CN102520100A - Auxiliary identification method of geographical origin of beef - Google Patents
Auxiliary identification method of geographical origin of beef Download PDFInfo
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- CN102520100A CN102520100A CN2011104145523A CN201110414552A CN102520100A CN 102520100 A CN102520100 A CN 102520100A CN 2011104145523 A CN2011104145523 A CN 2011104145523A CN 201110414552 A CN201110414552 A CN 201110414552A CN 102520100 A CN102520100 A CN 102520100A
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Abstract
The invention discloses an auxiliary identification method of geographical origin of beef, comprising the following steps: (1) collecting a beef sample from any one region of Jilin, Nixia, Guizhou and Hebei; (2) carrying out fat extraction on the beef sample to obtain a crude fat sample; (3) carrying out methyl-esterification reaction on the crude fat sample to obtain a fatty acid methyl ester sample; (4) measuring the percentage contents of alpha-linolenic acid, myristic acid, heptadecanoic acide, saturated fatty acid, and monounsaturated fatty acid in the fatty acid methyl ester sample, and respectively marking as X-(alpha-C18:3), X(C14:0), X(C17:0), X(SFA) and X(MUFA), wherein Y Jilin, Y Ningxia, Y Guilin and Y Hebei can be obtained according to discrimination modes (1)-(4), and the largest value of the Y Jilin, Y Ningxia, Y Guilin and Y Hebei represents where the beef sample comes from. According to the invention, the method is simple and practical, the whole accurate identification rate of the geographical origin of beef can reach more than 92 %, and the invention provides a scientific method for establishing and improving a beef quality safety traceability system.
Description
Technical field
The present invention relates to a kind of method of auxiliary discriminating beef origin, particularly a kind of method of the discriminating beef origin based on the fatty acid analysis method.
Background technology
In order to reduce the heavy losses that wait some eqpidemic disease epidemic situations to bring such as " rabid ox disease ", guarantee food security, each developed country implements food safety trace back system one after another in the world.The laws and regulations on the management No.178 of European Union (2002) required from January 1st, 2005, and all food of in European Union's scope, selling can both be followed the tracks of and review, otherwise do not allow list marketing.FDA (Food and Drug Adminstration) (FDA) required in the domestic and foreign food department that is engaged in food production, processing, packing or grasp crowd or animal consumption of the U.S. before 12 days Dec in 2003; Must register to FDA, follow the tracks of and review so that carry out food security.In addition, the sign of reviewing of food also becomes the choice criteria that consumer in the world buys food.The federal office of Switzerland's public health recent statistics shows that 82% consumer representes that the source, the place of production of food is that they buy the main foundation of selecting food, and wherein 71% consumer pays close attention to the source, the place of production of meat products." agricultural product geographical sign management method " approved in the 15th standing meeting of Ministry of Agriculture's on Dec 6th, 2007, and goes into effect from February 1st, 2008.
International reports utilize the fatty acid compositional analysis technology that food is carried out the research of tracing to the source in the place of production.Wherein utilize the fatty acid compositional analysis technology more to the research that plant source food carries out tracing to the source in the place of production, such as olive oil, coffee, grape wine, honey etc., and it is better to differentiate effect.Aspect meat, mainly be to study cowboying feed difference etc. in the bibliographical information through fatty acid composition and content analysis, lack beef origin fatty acid fingerprint analysis technical method.
Summary of the invention
The method that the purpose of this invention is to provide a kind of auxiliary discriminating beef origin.
