CN102680608B - Method for identifying abandoned kitchen oil - Google Patents
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- CN102680608B CN102680608B CN 201210195450 CN201210195450A CN102680608B CN 102680608 B CN102680608 B CN 102680608B CN 201210195450 CN201210195450 CN 201210195450 CN 201210195450 A CN201210195450 A CN 201210195450A CN 102680608 B CN102680608 B CN 102680608B
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Abstract
The invention discloses a method for identifying abandoned kitchen oil, which comprises the following steps: weighting an oil sample; adding acetonitrile as an extracting agent for extracting the oil sample; concentrating extract liquor and then fixing column, thereby obtaining a sample solution; analyzing by injecting the sample solution into a liquid chromatography-mass spectrum/mass spectrum instrument, taking capsaicine and dihydro-capsaicine as standard substances, taking chromatographic peak retention time and two monitored ion pairs monitored through mass spectrum multi-reaction of thestandard substances as nature-determining basis, and confirming if the sample solution contains capsaicine and dihydro-capsaicine; and if yes, confirming the oil sample as the abandoned kitchen oil. The method provided by the invention is simple and practical, is an effective method for identifying the abandoned kitchen oil in the areas (such as Sichuan, Chongqing, Guizhou, Hunan, Hubei, and the like) where people like peppery food, is applied to the field of food detection and is obvious in social benefit and economic benefit.
Description
Technical field:
The invention belongs to organism analyzing and testing field, be specifically related to a kind of discrimination method of the kitchen abendoned oil of eating.
Background technology:
Meal kitchen abendoned oil is made a general reference all kinds of discarded edible oils inferior, mainly is meal kitchen waste oil and refines the oil of back output by Animal Skin inferior, meat, internal organ processing.Meal kitchen waste oil mainly comprises swill oil (hogwash fat) and the old oil of frying.The meal kitchen abendoned oil of narrow sense refers to the greasy floating thing fished in the trench oil interceptors such as hotel, restaurant be pitchy liquid paste, and the sour foul odour is arranged; Swill oil refers to the upper strata oil slick after leftovers, leftovers (common name swill) are collected, and is golden yellow to kermesinus through refining, and the sour flavor is arranged; Fry old oil and refer at high temperature be used for repeatedly fried food, bad changes such as oxidative polymerization take place, can not continue the grease that eats again.
At present, still can't accurately identify meal kitchen abendoned oil both at home and abroad.For this reason, health ministry has successively been collected meal kitchen abendoned oil detection method 4 times, and 3 times the method for collecting all be facts have proved infeasible.In November, 2011, meal kitchen abendoned oil detection method is openly collected at Ministry of Public Health state food security risk assessment center the 4th in China.
In the area of liking hot and spicy food crowd massing, as Sichuan, Chongqing, Guizhou, Hunan, Hubei and other places, be unable to do without capsicum in the diet.In cooking process or chafing dish, seasoning, edible oil and fat contact with capsicum or chilli oil, and effective constituent capsicim and Dihydrocapsaicin in the capsicum can be brought in the grease; And capsicim and Dihydrocapsaicin are liposoluble substances, and be clean even meal kitchen abendoned oil also is difficult to remove through purified treatment, can remain in the abendoned oil of meal kitchen.
Summary of the invention:
In order to solve the deficiency of existing meal kitchen abendoned oil detection technique, the purpose of this invention is to provide a kind of discrimination method of the kitchen abendoned oil of eating.
The present invention measures in the oil sample whether have exogenous trace impurity composition capsicim and Dihydrocapsaicin by liquid chromatography/tandem mass spectrometry, and indicates composition with these two kinds of compositions as the feature of differentiating meal kitchen abendoned oil, thereby realizes purpose of the present invention.
The discrimination method of meal of the present invention kitchen abendoned oil is characterized in that, may further comprise the steps:
(1) take by weighing oil sample, add acetonitrile and as extractant oil sample is extracted, extract concentrates the back constant volume, obtains sample solution;
(2) sample solution is injected liquid chromatography-mass spectrography/mass spectrometer analysis, with capsicim and Dihydrocapsaicin as standard items, two couples monitoring ion pair with the chromatographic peak retention time of standard items and mass spectrum multiple-reaction monitoring (MRM) is qualitative foundation, identifies in the sample solution whether contain capsicim and Dihydrocapsaicin; If contain capsicim or Dihydrocapsaicin in the sample solution, judge that this oil sample is meal kitchen abendoned oil.
