CN103713066A - Method for extracting and detecting vanillin, methyl vanillin, ethyl vanillin and coumarin in milk powder - Google Patents
Method for extracting and detecting vanillin, methyl vanillin, ethyl vanillin and coumarin in milk powder Download PDFInfo
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- CN103713066A CN103713066A CN201410005844.5A CN201410005844A CN103713066A CN 103713066 A CN103713066 A CN 103713066A CN 201410005844 A CN201410005844 A CN 201410005844A CN 103713066 A CN103713066 A CN 103713066A
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- CBOQJANXLMLOSS-UHFFFAOYSA-N ethyl vanillin Chemical compound CCOC1=CC(C=O)=CC=C1O CBOQJANXLMLOSS-UHFFFAOYSA-N 0.000 title claims abstract description 76
- WJUFSDZVCOTFON-UHFFFAOYSA-N veratraldehyde Chemical compound COC1=CC=C(C=O)C=C1OC WJUFSDZVCOTFON-UHFFFAOYSA-N 0.000 title claims abstract description 76
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 title claims abstract description 39
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 229940073505 ethyl vanillin Drugs 0.000 title claims abstract description 38
- 239000000843 powder Substances 0.000 title claims abstract description 30
- 239000008267 milk Substances 0.000 title claims abstract description 27
- 210000004080 milk Anatomy 0.000 title claims abstract description 26
- 235000013336 milk Nutrition 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 18
- 229960000956 coumarin Drugs 0.000 title abstract 2
- 235000001671 coumarin Nutrition 0.000 title abstract 2
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 title abstract 2
- 235000012141 vanillin Nutrition 0.000 title abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 87
- 239000000243 solution Substances 0.000 claims abstract description 44
- 238000000605 extraction Methods 0.000 claims abstract description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000000523 sample Substances 0.000 claims abstract description 25
- 238000001514 detection method Methods 0.000 claims abstract description 18
- 239000012488 sample solution Substances 0.000 claims abstract description 15
- 239000002904 solvent Substances 0.000 claims abstract description 14
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 238000000874 microwave-assisted extraction Methods 0.000 claims abstract description 6
- 238000000746 purification Methods 0.000 claims abstract description 6
- 239000002304 perfume Substances 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 4
- PQMOXTJVIYEOQL-UHFFFAOYSA-N Cumarin Natural products CC(C)=CCC1=C(O)C(C(=O)C(C)CC)=C(O)C2=C1OC(=O)C=C2CCC PQMOXTJVIYEOQL-UHFFFAOYSA-N 0.000 claims description 35
- FSOGIJPGPZWNGO-UHFFFAOYSA-N Meomammein Natural products CCC(C)C(=O)C1=C(O)C(CC=C(C)C)=C(O)C2=C1OC(=O)C=C2CCC FSOGIJPGPZWNGO-UHFFFAOYSA-N 0.000 claims description 35
- 235000013599 spices Nutrition 0.000 claims description 29
- 238000012360 testing method Methods 0.000 claims description 25
- 230000000155 isotopic effect Effects 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 239000012086 standard solution Substances 0.000 claims description 9
- 239000003480 eluent Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 5
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- 239000007787 solid Substances 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
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- 239000012982 microporous membrane Substances 0.000 claims description 3
- 230000002000 scavenging effect Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000003643 water by type Substances 0.000 claims description 3
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- 238000012545 processing Methods 0.000 claims description 2
- 239000004526 silane-modified polyether Substances 0.000 claims description 2
- 239000012453 solvate Substances 0.000 claims description 2
- 235000013350 formula milk Nutrition 0.000 abstract description 11
- 238000011084 recovery Methods 0.000 abstract description 7
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 abstract description 6
- 238000010828 elution Methods 0.000 abstract description 5
- 238000002414 normal-phase solid-phase extraction Methods 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 4
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Abstract
The invention discloses a method for extracting and detecting vanillin, methyl vanillin, ethyl vanillin and coumarin in milk powder. The method comprises the following steps: performing preparation, purification, detection and recovery rate experiment on a to-be-tested sample; adding a methanol/water solution into the infant milk powder; performing microwave extraction to obtain a to-be-tested solution; centrifuging the to-be-tested solution, and performing solid-phase extraction; performing leaching through water and a methanol/water solution and elution through an elution solvent in sequence to obtain an elution solution; evaporating for concentrating the elution solution, then diluting the concentrated elution solution through a methanol solvent, and filtering to obtain a to-be-tested sample solution; preparing a standard working solution with stepped concentration, and feeding the to-be-tested sample solution and the standard working solution respectively into a gas chromatography-mass spectrometer/mass spectrometer (GC-MS/MS) to obtain a working curve and a mass spectrum, and performing qualitative and quantitative analysis according to the working curve and the mass spectrum; and weighing the infant milk powder with four perfumes with determined content, sequentially measuring the concentration of the four perfumes according to the experimental steps in sequence, and calculating measurement values and determination values to obtain the recovery rate of 83.6-107.8 percent. The method has the advantages of high extraction efficiency, accurate qualitative and quantitative determination, high sensitivity and accurate result.
