CN110501410A - The electrochemical method of total ceramide content in a kind of quick detection Chinese prickly ash pericarp - Google Patents
The electrochemical method of total ceramide content in a kind of quick detection Chinese prickly ash pericarp Download PDFInfo
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Abstract
The invention belongs to field of food detection, are related to a kind of electrochemical method for quickly detecting total ceramide content in Chinese prickly ash pericarp；Step are as follows: by the dry broken sieving of Chinese prickly ash fruit peel powder, dry powder obtains fagaramide extracting solution through dehydrated alcohol extraction, washery slag and liquid, dilution constant volume；Extracting solution is mixedly configured into fagaramide detection working solution with electrolyte solution, wherein electrolyte solution is the phosphate buffer containing ethyl alcohol；The differential pulse voltammetry curve of measurement detection working solution during reference substance hydroxyl-alpha-sanshool is repeatedly added, draw Multiple Standard Addition Method working curve diagram, the total ceramide content in detection solution is calculated, conversion obtains the total ceramide content in Chinese prickly ash peel sample, i.e. hydroxyl-alpha-sanshool equivalent value；The present invention solves the problems, such as that existing Chinese prickly ash total ceramide quantitative detecting method cannot achieve quick, inexpensive, accuracy and get both, this detection method can carry out quick, portable, accurate detection to total ceramide content in Chinese prickly ash pericarp.
Description
Technical field
The invention belongs to field of food detection, are related to a kind of electrochemical method for quickly detecting total ceramide content in Chinese prickly ash.
Background technique
Chinese prickly ash is one of Chinese tradition " eight big flavouring ", is liked deeply by its people because it is with unique numb taste.Cause
The main component of Chinese prickly ash fiber crops taste is chain polyunsaturated fatty acid amide, is referred to as fagaramide, wherein hydroxyl-alpha-sanshool
(Hydroxyl- α-sanshool, α-SOH), hydroxy-beta-sanshool (Hydroxyl- α-sanshool, α-SOH), hydroxyl-γ-
Sanshool (Hydroxyl- α-sanshool, α-SOH), hydroxyl-ε-sanshool (Hydroxyl- α-sanshool, α-SOH) are
Most representative amide substance, accounts for 98% or more of total ceramide content.
The height of fagaramide content be measure Chinese prickly ash quality an important indicator and Chinese prickly ash quality grading it is important
Foundation.The instrument detection method that fagaramide content can accurately be measured at present is high performance liquid chromatography (HPLC) or efficient liquid
Phase chromatographic tandem mass spectrography (HPLC-MS/MS)；A kind of utilization hplc device inspection is disclosed in patent document 201711217038.4
The method for surveying the fagaramide content in Chinese prickly ash sample, but high performance liquid chromatograph device is expensive, operation sequence is more complex, inspection
Survey environmental requirement it is high and can not portable use, these limitations hinder the large-scale application of this method in the industry.
It includes: mentioning for total ceramide medicinal extract that existing literature, which discloses a kind of method for rapidly detecting content of numb-taste components,
It takes；Amide is changed into ammonium nitrogen；Ammonium nitrogen assay.This method belongs to a kind of titration method of indirect determination fagaramide,
Wherein total ceramide extracts and amide was changed into the pretreatment process total time-consuming such as ammonium nitrogen up to 2-6 hours；And titration operation journey
Sequence is many and diverse, and titration end-point is easily judged by accident.Currently, there has been no published method can be achieved at the same time fagaramide content it is quick,
Portable, accurate, inexpensive detection.
Summary of the invention
The technical problem to be solved by the present invention is to complicated for operation for fagaramide method in detection Chinese prickly ash sample at present, at
The problems such as this is higher, provide it is a kind of it is simple, convenient quickly, testing cost it is low, can be realized to Chinese prickly ash total in Chinese prickly ash raw material
Amide content carries out the electrochemical method of quick, accurate, portable detection.