In the step (1), described oil sample is preferably 1g: 4ml with the ratio of the consumption of acetonitrile.
In the step (1), described extraction is extraction under the vortex vibration preferably, the time of vortex vibration is 5min, temperature is room temperature, mixed solution behind the vortex is through centrifuging and taking supernatant extract, surplus solution adds acetonitrile again and extracts 1 time under similarity condition, and merging supernatant extract after 0.22 μ m filter membrane, obtains sample solution after concentrating.
In the step (1), described centrifugal preferably at the centrifugal 10min of 4000r/min; Described concentrating is to blow concentrated at 50 ℃ of water-bath nitrogen.
In the step (2), described liquid chromatography-mass spectrography/mass spectrometer analysis, its chromatographic condition preferably: Thermo BDS Hypersil C8 (100mm * 2.1mm, 2.4 μ m) chromatographic column; Column temperature: 30 ℃; Flow velocity: 0.3mL/min; Sample size: 10 μ L; Phase: the A that flows is the aqueous solution that contains volume ratio 0.4% formic acid, and B is the acetonitrile solution that contains volume ratio 0.4% formic acid; Gradient elution program: 0 ~ 2min, 22% ~ 80%B; 2 ~ 2.5min, 80%B; 2.5 ~ 2.51min, 80% ~ 100%B; 2.51 ~ 3.5min, 100%B; 3.5 ~ 3.51min, 100% ~ 22%B keeps 3min.
In the step (2), described liquid chromatography-mass spectrography/mass spectrometer analysis, its mass spectrum condition is: electric spray ion source (ESI); The positive ion scan pattern; Multiple-reaction monitoring (MRM) acquisition mode; Dry gas temperature: 350 ℃; Dry gas flow: 10.0L/min; Atomization gas pressure: 276kPa; Capillary voltage: 4kV; MS1 and MS2 are the unit resolution rate.
In the step (2), two couple of described mass spectrum multiple-reaction monitoring (MRM) monitoring ion pair is: capsicim is m/z306〉122 and 306〉137, both relative abundance ratios are 24.3: 100; Dihydrocapsaicin is m/z 308〉122 and 308〉137, both relative abundance ratios are 26.7: 100.
In the step (2), the retention time in the oil sample spectrogram, with the retention time deviation of corresponding compound in capsicim or the Dihydrocapsaicin standard solution spectrogram within ± 2.5%; And in the sample spectrogram in the relative abundance of the qualitative ion pair of each component and the standard solution spectrogram relative abundance of capsicim or the qualitative ion pair of Dihydrocapsaicin compare, deviation is no more than ± 25%, then can be judged to be and contain capsicim or Dihydrocapsaicin in the sample; As long as contain one of capsicim or Dihydrocapsaicin in the sample, can determine that sample is meal kitchen abendoned oil.
The present invention has following advantage and effect with respect to prior art:
(1) this method adopts liquid chromatography/tandem mass spectrum (LC-MS/MS) technology to set up capsicim in the grease and the assay method of Dihydrocapsaicin, with capsicim and Dihydrocapsaicin as the feature indication composition of differentiating meal kitchen abendoned oil; Be applicable to the discriminating of meal kitchen abendoned oil.
(2) this method is simple and practical, is the effective ways of meal kitchen abendoned oil of differentiating the crowd's that likes hot and spicy food area (as Sichuan, Chongqing, Guizhou, Hunan, Hubei and other places), can be applicable to the food inspection field, has very obvious social and economic benefit.
Description of drawings:
Fig. 1 is the liquid chromatography/tandem mass spectrum MRM figure of capsicim and Dihydrocapsaicin standard specimen;
Fig. 2 is the liquid chromatography/tandem mass spectrum MRM figure of doubtful meal kitchen abendoned oil sample.
Embodiment:
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
The discriminating of certain doubtful meal kitchen abendoned oil sample may further comprise the steps:
(1) accurately takes by weighing even matter oil sample (certain doubtful meal kitchen abendoned oil sample) 5.00g (being accurate to 0.01g) and place 50mL teflon centrifuge tube, add the 10mL acetonitrile, vortex extracts 5min, temperature is room temperature, at the centrifugal 10min of 4000r/min, get whole supernatants in the 100mL beaker then; Add the 10mL acetonitrile again to residue, repeat to extract once, the centrifugal 10min of 4000r/min gets supernatant, merges 2 times supernatant, blows in 50 ℃ of water-bath nitrogen to be concentrated into 1mL, and it is standby to get sample solution behind the 0.22 μ m filter membrane excessively.