Description
Technical field
The invention belongs to food inspection field, relate to microwave abstracting-organophosphorous pesticides-tandem mass spectrometry (GC-MS/MS), the extraction and the detection method that specifically refer to vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder, be particularly useful for baby milk powder.Background technology
Spices comprises that vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin are conventional household chemicals spices, is also important food additives, is widely used as the fumet of chocolate, ice cream, cake.But heavy dose of spices such as vanillic aldehyde that use will be harmful to health.Due to the quality good or not of baby formula milk powder, frequently cause in recent years food safety affair.Therefore, set up that to measure accurately, reliably the content of vanillic aldehyde in baby formula milk powder, methyl vanillin, Ethyl vanillin and cumarin extremely important.
In baby formula milk powder, the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin residual component mainly comprises that from baby formula milk powder, extracting residual vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin makes sample, and this sample is carried out to two steps of test analysis, wherein sample is prepared most importantly, directly affects follow-up test precision of analysis.Analytical approach can be divided into qualitative analysis, quantitative test and qualitative and quantitative analysis technology.Quantitative test generally adopts chromatographic technique and hydrolysis and condensation, and chromatographic technique has the methods such as gas chromatography and liquid chromatography, and hydrolysis and condensation has the methods such as liquid chromatograph mass spectrography and gas chromatography-mass spectrography; Qualitative and quantitative analysis will have two kinds of technology concurrently and be integrated, testing result will provide structural information and the quantity information of object simultaneously, according to the regulation of the optional execution bill 2002/657/EC of EU Committee, forbidden drugs is confirmed to the method detecting the information of configuration aspects must be able to be provided, and will reach 4 confirmation points of this bill regulation.
At present, owing to can following without relevant criterion, spices detection method in relevant baby formula milk powder in spices or other materials has liquid phase chromatography, vapor-phase chromatography, Liquid Chromatography-Tandem Mass Spectrometry, capillary electrophoresis, but does not also detect the microwave abstracting-organophosphorous pesticides-tandem mass spectrometry of vanillic aldehyde in baby formula milk powder, methyl vanillin, Ethyl vanillin and cumarin at present simultaneously.
Summary of the invention
The object of the invention is the shortcoming and defect existing in order to overcome prior art, and provide extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder, the method is in conjunction with microwave abstracting-organophosphorous pesticides-tandem mass spectrometry, can measure vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in milk powder simultaneously, it is accurate, quick, highly sensitive that the method has advantages of, for the fast detecting of milk powder provides reliable analytical approach.
