CN106546679A - The liquid phase chromatography analytical method of acrylamide in a kind of detection fried food - Google Patents
The liquid phase chromatography analytical method of acrylamide in a kind of detection fried food Download PDFInfo
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- CN106546679A CN106546679A CN201610966508.6A CN201610966508A CN106546679A CN 106546679 A CN106546679 A CN 106546679A CN 201610966508 A CN201610966508 A CN 201610966508A CN 106546679 A CN106546679 A CN 106546679A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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Abstract
The present invention relates to a kind of liquid phase chromatography analytical method for detecting acrylamide in fried food, take the fried food after grinding and add acrylamide standard solution, plus normal hexane is vortexed and shakes, petroleum ether layer, plus deionized water are discarded, be vortexed concussion, constant temperature oscillation, ultrasonic vibration Jia Karui reagent Cs arrezI and CarrezII remove protein again, high speed centrifugation, the good SPE pillars of supernatant overactivation, filters to obtain need testing solution；Need testing solution is analyzed using liquid chromatography.Deionized water heating extraction of the present invention, supersound extraction acrylamide, extraction ratio 98.2%, RSD=2.83%；Deionized water safety non-toxic, it is cheap；Detection limit and quantitative limit are low, and detection is limited to 0.002 μ gmL 1, is quantitatively limited to 0.005 μ gmL 1.The inventive method is quick, accurate, efficient, safety, and accurately the content of acrylamide in fried food can be detected.
Description
Technical field
The present invention relates in a kind of fried food acrylamide analytical technology, and in particular in a kind of detection fried food
The liquid phase chromatography analytical method of acrylamide.
Background technology
Acrylamide has stronger permeability, can be rapidly entered by respiratory tract, digestive tract and skin and mucosa etc.
Cause poisoning in human body.Zoopery and the experiment of long-term exposed amount show that it mainly has neurotoxicity, genotoxicity, heredity
Toxicity and teratogenecity etc..Early in 1994, acrylamide is just classified as 2A groups by IARC, and " mankind were possible carcinogenic
Thing ".
The method of acrylamide is detected at present in the world mainly by Solid-Phase Extraction column purification and13C or2H labelling propylene
Amide is used in conjunction (HPLC- as the gas chromatography-mass spectrum (GC-MS) and High Performance Liquid Chromatography/Mass Spectrometry of the isotopic dilution of internal standard substance
MS/MS) method is measured.Both approaches use Mass Spectrometer Method, and quantitative accurate, detection limit is low but price is prohibitively expensive, less
Suitable common lab is promoted the use of, and GC-MS early stages need to carry out latter bromide derivative, and the sample treatment operation cycle is longer, is derived
Change reagent toxicity larger, while LC-MS/MS causes the impurity in food to coexist due to the weak retention of acrylamide when detecting
The summit that goes out of thing is covered, and the ion for having identical mass-to-charge ratio with acrylamide is produced in Mass Spectrometer Method, so as to affect detection
Precision.In the prior art, Acrylamide in Foods is determined with reversed-phase high-performance liquid chromatography and Normal-phase HPLC
Content, detection limit respectively reaches 4.0 μ gkg-1 and 10.0 μ gL-1, but pre-treatment is complex, and normal-phase chromatography exists
Laboratory is promoted less.
Existing GB analyzing detecting method (GB5009.204-2014), step are complicated, need more than 10 minutes, and time-consuming,
And need derivative；Using standard addition method, 1 sample can only be processed 1 time；10 μ g/kg are limited to quantitatively.
CN104122453 discloses a kind of method for quick of the acrylamide based on dielectric property, including following step
Suddenly：Detected sample is broken into pieces and is placed in deionized water, be refluxed 1～4h, filter, obtain filtrate；Determine filtrate 3 × 108
Loss factor under～1010Hz frequency conditions；The standard curve of the loss factor of acrylamide standard solution under same frequency
Or dependent equation, acrylamide concentration in filtrate is obtained, the content of acrylamide in sample is calculated.Detection time of the present invention is short,
Method is simple to operation, for the Detection results of detection low concentration acrylamide content are better than the detection side using liquid chromatograph
Method, has wide practical use in terms of the content of acrylamide in detection fried food.