A kind of technical solution of present invention solution above-mentioned technical problem are as follows: electrification for quickly detecting total ceramide content in Chinese prickly ash
Method, the specific steps are as follows:
(1) Chinese prickly ash pericarp preparation of samples；
It removes and wilts or the drying Chinese prickly ash pericarp of brown stain, crush, screened through sieve, obtain zanthoxylum powder, it is cold under the conditions of -20 DEG C
It hides to be detected；
(2) preparation of mother liquor is detected；
Step (1) resulting zanthoxylum powder is weighed, dehydrated alcohol A is added in quality W, and concussion shakes up, carries out under normal temperature condition
Ultrasound extraction, obtains crude extract；Crude extract is centrifuged, supernatant is collected；It collects filter residue and dehydrated alcohol B is separately added, shake
It is centrifuged again afterwards, collects supernatant；It so repeats, mixes the supernatant repeatedly collected, as detection mother liquor, record total volume V,
Refrigeration is stand-by；Entire step operates under the conditions of being protected from light；
(3) differential pulse voltammetry is analyzed；
Hydroxyl-alpha-sanshool standard of certain volume is added dropwise to sample detection solution according to the principle of Multiple Standard Addition Method
Solution (α-SOH), followed by the differential pulse voltammetry curve of fagaramide substance in three-electrode system detection mixed solution；
The group of configuration electrolysis detection solution, electrolysis detection solution becomes the mixed solution of dehydrated alcohol and PBS buffer solution；It inhales
The detection mother liquor for taking step (2) to be prepared, volume are denoted as V1, with electrolysis detection solution dilution, sample detection solution A is obtained,
Volume is denoted as V2, utilize the differential pulse voltammetry curve of fagaramide substance in three-electrode system detection mixed solution；
Second of absorption detection mother liquor, volume V1, being diluted to volume with electrolytic solution is V2, is prepared again to be detected molten
Then α-SOH titer is added in liquid, volume is added and is denoted as V3, obtains mixed solution, detects mixed solution using three-electrode system
The differential pulse voltammetry curve of middle fagaramide substance；
Third time draws detection mother liquor, and volume V1, being diluted to volume with electrolytic solution is V2, is prepared again to be detected molten
α-SOH the titer of 2 times of V3 volumes is added in liquid, utilizes the difference of fagaramide substance in three-electrode system detection mixed solution
Pulse Voltammetry curve；
4th absorption detects mother liquor, and volume V1, being diluted to volume with electrolytic solution is V2, is prepared again to be detected molten
α-SOH the titer of 3 times of V3 volumes is added in liquid, utilizes the difference of fagaramide substance in three-electrode system detection mixed solution
Pulse Voltammetry curve；The rest may be inferred, repetitive operation；
Finally using the mass concentration of α-SOH titer as abscissa, corresponding peak current is that ordinate does linear regression,
Corresponding differential pulse voltammetry peak current is that ordinate does linear regression curves, and linear regression curves are absolute with abscissa intersection point
Value, as the total ceramide content in sample detection solution, are denoted as m；
(4) total ceramide content calculates；
The total ceramide content in sample detection solution obtained according to step (3), is further obtained by following formula scales
Detect the total ceramide content in mother liquor, i.e. α-SOH equivalent content:
In the formula, M: total ceramide content in Chinese prickly ash sample, μ g/g；M: in sample detection solution total ceramide content (according to
Calculated according to Multiple Standard Addition Method working curve), μ g α-SOH equivalent/mL；V1: the detection used when preparing sample detection solution
Mother liquor volume；V2: the volume of sample detection solution；V: detection mother liquor total volume, mL；W: zanthoxylum powder quality, g；A: systematic error
Compensation coefficient.
Preferably, in step (1), the mesh number of the sieve is 20~40 mesh.
Preferably, in step (2), the amount ratio of the zanthoxylum powder and dehydrated alcohol A are 5-25g:25-100mL；It is described super
Sound extraction time is 20~40min.
Preferably, in step (2), the centrifugal condition is equal are as follows: 2000~3000r/min of revolving speed, time are 3~8min.
Preferably, in step (2), the amount ratio of the filter residue and dehydrated alcohol B are 1g:(1-6) mL.
