preparing the sample detection solution; v2: the volume of sample detection solution; v: detecting the total volume of the mother solution, mL; w: pepper powder mass, g; a: and (5) correcting the system error.
Preferably, in the step (1), the mesh number of the screen is 20-40 meshes.
Preferably, in the step (2), the dosage ratio of the pepper powder to the absolute ethyl alcohol A is 5-25 g: 25-100 mL; the ultrasonic leaching time is 20-40 min.
Preferably, in step (2), the centrifugation conditions are all as follows: the rotating speed is 2000-3000 r/min, and the time is 3-8 min.
Preferably, in the step (2), the dosage ratio of the filter residue to the absolute ethyl alcohol B is 1 g: (1-6) mL.
Preferably, in the step (3), the volume ratio of the absolute ethyl alcohol to the PBS buffer solution in the electrolysis detection solution is 1-3: 7-9; the concentration of PBS buffer was 0.1M.
Preferably, in the step (3), the volume V of the mother liquor is detected1Volume V of sample detection solution2The ratio of (0.03-0.5): 10.
preferably, in the step (3), the volume V of the mother liquor is detected1Volume V of standard solution of alpha-SOH added3The ratio of (1) to (2) is (3-12): 1; the concentration of the alpha-SOH standard solution is 1.0-10.0 g/L.
Preferably, in the step (3), the three-electrode system comprises a working electrode (glassy carbon electrode), a reference electrode (Ag/AgCl electrode) and an auxiliary electrode (platinum wire electrode); the detection condition parameters of the differential pulse voltammetry are as follows: forward scan, +0.6V- + 1.2V; the potential is stepped by 4 mV; amplitude 0.05V; the pulse width is 0.05 s; sampling width 0.0167 s; the pulse period is 0.5 s.
Preferably, in the step (3), the number of times of the repeated operation is 4-6.
Has the advantages that:
(1) the invention establishes the electrochemical detection method of the total content of the zanthoxylum bungeanum amide in the zanthoxylum peel for the first time, and compared with the existing zanthoxylum bungeanum amide detection method at home and abroad at present, the instrument used by the method has the characteristics of portability, low cost and simple operation.
(2) The bare glassy carbon electrode used in the electrochemical detection method does not need to carry out complex modification treatment on the electrode, and is simple and convenient to operate.
(3) The electrochemical detection method disclosed by the invention has high detection accuracy, the error rate of the detection result of the electrochemical detection method and the HPLC-MS method is lower than 6%, the detection speed is high (differential pulse voltammetry scanning is completed within 2 minutes), and the method can be used for quickly and accurately quantifying the total amide of the pepper.
(4) The electrochemical detection method provided by the invention is expected to be developed into a substitute method of HPLC-MS, and has important significance for detection and analysis of the total amides of the peppers and the products thereof.
Drawings
FIG. 1 is a differential pulse voltammogram for detecting the content of Zanthoxylum bungeanum amide by a standard addition method.
FIG. 2 is a schematic diagram of a standard addition process working curve calculation.
FIG. 3 is a graph showing a curve obtained by fitting the values measured by high performance liquid chromatography to the values measured by electrochemical method.
Detailed Description
In order to more clearly illustrate the present invention, the present invention is further described below with reference to specific examples, but it should be understood that these examples are for illustrative purposes only and should not be construed as limiting the practice of the present invention.
1. Experimental samples, Primary reagents and instruments
Sample preparation: 7 kinds of dried peppers with clear producing areas (all harvested locally) are adopted as detection objects, and the dried peppers are respectively: sample No. 1, first-grade Zanthoxylum piperitum (Sichuan Hanyuan); sample No. 2, Jiangjin green pricklyash peel (Chongqing Jiangjin); sample No. 3, JINYANGQINGZHAO (Sichuan Jinyang); sample No. 4, Shao Tong zanthoxylum bungeanum (Yunnan Shao Tong); no. 5 sample, Fu jiao first (Gansu Linxia); sample No. 6, first-grade red pepper (Fuping Shaanxi); no. 7, first-grade red bell pepper (Shandong Laiwu).
Reagent: the hydroxyl-alpha-sanshool (alpha-SOH), the hydroxyl-beta-sanshool (beta-SOH) and the hydroxyl-gamma-sanshool (gamma-SOH) standard products are purchased from Kyomai Desheng technology Co., Ltd, and the purity is more than 98%. The water used in the experimental process is deionized water, and the experimental reagents are analytically pure.
