CN108896674A - The detection method of polycyclic aromatic hydrocarbon in a kind of meat products - Google Patents
The detection method of polycyclic aromatic hydrocarbon in a kind of meat products Download PDFInfo
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- CN108896674A CN108896674A CN201810726684.1A CN201810726684A CN108896674A CN 108896674 A CN108896674 A CN 108896674A CN 201810726684 A CN201810726684 A CN 201810726684A CN 108896674 A CN108896674 A CN 108896674A
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Abstract
The invention belongs to harmful substance detection technique fields in food, and in particular to the detection method of polycyclic aromatic hydrocarbon in a kind of meat products.Naturally dry and broken meat mud after meat products is thawed, ultrasonic extraction after solvent vibrates is added, it is purified after extracting solution centrifugation by florisil silica solid phase extraction column, using gel chromatography column purification, finally by the content of the polycyclic aromatic hydrocarbon in gas chromatograph-mass spectrometer (GC-MS) (GC-MS) detection scavenging solution.Detection method precision is high, not vulnerable to impurity interference, highly reliable, can detect the content of 18 kinds of polycyclic aromatic hydrocarbons in meat products simultaneously.
Description
Technical field
The invention belongs to harmful substance detection technique field in food, a kind of inspection of polycyclic aromatic hydrocarbon in meat products is specifically designed
Survey method.
Background technique
Polycyclic aromatic hydrocarbon (Polycyclic Aromatic Hydrocarbons, PAHs) is the benzene with two or more
Cyclopolymerization is bonded and as many as formation ring carbon hydrogen compound, the generally product of imperfect combustion or high temperature pyrolysis, main source
Natural origin and artificial burning behavior source, combustion of the PAHs from volcano eruption, forest and prairie fire in nature can be divided into
Burning activity and formed, but the amount for increasing PAHs in natural environment of the activity of the mankind rapidly, the discharge amount of nature without
Compared with method PAHs amount caused by the mankind.PAHs caused by mankind's burning behavior mostlys come from the burning of fossil fuel, such as
Exhaust gas discharge, coke refining industry, pitch manufacturing industry and all industrial equipments for using fossil fuel of motor vehicles etc..PAHs
It is typically found in the petroleum chemicals such as creosote, tar, oil refining, dyestuff, lubricating oil, antirust oil, rubber, plastics.Toxicity, ecology
Toxicity and the environmental science committee (CSTEE) confirm through scientific research, the fat-soluble height of polycyclic aromatic hydrocarbon, not degradable and easily in biology
Cylinder accumulation has carcinogenic, teratogenesis and mutagenic effect, has huge potential hazard to human health and ecological environment.
Polycyclic aromatic hydrocarbon includes more than 150 compound such as naphthalene, anthracene, phenanthrene, pyrene.Polycyclic aromatic hydrocarbon is distributed widely in natural environment, such as：It is empty
Gas and water, deposit, soil, vegetables and animal tissue etc., even people warm oneself indoors, cook or all can when smoking
Polycyclic aromatic hydrocarbon is generated due to hydrocarbon partial imperfect combustion.Such compound has teratogenesis, carcinogenic and mutagenicity,
And carcinogenicity increases with the increase of phenyl ring number, and the common polycyclic aromatic hydrocarbon with carcinogenesis is mostly four to six rings
Fused ring compound.Polycyclic aromatic hydrocarbon in the environment it is most of be in the form of ADSORPTION STATE and emulsification state exist, once into environment, just by
To the influence of various nature intrinsic procedures, change.It is reacted by complicated physical migration, chemistry and bioconversion,
Constantly change in the systems such as atmosphere, water body, soil, organism, changes distribution situation.It is in different conditions, not homologous ray
Polycyclic aromatic hydrocarbon then shows different variation behaviors.After polycyclic aromatic hydrocarbon enters atmosphere, chemical reaction, depositing dust, rainfall, drop can be passed through
The processes such as snow enter in soil and water body, are further absorbed and enter in various meat products by poultry etc..