Preferably, in step (3), electrolysis detection solution in the volume ratio of dehydrated alcohol and PBS buffer solution be 1~3:7~
9；The concentration of PBS buffer solution is 0.1M.
Preferably, in step (3), the volume V of the detection mother liquor1With the volume V of sample detection solution2The ratio between be
(0.03-0.5): 10.
Preferably, in step (3), the volume V of the detection mother liquor1With the volume V of the α-SOH titer of addition3The ratio between
For (3-12): 1；α-SOH the concentration of standard solution is 1.0~10.0g/L.
Preferably, in step (3), three-electrode system group becomes working electrode (glass-carbon electrode), reference electrode (Ag/AgCl
Electrode), auxiliary electrode (platinum electrode)；The testing conditions parameter of differential pulse voltammetry are as follows: forward scan ,+0.6V-+1.2V；
Potential step 4mV；Amplitude 0.05V；Pulsewidth 0.05s；Sampling width 0.0167s；Pulse period 0.5s.
Preferably, in step (3), the number of the repetitive operation is 4~6 times.
The utility model has the advantages that
(1) present invention establishes the electrochemical detection method of total fagaramide content in Chinese prickly ash pericarp for the first time, with current state
Inside and outside existing fagaramide detection method is compared, and instrument used in this method has spy portable, inexpensive, easy to operate
Point.
(2) bare glassy carbon electrode used in electrochemical detection method of the present invention, without carrying out complexity to electrode
Moditied processing, it is easy to operate.
(3) electrochemical detection method detection accuracy of the present invention is high, the error rate with HPLC-MS method measurement result
Lower than 6%, detection speed is fast (differential pulse voltammetry scanning is completed in 2 minutes), can be used for carrying out Chinese prickly ash total ceramide fast
Fast accurate quantitative analysis.
(4) electrochemical detection method of the present invention is expected to the alternative that development is HPLC-MS, to Chinese prickly ash and its system
The detection and analysis of Chinese prickly ash total ceramide are of great significance in product.
Detailed description of the invention
Fig. 1 is the differential pulse voltammetry figure that standard addition method detects fagaramide content.
Fig. 2 is that standard addition method working curve calculates schematic diagram.
Fig. 3 is high effective liquid chromatography for measuring value and the matched curve of electrochemical process measured value.
Specific embodiment
In order to illustrate more clearly of the contents of the present invention, the present invention is made further below with reference to specific embodiment
It is bright, however, it should be noted that these embodiments are only to illustrate to be used, and be not necessarily to be construed as the limitation that the present invention is implemented.
1, laboratory sample, main agents and instrument
Sample: using the drying Chinese prickly ash of 7 kinds of places of production clear (being local harvesting) as test object, they are respectively as follows: 1
Number sample, southern green pepper level-one (Hanyuan County, sichuan Province)；No. 2 samples, Jiangjin pericarpium zanthoxyli schinifolii (Chongqing Jiangjin)；No. 3 samples, Jinyang pericarpium zanthoxyli schinifolii (Sichuan gold
Sun)；No. 4 samples, Shao lead to pericarpium zanthoxyli schinifolii (Yunnan Shao is logical)；No. 5 samples lie prostrate green pepper level-one (Gansu Linxia)；No. 6 samples, level-one pericarpium zanthoxyli bungeani (Shaanxi
Fuping)；No. 7 samples, level-one pericarpium zanthoxyli bungeani (Laiwu in Shandong province).
Reagent: hydroxyl-alpha-sanshool (α-SOH), hydroxy-beta-sanshool (β-SOH), hydroxyl-γ-sanshool (γ-SOH)
Standard items are purchased in Chengdu Mai Desheng Science and Technology Ltd., and purity is greater than 98%.Water used in experimentation is deionization
Water, experiment reagent are that analysis is pure.
Instrument: the three-electrode system of electrochemical workstation (CHI660E, Shanghai Chen Hua Instrument Ltd.) is: work electricity
Extremely 3mm diameter glass carbon disk electrode (model C HI104), reference electrode are Ag/AgCl electrode (model C HI111), auxiliary electrode
For platinum electrode (model C HI115).