For achieving the above object, technical scheme of the present invention is to comprise the following steps:
(1) preparation of testing sample: take milk powder, preparation Isotopic Internal Standard solution adds and extracts solvent and Isotopic Internal Standard solution in milk powder, is installed on after uniform stirring in Microwave Extraction Apparatus, after microwave abstracting, adds extraction solvent constant volume and obtains liquid to be checked;
(2) purification of testing sample: liquid to be checked prepared by step () is through the centrifugal liquid that is purified, fixedly phase extraction column carries out pre-service, get the upper extraction column of scavenging solution, through water and methanol/water solution drip washing, remove solid impurity, in methanol/water solution, the volume ratio of first alcohol and water is 1:9, with the vanillic aldehyde on eluting solvent wash-out extraction column, methyl vanillin, Ethyl vanillin and 4 kinds of spices of cumarin, obtain eluent again, by after eluent enrichment method, with methanol solvate, dilute and obtain testing sample solution after 0.22 μ m filtering with microporous membrane;
(3) detection of testing sample: preparation vanillic aldehyde, methyl vanillin, the standard operation solution of 4 kinds of spice solution ladder concentration of Ethyl vanillin and cumarin, the difference sample introduction on gas chromatography-tandem mass spectrometer by the testing sample solution in step (two) and standard operation solution, acquisition is with vanillic aldehyde in testing sample solution, methyl vanillin, the ratio of the concentration of Ethyl vanillin and 4 kinds of spices of cumarin and the concentration of isotopic number inner mark solution is horizontal ordinate, again with vanillic aldehyde in testing sample solution, methyl vanillin, the working curve that the ratio of the peak area of Ethyl vanillin and 4 kinds of spices of cumarin and the peak area of isotopic number inner mark solution is ordinate, also have and obtain the vanillic aldehyde that can determine parent ion, methyl vanillin, 4 kinds of spices of Ethyl vanillin and cumarin and isotopic one-level mass spectrogram, determine after parent ion, adopt daughter ion scanning to obtain second order ms figure, then from second order ms figure, choose quota ion and qualitative ion, then with working curve, quota ion is carried out quantitatively calculating and obtain vanillic aldehyde in testing sample solution, methyl vanillin, 4 kinds of concentration of perfume of Ethyl vanillin and cumarin.
Further arrange is that extraction solvent in described step () is wherein a kind of in methanol/water solution, ethanol/water solution, methyl alcohol, ethanol.Preferably, the component volume ratio of methanol/water solution is 1:1; The component volume ratio of ethanol/water solution is also 1:1.
Further arranging is that described methanol aqueous solution concentration is 25% ~ 90%.
Further setting is that the microwave abstracting temperature in described step () is 60～80 ℃.
Further setting is that the Isotopic Internal Standard solution in described step () is
13c
6-vanillic aldehyde,
13c-methyl vanillin and D
4-cumarin inner mark solution.
Further arrange be extraction in described step (two) fixing be Waters Oasis HLB SPE post mutually, and by methyl alcohol, water, SPE post is carried out to activation processing.
Further setting is that the eluting solvent in described step (two) is methyl alcohol.
Principle and advantage of the present invention:
Adopt such scheme, first, the present invention passes through microwave-assisted extraction technique, take methanol/water, ethanol/water, methyl alcohol, ethanol as extracting solvent, in sample, add appropriate Isotopic Internal Standard solution and extract solvent soaking, after vortex vibration, then by uniform temperature, time and power, extract and make liquid to be checked in Microwave Extraction Apparatus.If adopt traditional ultrasonic extraction method of not heating, the stability that is conducive to spices, ultrasonication only applies the bonding action that is not enough to destroy spices and albumen, the intermolecular formation of fatty isopolarity, cause the spices stripping of dissociating not thorough, affect extraction efficiency, cause final detection precision of analysis and reappearance bad.The present invention adopts microwave abstracting, because microwave energy is to destroy bonding action by inner homogeneous heating acceleration molecular motion.Therefore in the situation that power, temperature, time, pressure are controlled, microwave abstracting can make some organic components in solid or semi-solid sample effectively separated with base matter, thereby improves extraction efficiency, and can also reappear well recovery of extraction.Compare with traditional ultrasonic extraction, extraction of ocean eddies, extracting method extraction effect of the present invention is good, and the recovery can reach 83.6～107.8%; Far above the recovery 74.6-83.2% of ultrasonic extraction and the recovery 72.8-80.9% of extraction of ocean eddies.And that extracting method of the present invention also has is quick, energy-conservation, save solvent, pollute littlely, and is conducive to extract the advantage of determination system of thermal unstable material.