The method of a kind of auxiliary discriminating beef origin provided by the invention comprises the steps:
(1) beef sample is gathered in arbitrary region from Jilin, Ningxia, Guizhou and Hebei;
(2) said beef sample is carried out fat-extraction and obtain the crude fat sample;
(3) said crude fat sample is carried out esterification reaction of organic acid and obtain the fatty acid methyl ester sample;
(4) detect the percentage composition of alpha-linolenic acid, myristic acid, Heptadecanoic acide, saturated fatty acid and monounsaturated fatty acids in the said fatty acid methyl ester sample, be labeled as respectively: X (α-C18:3), X (C14:0), X (C17:0), X (SFA) and X (MUFA); Can obtain Y according to discrimination model (1)～(4)
Jilin, Y
Ningxia, Y
GuizhouAnd Y
Hebei, then said beef sample derives from Y
Jilin, Y
Ningxia, Y
GuizhouAnd Y
HebeiThe maximum region of middle numerical value;
Y
Jilin=-6355.908+160.769 * X (α-C18:3)-32.558 * X (C14:0)+160.152 * X (C17:0)+126.983 * X (SFA)+141.307 * X (MUFA) (1)
Y
Ningxia=-6373.168+161.009 * X (α-C18:3)-31.602 * X (C14:0)+173.638 * X (C17:0)+126.696 * X (SFA)+141.651 * X (MUFA) (2)
Y
Guizhou=-6457.754+163.852 * X (α-C18:3)-32.300 * X (C14:0)+154.604 * X (C17:0)+127.913 * X (SFA)+142.540 * X (MUFA) (3)
Y
Hebei=-6191.874+159.260 * X (α-C18:3)-30.271 * X (C14:0)+170.184 * X (C17:0)+124.938 * X (SFA)+139.333 * X (MUFA (4).
In the above-mentioned method; Beef sample can be meat cubelets described in the step (1); Like the square meat cubelets of 1cm; Said method also comprises carries out cryodesiccated step with said meat cubelets, and said cryodesiccated step comprises placing carries out freezing in advance 12h～24h under-20 ℃～-80 ℃, places the step of vacuum freezing drying oven dry 24h～48h under-30 ℃～-50 ℃ again.
In the above-mentioned method, beef sample can be muddy flesh described in the step (1), and said method also can comprise the step that said muddy flesh is placed 60 ℃～70 ℃ of dry 36h～48h, specifically can be 60 ℃ of following dry 36h or 70 ℃ of following dry 48h.
In the above-mentioned method, can comprise also in the step (2) that it is 100 orders～200 purpose steps that said beef sample is crushed to granularity, like 100 orders or 200 orders.
In the above-mentioned method, fat-extraction described in the step (2) can be extracted through Suo Shi and carry out, and the solvent that said Suo Shi extracts is that boiling range is 30 ℃～60 ℃ a sherwood oil, and the time can be 18h～26h, specifically can be 18h or 26h.
In the above-mentioned method, esterification reaction of organic acid can carry out in the sulfuric acid methanol solution described in the step (3); The volumn concentration of sulfuric acid can be 1% in the said sulfuric acid methanol solution.
In the above-mentioned method, the temperature of esterification reaction of organic acid can be 60 ℃～80 ℃ described in the step (3), specifically can be 60 ℃ or 70 ℃, and the time can be 50min～80min, specifically can be 30min or 80min.
In the above-mentioned method, the method that detects described in the step (4) can be vapor-phase chromatography.
Method provided by the invention is simple, practical, and the whole correct resolution that beef origin is originated can reach more than 92%; The present invention can be foundation and the perfect scientific approach that provides that the beef quality safety is reviewed system.
Embodiment
Employed experimental technique is conventional method like no specified otherwise among the following embodiment.
Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.
The evaluation of embodiment 1, beef origin
(1) beef sample collection, rubbing, drying, pulverizing, fat-extraction
From the Anshun City, Guizhou Province, Tongxin County two places in Ningxia gather the beef sample of 27 oxen, beef sample rubs meat mud through meat grinder, at 70 ℃ of freeze-day with constant temperature 48h, using comminutor to be ground into granularity is that 100 purposes are Powdered; Placing beaker to add boiling range powdered beef is 30～60 ℃ sherwood oil lixiviate, centrifugal and filter three times; Lixiviate 6h for the first time, lixiviate for the second time spend the night (about 12h), lixiviate 8h for the third time; Supernatant is placed Rotary Evaporators; Condition recover sherwood oil in that 35 ℃ of following vacuum tightnesss are 0.08MPa obtains the crude fat sample, and is subsequent use.
(2) methyl esterification of fatty acid
Take by weighing 50mg crude fat sample, place 25mL tool plug test tube, the volumn concentration that adds sulfuric acid is 1% sulfuric acid methanol solution 2mL; In 70 ℃ of water-baths, heat 30min, add the 2mL normal hexane then, cover glass stopper and fiercely jolt 30s; Adding distil water is settled to 25mL again, leaves standstill to clarification; Take out supernatant liquor (hexane solution that contains methyl esters), place beaker, add the 1mL normal hexane in the test tube again and wash once, merge supernatant liquor in beaker, adding 1g NaHSO
4, leave standstill after jolting 10s, treat the salt post precipitation, shift in upper solution to the 10mL volumetric flask and constant volume, do stratographic analysis fully.