Embodiment 1:
(1) Chinese prickly ash pericarp preparation of samples；
The drying Chinese prickly ash pericarp (back abbreviation Chinese prickly ash) for removing wilting or brown stain, crushed 30 meshes, obtains zanthoxylum powder ,-
It is refrigerated under the conditions of 20 DEG C to be detected；
(2) preparation of mother liquor is detected；
Zanthoxylum powder (W) 5g that step (1) obtains is accurately weighed in 50mL brown stuffed conical flask, the anhydrous second of 25mL is added
Alcohol, concussion shake up, and (25 ± 2 DEG C) ultrasound extraction 30min of room temperature, obtain crude extract under water bath with thermostatic control assistance, further will be thick
Extract centrifugation, 2000r/min are centrifuged 5min；Supernatant pours into 200mL brown volumetric flask (funnel auxiliary), collects filter residue and separately adds
Enter 25mL dehydrated alcohol, whirlpool shakes 1min, 5min is centrifuged under 2000r/min, collects supernatant, be so repeated 3 times, and merges 3
3 merging supernatants are settled to 200mL with dehydrated alcohol by secondary supernatant, shake up spare, and as detection mother liquor, record is overall
Product V (200mL)；
(3) differential pulse voltammetry is analyzed；
Hydroxyl-alpha-sanshool standard of certain volume is added dropwise to sample detection solution according to the principle of Multiple Standard Addition Method
Solution (α-SOH), followed by the differential pulse voltammetry curve of fagaramide substance in three-electrode system detection mixed solution；
Configuration electrolysis detection solution, consisting of: ethyl alcohol: PBS (pH=2) (1:9v/v)；The concentration of PBS buffer solution is
0.1M, draws the detection mother liquor that 30 μ L steps (2) are prepared, and volume is denoted as V1(30 μ L), it is fixed with electrolysis detection solution dilution
Hold to 10mL, obtain sample detection solution A, volume is denoted as V2(10mL) utilizes Chinese prickly ash in three-electrode system detection mixed solution
The differential pulse voltammetry curve of amide material；
Detection mother liquor and electrolysis detection solution mixing are drawn for the first time, prepare sample detection solution B again, 10 μ L are added
α-SOH titer is added volume and is denoted as V3 (10 μ L), and the α-SOH concentration of standard solution of addition is 2.6g/L, utilizes three-electrode system
Detect the differential pulse voltammetry curve of fagaramide substance in mixed solution；Second of absorption detection mother liquor and electrolysis detection solution
Sample detection solution C is prepared in mixing again, and the α-SOH titer of 20 μ L is added, using in three-electrode system detection mixed solution
The differential pulse voltammetry curve of fagaramide substance；Third time draws detection mother liquor and electrolysis detection solution mixing, prepares again
α-SOH the titer of 30 μ L is added in sample detection solution D, utilizes fagaramide substance in three-electrode system detection mixed solution
Differential pulse voltammetry curve；4th absorption detects mother liquor and electrolysis detection solution mixing, prepares sample detection solution again
α-SOH the titer of 40 μ L is added in E, is lied prostrate using the differential pulse of fagaramide substance in three-electrode system detection mixed solution
Pacify curve；Wherein sample detection solution A, sample detection solution B, sample detection solution C, sample detection solution D and sample detection
The volume and concentration of solution E are all the same, are the detection mother liquor drawing 30 μ L steps (2) and being prepared, detect solution with electrolysis
Dilution is settled to 10mL；It is superimposed the differential pulse voltammetry curve of electrolytic solution control, sample detection solution and four mark-ons, is obtained
To the differential pulse voltammetry figure of standard addition method detection Chinese prickly ash total ceramide content, as shown in Figure 1, as shown in Figure 1, with mark-on
Several increases, corresponding peak current are consequently increased.
