CN106053634A - Method for detecting N-dimethyl nitrosamine in cooked meat products - Google Patents
Method for detecting N-dimethyl nitrosamine in cooked meat products Download PDFInfo
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- CN106053634A CN106053634A CN201610345727.2A CN201610345727A CN106053634A CN 106053634 A CN106053634 A CN 106053634A CN 201610345727 A CN201610345727 A CN 201610345727A CN 106053634 A CN106053634 A CN 106053634A
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- G—PHYSICS
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Abstract
The invention discloses a method for detecting N-dimethyl nitrosamine in cooked meat products. The detection method comprises the following steps: preparing a standard solution, preparing a sample solution to be measured, carrying out qualitative detection on the sample solution, and carrying out quantitative detection on the sample solution. The sample preparation method is convenient, fast, and efficient, a GC-MS/MS analyzer is used, a standard curve is established through an external standard method, and finally the content of N-dimethyl nitrosamine is detected and calculated. Compared with the conventional gas chromatography-mass spectrograph (GC-MS), the steps such as water steam steaming, water bath concentration, and the like are eliminated, the operation becomes simpler, the needed instruments are reduced, the detection limit becomes lower, and the accuracy is improved.
Description
Technical field
The invention belongs to detection technique field, be specifically related to N-N-nitrosodimethylamine detection method in a kind of cooked meat product.
Background technology
N-N-nitrosodimethylamine is widely present in the environment.Up to 0.1-300 μ g/kg in the dynamic herbivore product such as fish, meat.?
Nicotiana tabacum L., vegetable, frumentum all can detect, belong to high poison, it has been determined that for animal carcinogen, absorb rapidly through digestive tract, respiratory tract, warp
Skin absorption is slow, and target organ is mainly liver and kidney.
Containing N dimethylamine nitroso-group, N dimethylamine nitroso-group etc. in the food such as drinks, meat and Fish curing food, bean product
Deng.Nitrosamine compound has certain acute toxicity, strong carcinogenecity, mainly causes hepatic necrosis hemorrhage, and its carcinogenecity is more people
Paid close attention to, the nitrosamine that hitherto it is found that has kind more than 300；Wherein N-N-nitrosodimethylamine be great representational material it
One.The source of the nitrosamine in food is as follows: the effect of salt nitrite when (1) pickles, such as cured fish, bacon etc.；(2) adding
During heated drying, in air, nitrogen is oxidized to the effect of nitrogen oxides, such as medicated beer, milk powder, bean product.In recent years for volatility
Nitrosamine detection display Food & Drink and meat, drinks present in N dimethylamine nitrosamine at μ g/kg or μ g/L number
Magnitude, so needing a kind of high sensitivity of foundation and the selective method of height to detect trace N dimethylamine nitrosamine.
China once specified the N dimethylamine content of nitrosamines in meat should 3 μ g/kg, at present, GB/T5009.26-
Gas chromatograph-mass spectrometer method in the instrument detection method of 2003 regulations, N-N-nitrosodimethylamine separating step is loaded down with trivial details, instrument cost
High；Up-to-date national food safety standard " mensuration of N-nitrosamine compound in food " exposure draft three kinds of methods of employing:
First method be gas chromatogram-nitrogen chemical reflective detector method, the second method be gas chromatogram-thermal energy analyzer method, the 3rd method be gas
Phase chromatograph high resolution mass spectrometry, these method disadvantage are exactly that the specific requirements to instrument is high, and detectable substance separating step is numerous
Trivial, time cost is high.
Summary of the invention
For present situation, the present invention provides N-N-nitrosodimethylamine detection method in a kind of cooked meat product, it is possible to reduces and separates
Step, reduces time cost low, improves accuracy rate.