It is current to have paper chromatography, thin-layer chromatography, packed column gas chromatography, capillary column in polycyclic aromatic hydrocarbon analysis method for measuring
The methods of gas-chromatography, gas chromatography-mass spectrum, high performance liquid chromatography.The side such as paper chromatography, thin-layer chromatography, packed column gas chromatography
Method can be carried out quantitative analysis, but separation efficiency and analysis speed are poor.Capillary Column GC and high performance liquid chromatography
Method has very high separating degree, and high sensitivity, the range of linearity is wide, and quantitative precision is good, but qualitative main according to reservation
Time.The extracting method of PAHs has following several method：Soxhlet extraction, ultrasonic extraction method, microwave abstracting, accelerated solvent extraction
Method, supercritical fluid extraction and collaboration extractive technique.Under normal circumstances, PAHs extraction solvent used is non-selective,
In extracting solution other than containing PAHs, inevitably contain a certain amount of non-aromatic hydrocarbon impurity, these impurity may interfere with
The quantitative analysis of PAHs, therefore extracting solution has to be purified before carrying out quantitative analysis, purification effect quality is directly closed
It is to qualitative and quantitative accuracy.General purification process is column chromatography, and common Adsorption agent has following 3 kinds：
Silica gel, neutral alumina and Fu Luoli soil.Wherein silica gel be PAHs analysis purifying in be most widely used；Neutral alumina by
Less desirable in the removal effect to alkane, general and silica gel is made into mixing silicagel column with certain ratio column；Fu Luoli soil is net
Change method is easy to operate, favorable reproducibility, and it is good to isolate and purify effect for the PAHs of high molecular weight.Currently, in natural fuel oil
The measuring method of PAHs generallys use combined gas chromatography mass spectrometry, and concrete operation step is first to dissolve sample with hexamethylene,
After being extracted with dimethyl sulfoxide, sodium chloride solution is added, then be stripped with hexamethylene.After hexamethylene alkane extract is washed, nitrogen is used
Air-blowing is to close dry, after n-hexane dissolution, then with Solid Phase Extraction column purification, after concentrated constant volume, uses gas chromatography-mass spectrography
Instrument (GC-MS) measurement, inner mark method ration.
Summary of the invention
The object of the present invention is to provide a kind of detection methods of polycyclic aromatic hydrocarbon in meat products, dry in the air naturally after meat products is thawed
Dry doubling is crushed meat mud, and ultrasonic extraction after solvent oscillation is added, passes through florisil silica solid phase extraction column after extracting solution centrifugation
Purification, using gel chromatography column purification, finally by more in gas chromatograph-mass spectrometer (GC-MS) (GC-MS) detection scavenging solution
The content of cycloaromatics.Detection method precision is high, not vulnerable to impurity interference, highly reliable, can detect in meat products simultaneously
The content of 18 kinds of polycyclic aromatic hydrocarbons.
The technical scheme is that：
The detection method of polycyclic aromatic hydrocarbon, includes the following steps in a kind of meat products：
(1) meat products is thawed, then naturally dry, by the meat products after drying be crushed meat mud, be added n-hexane and
Methylene chloride mixed solvent, ultrasonic extraction after oscillation, after extracting solution is centrifuged, n-hexane and methylene chloride is added in filter residue again
Mixed solvent ultrasonic extraction is simultaneously centrifuged, and is merged the liquid concentration after being centrifuged twice to doing, is re-dissolved to obtain to net with n-hexane
Change liquid；
(2) florisil silica solid phase extraction column is activated with n-hexane, then by liquid loading to be clean, after collection and purification
Solution, use n-hexane and acetone mixed solvent as elution and collect eluent, by the purified solution of collection
It is concentrated to dryness after merging with eluent, is then re-dissolved with n-hexane；
(3) gel chromatographic columns are eluted using n-hexane and methylene chloride mixed solvent, after having eluted, be added in step (2)
The solution that re-dissolves of n-hexane, n-hexane and the elution of methylene chloride mixed solvent is then added, collects leacheate, is concentrated into
It is dry, it is then re-dissolved to obtain prepare liquid with n-hexane；
(4) above-mentioned prepare liquid is detected to the content of wherein polycyclic aromatic hydrocarbon using gas chromatograph-mass spectrometer (GC-MS).