Finally using the mass concentration of α-SOH titer as abscissa, corresponding differential pulse voltammetry peak current is ordinate
Linear regression curves are done, the working curve of standard addition method is obtained, as shown in Fig. 2, according to linear regression curves in figure and horizontal seat
The absolute value for marking intersection point, obtains the total ceramide content in sample detection solution, is denoted as m；
Electrochemical analyser using three-electrode system (working electrode (glass-carbon electrode), reference electrode (Ag/AgCl electrode),
Auxiliary electrode (platinum electrode)).The testing conditions parameter of differential pulse voltammetry are as follows: forward scan ,+0.6V-+1.2V；Current potential
Step 4mV；Amplitude 0.05V；Pulsewidth 0.05s；Sampling width 0.0167s；Pulse period 0.5s.
(4) total ceramide content calculates；
The total ceramide content in sample detection solution obtained according to step (3), is further obtained by following formula scales
Detect the total ceramide content in mother liquor, i.e. α-SOH equivalent content:
In the formula, M: total ceramide content in Chinese prickly ash sample, μ g/g；M: total ceramide content (foundation in sample detection solution
Multiple Standard Addition Method working curve calculates), μ g α-SOH equivalent/mL；V1: the detection used when preparing sample detection solution is female
Liquid product；V2: the volume of sample detection solution；V: detection mother liquor total volume, mL；W: zanthoxylum powder quality, g；A: systematic error is rectified
Positive coefficient.
(5) testing result is verified；
Seven kinds of Chinese prickly ash detection mother liquors are measured using HPLC-MS method and calculate the total ceramide content in Chinese prickly ash sample, to test
Demonstrate,prove the accuracy of the method for the present invention measurement result.
Hydroxy-beta-sanshool (β-SOH), hydroxyl-γ-sanshool (γ-SOH) in Chinese prickly ash sample are calculated with external standard method respectively
Content；Hydroxyl-alpha-sanshool (α-SOH), hydroxyl-ε-sanshool (ε-SOH) content in Chinese prickly ash sample are calculated with internal standard method, specifically
Step are as follows: β-SOH and γ-SOH mark product are dissolved with methanol respectively first, be configured to a series of solution of various concentrations and on
Machine is tested, and is required to 0.22 μm of organic system filter membrane before all solution examination with computer；To mark product concentration as abscissa, peak face is corresponded to
Product is that ordinate draws standard curve；Next takes detection mother liquor 0.5mL obtained in step (2), and volume is denoted as V4, dilute with methanol
It releases and is settled to 10mL, volume is denoted as V5；Examination with computer after 1mL is filtered is drawn, is measured in parallel 3 times, is averaged.It detects in mother liquor
β-SOH and γ-SOH material concentration are calculated according to standard curve, are denoted as m1 and m2, and convert by formula (a1) and (a2)
Obtain the β-SOH and γ-SOH content in Chinese prickly ash sample.
In formula: M: β-SOH content in Chinese prickly ash sample, μ g/g；M1: (establishing criteria is bent for β-SOH concentration in examination with computer sample
Line computation obtains), μ g/mL；(V5/V4): detection mother liquor extension rate, 20；V: detection mother liquor volume, 20mL；W: sample quality,
5g。
In formula: M: γ-SOH content in Chinese prickly ash sample, μ g/g；M2: γ-SOH concentration (establishing criteria in examination with computer sample
Curve is calculated), μ g/mL；(V5/V4): detection mother liquor extension rate, 20；V: detection mother liquor volume, 200mL；W: sample matter
Amount, 5g.