For solving above-mentioned technical problem, the present invention adopts the following technical scheme that
N-N-nitrosodimethylamine detection method in a kind of cooked meat product, described detection method comprises the steps:
(1) standard solution of variable concentrations is prepared: first prepare standard reserving solution, weigh N-N-nitrosodimethylamine standard
Product, become the storing solution that concentration is 1000mg/L of N-N-nitrosodimethylamine, keep in Dark Place in 4 DEG C of refrigerators with acetontrile；So
After become concentration to be 0.5ng/mL, 1ng/mL, 2.5ng/mL, 4ng/mL, 5ng/mL standard reserving solution acetontrile standard
Product curve point is standby；
(2) preparation of testing sample solution:
A, weigh testing sample 5.00g, add the solution 10mL containing 1% acetic acid in acetonitrile, under 2500r/min rotating speed
Whirlpool is mixed whirlpool in agitator and is mixed 5min；
B, add 5g anhydrous magnesium sulfate, 1g sodium chloride and 1g disodium citrate after whirlpool mix 3min, then at 5000r/min
Supernatant is collected after centrifugal 5min under rotating speed；
C, absorption 6mL supernatant move into equipped with 0.5gPSA, 0.5gC18With in the 15mL centrifuge tube of 1g anhydrous magnesium sulfate, whirlpool
Mixed 3min, 5000r/min collect supernatant after being centrifuged 5min again；
D, again take supernatant, with after the 0.22 organic membrane filtration of μm as testing sample solution；
(3) testing sample is qualitative and detection by quantitative:
A, take 1.0 μ L testing sample solution sample introductions, detect with triple level Four bar gas chromatograph-mass spectrometer (GC-MS) analyses,
To the total ions chromatogram of testing sample solution, quota ion to chromatogram and qualitative ion pair relative abundance chromatogram；
B, the N-N-nitrosodimethylamine standard solution of variable concentrations step (1) prepared, by triple level Four bar gas phases
Chromatograph-mas spectrometer analysis detects, and testing conditions is identical with testing sample solution, it is thus achieved that the total ion current color of standard solution
Spectrogram, quota ion chromatogram and qualitative ion pair relative abundance chromatogram；
C, with the quota ion of N-N-nitrosodimethylamine, to making with the chromatographic peak of qualitative ion pair, to obtain N-dimethyl sub-
Nitramine standard curve；
D, according to the quota ion of N-N-nitrosodimethylamine in testing sample solution to the chromatographic peak with qualitative ion pair, knot
Standardization curve, is calculated and treats the concentration C of N-N-nitrosodimethylamine in sample measuring liquid, and be calculated cold cuts system as follows
The content X of N-N-nitrosodimethylamine in product, cubage formula is:
X=C*V/ (M*1000)
The content of N-N-nitrosodimethylamine during wherein X is testing sample, unit is mg/kg；C is N-diformazan in testing sample
The concentration of base nitrosamine, unit is ng/mL；V is testing sample volume of dissolution, and unit is mL；M is testing sample quality, unit
For g.
Preferably, described quota ion is to for 74 → 42.1, and qualitative ion pair is 73.9 → 42.1.
Preferably, the parameter used during triple level Four bar gas chromatograph-mass spectrometer (GC-MS) analyses detection is respectively as follows:
A, chromatographic condition
Chromatographic column is Agilent HP-INOWAX capillary chromatographic column, injector temperature 250 DEG C, Splitless injecting samples, carrier gas
(helium >=99.999%) flow velocity 0.887mL/min, sample size 1 μ L；Temperature programming: initial temperature 60 DEG C keeps 1min, with 40
DEG C/min is raised to 120 DEG C, it is raised to 250 DEG C with 5 DEG C/min the most again and keeps 3min.
B, Mass Spectrometry Conditions
Ion source: 230 DEG C, level Four bar: 150 DEG C, transmission line: 250 DEG C, scan pattern MRM, ionizing energy 70eV, solvent
Postpone 4.5min.
Wherein, above-mentioned testing sample is that the decision method of N-N-nitrosodimethylamine is: testing sample and N-dimethyl nitrous
Amine standard substance retention time is identical, identical quota ion to for 74 → 42.1, qualitative ion pair be 73.9 → 42.1 conditions
Under, both quota ions to and qualitative ion pair relative abundance ratio between 80%-120%, it is determined that for testing sample be
N-N-nitrosodimethylamine.
The invention has the beneficial effects as follows:
Compared with the gas chromatograph-mass spectrometer method being previously mentioned in GB/T5009.26-2003, save vapor distillation and water
The steps such as bath concentration, operation is simpler, and required instrument reduces；And detection limit is lower, accuracy is higher.
Accompanying drawing explanation
Fig. 1 is the total ions chromatogram of sample solution；
Fig. 2 is that the quota ion of sample solution is to chromatogram；
Fig. 3 is the qualitative ion pair relative abundance chromatogram of sample solution；
Fig. 4 is the total ions chromatogram of standard solution；
Fig. 5 is that the quota ion of standard solution is to chromatogram；
Fig. 6 is the qualitative ion pair relative abundance chromatogram of standard solution；
Fig. 7 is N-N-nitrosodimethylamine standard curve.