Preferably, in step (1) and (3) n-hexane and methylene chloride in the mixed solvent n-hexane and methylene chloride volume
Than being 1:1.
Preferably, duration of oscillation is 15min~30min in step (1).
Preferably, supersonic frequency is 50HZ~80HZ in step (1), and ultrasonic time is 40min~60min.
Preferably, the revolving speed being centrifuged in step (1) is 4000r/min~5000r/min, centrifugation time be 5min~
10min。
Preferably, the volume ratio of n-hexane and acetone is 4 in n-hexane and acetone mixed solvent in step (2):1.
Preferably, the temperature in step (1) to (3) when concentration is 30 ± 1 DEG C.
Preferably, the analysis parameter of chromatograph-mas spectrometer is in step (4)：
Analytical column：DB-5MS(30m×0.25um×0.25um)
Sample introduction mode：Split ratio is 5:1
Injector temperature：290℃
Partial flow rate：5ml/min
Temperature gradient：60 DEG C of initial temperature, retain 0min；120 DEG C are warming up to the speed of 30 DEG C/min, retains 0min；
250 DEG C are warming up to the speed of 10 DEG C/min, retains 0min；310 DEG C are warming up to the speed of 10 DEG C/min, retains 2.5min.
Preferably, above-mentioned polycyclic aromatic hydrocarbon includes：Naphthalene, 1- methyl naphthalene, 2- methyl naphthalene, acenaphthylene, acenaphthene, fluorenes, phenanthrene, anthracene, fluoranthene,
Pyrene, benzo [a] anthracene, bend, benzo [b] fluoranthene, benzo [k] fluoranthene, benzo [a] pyrene, indenes benzene [1,2,3-cd] pyrene, dibenzo [a,
H] 18 kinds of anthracene, benzo [g, h, i] substances.
Beneficial effects of the present invention are as follows：
Naturally dry and broken meat mud, are added ultrasonic extraction after solvent oscillation, extract after the present invention thaws meat products
It is purified after liquid centrifugation by florisil silica solid phase extraction column, using gel chromatography column purification, finally by gas-chromatography-
Mass spectrometer (GC-MS) detects the content of the polycyclic aromatic hydrocarbon in scavenging solution.Detection method precision is high, not vulnerable to impurity
It interferes, is highly reliable, the content of 18 kinds of polycyclic aromatic hydrocarbons in meat products can be detected simultaneously.
Specific embodiment
With reference to embodiment, technical solution of the present invention is described in further detail, but do not constituted pair
Any restrictions of the invention.