It accurately weighs zanthoxylum powder (W) 5g the and 2mg β-SOH standard items that step (1) obtains and has plug taper in 50mL brown
In bottle, β-SOH standard items quality is added and is denoted as n；25mL dehydrated alcohol is added, concussion shakes up, the room temperature under water bath with thermostatic control assistance
(25 ± 2 DEG C) ultrasound extraction 30min, obtain crude extract, are further centrifuged crude extract, 2000r/min is centrifuged 5min；Supernatant
It pouring into 200mL brown volumetric flask (funnel auxiliary), collects filter residue and 25mL dehydrated alcohol is separately added, whirlpool shakes 1min,
It is centrifuged 5min under 2000r/min, collects supernatant, is so repeated 3 times, merges 3 supernatants, by 3 merging supernatant nothings
Water-ethanol is settled to 200mL, shake up it is spare, as add internal standard detect mother liquor；0.5mL is taken to add internal standard detection mother liquor and step respectively
(2) obtained to detect mother liquor, it is settled to 10mL with methanol dilution, respectively examination with computer after absorption 1mL filtering, in record detection mother liquor
The peak area (A1) and ε-SOH peak area (A2) of α-SOH, and record detects mother liquor and internal standard detection mother liquor is added to be measured respectively
β-SOH peak area, calculating its difference is Δ A, and the α-SOH that detection mother liquor is calculated in Chinese prickly ash sample by formula (b1) contains
Amount, the ε-SOH content in Chinese prickly ash sample is calculated by formula (b1)；
In formula: M: α-SOH content in Chinese prickly ash sample, μ g/g；N: the content of addition β-SOH internal standard compound, 2000 μ g；A1: α-
SOH peak area；Δ A: being added internal standard and the difference for the β-SOH peak area that internal standard is measured is not added；W: sample quality, 5g.
In formula: M: ε-SOH content in Chinese prickly ash sample, μ g/g；N: the content of addition β-SOH internal standard compound, 2000 μ g；A2: ε-
SOH peak area；Δ A: being added internal standard and the difference for the β-SOH peak area that internal standard is measured is not added；W: sample quality, 5g.
α-SOH, β-SOH, γ-SOH and ε-SOH content summation are total in Chinese prickly ash sample in calculated Chinese prickly ash sample
Amide content.
DPV method (differential pulse voltammetry) and the Chinese prickly ash total ceramide content of material that HPLC-MS method measures are as shown in table 1.With
Electrochemical measurement is abscissa, using HPLC-MS measurement result as ordinate, carries out linear fit, obtains HPLC-MS and electricity
The matched curve of chemical determination value, as shown in figure 3, from the figure 3, it may be seen that being deposited between DPV method measured value and HPLC-MS method measured value
In systematic error, line style relationship meets y=2.6385x (R2=0.9955), i.e., contained using total ceramide in DPV method detection Chinese prickly ash
When amount, compensation coefficient 2.6385 should be used to be corrected its measured value, obtain DPV calibration of the output results value.HPLC-MS method and DPV
Method measurement result is as shown in table 1；Relative error between the HPLC-MS measured value and DPV calibration of the output results value of different Chinese prickly ash samples
Range is below 6% in 0.02%-5.87%.The result shows that: it can be obtained well in Chinese prickly ash detection mother liquor using DPV method
Chinese prickly ash total ceramide content, result is approximate with the result height of HPLC-MS.
1 HPLC-MS method of table is compared with DPV method measurement result
(6) the Chinese prickly ash total ceramide rate of recovery and precision mark-on reclaims result
1,4, No. 7 Chinese prickly ash sample is chosen, α-SOH is added during ultrasonic extraction and marks product, the content difference of α-SOH is added
For 13.15mg, 26.30mg, 39.45mg, the detection mother containing Chinese prickly ash total ceramide is prepared by the step (2) in embodiment, (3)
Liquid and the detection mother liquor that product are marked containing α-SOH.Detection 500 μ L of mother liquor is pipetted respectively, is settled to electrolysis detection solution dilution
10mL calculates the total ceramide concentration in extracting solution according still further to the step (4) in embodiment, and the results are shown in Table 2:
2 recovery testu result of table
As seen from the results in Table 2, the rate of recovery is between 94% to 108%, and relative standard deviation is below 4.0%, data knot
Fruit shows that detection method of the invention is reliable.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (9)
1. the electrochemical method of total ceramide content in a kind of quickly detection Chinese prickly ash pericarp, which comprises the following steps:
(1) the drying Chinese prickly ash pericarp for removing wilting or brown stain, crushes, screens through sieve, obtain zanthoxylum powder；