(3) content of fatty acid, composition detect
With the methyl esterification of fatty acid sample, be diluted to 500ppm, adopt auto injection method sample introduction, be furnished with hydrogen flame ionization detector (FID) with Varian GC-450 gas chromatograph and detect, replication 3 times.
GC conditions: chromatographic column: HP-5 capillary column (0.25mm * 30m * 0.25um CP-SIL-5CB U.S. Varian); The temperature programme condition is: 100 ℃ of initial temperature, behind the maintenance 2min, be warming up to 250 ℃ with 5 ℃/min speed, and keep 5min; Injector temperature is 280 ℃; Adopting split ratio is 1: 20 split sampling; Carrier gas is 99.999%N
2, flow velocity is 1.0ml/min.
Quantitative test: adopt external standard method, the mark article are made into the titer of variable concentrations, through stratographic analysis production standard curve, the peak area according to each fatty acid carries out quantitatively then.
The percentage composition such as the table 1 of the beef sample five indices of Anshun, Guizhou and Ningxia Tongxin County.
The beef sample of Anshun, table 1 Guizhou and Ningxia Tongxin County
Saturated fatty acid SFA in the table 1 in each sample comprises tetradecanoic acid, hexadecanoic acid and octadecanoid acid; Monounsaturated fatty acids MUFA comprises gaidic acid and 16 carbon enoic acids.
No. 1 sample with Guizhou and area, Ningxia is an example respectively, and the percentage composition of Anshun, Guizhou county No. 1 beef appearance five indices is brought in discrimination model (1)～(4), can get Y
Guizhou=6371.142＞Y
Ningxia=6361.837, so beef sample belongs to Guizhou province;
The percentage composition of No. 1 beef sample five indices that the Ningxia Tongxin County is regional is brought in discrimination model (1)～(4), can get Y
Ningxia=6388.93＞Y
Guizhou=6379.234, so beef sample belongs to the area, Ningxia.
Successively the five indices of 25 samples in the table 1 is brought in the discrimination model, judged, have 1 sample to be misjudged in 27 samples as a result, whole correct decision rate is 96.3%.
The evaluation of embodiment 2, beef origin
(1) beef sample collection, rubbing, drying, pulverizing, fat-extraction
The beef sample of 34 oxen is gathered in Yushu City, two places, Zhangbei County, Hebei province from Jilin Province, and beef sample rubs through meat grinder, at 60 ℃ of freeze-day with constant temperature 36h; Using comminutor to be ground into granularity is that 200 purposes are Powdered, with powdered beef place beaker add boiling range be 30～60 ℃ the sherwood oil lixiviate, centrifugal, filter the 3h of lixiviate for the first time three times; Lixiviate for the second time spend the night (about 12h); Lixiviate 3h places Rotary Evaporators with supernatant for the third time, at 35 ℃ of following vacuum tightnesss condition recover sherwood oil that is 0.08MPa; Obtain the crude fat sample, subsequent use.
(2) methyl esterification of fatty acid
Take by weighing 50mg crude fat sample, place 25mL tool plug test tube, the volumn concentration that adds sulfuric acid is 1% sulfuric acid methanol solution 2mL; In 60 ℃ of water-baths, heat 80min, add the 2mL normal hexane then, cover glass stopper and fiercely jolt 30s; Adding distil water is settled to 25mL again, leaves standstill to clarification.Take out supernatant liquor (hexane solution that contains methyl esters), place beaker, add the 1mL normal hexane in the test tube again and wash once, merge supernatant liquor in beaker, adding 1g NaHSO
4, leave standstill after jolting 10s, treat the salt post precipitation, shift in upper solution to the 10mL volumetric flask and constant volume, do stratographic analysis fully.
(3) content of fatty acid, composition detect
With the methyl esterification of fatty acid sample, be diluted to 500ppm, adopt auto injection method sample introduction, be furnished with hydrogen flame ionization detector (FID) with Varian GC-450 gas chromatograph and detect, replication 3 times.