The instrument comprises the following steps: the three-electrode system of the electrochemical workstation (CHI660E, shanghai chen instruments ltd) was: the working electrode is a 3mm diameter glassy carbon disk electrode (model CHI104), the reference electrode is an Ag/AgCl electrode (model CHI111), and the auxiliary electrode is a platinum wire electrode (model CHI 115).
Example 1:
(1) preparing a zanthoxylum peel sample;
removing wilting or browning dried pericarpium Zanthoxyli (hereinafter abbreviated as fructus Zanthoxyli), pulverizing, sieving with 30 mesh sieve to obtain fructus Zanthoxyli powder, and refrigerating at-20 deg.C to be detected;
(2) detecting the preparation of mother liquor;
accurately weighing 5g of the pepper powder (W) obtained in the step (1) into a 50mL brown conical flask with a plug, adding 25mL of absolute ethyl alcohol, shaking uniformly, carrying out ultrasonic extraction for 30min at room temperature (25 +/-2 ℃) under the assistance of a constant-temperature water bath to obtain a crude extract, further centrifuging the crude extract, and centrifuging for 5min at 2000 r/min; pouring the supernatant into a 200mL brown volumetric flask (assisted by a funnel), collecting filter residues, adding 25mL of absolute ethyl alcohol, shaking in a vortex mode for 1min, centrifuging at 2000r/min for 5min, collecting the supernatant, repeating the operation for 3 times, combining the supernatants for 3 times, fixing the volume of the combined supernatants to 200mL by using the absolute ethyl alcohol for 3 times, shaking uniformly for later use as a detection mother solution, and recording the total volume V (200 mL);
(3) differential pulse voltammetry analysis;
dripping a certain volume of hydroxy-alpha-sanshool standard solution (alpha-SOH) into a sample detection solution according to the principle of a multiple standard addition method, and then detecting a differential pulse voltammetry curve of a pepper amide substance in a mixed solution by using a three-electrode system;
preparing an electrolytic detection solution, which comprises the following components: ethanol: PBS (pH 2) (1: 9 v/v); the concentration of PBS buffer solution is 0.1M, 30. mu.L of the detection mother solution prepared in the step (2) is sucked, and the volume is recorded as V1(30 mu L), diluting the solution to a constant volume of 10mL by using an electrolytic detection solution to obtain a sample detection solution A, and recording the volume as V2(10mL), detecting a differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system;
sucking and detecting mother liquor for the first time and mixing the detection mother liquor with an electrolysis detection solution, preparing a sample detection solution B again, adding 10 mu L of alpha-SOH standard solution, wherein the adding volume is recorded as V3(10 mu L), the concentration of the added alpha-SOH standard solution is 2.6g/L, and detecting a differential pulse voltammetry curve of the Zanthoxylum bungeanum amide substance in the mixed solution by using a three-electrode system; absorbing the detection mother solution for the second time and mixing the detection mother solution with the electrolysis detection solution, preparing a sample detection solution C again, adding 20 mu L of alpha-SOH standard solution, and detecting the differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system; thirdly, absorbing and mixing the detection mother solution and the electrolytic detection solution, preparing a sample detection solution D again, adding 30 mu L of alpha-SOH standard solution, and detecting the differential pulse voltammetry curve of the pepper amide substance in the mixed solution by using a three-electrode system; absorbing the detection mother solution and the electrolytic detection solution for the fourth time, mixing, preparing a sample detection solution E again, adding 40 mu L of alpha-SOH standard solution, and detecting the differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system; wherein the volumes and the concentrations of the sample detection solution A, the sample detection solution B, the sample detection solution C, the sample detection solution D and the sample detection solution E are the same, and are all 30 microliter of detection mother liquor prepared in the step (2), and the detection mother liquor is diluted by an electrolytic detection solution to be 10 mL; and (3) superposing the control of the electrolytic solution, the sample detection solution and the quadruply-labeled differential pulse voltammetry curve to obtain a differential pulse voltammogram for detecting the total amide content of the pepper by the standard addition method, as shown in figure 1, as can be seen from figure 1, the corresponding peak current is increased along with the increase of the labeling times.