The content of the invention
The purpose of the present invention is for above-mentioned present situation, it is desirable to provide a kind of energy is quick, accurate, efficient, safety is to fried food
In product, the content of acrylamide carries out the liquid phase chromatography analytical method for detecting acrylamide in accurately detection fried food.
The implementation of the object of the invention is, the liquid phase chromatography analytical method of acrylamide in a kind of detection fried food,
Concretely comprise the following steps：
1) weigh the fried food after 2.0g grinds to be placed in 20mL politef centrifuge tubes, add acrylamide
Standard solution, stands 10min；
2) to step 1) obtained by stand liquid in add 10mL normal hexane to be vortexed concussion 1min, and stand 10min defats, discard
Petroleum ether layer, 40 ± 1 DEG C of nitrogen are dried up, and the above-mentioned normal hexane skimming processes of repetition are once；
3) to Jing step 2) 14mL deionized waters are added in sample after defat, it is vortexed concussion half a minute, is then placed in perseverance
Warm agitator is taken out after 30min is shaken at 60 DEG C, resulting solution ultrasonic vibration 20min again；
4) to Jing steps 3) obtained by shake liquid in add auspicious reagent C arrezI of card and each 0.5mL of CarrezII, stand
10min, removes protein, in 4 DEG C, 11000rpm high speed centrifugation 20min；Take supernatant 2mL, the good SPE pillars of overactivation,
Front 2mL effluent is discarded, after filtering after collection, need testing solution is obtained；
5) by step 4) in need testing solution, be analyzed using liquid chromatography；
Chromatographic column used is：WatersT3；
The high efficiency liquid phase chromatographic analysis method condition is：Mobile phase：Methanol：0.1% aqueous formic acid=5:95；Flow velocity：
1.0ml/min；Detection column temperature：30℃；Detection wavelength：205nm；Sample size：20μL.
Compared with prior art, the invention has the advantages that：
1st, deionized water extracts acrylamide, and extraction ratio is higher, up to 98.2%, RSD=2.83%；Simultaneously water security without
Poison, it is cheap；
2nd, mobile phase ratio than considerable influence, flow velocity and sample size will unite to chromatographic condition, by experiment
To determine final condition, acrylamide can be by good eluting, and peak shape baseline is steady, without tailing peak；
3rd, compare existing GB analyzing detecting method (GB5009.204-2014) quantitative limit low, only 5 μ g/kg；Using interior
Mark method, can process multiple samples, efficiency high simultaneously；Used time is only 5 minutes, and detection is quick；
4th, adopt first defat, purify again, during purification, employ the auspicious reagent of card (CarrezI and CarrezII) and SPE
(SupercleanTM ENVITM- 18) solid phase pillar, can adsorb the little molecule pigment of polarity, and the acrylamide big to polarity
Without absorption, detergent power can be brought up to by Solid-Phase Extraction；While avoiding the presence of the impurity such as the protein in sample, and lead
Cause emulsion to be produced in layering interfaces, it is impossible to remove oil impurities well, affect the precision for determining.
Description of the drawings
Comparisons of the Fig. 1 for different solvents extraction ratio,
Fig. 2 is acrylamide standard solution chromatogram,
Fig. 3 is acrylamide HPLC collection of illustrative plates in pure sample product,
Fig. 4 is acrylamide HPLC collection of illustrative plates in sample mark-on.
Specific embodiment
The present invention takes the fried food after grinding and adds acrylamide standard solution, plus normal hexane to be vortexed and shake, and abandons
Petroleum ether layer, plus deionized water are removed, be vortexed concussion, constant temperature oscillation, then ultrasonic vibration Jia Karui reagent Cs arrezI and CarrezII
Protein is removed, high speed centrifugation, the good SPE pillars of supernatant overactivation filter to obtain need testing solution；Need testing solution can profit
It is analyzed with liquid chromatography.
Described fried food is commercially available French fries, fried drumstick, sweet potato cake, instant noodles, pan-fried dumpling or fritters of twisted dough.