Secondly, the present invention also carries out purified treatment to the liquid to be checked being obtained by microwave abstracting, and this purified treatment comprises the process of purifying and enrichment.First step purifying: first by liquid to be checked through centrifugal rough purification, then the scavenging solution after centrifugal is passed through through water and methanol/water solution drip washing, methanol-eluted fractions successively by the method for Solid phase extraction separation; Second step enrichment: by the concentrated evaporate to dryness of the liquid to be checked of purifying in the first step, then add dissolve with methanol solution, through 0.22 μ m filtering with microporous membrane, the liquid to be checked being purified.Liquid to be checked after this purification can not pollute detecting instrument.In the present invention, the SPE post of Solid-Phase Extraction is Waters Oasis HLB SPE post, because the polarity size of spices and other impurity there are differences, on solid-phase extraction column, absorption produces and retains difference, then by preferred drip washing condition and elution requirement, reaches the object of purification enrichment spices.
Again, the present invention is to analyze by gas chromatography-tandem mass spectrometer (GC-MS/MS) for the qualitative and quantitative detection analysis of vanillic aldehyde, methyl vanillin, Ethyl vanillin and coumarinoids in baby formula milk powder.First configure one group of step-like standard operation solution, then the difference sample introduction on gas chromatography-tandem mass spectrometer by standard operation liquid and liquid to be checked.The chromatographic peak retention time occurring in liquid to be checked is consistent with standard operation solution, be less than ± 2.5 % of permissible variation, the relative abundance of the standard operation solution that the relative abundance of the corresponding mass spectrometry ion of this chromatographic peak is suitable with concentration is consistent, and relative abundance deviation is no more than regulation, can determines in this liquid to be checked and contain spices.In addition, in mass spectrum quantitative test, having two kinds of methods is respectively internal standard method and external standard method.Internal standard method is that a certain amount of isotope labeling material is made to internal standard compound, joins in the sample of accurate weighing, according to the ratio of the mass ratio of tested sample and internal standard compound and corresponding chromatographic peak area thereof, calculates the content of tested component.Select internal standard compound to have 4 requirements: 1. internal standard compound should be non-existent pure material in this sample; 2. it must be dissolved in sample completely, and completely separated with the chromatogram peak energy of each component in sample; 3. add the amount of internal standard compound should be close to tested component; 4. the position of chromatographic peak should be close with the position of the chromatographic peak of tested component, or in the middle of several tested component chromatographic peaks.External standard method is to compare material by the sterling of component to be measured, with the response signal of component to be measured in control substance of plant drug and sample, compares and carries out quantitative method.Because internal standard method is to calculate by measuring the relative value of the peak area of internal standard compound and tested component, thereby eliminated to a certain extent the caused error of variation of operating conditions etc., its measurement result is comparatively accurate.Although and external standard method is easy, it is very accurate that sample size requires, and be strict controlled under the operating conditions identical with reference material and carry out, otherwise cause analytical error, can not get measurement result accurately.
Below in conjunction with specification drawings and specific embodiments, the present invention is described further.
Accompanying drawing explanation
Fig. 1 is 4 kinds of spices and the 3 kinds of Isotopic Internal Standard structural drawing in the specific embodiment of the invention; In Fig. 1, A: vanillic aldehyde, B: methyl vanillin, C: Ethyl vanillin, D: cumarin E:
13c
6-vanillic aldehyde, F:
13c-methyl vanillin, G:D
4-cumarin;
Fig. 2 is 10 μ g kg in the specific embodiment of the invention
-1milk powder mark-on MRM chromatogram; In Fig. 2, each little figure is corresponding respectively: vanillic aldehyde (A),
13c
6-vanillic aldehyde (B), Ethyl vanillin (C), methyl vanillin (D),
13c-methyl vanillin (E), cumarin (F), D
4-cumarin (G);
Fig. 3 is 4 kinds of spices and 3 kinds of Isotopic Internal Standard GC-MS/MS scanning second order ms figure in the specific embodiment of the invention; In Fig. 3, each little figure is corresponding respectively: vanillic aldehyde (A),
13c
6-vanillic aldehyde (B), Ethyl vanillin (C), methyl vanillin (D),
13c-methyl vanillin (E), cumarin (F), D
4-cumarin (G);
Fig. 4 is that in the specific embodiment of the invention, 4 kinds of spices and 3 kinds of Isotopic Internal Standard GC-MS/MS scanning second order ms figure resolve collection of illustrative plates; In Fig. 4, each little figure is corresponding respectively: vanillic aldehyde (A),
13c
6-vanillic aldehyde (B), Ethyl vanillin (C), methyl vanillin (D),
13c-methyl vanillin (E), cumarin (F), D
4-cumarin (G);
Fig. 5 is interior mark typical curve in the specific embodiment of the invention, and in Fig. 5, each little figure is corresponding respectively: vanillic aldehyde (A), methyl vanillin (B), Ethyl vanillin (C), cumarin (D);
Embodiment
Below by embodiment, the present invention is specifically described; only be used to further illustrate the present invention; can not be interpreted as limiting the scope of the present invention, the technician in this field can make some nonessential improvement and adjustment to the present invention according to the content of foregoing invention.