Detailed description of the invention
By specific embodiment, the present invention will be described in detail below.These embodiments are provided to be able to more
Thoroughly understand the present invention, and complete for the scope of the present invention can be conveyed to those skilled in the art.
" comprising " or " including " as mentioned by the middle of description and claim in the whole text is an open language, therefore should
It is construed to " comprise but be not limited to ".Description subsequent descriptions is to implement the better embodiment of the present invention, and right described description is
For the purpose of the rule of description, it is not limited to the scope of the present invention.Protection scope of the present invention is when regarding appended power
Profit requires that defined person is as the criterion.Below by way of specific experiment, the present invention is further expalined.
One, reagent and instrument
Chromatographic column: Agilent GC-MS 7890B-7000C, centrifuge: SIGMA 3K15, whirlpool mixes agitator: Germany
Heidolph, liquid-transfering gun: Germany eppendorf, whirlpool mixes agitator: VORTEXGEIE2, ultrasonator, 100,000/sky
It is flat: METTER TOLEDO XS205, ten thousand/balance: METTER TOLEDO ME204E, PSAAgela, C18Agela,
0.22 μm organic filter membrane Agela, N-dimethyl nitrous ammonium (chromatographically pure), 50mL centrifuge tube, cooked meat product (supermarket purchase).
Two, the preparation of standard solution
Standard reserving solution: weigh N-dimethyl nitrous ammonium standard substance, become the storing solution of 1000mg/L with acetontrile, in 4
DEG C refrigerator keeps in Dark Place；
Standard solution: standard reserving solution dilution in acetonitrile to concentration is respectively 0.5ng/mL, 1ng/mL, 2.5ng/
The standard solution of mL, 4ng/mL and 5ng/mL is standby.
Three, the preparation of testing sample solution
A, weigh testing sample 5.00g, add the solution 10mL of acetic acid containing 1% in acetonitrile, under 2500r/min rotating speed
Whirlpool is mixed whirlpool in agitator and is mixed 5min；
B, add 5g anhydrous sodium sulfate, 1g sodium chloride and 1g disodium citrate after whirlpool mix 3min, then at 5000r/min
Supernatant is collected after centrifugal 5min under rotating speed；
C, absorption 6mL supernatant move into equipped with 0.5gPSA, 0.5g C18With in the 15mL centrifuge tube of 1g anhydrous magnesium sulfate, whirlpool
Mixed 3min, 5000r/min collect supernatant after being centrifuged 5min again；
D, again take supernatant, with after the 0.22 organic membrane filtration of μm as testing sample solution.
Four, sample is qualitative and detection by quantitative
Standard solution and sample solution are entered according to the running parameter of following triple level Four bar gas chromatograph-mass spectrometer (GC-MS)s
Row measures.
The parameter used during triple level Four bar gas chromatograph-mass spectrometer (GC-MS) analyses detection is respectively as follows:
A, chromatographic condition
Chromatographic column is Agilent HP-INOWAX capillary chromatographic column, injector temperature 250 DEG C, Splitless injecting samples, carrier gas
(helium >=99.999%) flow velocity 0.887mL/min, sample size 1 μ L；Temperature programming: initial temperature 60 DEG C keeps 1min, with 40
DEG C/min is raised to 120 DEG C, it is raised to 250 DEG C with 5 DEG C/min the most again and keeps 3min.
B, Mass Spectrometry Conditions
Ion source: 230 DEG C, level Four bar: 150 DEG C, transmission line: 250 DEG C, scan pattern MRM, ionizing energy 70eV, solvent
Postpone 4.5min.
C, table 1 are standard appearance time (retention time) and ion pair, impact energy scale
Table 1 standard appearance time and ion pair, impact energy scale
| Compound name | Retention time | Ion pair | Collision energy |
| N-dimethyl nitrous ammonium | 8.260 | 74→42.1 | 15 |
| N-dimethyl nitrous ammonium | 8.260 | 73.9→42.1 | 20 |
First identical with N-dimethyl nitrous ammonium standard substance retention time according to testing sample, quota ion to for 74 →
42.1, under the conditions of qualitative ion pair is 73.9 → 42.1, both quota ions exist to qualitative ion pair relative abundance ratio
Between 80%-120%, it is determined that for same compound.