The SPE pillars for having activated are using front needing with 2mL methanol and 2mL deionized water balances.
Because acrylamide is used as a kind of highly polar material, the extremely weak (k' of the reservation in common reversed phase chromatographic column<2.0),
Mix with solvent peak so as to cause appearance time too fast, be not readily separated, common C18 posts cannot meet analysis needs.This Shen
The different chromatographic columns of Jing researchs of asking someone find, due to WatersT3 its special stationary phase material can be resistant to Gao Shuixiang,
Adapt to the reservation of polar compound and separate, so as to meet the needs of detection, therefore the chromatographic column of the present invention elects Waters asT3。
(2) present invention research different solvents and method, finally establish acryloyl in extraction with aqueous solution fried food
Amine, water are higher as extractant extraction ratio, are the optimum solvents extracted as acrylamide, while water security is nontoxic, price is low
It is honest and clean, by constantly groping to finally found that using heating and supersound extraction acrylamide, extraction ratio highest (98.2%, RSD=
2.83%).
The applicant has tested the experiment of different solvents extraction ratio, and experimental result is shown in Fig. 1.It can be seen from figure 1 that deionization
Water, extraction ratio (98.2%, RSD=2.83%) high with the extraction ratio of methanol, deionized water+methanol.Therefore the final decision of the present invention
The optimum solvent that ionized water is extracted as acrylamide；Deionized water safety non-toxic, cheap simultaneously.
With specific embodiment in detail the present invention is described in detail below.
Embodiment 1,
1) the commercially available French fries after 2.0g grinds are weighed to be placed in 20mL politef centrifuge tubes, 3 levels are added
Acrylamide standard solution, stand 10min；
2) to step 1) obtained by stand liquid in add 10mL normal hexane to be vortexed concussion 1min to stand 10min, discard oil
Ether layer, 40 ± 1 DEG C of nitrogen are dried up, and the above-mentioned normal hexane skimming processes of repetition are once；
3) to Jing step 2) 14mL deionized waters are added in sample after defat, it is vortexed concussion half a minute, is then placed in perseverance
Warm agitator is taken out after 30min is shaken at 60 DEG C, resulting solution ultrasonic vibration 20min again；
4) to Jing steps 3) obtained by shake liquid in add auspicious reagent C arrezI of card and each 0.5mL of CarrezII, stand
10min takes supernatant 2mL to remove protein in 4 DEG C, 11000rpm high speed centrifugation 20min, and good SPE is little for overactivation
Post, discards front 2mL effluent, after filtering, obtains need testing solution after collection；
5) by step 4) in need testing solution, be analyzed using liquid chromatography；
The chromatographic column is：WatersT3；
The high efficiency liquid phase chromatographic analysis method condition is：Mobile phase：Methanol：0.1% aqueous formic acid=5:95；Flow velocity：
1.0ml/min；Detection column temperature：30℃；Detection wavelength：205nm；Sample size：20μL.
Embodiment 2, with embodiment 1, except for the difference that,
1) weigh the commercially available fried drumstick after 2.0g grinds to be placed in 20mL politef centrifuge tubes, add 3 water
Flat acrylamide standard solution, stands 10min.
Embodiment 3, with embodiment 1, except for the difference that,
1) weigh the commercially available sweet potato cake after 2.0g grinds to be placed in 20mL politef centrifuge tubes, add 3 water
Flat acrylamide standard solution, stands 10min.
Embodiment 4, with embodiment 1, except for the difference that,
1) the commercially available instant noodles after 2.0g grinds are weighed to be placed in 20mL politef centrifuge tubes, 3 water are added
Flat acrylamide standard solution, stands 10min.
Embodiment 5, with embodiment 1, except for the difference that,
1) weigh the commercially available pan-fried dumpling after 2.0g grinds to be placed in 20mL politef centrifuge tubes, add 3 levels
Acrylamide standard solution, stand 10min.
Embodiment 6, with embodiment 1, except for the difference that,
1) the commercially available fritters of twisted dough after 2.0g grinds are weighed to be placed in 20mL politef centrifuge tubes, 3 levels are added
Acrylamide standard solution, stand 10min.
