CN111393337A - Method for extracting carmine from meat product - Google Patents
Method for extracting carmine from meat product Download PDFInfo
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- CN111393337A CN111393337A CN202010142440.6A CN202010142440A CN111393337A CN 111393337 A CN111393337 A CN 111393337A CN 202010142440 A CN202010142440 A CN 202010142440A CN 111393337 A CN111393337 A CN 111393337A
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- carmine
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- petroleum ether
- sample
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- 238000000034 method Methods 0.000 title claims abstract description 30
- 235000012730 carminic acid Nutrition 0.000 title claims abstract description 25
- 235000013622 meat product Nutrition 0.000 title claims abstract description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000003208 petroleum Substances 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 238000002156 mixing Methods 0.000 claims abstract description 13
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- 238000007710 freezing Methods 0.000 claims abstract description 4
- 230000008014 freezing Effects 0.000 claims abstract description 4
- 239000006228 supernatant Substances 0.000 claims abstract description 3
- 238000000605 extraction Methods 0.000 claims description 10
- 238000001514 detection method Methods 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 4
- 238000000464 low-speed centrifugation Methods 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 8
- 239000000523 sample Substances 0.000 description 21
- 239000000126 substance Substances 0.000 description 6
- 235000013305 food Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 150000004676 glycans Chemical class 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 239000001040 synthetic pigment Substances 0.000 description 3
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 3
- 239000003086 colorant Substances 0.000 description 2
- IINNWAYUJNWZRM-UHFFFAOYSA-L erythrosin B Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 IINNWAYUJNWZRM-UHFFFAOYSA-L 0.000 description 2
- 235000012732 erythrosine Nutrition 0.000 description 2
- 239000004174 erythrosine Substances 0.000 description 2
- 229940011411 erythrosine Drugs 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 235000012731 ponceau 4R Nutrition 0.000 description 2
- 239000004175 ponceau 4R Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- -1 scarlet Chemical compound 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 235000009328 Amaranthus caudatus Nutrition 0.000 description 1
- 240000001592 Amaranthus caudatus Species 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 235000012735 amaranth Nutrition 0.000 description 1
- 239000004178 amaranth Substances 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 235000012970 cakes Nutrition 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 235000015177 dried meat Nutrition 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- NYGZLYXAPMMJTE-UHFFFAOYSA-M metanil yellow Chemical group [Na+].[O-]S(=O)(=O)C1=CC=CC(N=NC=2C=CC(NC=3C=CC=CC=3)=CC=2)=C1 NYGZLYXAPMMJTE-UHFFFAOYSA-M 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 235000020995 raw meat Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C303/00—Preparation of esters or amides of sulfuric acids; Preparation of sulfonic acids or of their esters, halides, anhydrides or amides
- C07C303/42—Separation; Purification; Stabilisation; Use of additives
- C07C303/44—Separation; Purification
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
Abstract
The invention provides a method for extracting carmine from meat products, which is characterized by comprising the following steps: 1) and g: and ml is calculated as 1: mixing the sample with water according to a proportion of 5-10, heating to 60-80 ℃, shaking, mixing uniformly, extracting, centrifuging at a low speed, and collecting supernatant centrifugate; 2) mixing the upper layer centrifugate with petroleum ether, uniformly mixing, freezing and centrifuging to obtain the middle layer liquid; the volume ratio of the upper layer of the centrifugal liquid to the petroleum ether is 3-5: 1; 3) removing residual petroleum ether in the middle layer liquid by water bath to obtain carmine extracting solution. The method provided by the invention is economical, simple and convenient, is easy to operate, greatly saves the reagent dosage and the operation time, and is suitable for batch sample operation.
Description
Technical Field
The invention relates to the technical field of food, in particular to a method for extracting and purifying carmine in meat products.