(2) step (1) resulting zanthoxylum powder is weighed, dehydrated alcohol A is added in quality W, and concussion shakes up, carries out under normal temperature condition
Ultrasound extraction, obtains crude extract；Crude extract is centrifuged, supernatant is collected；It collects filter residue and dehydrated alcohol B is separately added, shake
It is centrifuged again afterwards, collects supernatant；It so repeats, mixes the supernatant repeatedly collected, as detection mother liquor, record total volume V,
Refrigeration is stand-by；Entire step operates under the conditions of being protected from light；
(3) configuration electrolysis detects solution first, and the group of electrolysis detection solution becomes dehydrated alcohol and the mixing of PBS buffer solution is molten
Liquid；The detection mother liquor that aspiration step (2) is prepared, volume are denoted as V1, with electrolysis detection solution dilution, it is molten to obtain sample detection
Liquid A, volume are denoted as V2, utilize the differential pulse voltammetry curve of fagaramide substance in three-electrode system detection mixed solution；
Second of absorption detection mother liquor, volume V1, being diluted to volume with electrolytic solution is V2, prepares solution to be detected again,
Then α-SOH the titer that volume is V3 is added, obtains mixed solution, utilizes Chinese prickly ash acyl in three-electrode system detection mixed solution
The differential pulse voltammetry curve of amine substance；
Third time draws detection mother liquor, and volume V1, being diluted to volume with electrolytic solution is V2, prepares solution to be detected again,
α-SOH the titer of 2 times of V3 volumes is added, utilizes the differential pulse of fagaramide substance in three-electrode system detection mixed solution
Volt-ampere curve；
4th absorption detects mother liquor, and volume V1, being diluted to volume with electrolytic solution is V2, prepares solution to be detected again,
α-SOH the titer of 3 times of V3 volumes is added, utilizes the differential pulse of fagaramide substance in three-electrode system detection mixed solution
Volt-ampere curve；The rest may be inferred, repetitive operation；
Finally using the mass concentration of α-SOH titer as abscissa, corresponding peak current is that ordinate does linear regression, corresponding
Differential pulse voltammetry peak current be that ordinate does linear regression curves, the absolute value of linear regression curves and abscissa intersection point,
Total ceramide content as in sample detection solution, is denoted as m；
(4) the total ceramide content in sample detection solution obtained according to step (3), is further examined by following formula scales
Survey the total ceramide content in mother liquor, i.e. α-SOH equivalent content:
In the formula, M: total ceramide content in Chinese prickly ash sample, μ g/g；M: the total ceramide content in sample detection solution, μ g/mL；
V1: the detection mother liquor volume used when preparing sample detection solution；V2: the volume of sample detection solution；V: detection mother liquor is overall
Product, mL；W: zanthoxylum powder quality, g；A: systematic error compensation coefficient.
2. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (1), the mesh number of the sieve is 20~40 mesh.
3. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (2), the amount ratio of the zanthoxylum powder and dehydrated alcohol A are 5-25g:25-100mL；The ultrasound extraction time
For 20~40min.
4. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (2), the amount ratio of the filter residue being collected after centrifugation and dehydrated alcohol B are 1g:1-6mL；The centrifugal condition
Are as follows: 2000~3000r/min of revolving speed, time are 3~8min.
5. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (3), the volume ratio of dehydrated alcohol and PBS buffer solution is 1~3:7~9 in the electrolysis detection solution.
6. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (3), the volume V of the detection mother liquor1With the volume V of sample detection solution2The ratio between be 0.03-0.5:10.
7. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (3), the volume V of the detection mother liquor1With the volume V of the α-SOH titer of addition3The ratio between be 3-12:1；
α-SOH the concentration of standard solution is 1.0~10.0g/L.
8. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (3), the three-electrode system group becomes glass-carbon electrode, Ag/AgCl electrode and platinum electrode；Differential pulse volt
The testing conditions parameter of peace method are as follows: forward scan ,+0.6V-+1.2V；Potential step 4mV；Amplitude 0.05V；Pulsewidth 0.05s；It adopts
Sample width 0.0167s；Pulse period 0.5s.
9. the electrochemical method of total ceramide content, feature in a kind of quickly detection Chinese prickly ash pericarp according to claim 1
It is, in step (3), the number of the repetitive operation is 4~6 times.
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