Embodiment 1
Comprise the following steps:
(1) preparation of testing sample: take baby formula milk powder 1.0g, add 0.5mL
13c
6-vanillic aldehyde,
13c-methyl vanillin and D
4in-cumarin, (concentration is 400 ng mL to mark mixed solution
-1), then add 15mL methanol/water mixed solution, and after uniform stirring, be installed in Microwave Extraction Apparatus, after microwave abstracting, extraction temperature is 70
?c, takes out standingly, gets supernatant liquor, is settled to 50mL volumetric flask obtains liquid to be checked with pure water;
(2) Solid phase extraction: get HLB SPE post, successively with first alcohol and water activation SPE post, get 4～6mL liquid to be clean to SPE post, use successively again 4～6mL water and 4～6mL methanol/water mixed solution (v/v, 1/9) drip washing SPE post, SPE post vacuum is drained, with the methyl alcohol of 6～8mL, SPE post is carried out to wash-out, obtain eluent, eluent is concentrated after, obtain testing sample solution after diluting miillpore filter with methyl alcohol;
(3) configuration of standard solution: get vanillic aldehyde, methyl vanillin, Ethyl vanillin, cumarin,
13c
6-vanillic aldehyde,
13c-methyl vanillin and D
4-cumarin, with methanol/water mixed solution, be mixed with respectively single mark storing solution that mass concentration is 200 mg/L, measure while using, with methyl alcohol, dilute above-mentioned standard reserving solution, vanillic aldehyde, methyl vanillin, Ethyl vanillin, cumarin are mixed with respectively to 1.0,2.0,4.0, the hybrid standard working solution of 10.0,20.0 ng/mL contains in each standard solution simultaneously
13c
6-vanillic aldehyde,
13c-methyl vanillin and D
4-cumarin inner mark solution is 10ng/mL;
(4) detection of testing sample: the difference sample introduction on gas chromatography-tandem mass spectrometer by the testing sample solution in step (two) (three) and standard operation solution, it is horizontal ordinate that the ratio of the concentration of each spices and the concentration of isotopic number inner mark solution in standard operation solution is take in acquisition, take again each spices in standard operation solution peak area and the ratio of the peak area of the isotopic number inner mark solution working curve that is ordinate, also have and obtain spices and the isotopic one-level mass spectrogram that can determine parent ion, determine after parent ion, adopt daughter ion scanning to obtain second order ms figure, then from second order ms figure, choose quota ion and qualitative ion, then with working curve, quota ion is carried out quantitatively calculating and obtain each concentration of perfume in testing sample solution.