The measurement result of each embodiment acrylamide content see the table below：
From upper table, using acrylamide content value and the GB analysis detection (GB5009.204- of present invention detection
2014) value compares, and error (%) is 0.46-2.24, illustrates that the detection of the present invention is quick, accurate, efficient.
The acrylamide standard solution chromatogram of Fig. 2, acrylamide HPLC collection of illustrative plates in the pure sample product of Fig. 3, the sample of Fig. 4 add
In mark, acrylamide HPLC collection of illustrative plates explanations this method can reduce sample pre-treatments error.
Claims (3)
1. it is a kind of detection fried food in acrylamide liquid phase chromatography analytical method, it is characterised in that：Comprise the following steps that：
1) weigh the fried food after 2.0g grinds to be placed in 20mL politef centrifuge tubes, add acrylamide standard
Product solution, stands 10min；
2) to step 1) obtained by stand liquid in add 10mL normal hexane to be vortexed concussion 1min, and stand 10min defats, discard oil
Ether layer, 40 ± 1 DEG C of nitrogen are dried up, and the above-mentioned normal hexane skimming processes of repetition are once；
3) to Jing step 2) 14mL deionized waters are added in sample after defat, it is vortexed concussion half a minute, is then placed in constant temperature and shakes
Swing after device shakes 30min at 60 DEG C and take out, resulting solution ultrasonic vibration 20min again；
4) to Jing steps 3) obtained by shake liquid in add auspicious reagent C arrezI of card and each 0.5mL of CarrezII, stand 10min, remove
Deproteinization matter, in 4 DEG C, 11000rpm high speed centrifugation 20min；Take supernatant 2mL, the good SPE pillars of overactivation, before discarding
2mL effluent, after filtering, obtains need testing solution after collection；
5) by step 4) in need testing solution, be analyzed using liquid chromatography；
Chromatographic column used is：WatersT3；
The high efficiency liquid phase chromatographic analysis method condition is：Mobile phase：Methanol：0.1% aqueous formic acid=5:95；Flow velocity：
1.0ml/min；Detection column temperature：30℃；Detection wavelength：205nm；Sample size：20μL.
2. it is according to claim 1 it is a kind of detection fried food in acrylamide liquid phase chromatography analytical method, its feature
It is：Step 1) described in fried food be commercially available French fries, fried drumstick, sweet potato cake, instant noodles, decoct dumpling or fritters of twisted dough.
3. it is according to claim 1 it is a kind of detection fried food in acrylamide liquid phase chromatography analytical method, its feature
It is：Step 4) in the SPE pillars that activated using front needing with 2mL methanol and 2mL deionized water balances.
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Cited By (6)
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| CN106770842A (en) * | 2017-03-31 | 2017-05-31 | 安徽国科检测科技有限公司 | The assay method of acrylamide in a kind of leisure food |
| CN110208439A (en) * | 2019-07-16 | 2019-09-06 | 中国热带农业科学院农产品加工研究所 | A kind of rapid detection method of acrylamide |
| CN111398481A (en) * | 2020-04-22 | 2020-07-10 | 上海汽车集团股份有限公司 | Analysis test method of perfluorosulfonyl fluoroolefin ether |
| CN113791156A (en) * | 2021-09-18 | 2021-12-14 | 昆明品世食品有限公司 | Food safety detection method |
| CN114544802A (en) * | 2022-01-21 | 2022-05-27 | 河北科技大学 | High performance liquid chromatography analysis method of acrylamide resin monomer |
| CN115078592A (en) * | 2022-07-15 | 2022-09-20 | 广东广纳安疗科技有限公司 | Detection method and application of acrylamide monomer |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115078592A (en) * | 2022-07-15 | 2022-09-20 | 广东广纳安疗科技有限公司 | Detection method and application of acrylamide monomer |
| CN115078592B (en) * | 2022-07-15 | 2024-01-23 | 广东广纳安疗科技有限公司 | Detection method of acrylamide monomer and application thereof |
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