Finally, taking the mass concentration of the alpha-SOH standard solution as an abscissa and the corresponding differential pulse voltammetry peak current as an ordinate to make a linear regression curve to obtain a working curve of the standard addition method, as shown in FIG. 2, obtaining the total amide content in the sample detection solution as m according to the absolute value of the intersection point of the linear regression curve and the abscissa in the graph;
the electrochemical analyzer employs a three-electrode system (working electrode (glassy carbon electrode), reference electrode (Ag/AgCl electrode), auxiliary electrode (platinum wire electrode)). The detection condition parameters of the differential pulse voltammetry are as follows: forward scan, +0.6V- + 1.2V; the potential is stepped by 4 mV; amplitude 0.05V; the pulse width is 0.05 s; sampling width 0.0167 s; the pulse period is 0.5 s.
(4) Calculating the total amide content;
and (4) according to the total amide content in the sample detection solution obtained in the step (3), further converting the total amide content in the detection mother liquor according to the following formula, namely the content of alpha-SOH equivalent:
in this formula, M: the total amide content in the pepper sample is mu g/g; m: detecting the total amide content (calculated according to a working curve of a multi-time standard addition method) in the solution by using a sample, wherein the content is mu g of alpha-SOH equivalent/mL; v1: the volume of the detection mother liquor used when preparing the sample detection solution; v2: the volume of sample detection solution; v: detecting the total volume of the mother solution, mL; w: pepper powder mass, g; a: and (5) correcting the system error.
(5) Verifying the detection result;
the seven pepper detection mother solutions are measured by an HPLC-MS method, and the total amide content in pepper samples is calculated, so that the accuracy of the measurement result of the method is verified.
Respectively calculating the contents of hydroxy-beta-sanshool (beta-SOH) and hydroxy-gamma-sanshool (gamma-SOH) in the pepper sample by an external standard method; calculating the contents of hydroxy-alpha-sanshool (alpha-SOH) and hydroxy-epsilon-sanshool (epsilon-SOH) in the pepper sample by using an internal standard method, and specifically comprising the following steps: firstly, dissolving beta-SOH and gamma-SOH standard products by using methanol respectively, preparing a series of solutions with different concentrations, and testing the solutions on a machine, wherein all the solutions need to pass through a 0.22 mu m organic filter membrane before the testing on the machine; taking the concentration of the standard as a horizontal coordinate and taking the corresponding peak area as a vertical coordinate to draw a standard curve; taking 0.5mL of detection mother liquor obtained in the step (2), wherein the volume is recorded as V4, diluting with methanol to a constant volume of 10mL, and the volume is recorded as V5; sucking 1mL of filtered solution, testing the filtered solution on a machine, performing parallel measurement for 3 times, and taking an average value. And (3) detecting the concentration of the beta-SOH and gamma-SOH substances in the mother liquor, calculating according to a standard curve, marking as m1 and m2, and converting by formulas (a1) and (a2) to obtain the content of the beta-SOH and the gamma-SOH in the pepper sample.
In the formula: m: the content of beta-SOH in the pepper sample is mu g/g; m 1: the concentration of beta-SOH (calculated according to a standard curve) in the sample is tested on the machine, and the concentration is microgram/mL; (V5/V4): detecting the dilution multiple of the mother solution, 20; v: detecting the volume of the mother solution, 20 mL; w: sample mass, 5 g.
In the formula: m: the gamma-SOH content in the pepper sample is mu g/g; m 2: the concentration of gamma-SOH (calculated according to a standard curve) in the sample is tested on the machine, and the concentration is mu g/mL; (V5/V4): detecting the dilution multiple of the mother solution, 20; v: detecting the volume of the mother solution, 200 mL; w: sample mass, 5 g.
Accurately weighing 5g of the pepper powder (W) obtained in the step (1) and 2mg of a beta-SOH standard substance into a 50mL brown conical flask with a plug, and adding the beta-SOH standard substance with the quality number n; adding 25mL of absolute ethyl alcohol, shaking uniformly, performing ultrasonic extraction at room temperature (25 +/-2 ℃) for 30min with the assistance of constant-temperature water bath to obtain a crude extract, further centrifuging the crude extract, and centrifuging at 2000r/min for 5 min; pouring the supernatant into a 200mL brown volumetric flask (assisted by a funnel), collecting filter residues, adding 25mL of absolute ethyl alcohol, shaking in a vortex mode for 1min, centrifuging at 2000r/min for 5min, collecting the supernatant, repeating the operation for 3 times, combining the supernatants for 3 times, fixing the volume of the combined supernatants to 200mL by using the absolute ethyl alcohol for 3 times, and shaking uniformly for later use as an internal standard detection mother solution; respectively taking 0.5mL of detection mother liquor added with an internal standard and the detection mother liquor prepared in the step (2), diluting with methanol to a constant volume of 10mL, sucking 1mL of the detection mother liquor respectively, performing an on-machine test, recording peak areas (A1) and epsilon-SOH peak areas (A2) of alpha-SOH in the detection mother liquor, recording beta-SOH peak areas measured by the detection mother liquor and the detection mother liquor added with the internal standard, calculating the difference value to be delta A, calculating the alpha-SOH content in the pepper sample by the detection mother liquor through a formula (b1), and calculating the epsilon-SOH content in the pepper sample through a formula (b 1);
in the formula: m: the alpha-SOH content in the pepper sample is mu g/g; n: adding the content of the beta-SOH internal standard substance, 2000 mug; a1: alpha-SOH peak area; Δ A: the difference between the peak areas of the beta-SOH measured with and without the addition of the internal standard; w: sample mass, 5 g.