Embodiment 1
The detection method of polycyclic aromatic hydrocarbon, includes the following steps in a kind of meat products：
(1) meat products is thawed, then naturally dry, the meat products after drying is crushed meat mud, 4g muddy flesh is weighed and adds
Enter into 50mL polytetrafluoroethylene (PTFE) centrifuge tube, it is 1 that 20mL volume ratio, which is added,:1 n-hexane and methylene chloride mixed solvent, oscillation
After 15min under 60HZ frequency ultrasonic extraction 50min, then centrifuge tube is placed in a centrifuge under the revolving speed of 4500r/min from
Heart 5min, filters to take supernatant；Above-mentioned n-hexane is added in filter residue again and the mixed solvent similarity condition ultrasound of methylene chloride mentions
Simultaneously centrifugal filtration is taken, merges filtered liquid twice, is placed in Rotary Evaporators rotary evaporation under the conditions of 30 DEG C and is concentrated into
It is dry, 2mL n-hexane is then added and re-dissolves to obtain liquid to be clean；
(2) florisil silica solid phase extraction column is activated with 5mL n-hexane, then by 2mL liquid loading to be clean,
Solution after collection and purification is 4 with 15mL volume ratio:1 n-hexane and acetone mixed solvent as elution and is collected
Purified solution and the eluent merging of collection are placed in Rotary Evaporators rotary evaporation under the conditions of 30 DEG C by eluent
It is concentrated to dryness, is then re-dissolved with 5mL n-hexane；
(3) cleaned gel is loaded into chromatographic column, filling height is 20cm, is 1 with 100mL volume ratio:1 just
Hexane and methylene chloride mixed solvent elute GPC column, and the 5mL sample re-dissolved in step (2) is transferred on GPC column, is added
Entering 100mL volume ratio is 1:1 n-hexane and methylene chloride mixed solvent elution, collects eluent, is placed on Rotary Evaporators
Rotary evaporation is concentrated to dryness under the conditions of 30 DEG C, then re-dissolves to obtain prepare liquid with 1mL n-hexane；
(4) analytical parameter setup of gas chromatograph-mass spectrometer (GC-MS) is：
Analytical column：DB-5MS(30m×0.25um×0.25um)
Sample introduction mode：Split ratio is 5:1
Injector temperature：290℃
Partial flow rate：5ml/min
Temperature gradient：60 DEG C of initial temperature, retain 0min；110 DEG C are warming up to the speed of 30 DEG C/min, retains 2min；
250 DEG C are warming up to the speed of 8 DEG C/min, retains 5min；310 DEG C are warming up to the speed of 10 DEG C/min, retains 2.5min.
(5) 18 kinds of (naphthalene, 1- methyl naphthalene, 2- methyl naphthalene, acenaphthylene, acenaphthene, fluorenes, phenanthrene, anthracene, fluoranthene, pyrene, benzos [a] are weighed respectively
Anthracene is bent, benzo [b] fluoranthene, benzo [k] fluoranthene, benzo [a] pyrene, indenes benzene [1,2,3-cd] pyrene, dibenzo [a, h] anthracene, benzo
[g, h, i]) polycyclic aromatic hydrocarbon standard substance 0.1000g (being accurate to 0.1mg), it is settled to respectively after being dissolved with methylene chloride
In 100mL volumetric flask, the Standard Stock solutions that concentration is 1000mg/L are obtained；Above-mentioned 18 kinds of polycyclic aromatic hydrocarbon marks are accurately pipetted respectively
Quasi- stock solution 1mL is settled to scale with n-hexane in 25mL volumetric flask, obtains the hybrid standard deposit that concentration is 40mg/L
Solution；Take a certain amount of mixing standard reserving solution, diluted with n-hexane, be configured to respectively concentration be 2ug/L, 5ug/L, 10ug/L,
20u/L, 50ug/L standard working curve solution.Under the chromatographic condition of setting, from low to high by concentration by standard working solution
It is successively measured, using the peak area of obtained chromatographic peak as ordinate, the sample introduction concentration of corresponding each substance is abscissa work
Figure draws standard working curve.Addition concentration when with every kind of compound signal-to-noise ratio (S/N)=3 is determined as method detection limit, with
Addition concentration when signal-to-noise ratio (S/N)=10 is determined as method quantitative limit.The titles of 18 kinds of obtained polycyclic aromatic hydrocarbons, when retaining
Between, mass spectrometry parameters it is as shown in table 1 below；The standard curve of 18 kinds of polycyclic aromatic hydrocarbons is linear and quantitative limit is as shown in table 2 below.