Background
Carmine, also known as edible Red No. 102 (Japan), edible Red No. 7, ponceau 4R, scarlet, and brilliant scarlet, is an aqueous azo-based colorant. The chemical name is 1- (4 '-sulfonic group-1' -naphthylazo) -2-naphthol-6, 8-disulfonic acid trisodium salt, which is an isomer of amaranth. Carmine (Ponceau 4R) is a monoazo synthetic pigment which is widely used and used in the largest amount in China at present. In recent years, it has been reported that carmine and Sudan I, which is forbidden in the European Union standards, belong to azo pigments, and that azo compounds are metabolized in vivo to produce an aromatic amine compound, which is a mutagenic precursor. The food range and the dosage of the artificial synthetic pigment carmine added in China are specified by national standards, the carmine can be used for coloring foods such as fruit juice beverages, prepared wines, carbonated beverages, candies, cakes, ice creams, yoghourt and the like, but cannot be used for foods such as jerky, dried meat products, aquatic products and the like, and the artificial synthetic pigment carmine is mainly used for preventing lawless people from hiding the appearance of poor raw meat such as deteriorated meat by excessively using the pigment to deceive consumers.
At present, the content of the carmine in the food is mainly detected by high performance liquid chromatography, thin layer chromatography, oscillography and the like, but before detection, a sample needs to be pretreated to extract and purify the carmine. In GB/T9695.6-2008 determination Standard of carmine colorant for meat products, a method for extracting and purifying carmine from meat products is provided, wherein a sample is treated by petroleum ether, and then extracted after grease is removed. The operation is complex, the time consumption is long, the reagent dosage is large, and especially the dosage of the organic reagent petroleum ether is huge.
Disclosure of Invention
In view of the above needs, the present invention provides an extraction method that is economical, simple, easy to operate, and uses a small amount of reagents. The method is applied to the extraction and purification of the carmine in the meat product, so that the use amount of organic reagents is greatly reduced, and the working efficiency is improved.
The technical scheme provided by the invention is as follows:
a method for extracting carmine from meat product comprises:
1) and g: and ml is calculated as 1: mixing the sample with water according to a proportion of 5-10, heating to 50-70 ℃, shaking, mixing uniformly, extracting, centrifuging at a low speed, and collecting supernatant centrifugate;
2) mixing the upper layer centrifugate with petroleum ether, uniformly mixing, freezing and centrifuging to obtain a middle layer liquid; the volume ratio of the upper layer of the centrifugal liquid to the petroleum ether is 3-5: 1;
3) and removing residual petroleum ether in the middle layer liquid to obtain a carmine extracting solution.
In one embodiment according to the invention, step 1) further comprises subjecting the sample to a comminution treatment prior to extraction.
In one embodiment of the invention, the sample is mixed with water in the step 1) and then heated in a water bath for 5-15 min.
In one embodiment according to the invention, step 1) further comprises repeating the extraction 2 times.
In one embodiment according to the invention, the low speed centrifugation in step 1) is at a rotation speed of 2200r/min to 3800 r/min.
In one embodiment according to the invention, the petroleum ether boiling range in step 2) is from 30 to 60 ℃.
In one embodiment according to the invention, the refrigerated centrifugation in step 2) is carried out at a rotation speed of 2 to 6 ℃ and 8000 to 10000 r/min.
In one embodiment according to the invention, step 3) is carried out by means of a water bath and/or nitrogen flushing.
In one embodiment according to the invention, the temperature of the water bath is 60-80 ℃; preferably 60 deg.c.
The invention also provides application of the method in detecting the carmine content in the meat product.
Compared with the extraction and purification method in the national standard, the method provided by the invention is economic, simple and convenient and easy to operate. The dosage of the reagent is small, and particularly the dosage of the organic reagent petroleum ether is less than one tenth of the dosage of the standard method.
Drawings
FIG. 1 shows a chromatogram of a carmine standard (retention time 5.003 min);
FIG. 2 shows a spectrum of a carmine standard;
FIG. 3 shows a chromatographic overlay of a sample and a labeled sample.