The preparation of standard solution: be that solvent is prepared capsicim and the Dihydrocapsaicin standard inventory solution that mass concentration is 1000mg/L respectively with methyl alcohol, put 4 ℃ of preservations of brown volumetric flask lucifuge, facing the time spent, to become mass concentration with the initial flow phase dilution be the standard solution of 1.0 ~ 100 μ g/L, behind 0.22 μ m membrane filtration as working fluid.
(2) chromatogram and mass spectrum condition: Thermo BDS Hypersil C8 post (100mm * 2.1mm, 2.4 μ m) chromatographic column; Column temperature: 30 ℃; Flow velocity: 0.3mL/min; Sample size: 10 μ L; Flow phase: A for containing the aqueous solution of 0.4% (volume ratio) formic acid, and B is for containing the acetonitrile of 0.4% (volume ratio) formic acid; Gradient elution program: 0 ~ 2min, 22% ~ 80%B; 2 ~ 2.5min, 80%B; 2.5 ~ 2.51min, 80% ~ 100%B; 2.51 ~ 3.5min, 100%B; 3.5 ~ 3.51min, 100% ~ 22%B keeps 3min.The mass spectrum condition is: electric spray ion source (ESI); The positive ion scan pattern; Multiple-reaction monitoring (MRM) acquisition mode; Dry gas temperature: 350 ° of C; Dry gas flow: 10.0L/min; Atomization gas pressure: 276kPa; Capillary voltage: 4kV; MS1 and MS2 are the unit resolution rate.
(3) assay method: each 10 μ L injection liquid chromatography-series connection quadrupole mass spectrometer of extracting sample solution and standard solution is measured according to above-mentioned chromatogram and mass spectrum condition respectively respectively.
Standard items wherein: the MRM mass spectrum parameter of capsicim and Dihydrocapsaicin sees Table 1, LC-MS/MS MRM chromatogram as shown in Figure 1.
Table 1: multiple-reaction monitoring (MRM) mass spectrum parameter
*. for quota ion right.
Fig. 1 is the LC-MS/MS MRM chromatogram of capsicim, Dihydrocapsaicin standard solution.The retention time 3.83min of capsicim, the qualitative ion pair m/z 306 of MRM〉122 and 306〉137, its relative abundance ratio is 24.3: 100; The retention time 4.00min of Dihydrocapsaicin, the qualitative ion pair m/z 308 of MRM〉122 and 308〉137, its relative abundance ratio is 26.7: 100.
The total ions chromatogram of the sample solution of doubtful meal kitchen abendoned oil is seen Fig. 2, and the time of finding in the spectrogram to a hook at the end is 3.88min and two chromatographic peaks of 4.05min, respectively with capsicim and Dihydrocapsaicin standard solution spectrogram in corresponding; And MRM ion pair m/z 306〉122 and 306〉137, the m/z 308 of these 2 chromatographic peaks in the sample spectrogram〉relative abundance of capsicim or the qualitative ion pair of Dihydrocapsaicin is consistent in 122 and 308〉137 relative abundance and the standard solution spectrogram, and judge in the sample and contain capsicim and Dihydrocapsaicin.
(4) contain capsicim and Dihydrocapsaicin in per sample, infer this sample and be meal kitchen abendoned oil, by confirming that this sample is from the meal kitchen abendoned oil in the crowd's of liking hot and spicy food area really, this shows that the sample that is accredited as meal kitchen abendoned oil through the present invention is meal kitchen abendoned oil really.
Embodiment 2:
Get the many batches of areas that come from the crowd of liking hot and spicy food respectively, meal kitchen abendoned oil as Sichuan, Chongqing, Guizhou, Hunan, Hubei and other places, method according to embodiment 1 detects, and all detects capsicim or Dihydrocapsaicin or both and detect in these meal kitchen abendoned oils.
Comparative Examples 1:
Get a collection of oil sample that turns out to be non-meal kitchen abendoned oil, detect according to the method for embodiment 1, testing result shows and does not have capsicim or Dihydrocapsaicin in the oil sample.