GC conditions: chromatographic column: HP-5 capillary column (0.25mm * 30m * 0.25um CP-SIL-5CB U.S. Varian); The temperature programme condition is: 100 ℃ of initial temperature, behind the maintenance 2min, be warming up to 250 ℃ with 5 ℃/min speed, and keep 5min; Injector temperature is 280 ℃; Adopting split ratio is 1: 20 split sampling; Carrier gas is 99.999%N
2, flow velocity is 1.0ml/min.
Quantitative test: adopt external standard method, the mark article are made into the titer of variable concentrations, through stratographic analysis production standard curve, the peak area according to each fatty acid carries out quantitatively then.
The percentage composition such as the table 2 of the beef sample five indices of yushu city and Zhangbei County, Hebei.
The beef sample of table 2 yushu city and Zhangbei County, Hebei
Saturated fatty acid SFA in the table 2 in each sample comprises tetradecanoic acid, hexadecanoic acid and octadecanoid acid; Monounsaturated fatty acids MUFA comprises gaidic acid and 16 carbon enoic acids.
No. 1 sample with Jilin and Hebei area is an example, brings the percentage composition of No. 1 beef sample five indices in yushu city into discrimination model, can get Y
Jilin=6396.084＞Y
Hebei=6391.287, so beef sample belongs to Jilin Area;
Bring the percentage composition of No. 1 beef sample five indices of Zhangbei County, Hebei into discrimination model, can get Y
Hebei=6111.448＞Y
Jilin=6105.106, so beef sample belongs to the area, Hebei.
Successively the five indices of 32 samples in the table 2 is brought in the discrimination model, judged, have 2 samples to be misjudged in 34 samples as a result, whole correct decision rate is 94.1%.
Claims (8)
1. an auxiliary method of differentiating beef origin comprises the steps:
(1) beef sample is gathered in arbitrary region from Jilin, Ningxia, Guizhou and Hebei;
(2) said beef sample is carried out fat-extraction and obtain the crude fat sample;
(3) said crude fat sample is carried out esterification reaction of organic acid and obtain the fatty acid methyl ester sample;
(4) detect the percentage composition of alpha-linolenic acid, myristic acid, Heptadecanoic acide, saturated fatty acid and monounsaturated fatty acids in the said fatty acid methyl ester sample, be labeled as respectively: X (α-C18:3), X (C14:0), X (C17:0), X (SFA) and X (MUFA); Can obtain Y according to discrimination model (1)～(4)
Jilin, Y
Ningxia, Y
GuizhouAnd Y
Hebei, then said beef sample derives from Y
Jilin, Y
Ningxia, Y
GuizhouAnd Y
HebeiThe maximum region of middle numerical value;
Y
Jilin=-6355.908+160.769 * X (α-C18:3)-32.558 * X (C14:0)+160.152 * X (C17:0)+126.983 * X (SFA)+141.307 * X (MUFA) (1)
Y
Ningxia=-6373.168+161.009 * X (α-C18:3)-31.602 * X (C14:0)+173.638 * X (C17:0)+126.696 * X (SFA)+141.651 * X (MUFA) (2)
Y
Guizhou=-6457.754+163.852 * X (α-C18:3)-32.300 * X (C14:0)+154.604 * X (C17:0)+127.913 * X (SFA)+142.540 * X (MUFA) (3)
Y
Hebei=-6191.874+159.260 * X (α-C18:3)-30.271 * X (C14:0)+170.184 * X (C17:0)+124.938 * X (SFA)+139.333 * X (MUFA) (4).
2. method according to claim 1; It is characterized in that: beef sample is meat cubelets described in the step (1); Said method also comprises carries out cryodesiccated step with said meat cubelets; Said cryodesiccated step comprises placing carries out freezing in advance 12h～24h under-20 ℃～-80 ℃, place the step of vacuum freezing drying oven dry 24h～48h under-30 ℃～-50 ℃ again.
3. method according to claim 1 is characterized in that: beef sample is a muddy flesh described in the step (1), and said method also comprises the steps that said muddy flesh placed 60 ℃～70 ℃ following dry 36h～48h.
4. according to arbitrary described method among the claim 1-3, it is characterized in that: comprise also in the step (2) that it is 100 orders～200 purpose steps that said beef sample is crushed to granularity.