Detailed Description
The following examples are intended to illustrate the present application but are not intended to limit the scope of the present application.
Specific embodiments of the present application will be described in more detail below. These embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the present application, but is made for the purpose of illustrating the general principles of the application and not for the purpose of limiting the scope of the application. The protection scope of the present application shall be subject to the definitions of the appended claims.
Unless otherwise specified, the reagents used in the present invention are commercially available.
Example 1 extraction of carmine from meat product samples
1) Crushing:
the sample is prepared into the meat paste which is convenient to be uniformly mixed with the extracting solution and is fully extracted.
2) Extraction:
accurately weighing 2.00g (accurate to 0.001g) of sample in a 50m L centrifuge tube, adding primary water to a constant volume of 15m L;
heating in water bath at 60 ℃ for 10min, vortex extracting for 5min, centrifuging at 3000r/min for 3min at normal temperature, taking the upper layer liquid, repeatedly adding water to 15 or 10m L, extracting for 2 times, heating to enable vortex extraction of the sample to be carried out at a higher temperature, wherein the higher temperature is favorable for destroying hydrogen bonds among partial molecules and in the molecules, so that the viscosity in the extraction process is reduced, and the to-be-detected substances in the sample are favorably dispersed in the extracting solution more uniformly.
(3) Adding 10m L petroleum ether (boiling range is 30-60 ℃) into the upper-layer centrifugate, fully mixing uniformly, and freezing and centrifuging at 4 ℃ at 9000r/min for 5 min;
(4) the method is characterized in that a middle clear liquid layer is taken out and put in a new 50m L centrifugal tube, a small amount of petroleum ether (boiling range is 30-60 ℃) is possibly brought in, the centrifugal tube is placed in a 60 ℃ water bath for heating for 10min, residual petroleum ether can be removed, 1 step can be carried out to remove oil and fat-soluble protein in a sample, and part of water-soluble protein, polysaccharide substances and other impurities, wherein the petroleum ether is a weak polar solvent and is insoluble in water, and the relative density (water ═ 1) is between 0.64 and 0.66, so that even if 2g of the sample is totally oil and fat and is dispersed in 10m L petroleum ether (the volume is constant), the relative density is still less than 1, and multiple experimental phenomena prove the result, the low temperature of 4 ℃ can reduce the solubility of the water-soluble protein, the polysaccharide substances and other impurities in water, the sample is frozen and centrifuged for 5min at 9000r/min, the layering of the solution and the weak polar solution can be accelerated, meanwhile, insoluble substances are added, the relatively small density moves upwards, the relatively large density moves downwards, so that the oil and the water-soluble protein, the fat-soluble protein, the polysaccharide substances.
Example 2 purification of extract:
(1) purification step of the Standard method (GB/T9695.6-2008).
(2) SPE column method, the item containing erythrosine and using PWAX SPE column (150mg/6m L), and the item containing no erythrosine and using polyamide SPE column (1g/6m L).
The specific steps are operated according to the instructions of corresponding manufacturers, eluent is collected, water bath nitrogen blow drying is carried out at 50 ℃, water is added, the volume is fixed to 1m L, the mixture is filtered through a special pigment film with the diameter of 0.45 mu m, and the mixture is to be detected.
Example 3 comparison of the invention with the national Standard method
The samples were tested according to the chromatographic method in the national standard and the results are shown in FIGS. 1-3.
The selection of the labeling points is that the detection limit of the GB 9695.6-2008 method is 0.05mg/kg (sample weighing is 5.0 g-10.0 g; the final volume of the sample liquid obtained after the sample is processed is 10.0m L), and the corresponding concentration of the sample liquid is 0.05 mg/L. for better confirmation of the feasibility of the method, relatively accurate detection results are required, so 2 points with detection limits increased by 3 times and 10 times in the final concentration of the sample liquid, namely 0.15 mg/L and 0.50 mg/L, are selected.