Table 1 18 kinds of polycyclic aromatic hydrocarbon titles, retention time, mass spectrometry parameters
| Serial number | Title | Retention time | Ion |
| 1 | Naphthalene | 8.95 | 128.1,127.1,129.1,102.1 |
| 2 | 1- methyl naphthalene | 9.99 | 142.1,141.1,139,115.1 |
| 3 | 2- methyl naphthalene | 10.23 | 142.1,141.1,139,115.1 |
| 4 | Acenaphthylene | 12.11 | 152.1,153.1,151.1,150.1 |
| 5 | Acenaphthene | 12.58 | 154.1,153.1,152,76.1 |
| 6 | Fluorenes | 14.35 | 166.1,165.1,167.1,139 |
| 7 | It is luxuriant and rich with fragrance | 18.59 | 178.1,179.1,176.1,152.1 |
| 8 | Anthracene | 18.99 | 178.1,179.1,176.1,152.1 |
| 9 | Fluoranthene | 25.37 | 202.1,203.1,200.1,101.1 |
| 10 | Pyrene | 27.11 | 202.1,203.1,200.1,101.1 |
| 11 | Benzo [a] anthracene | 34.24 | 228.1,226.1,229.1,227.1 |
| 12 | It bends | 35.45 | 228.1,226.1,229.1,227.1 |
| 13 | Benzo [b] fluoranthene | 38.23 | 252.1,253.1,250.1,126.1 |
| 14 | Benzo [k] fluoranthene | 38.86 | 252.1,253.1,250.1,126.1 |
| 15 | Benzo [a] pyrene | 39.35 | 252.1,253.1,250.1,126.1 |
| 16 | Indenes benzene [1,2,3-cd] pyrene | 43.23 | 276.1,277.1,274.1,138 |
| 17 | Dibenzo [a, h] anthracene | 42.45 | 278.1,276.1,279.1,139.1 |
| 18 | Benzo [g, h, i] | 43.69 | 276.1,274.1,277.1,138.1 |
The standard curves of 2 18 kinds of polycyclic aromatic hydrocarbons of table is linear and quantitative limit
| Serial number | Title | Range of linearity ug/L | R | Quantitative limit ug/L |
| 1 | Naphthalene | 2-50 | 0.9995 | 0.8 |
| 2 | 1- methyl naphthalene | 2-50 | 0.9994 | 0.8 |
| 3 | 2- methyl naphthalene | 2-50 | 0.9995 | 0.8 |
| 4 | Acenaphthylene | 2-50 | 0.9992 | 0.8 |
| 5 | Acenaphthene | 2-50 | 0.9992 | 0.8 |
| 6 | Fluorenes | 2-50 | 0.9993 | 0.8 |
| 7 | It is luxuriant and rich with fragrance | 2-50 | 0.9996 | 0.8 |
| 8 | Anthracene | 2-50 | 0.9995 | 0.8 |
| 9 | Fluoranthene | 2-50 | 0.9992 | 0.8 |
| 10 | Pyrene | 2-50 | 0.9997 | 0.8 |
| 11 | Benzo [a] anthracene | 2-50 | 0.9992 | 0.8 |
| 12 | It bends | 2-50 | 0.9993 | 0.8 |
| 13 | Benzo [b] fluoranthene | 2-50 | 0.9994 | 0.8 |
| 14 | Benzo [k] fluoranthene | 2-50 | 0.9995 | 0.8 |
| 15 | Benzo [a] pyrene | 2-50 | 0.9995 | 0.8 |
| 16 | Indenes benzene [1,2,3-cd] pyrene | 2-50 | 0.9995 | 0.8 |
| 17 | Dibenzo [a, h] anthracene | 2-50 | 0.9996 | 0.8 |
| 18 | Benzo [g, h, i] | 2-50 | 0.9996 | 0.8 |
By the data of above-mentioned table 2 it is found that detection method has preferable curve linear relationship, have lower
It is quantitative.