5. according to arbitrary described method among the claim 1-4, it is characterized in that: fat-extraction described in the step (2) is extracted through Suo Shi and is carried out, and the solvent that said Suo Shi extracts is that boiling range is 30 ℃～60 ℃ a sherwood oil, and the time is 18h～24h.
6. according to arbitrary described method among the claim 1-5, it is characterized in that: esterification reaction of organic acid carries out in the sulfuric acid methanol solution described in the step (3); The volumn concentration of sulfuric acid is 1% in the said sulfuric acid methanol solution.
7. according to arbitrary described method among the claim 1-6, it is characterized in that: the temperature of esterification reaction of organic acid described in the step (3) is 60 ℃～80 ℃, and the time is 50min～80min.
8. according to arbitrary described method among the claim 1-7, it is characterized in that: the method that detects described in the step (4) is a vapor-phase chromatography.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102749396A (en) * | 2012-07-17 | 2012-10-24 | 陕西科技大学 | Method for detecting fatty acid content by gas chromatography-mass spectrometry (GC-MS) |
| CN103399046A (en) * | 2013-08-09 | 2013-11-20 | 中国农业科学院农产品加工研究所 | Method for auxiliary discrimination of production areas of pears |
| CN105388207A (en) * | 2015-12-24 | 2016-03-09 | 中国农业科学院农产品加工研究所 | Method for identifying geographical indication agricultural product Zhongning lycium chinense |
| CN105606750A (en) * | 2016-01-06 | 2016-05-25 | 大连海事大学 | Aquatic product producing area tracing method based on fatty acid carbon stable isotopes |
| CN107356688A (en) * | 2017-07-07 | 2017-11-17 | 上海海洋大学 | A kind of method for distinguishing multiple geographical population jumbo flying squids using muscle essential fatty acid composition |
| CN109696501A (en) * | 2019-01-31 | 2019-04-30 | 中国农业科学院农产品加工研究所 | Meat or meat products feature volatile flavor substance discrimination method |
| CN109839489A (en) * | 2019-01-17 | 2019-06-04 | 北京农业质量标准与检测技术研究中心 | A method of beef origin is identified using electronic nose |
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| CN101793877A (en) * | 2009-11-26 | 2010-08-04 | 天津出入境检验检疫局动植物与食品检测中心 | Method for detecting fatty acid irradiated products in food |
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| JP2009204596A (en) * | 2008-02-29 | 2009-09-10 | Kose Corp | Analytical method of fatty acid content in sebum |
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Non-Patent Citations (15)
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102749396A (en) * | 2012-07-17 | 2012-10-24 | 陕西科技大学 | Method for detecting fatty acid content by gas chromatography-mass spectrometry (GC-MS) |
| CN102749396B (en) * | 2012-07-17 | 2013-10-23 | 陕西科技大学 | Method for detecting fatty acid content by gas chromatography-mass spectrometry (GC-MS) |
| CN103399046A (en) * | 2013-08-09 | 2013-11-20 | 中国农业科学院农产品加工研究所 | Method for auxiliary discrimination of production areas of pears |
| CN105388207A (en) * | 2015-12-24 | 2016-03-09 | 中国农业科学院农产品加工研究所 | Method for identifying geographical indication agricultural product Zhongning lycium chinense |
| CN105388207B (en) * | 2015-12-24 | 2018-06-29 | 中国农业科学院农产品加工研究所 | A kind of method for differentiating peaceful matrimony vine in geographical sign agricultural product |
| CN105606750A (en) * | 2016-01-06 | 2016-05-25 | 大连海事大学 | Aquatic product producing area tracing method based on fatty acid carbon stable isotopes |
| CN107356688A (en) * | 2017-07-07 | 2017-11-17 | 上海海洋大学 | A kind of method for distinguishing multiple geographical population jumbo flying squids using muscle essential fatty acid composition |
| CN109839489A (en) * | 2019-01-17 | 2019-06-04 | 北京农业质量标准与检测技术研究中心 | A method of beef origin is identified using electronic nose |
| CN109696501A (en) * | 2019-01-31 | 2019-04-30 | 中国农业科学院农产品加工研究所 | Meat or meat products feature volatile flavor substance discrimination method |
| CN109696501B (en) * | 2019-01-31 | 2021-12-07 | 中国农业科学院农产品加工研究所 | Method for identifying characteristic volatile flavor substances of meat or meat products |
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Application publication date: 20120627 |