In the formula: m: the content of epsilon-SOH in the pepper sample is mu g/g; n: adding the content of the beta-SOH internal standard substance, 2000 mug; a2: epsilon-SOH peak area; Δ A: the difference between the peak areas of the beta-SOH measured with and without the addition of the internal standard; w: sample mass, 5 g.
And the sum of the contents of alpha-SOH, beta-SOH, gamma-SOH and epsilon-SOH in the pepper sample is calculated to be the total amide content in the pepper sample.
The content of the total amide substances of the pepper, which is measured by a DPV method (differential pulse voltammetry) and an HPLC-MS method, is shown in Table 1. As shown in fig. 3, it is understood from fig. 3 that a system error exists between the DPV measurement value and the HPLC-MS measurement value, and the linear relationship corresponds to y 2.6385x (R is 2.6385 x) (as shown in fig. 3)20.9955), namely when the DPV method is adopted to detect the content of the total amide in the pepper, the measured value is corrected by using a correction coefficient 2.6385Positive, a DPV result correction is obtained. The results of HPLC-MS and DPV measurements are shown in Table 1; the relative error range between the HPLC-MS measured value and the DPV result correction value of different pepper samples is 0.02-5.87%, and is lower than 6%. The results show that: the DPV method can be used for well obtaining the content of the total amide in the mother liquor of the zanthoxylum bungeanum maxim, and the result is highly similar to that of HPLC-MS.
TABLE 1 comparison of the results of HPLC-MS and DPV methods
(6) Recovery rate and precision of total amide of Chinese prickly ash adding standard recovery result
Selecting No. 1, No. 4 and No. 7 pepper samples, adding alpha-SOH standard products in the ultrasonic extraction process, wherein the content of the added alpha-SOH is respectively 13.15mg, 26.30mg and 39.45mg, and preparing a detection mother solution containing pepper total amide and a detection mother solution containing the alpha-SOH standard products according to the steps (2) and (3) in the embodiment. Separately, 500. mu.L of each assay mother solution was transferred, diluted with an electrolytic assay solution to a volume of 10mL, and the total amide concentration in the extract was calculated according to step (4) of the embodiment, and the results are shown in Table 2:
TABLE 2 recovery test results with additional labeling
As can be seen from the results in Table 2, the recovery rate is between 94% and 108%, the relative standard deviation is lower than 4.0%, and the data result shows that the detection method of the invention is reliable.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. An electrochemical method for rapidly detecting the content of total amide in pepper peel is characterized by comprising the following steps:
(1) removing wilting or browning dried pericarpium Zanthoxyli, pulverizing, and sieving to obtain fructus Zanthoxyli powder;
(2) weighing the pepper powder obtained in the step (1), adding absolute ethyl alcohol A with the mass of W, shaking up, and performing ultrasonic extraction at normal temperature to obtain a crude extract; centrifuging the crude extract, and collecting supernatant; collecting filter residue, adding absolute ethyl alcohol B, shaking, centrifuging again, and collecting supernatant; repeating the steps, mixing the supernatant liquid collected for multiple times to obtain a detection mother liquid, recording the total volume V, and refrigerating for later use; the whole step is operated under the condition of keeping out of the sun;
(3) firstly, preparing an electrolytic detection solution, wherein the electrolytic detection solution is a mixed solution of absolute ethyl alcohol and a PBS buffer solution; absorbing the detection mother liquor prepared in the step (2), and recording the volume as V1Diluting with electrolytic detection solution to obtain sample detection solution A, and recording volume as V2Detecting a differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system;
absorbing the detection mother solution with the volume of V1 for the second time, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding an alpha-SOH standard solution with the volume of V3 to obtain a mixed solution, and detecting the differential pulse voltammetry curve of the Zanthoxylum bungeanum amide substance in the mixed solution by using a three-electrode system;
absorbing the detection mother solution with the volume of V1 for the third time, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding 2 times of the alpha-SOH standard solution with the volume of V3, and detecting the differential pulse voltammetry curve of the Zanthoxylum piperitum amide substance in the mixed solution by using a three-electrode system;