Referring to table 1, it is 4 kinds of spices and 3 kinds of isotope GC-MS/MS testing conditions
4 kinds of spices that are the embodiment of the present invention 1 referring to table 2 add the recovery, precision and quantitative limit
Claims (7)
1. the extraction of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin and a detection method in milk powder, is characterized in that comprising the following steps:
The preparation of testing sample: take milk powder, preparation Isotopic Internal Standard solution adds and extracts solvent and Isotopic Internal Standard solution in milk powder, is installed on after uniform stirring in Microwave Extraction Apparatus, after microwave abstracting, adds extraction solvent constant volume and obtains liquid to be checked;
The purification of testing sample: liquid to be checked prepared by step () is through the centrifugal liquid that is purified, fixedly phase extraction column carries out pre-service, get the upper extraction column of scavenging solution, through water and methanol/water solution drip washing, remove solid impurity, with the vanillic aldehyde on eluting solvent wash-out extraction column, methyl vanillin, Ethyl vanillin and 4 kinds of spices of cumarin, obtain eluent again, by after eluent enrichment method, with methanol solvate, dilute and obtain testing sample solution after 0.22 μ m filtering with microporous membrane;
The detection of testing sample: preparation vanillic aldehyde, methyl vanillin, the standard operation solution of 4 kinds of spice solution ladder concentration of Ethyl vanillin and cumarin, the difference sample introduction on gas chromatography-tandem mass spectrometer by the testing sample solution in step (two) and standard operation solution, acquisition is with vanillic aldehyde in testing sample solution, methyl vanillin, the ratio of the concentration of Ethyl vanillin and 4 kinds of spices of cumarin and the concentration of isotopic number inner mark solution is horizontal ordinate, again with vanillic aldehyde in testing sample solution, methyl vanillin, the working curve that the ratio of the peak area of Ethyl vanillin and 4 kinds of spices of cumarin and the peak area of isotopic number inner mark solution is ordinate, also have and obtain the vanillic aldehyde that can determine parent ion, methyl vanillin, 4 kinds of spices of Ethyl vanillin and cumarin and isotopic one-level mass spectrogram, determine after parent ion, adopt daughter ion scanning to obtain second order ms figure, then from second order ms figure, choose quota ion and qualitative ion, then with working curve, quota ion is carried out quantitatively calculating and obtain vanillic aldehyde in testing sample solution, methyl vanillin, 4 kinds of concentration of perfume of Ethyl vanillin and cumarin.
2. extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder according to claim 1, is characterized in that: the extraction solvent in described step () is wherein a kind of in methanol/water solution, ethanol/water solution, methyl alcohol, ethanol.
3. according to extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder in claim 2, it is characterized in that: described methanol aqueous solution concentration is 25% ~ 90%.
4. extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder according to claim 1, is characterized in that: the microwave abstracting temperature in described step () is 60～80 ℃.
5. extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder according to claim 1, is characterized in that: the Isotopic Internal Standard solution in described step () is
13c
6-vanillic aldehyde,
13c-methyl vanillin and D
4-cumarin inner mark solution.
6. extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder according to claim 1, it is characterized in that: it is Waters Oasis HLB SPE post mutually that the extraction in described step (two) is fixed, and by methyl alcohol, water, SPE post is carried out to activation processing.
7. extraction and the detection method of vanillic aldehyde, methyl vanillin, Ethyl vanillin and cumarin in a kind of milk powder according to claim 1, is characterized in that: the eluting solvent in described step (two) is methyl alcohol.