Qualitative and the quantitative concrete detection of D, testing sample is as follows:
A, take 1.0 μ L testing sample solution sample introductions, detect with triple level Four bar gas chromatograph-mass spectrometer (GC-MS) analyses,
To the total ions chromatogram (Fig. 1) of testing sample solution, quota ion to chromatogram (Fig. 2) and qualitative ion pair relative abundance
Chromatogram (Fig. 3)；
B, the N-N-nitrosodimethylamine standard solution of variable concentrations step (1) prepared, by triple level Four bar gas phases
Chromatograph-mas spectrometer analysis detects, and testing conditions is identical with testing sample solution, it is thus achieved that the total ion current color of standard solution
Spectrogram (Fig. 4), quota ion are to chromatogram (Fig. 5) and qualitative ion pair relative abundance chromatogram (Fig. 6).
C, with the quota ion of N-N-nitrosodimethylamine to 74 → 42.1 and the chromatographic peak of qualitative ion pair 73.9 → 42.1
Making obtains N-N-nitrosodimethylamine standard curve (Fig. 7)；And draw Y=6375.486881*X-967.980516, linearly
Coefficient R2=0.9954, detection is limited to 0.5ng/mL, is quantitatively limited to 1.5ng/mL, wherein: X represents that sample concentration, Y represent
Response value；
D, according to the quota ion of N-N-nitrosodimethylamine in testing sample solution to 74 → 42.1 and qualitative ion pair
The chromatographic peak of 73.9 → 42.1, combined standard curve, it is calculated and treats the concentration C of N-N-nitrosodimethylamine in sample measuring liquid, and press
Below equation is calculated the content X of N-N-nitrosodimethylamine in cooked meat product, and cubage formula is:
X=C*V/ (M*1000)；The content of N-N-nitrosodimethylamine during wherein X is testing sample, unit is mg/kg；C is
The concentration of N-N-nitrosodimethylamine in testing sample, unit is ng/mL；V is testing sample volume of dissolution, and unit is mL；M is to treat
Test sample quality, unit is g.
Number and national standard according to content determine whether detected cooked meat product belongs to certified products.
The foregoing is only embodiments of the invention, not thereby limit the scope of the claims of the present invention, every utilize this
Equivalent structure or equivalence flow process that bright description is made convert, or are directly or indirectly used in other relevant technology necks
Territory, is the most in like manner included in the scope of patent protection of the present invention.
Claims (5)
1. N-N-nitrosodimethylamine detection method in a cooked meat product, it is characterised in that described detection method includes walking as follows
Rapid:
(1) standard solution of variable concentrations is prepared: first prepare standard reserving solution, weigh N-N-nitrosodimethylamine standard substance,
Become the storing solution that concentration is 1000mg/L of N-N-nitrosodimethylamine with acetontrile, keep in Dark Place in 4 DEG C of refrigerators；Then will
Standard reserving solution acetontrile becomes concentration to be respectively the standard of 0.5ng/mL, 1ng/mL, 2.5ng/mL, 4ng/mL, 5ng/mL
Product solution for standby；
(2) preparation of testing sample solution:
A, weighing testing sample 5.00g, add the solution 10mL containing 1% acetic acid in acetonitrile, the whirlpool under 2500r/min rotating speed is mixed
In agitator, 5min is mixed in whirlpool；
B, add 5g anhydrous magnesium sulfate, 1g sodium chloride and 1g disodium citrate after whirlpool mix 3min, then at 5000r/min rotating speed
Under centrifuge in collect supernatant after centrifugal 5min；
C, absorption 6mL supernatant move into equipped with 0.5gPSA, 0.5gC18With in the 15mL centrifuge tube of 1g anhydrous magnesium sulfate, whirlpool is mixed
3min, collects supernatant in the centrifuge under 5000r/min rotating speed after being centrifuged 5min again；
D, again take supernatant, with after the 0.22 organic membrane filtration of μm as testing sample solution；
(3) testing sample is qualitative and detection by quantitative:
A, take 1.0 μ L testing sample solution sample introductions, detect with triple level Four bar gas chromatograph-mass spectrometer (GC-MS) analyses, treated
Survey the total ions chromatogram of sample solution, quota ion to chromatogram and qualitative ion pair relative abundance chromatogram；
B, the N-N-nitrosodimethylamine standard solution of the variable concentrations that step (1) is prepared, with triple level Four bar gas chromatograies-
GC-MS analysis detects, and testing conditions is identical with testing sample solution, it is thus achieved that the total ions chromatogram of standard solution,
Quota ion chromatogram and qualitative ion pair relative abundance chromatogram；
C, with the quota ion of N-N-nitrosodimethylamine the chromatographic peak with qualitative ion pair made and obtain N-N-nitrosodimethylamine
Standard curve；
D, according to the quota ion of N-N-nitrosodimethylamine in testing sample solution to the chromatographic peak with qualitative ion pair, in conjunction with mark
Directrix curve, is calculated and treats the concentration C of N-N-nitrosodimethylamine in sample measuring liquid, and be calculated as follows in cooked meat product
The content X of N-N-nitrosodimethylamine, cubage formula is:
X=C*V/ (M*1000)
The content of N-N-nitrosodimethylamine during wherein X is testing sample, unit is mg/kg；C is that in testing sample, N-dimethyl is sub-
The concentration of nitramine, unit is ng/mL；V is testing sample volume of dissolution, and unit is mL；M is testing sample quality, and unit is g.