(6) prepare liquid is prepared in step (3) and carries out gas chromatography-mass spectrometry analysis with the chromatographic condition in step (4),
Obtained chromatographic peak area is calculated with the curve in step (5), is obtained 18 kinds of the specific of polycyclic aromatic hydrocarbon in the meat products and is contained
Amount.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (9)
1. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products, which is characterized in that include the following steps：
(1) meat products is thawed, then naturally dry, the meat products after drying is crushed meat mud, n-hexane and dichloro is added
Methane blended solvent, ultrasonic extraction after oscillation, after extracting solution is centrifuged, the mixing of n-hexane and methylene chloride is added in filter residue again
Solvent supersonic is extracted and is centrifuged, and is merged the liquid concentration after being centrifuged twice to doing, is re-dissolved to obtain liquid to be clean with n-hexane；
(2) florisil silica solid phase extraction column is activated with n-hexane, it is molten after collection and purification then by liquid loading to be clean
Liquid uses n-hexane and acetone mixed solvent as elution and collects eluent, by the purified solution of collection and washes
De- liquid is concentrated to dryness after merging, and is then re-dissolved with n-hexane；
(3) gel chromatographic columns are eluted using n-hexane and methylene chloride mixed solvent, after having eluted, be added in step (2) just
Then the solution that hexane re-dissolves is added n-hexane and the elution of methylene chloride mixed solvent, collects leacheate, be concentrated to dryness,
Then it is re-dissolved to obtain prepare liquid with n-hexane；
(4) prepare liquid is detected to the content of wherein polycyclic aromatic hydrocarbon using gas chromatograph-mass spectrometer (GC-MS).
2. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that step (1) and
(3) volume ratio of n-hexane and methylene chloride in the mixed solvent n-hexane and methylene chloride is 1 in:1.
3. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that in step (1)
Duration of oscillation is 15min~30min.
4. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that in step (1)
Supersonic frequency is 50HZ~80HZ, and ultrasonic time is 40min~60min.
5. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that in step (1)
The revolving speed of centrifugation is 4000r/min~5000r/min, and centrifugation time is 5min~10min.
6. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that in step (2)
The volume ratio of n-hexane and acetone is 4 in n-hexane and acetone mixed solvent:1.
7. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that step (1) is extremely
(3) temperature in when concentration is 30 ± 1 DEG C.
8. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that in step (4)
The analysis parameter of chromatograph-mas spectrometer is：
Analytical column：DB-5MS；Sample introduction mode：Split ratio is 5:1；
Injector temperature：290℃；
Partial flow rate：5ml/min；
Temperature gradient：60 DEG C of initial temperature, retain 0min；120 DEG C are warming up to the speed of 30 DEG C/min, retains 0min；With 10
DEG C/speed of min is warming up to 250 DEG C, retain 0min；310 DEG C are warming up to the speed of 10 DEG C/min, retains 2.5min.
9. the detection method of polycyclic aromatic hydrocarbon in a kind of meat products according to claim 1, which is characterized in that the polycyclic virtue
Hydrocarbon includes：Naphthalene, 1- methyl naphthalene, 2- methyl naphthalene, acenaphthylene, acenaphthene, fluorenes, phenanthrene, anthracene, fluoranthene, pyrene, benzo [a] anthracene, in the wrong, benzo [b] is glimmering
18 kinds of anthracene, benzo [k] fluoranthene, benzo [a] pyrene, indenes benzene [1,2,3-cd] pyrene, dibenzo [a, h] anthracene, benzo [g, h, i] objects
Matter.
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| CN111289672A (en) * | 2020-03-06 | 2020-06-16 | 浙江省疾病预防控制中心 | Sample pretreatment method and determination method for detecting polycyclic aromatic hydrocarbon marker in traditional Chinese medicine product |
| CN111562334A (en) * | 2020-06-09 | 2020-08-21 | 劲牌有限公司 | Method for detecting polycyclic aromatic hydrocarbon residue in plant traditional Chinese medicinal materials and extracts |
| CN112834477A (en) * | 2020-09-28 | 2021-05-25 | 厦门市普识纳米科技有限公司 | Method for detecting polycyclic aromatic hydrocarbon in soil |
| CN115266960A (en) * | 2022-04-28 | 2022-11-01 | 南京农业大学 | Method for extracting and detecting fluoranthene from trace plant tissue sample |
| CN114965797B (en) * | 2022-06-22 | 2023-08-08 | 广东工业大学 | Rapid detection method for 24 polycyclic aromatic hydrocarbons in complex cooked food matrix |
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