Above-described embodiment is preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spiritual essence of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (6)
1. the discrimination method of kitchen abendoned oil of eating is characterized in that, may further comprise the steps:
(1) take by weighing oil sample, add acetonitrile and as extractant oil sample is extracted, extract concentrates the back constant volume, obtains sample solution;
(2) sample solution is injected liquid chromatography-mass spectrography/mass spectrometer analysis, with capsicim and Dihydrocapsaicin as standard items, two couples monitoring ion pair with the chromatographic peak retention time of standard items and mass spectrum multiple-reaction monitoring is qualitative foundation, identifies in the sample solution whether contain capsicim and Dihydrocapsaicin; If contain capsicim or Dihydrocapsaicin in the sample solution, judge that this oil sample is meal kitchen abendoned oil;
In the step (2), described liquid chromatography-mass spectrography/mass spectrometer analysis, its chromatographic condition is: Thermo BDS Hypersil C8 chromatographic column-100mm * 2.1mm, 2.4 μ m; Column temperature: 30 ℃; Flow velocity: 0.3mL/min; Sample size: 10 μ L; Phase: the A that flows is the aqueous solution that contains volume ratio 0.4% formic acid, and B is the acetonitrile solution that contains volume ratio 0.4% formic acid; Gradient elution program: 0～2min, 22%～80%B; 2～2.5min, 80%B; 2.5～2.51min, 80%～100%B; 2.51～3.5min, 100%B; 3.5～3.51min, 100%～22%B keeps 3min.
2. the discrimination method of meal according to claim 1 kitchen abendoned oil is characterized in that in the step (1), described oil sample is 1g:4ml with the ratio of the consumption of acetonitrile.
3. the discrimination method of meal according to claim 1 kitchen abendoned oil, it is characterized in that, in the step (1), described extraction is extraction under the vortex vibration, and the time of vortex vibration is 5min, and temperature is room temperature, mixed solution behind the vortex is through centrifuging and taking supernatant extract, surplus solution adds acetonitrile again and extracts 1 time under similarity condition, and merging supernatant extract after 0.22 μ m filter membrane, obtains sample solution after concentrating.
4. the discrimination method of meal according to claim 3 kitchen abendoned oil is characterized in that, in the step (1), described centrifugal be at the centrifugal 10min of 4000r/min; Described concentrating is to blow concentrated at 50 ℃ of water-bath nitrogen.
5. the discrimination method of meal according to claim 1 kitchen abendoned oil is characterized in that, in the step (2), and described liquid chromatography-mass spectrography/mass spectrometer analysis, its mass spectrum condition is: electric spray ion source; The positive ion scan pattern; The multiple-reaction monitoring acquisition mode; Dry gas temperature: 350 ℃; Dry gas flow: 10.0L/min; Atomization gas pressure: 276kPa; Capillary voltage: 4kV; MS1 and MS2 are the unit resolution rate.
6. the discrimination method of meal according to claim 1 kitchen abendoned oil, it is characterized in that, in the step (2), two couple of described mass spectrum multiple-reaction monitoring monitoring ion pair is: capsicim is m/z306〉122 and 306〉137, both relative abundances are than being 24.3:100; Dihydrocapsaicin is m/z308〉122 and 308〉137, both relative abundances are than being 26.7:100.
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103604911A (en) * | 2013-11-18 | 2014-02-26 | 江西绿杉油脂有限公司 | Quality test method of edible fat and oil |
| CN104383889B (en) * | 2014-11-21 | 2016-08-24 | 中国农业科学院油料作物研究所 | A kind of Dihydrocapsaicin polyclonal antibody adsorbent, immune affinity column and its preparation method and application |
| CN108169393A (en) * | 2016-12-07 | 2018-06-15 | 中国科学院大连化学物理研究所 | Portable high-efficiency liquid-chromatography apparatus and its application in gutter oil differentiates detection |
| CN107144558B (en) * | 2017-05-12 | 2022-01-18 | 吉林大学 | Method for identifying illegal cooking oil by using Raman spectrum technology |
| CN109142568A (en) * | 2018-08-10 | 2019-01-04 | 益海嘉里（兖州）粮油工业有限公司 | A method of utilizing gas chromatograph-mass spectrometer (GC-MS) quantitative detection Capsaicinoids |
| CN112098553B (en) * | 2020-09-11 | 2023-06-09 | 安徽中创食品检测有限公司 | Method for detecting capsaicin compounds in duck necks |
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