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| CN201410005844.5A CN103713066B (en) | 2014-01-03 | 2014-01-03 | Method for extracting and detecting vanillin, methyl vanillin, ethyl vanillin and coumarin in milk powder |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104655767A (en) * | 2015-03-20 | 2015-05-27 | 中国烟草总公司郑州烟草研究院 | Method for simultaneously determining vanillin, ethyl vanillin and coumarin in electronic cigarette solution |
| CN106841451A (en) * | 2017-02-24 | 2017-06-13 | 中国烟草总公司郑州烟草研究院 | The assay method of cumarin and its metabolin in mouse blood based on Ultra Performance Liquid Chromatography track trap high resolution mass spectrum |
| CN107677757A (en) * | 2017-09-08 | 2018-02-09 | 大连市食品检验所 | The method for determining vanillic aldehyde in food, methyl vanillin, Ethyl vanillin simultaneously |
| CN108645932A (en) * | 2018-05-16 | 2018-10-12 | 陕西科技大学 | A kind of method of violated addition essence and flavoring agent in rapid screening dilute cream |
| CN110031571A (en) * | 2019-05-23 | 2019-07-19 | 上海烟草集团有限责任公司 | The measuring method of vanillic aldehyde and Ethyl vanillin content in a kind of cigarette mainstream flue gas |
| CN112578038A (en) * | 2020-11-17 | 2021-03-30 | 沈阳化工大学 | Analysis and detection method for berberine hydrochloride bulk drug |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507833A (en) * | 2011-11-08 | 2012-06-20 | 韩超 | Method for determining vanillin, ethyl vanillin and coumarin in milk by solid phase extraction-liquid chromatogram-electrospray tandem mass spectrometry |
| CN102944616A (en) * | 2012-10-15 | 2013-02-27 | 青岛谱尼测试有限公司 | Method for simultaneous detection of vanillin and ethyl vanillin in milk powder |
-
2014
- 2014-01-03 CN CN201410005844.5A patent/CN103713066B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507833A (en) * | 2011-11-08 | 2012-06-20 | 韩超 | Method for determining vanillin, ethyl vanillin and coumarin in milk by solid phase extraction-liquid chromatogram-electrospray tandem mass spectrometry |
| CN102944616A (en) * | 2012-10-15 | 2013-02-27 | 青岛谱尼测试有限公司 | Method for simultaneous detection of vanillin and ethyl vanillin in milk powder |
Non-Patent Citations (7)
| Title |
|---|
| ANUJ SHARMA ET AL.: "Microwave- and ultrasound-assisted extraction of vanillin and its quantification by high-performance liquid chromatography in Vanilla planifolia", 《J. SEP. SCI.》 * |
| ARTURO LONGARES-PATRON ET AL.: "Focused microwaves-assisted extraction and simultaneous spectrophotometric determination of vanillin and p-hydroxybenzaldehyde from vanilla fragans", 《TALANTA》 * |
| LOWRI S. DE JAGER ET AL.: "Comparison of headspace-SPME-GC–MS and LC–MS for the detection and quantification of coumarin, vanillin, and ethyl vanillin in vanilla extract products", 《FOOD CHEMISTRY》 * |
| 冷中成 等: "乳制品中香兰素和乙基香兰素的溶剂萃取—气相色谱分析法", 《职业与健康》 * |
| 欧菊芳 等: "气相色谱法测定婴幼儿配方奶粉中香兰素和乙基香兰素", 《江西科学》 * |
| 罗冠中 等: "液相色谱串联四极杆质谱法同时测定婴幼儿配方食品中甲基香兰素和乙基香兰素", 《分析试验室》 * |
| 赵建国 等: "气相色谱质谱联用测定婴幼儿奶粉中香兰素和乙基香兰素", 《生命科学仪器》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104655767A (en) * | 2015-03-20 | 2015-05-27 | 中国烟草总公司郑州烟草研究院 | Method for simultaneously determining vanillin, ethyl vanillin and coumarin in electronic cigarette solution |
| CN106841451A (en) * | 2017-02-24 | 2017-06-13 | 中国烟草总公司郑州烟草研究院 | The assay method of cumarin and its metabolin in mouse blood based on Ultra Performance Liquid Chromatography track trap high resolution mass spectrum |
| CN107677757A (en) * | 2017-09-08 | 2018-02-09 | 大连市食品检验所 | The method for determining vanillic aldehyde in food, methyl vanillin, Ethyl vanillin simultaneously |
| CN107677757B (en) * | 2017-09-08 | 2019-11-08 | 大连市食品检验所 | The method for measuring vanillic aldehyde in food, methyl vanillin, Ethyl vanillin simultaneously |
| CN108645932A (en) * | 2018-05-16 | 2018-10-12 | 陕西科技大学 | A kind of method of violated addition essence and flavoring agent in rapid screening dilute cream |
| CN108645932B (en) * | 2018-05-16 | 2020-09-01 | 陕西科技大学 | Method for rapidly screening prohibited essence and spice added in cream |
| CN110031571A (en) * | 2019-05-23 | 2019-07-19 | 上海烟草集团有限责任公司 | The measuring method of vanillic aldehyde and Ethyl vanillin content in a kind of cigarette mainstream flue gas |
| CN110031571B (en) * | 2019-05-23 | 2021-09-07 | 上海烟草集团有限责任公司 | Method for measuring content of vanillin and ethyl vanillin in cigarette mainstream smoke |
| CN112578038A (en) * | 2020-11-17 | 2021-03-30 | 沈阳化工大学 | Analysis and detection method for berberine hydrochloride bulk drug |
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