The method is used for testing the actual sample and the standard sample, the recovery rate is 89.3 to 92.0 percent when the final concentration of the standard sample solution is 0.15 mg/L, and the recovery rate is 94.8 to 100.0 percent when the final concentration of the standard sample solution is 0.50 mg/L.
The comparison of the process with the national standard process is shown in table 1.
TABLE 1 COMPARATIVE TABLE
It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.
Claims (10)
1. A method for extracting carmine from meat products is characterized by comprising the following steps:
1) and g: and ml is calculated as 1: mixing the sample with water according to a proportion of 5-10, heating to 60-80 ℃, shaking, mixing uniformly, extracting, centrifuging at a low speed, and collecting supernatant centrifugate;
2) mixing the upper layer centrifugate with petroleum ether, uniformly mixing, freezing and centrifuging to obtain a middle layer liquid; the volume ratio of the upper layer of the centrifugal liquid to the petroleum ether is 3-5: 1;
3) removing residual petroleum ether in the middle layer liquid by water bath to obtain carmine extracting solution.
2. The method of claim 1, wherein step 1) further comprises subjecting the sample to a comminution process prior to extraction.
3. The method of claim 1, wherein the sample is mixed with water in step 1) and heated in a water bath for 5-15 min.
4. The method of claim 1, wherein step 1) further comprises repeating the extracting 2 times.
5. The method of claim 1, wherein the low speed centrifugation of step 1) is at a speed of 2200r/min to 3800 r/min.
6. The method of any one of claims 1-5, wherein the petroleum ether boiling range in step 2) is from 30 ℃ to 60 ℃.
7. The method of claim 6, wherein the step 2) of said cryocentrifugation is performed at a rotation speed of 2-6 ℃ and 8000-10000 r/min.
8. The method of claim 6, wherein step 3) is performed by a water bath method.
9. The method of claim 8, wherein the temperature of the water bath is 60-80 ℃; preferably 60 deg.c.
10. Use of the method according to any one of claims 1 to 9 for the detection of the carmine content in a meat product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010142440.6A CN111393337B (en) | 2020-03-04 | 2020-03-04 | Method for extracting carmine from meat product |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010142440.6A CN111393337B (en) | 2020-03-04 | 2020-03-04 | Method for extracting carmine from meat product |
Publications (2)
| Publication Number | Publication Date |
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| CN111393337A true CN111393337A (en) | 2020-07-10 |
| CN111393337B CN111393337B (en) | 2022-07-01 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104198414A (en) * | 2014-09-05 | 2014-12-10 | 北京智云达科技有限公司 | Method for detecting artificially synthesized edible pigments |
| CN104569273A (en) * | 2015-01-21 | 2015-04-29 | 华南理工大学 | HPLC-MS/MS detection method for 11 edible synthetic pigments in meat or meat products |
| CN106645484A (en) * | 2016-12-27 | 2017-05-10 | 广州广电计量检测股份有限公司 | Identification method of fake and shoddy meat floss products |
| CN106990183A (en) * | 2017-05-05 | 2017-07-28 | 蚌埠市疾病预防控制中心 | The method for chromatographic determination of synthetic coloring matter in a kind of food |
| CN109655567A (en) * | 2018-12-25 | 2019-04-19 | 沈阳出入境检验检疫局检验检疫综合技术中心 | A kind of method of acid stain in detection animal meat products |
-
2020
- 2020-03-04 CN CN202010142440.6A patent/CN111393337B/en not_active Expired - Fee Related
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| CN104198414A (en) * | 2014-09-05 | 2014-12-10 | 北京智云达科技有限公司 | Method for detecting artificially synthesized edible pigments |
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| CN106645484A (en) * | 2016-12-27 | 2017-05-10 | 广州广电计量检测股份有限公司 | Identification method of fake and shoddy meat floss products |
| CN106990183A (en) * | 2017-05-05 | 2017-07-28 | 蚌埠市疾病预防控制中心 | The method for chromatographic determination of synthetic coloring matter in a kind of food |
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