N-N-nitrosodimethylamine detection method in cooked meat product the most according to claim 1, it is characterised in that described quantitatively
Ion pair is 74 → 42.1, and qualitative ion pair is 73.9 → 42.1.
N-N-nitrosodimethylamine detection method in cooked meat product the most according to claim 1, it is characterised in that triple level Four
The parameter used during bar gas chromatograph-mass spectrometer (GC-MS) analysis detection is respectively as follows:
A, chromatographic condition
Chromatographic column is Agilent HP-INOWAX capillary chromatographic column, injector temperature 250 DEG C, Splitless injecting samples, flow rate of carrier gas
0.887mL/min, sample size 1 μ L；Temperature programming: initial temperature 60 DEG C keeps 1min, is raised to 120 DEG C with 40 DEG C/min, then
It is raised to 250 DEG C with 5 DEG C/min again and keeps 3min.
B, Mass Spectrometry Conditions
Ion source: 230 DEG C, level Four bar: 150 DEG C, transmission line: 250 DEG C, scan pattern MRM, ionizing energy 70eV, solvent delay
4.5min。
N-N-nitrosodimethylamine detection method in cooked meat product the most according to claim 3, it is characterised in that described carrier gas
Middle helium >=99.999%.
N-N-nitrosodimethylamine detection method in cooked meat product the most according to claim 1, it is characterised in that testing sample
Decision method for N-N-nitrosodimethylamine is: testing sample is identical, in phase with N-N-nitrosodimethylamine standard substance retention time
With quota ion to for 74 → 42.1, under the conditions of qualitative ion pair is 73.9 → 42.1, both quota ions are to qualitative
Ion pair relative abundance ratio is between 80%-120%, it is determined that be N-N-nitrosodimethylamine for testing sample.
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| CN107045020A (en) * | 2016-12-26 | 2017-08-15 | 广西壮族自治区食品药品检验所 | A kind of method of N content of nitrosamines in complete detection meat products |
| CN109856260A (en) * | 2018-12-29 | 2019-06-07 | 广电计量检测(南宁)有限公司 | The detection method of N- N-nitrosodimethylamine in a kind of meat product |
| CN111537634A (en) * | 2020-04-30 | 2020-08-14 | 湖南师范大学 | Method for detecting NDMA content in tini-class medicines |
| KR20220038965A (en) * | 2020-09-21 | 2022-03-29 | 중앙대학교 산학협력단 | Analytical method of nitrosodimethylamine in metformin drugs |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107045020A (en) * | 2016-12-26 | 2017-08-15 | 广西壮族自治区食品药品检验所 | A kind of method of N content of nitrosamines in complete detection meat products |
| CN107045020B (en) * | 2016-12-26 | 2019-11-15 | 广西壮族自治区食品药品检验所 | A kind of method of N- content of nitrosamines in complete detection meat products |
| CN109856260A (en) * | 2018-12-29 | 2019-06-07 | 广电计量检测(南宁)有限公司 | The detection method of N- N-nitrosodimethylamine in a kind of meat product |
| CN111537634A (en) * | 2020-04-30 | 2020-08-14 | 湖南师范大学 | Method for detecting NDMA content in tini-class medicines |
| CN111537634B (en) * | 2020-04-30 | 2022-05-27 | 湖南师范大学 | Method for detecting NDMA content in tini-class medicines |
| KR20220038965A (en) * | 2020-09-21 | 2022-03-29 | 중앙대학교 산학협력단 | Analytical method of nitrosodimethylamine in metformin drugs |
| KR102507398B1 (en) * | 2020-09-21 | 2023-03-07 | 중앙대학교 산학협력단 | Analytical method of nitrosodimethylamine in metformin drugs |
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| CN106053634B (en) | 2018-11-20 |
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