absorbing the detection mother solution with the volume of V1 for the fourth time, diluting the detection mother solution with an electrolytic solution to the volume of V2, preparing the solution to be detected again, adding 3 times of the alpha-SOH standard solution with the volume of V3, and detecting the differential pulse voltammetry curve of the pepper amide substances in the mixed solution by using a three-electrode system; repeating the operation by analogy;
finally, taking the mass concentration of the alpha-SOH standard solution as an abscissa, taking the corresponding peak current as an ordinate to perform linear regression, taking the corresponding differential pulse voltammetry peak current as an ordinate to perform linear regression curve, and taking the absolute value of the intersection point of the linear regression curve and the abscissa as the total amide content in the sample detection solution, and marking the absolute value as m;
(4) and (4) according to the total amide content in the sample detection solution obtained in the step (3), further converting according to the following formula to obtain the total amide content in the detection mother liquor, namely the equivalent content of alpha-SOH:
in this formula, M: the total amide content in the pepper sample is mu g/g; m: detecting the total amide content in the solution by using a sample, wherein the content is mu g/mL; v1: the volume of the detection mother liquor used when preparing the sample detection solution; v2: the volume of sample detection solution; v: detecting the total volume of the mother solution, mL; w: pepper powder mass, g; a: and (5) correcting the system error.
2. The electrochemical method for rapidly detecting the total amide content in the pericarpium zanthoxyli of claim 1, wherein in the step (1), the mesh number of the screen is 20-40 meshes.
3. The electrochemical method for rapidly detecting the total amide content in the peel of zanthoxylum bungeanum as claimed in claim 1, wherein in the step (2), the dosage ratio of the paprika powder to the absolute ethyl alcohol A is 5-25 g: 25-100 mL; the ultrasonic leaching time is 20-40 min.
4. The electrochemical method for rapidly detecting the content of the total amide in the pericarpium zanthoxyli is characterized in that in the step (2), the dosage ratio of the filter residue collected after centrifugation to the absolute ethyl alcohol B is 1 g: 1-6 mL; the centrifugation conditions were all: the rotating speed is 2000-3000 r/min, and the time is 3-8 min.
5. The electrochemical method for rapidly detecting the content of the total amides in the peel of zanthoxylum bungeanum according to claim 1, wherein in the step (3), the volume ratio of the absolute ethyl alcohol to the PBS buffer solution in the electrolytic detection solution is 1-3: 7 to 9.
6. The electrochemical method for rapidly detecting the total amide content in the peel of zanthoxylum bungeanum as claimed in claim 1, wherein in the step (3), the volume V of the mother solution is detected1Volume V of sample detection solution2The ratio of (A) to (B) is 0.03-0.5: 10.
7. the electrochemical method for rapidly detecting the total amide content in the peel of zanthoxylum bungeanum as claimed in claim 1, wherein in the step (3), the volume V of the mother solution is detected1Volume V of standard solution of alpha-SOH added3The ratio of (A) to (B) is 3-12: 1; the concentration of the alpha-SOH standard solution is 1.0-10.0 g/L.
8. The electrochemical method for rapidly detecting the total amide content in the pericarpium zanthoxyli peel according to claim 1, characterized in that in the step (3), the three-electrode system consists of a glassy carbon electrode, an Ag/AgCl electrode and a platinum wire electrode; the detection condition parameters of the differential pulse voltammetry are as follows: forward scan, +0.6V- + 1.2V; the potential is stepped by 4 mV; amplitude 0.05V; the pulse width is 0.05 s; sampling width 0.0167 s; the pulse period is 0.5 s.
9. The electrochemical method for rapidly detecting the total amide content in the peel of zanthoxylum bungeanum as claimed in claim 1, wherein in the step (3), the number of times of the repeated operation is 4-6.
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Inventor after: Zhang Di Inventor after: Zou Xiaobo Inventor after: Zhao Lei Inventor after: Shi Bolin Inventor after: Sun Xiaoxia Inventor before: Zou Xiaobo Inventor before: Zhao Lei Inventor before: Zhang Di Inventor before: Shi Bolin Inventor before: Sun Xiaoxia |