CN106501411A - A kind of method for quick of benzopyrene from edible oil - Google Patents
A kind of method for quick of benzopyrene from edible oil Download PDFInfo
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- CN106501411A CN106501411A CN201610960598.8A CN201610960598A CN106501411A CN 106501411 A CN106501411 A CN 106501411A CN 201610960598 A CN201610960598 A CN 201610960598A CN 106501411 A CN106501411 A CN 106501411A
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- edible oil
- benzopyrene
- organic solvent
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- acetonitrile
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- 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
Abstract
The invention discloses a kind of method for quick of benzopyrene from edible oil, including：（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, organic solvent is added, is shaken after jumping a queue；Supersound extraction, collected after centrifugation supernatant；Organic solvent is added in residual edible oil sample, and supersound extraction, centrifugation merge supernatant；Taking organic solvent adds solid-phase extraction column to be activated, and the supernatant of merging is transferred in the solid-phase extraction column after activation after the completion of activation, uses organic solvent eluting, collects eluent to measuring bottle, constant volume, crosses 0.22 m microporous filter membrane to sample injection bottle, standby；（2）The preparation of standard solution；（3）Detection：Successively benzopyrene standard serial solution and need testing solution injection HPLC are detected., using supersound extraction twice and solid-phase extraction column enriching and purifying, the method is easy to operate and safe, low cost for the present invention, extracts completely, and testing result is accurately, true and reliable.
Description
Technical field
The present invention relates to technical field of food detection, specifically a kind of method for quick of benzopyrene from edible oil.
Background technology
Benzopyrene is also known as benzo（а）Pyrene, english abbreviation BaP is organic compound that a class has obvious carcinogenesis.It
It is the polycyclic arene compound being combined into by a phenyl ring and a pyrene molecule.After testing, nonexpondable megistotherm
Oily, the fried food for overdoing can all produce B (a) P, and Reusability high temperature fries mode, the organic matter decomposes in food, warp
It is cyclized, is polymerized and forms the carcinogenic components such as B (a) P.
Often there is the exceeded event of benzopyrene in vegetable oil, 40 kinds of olive oil samples on Brazilian market in 1996 are tested
Go out containing benzopyrene, highest content is up to 164 micro- g kgs.July calendar year 2001《Xinhua Daily Telegraph》Report Hispanic olive oil
Middle BaP content overproofs, are mainly considered the olive oil of secondary refinement.In October, 2006,《Consumption daily paper》With《Foochow daily paper》Report
The Oleum sesami BaP of the ground such as road Hubei, Anhui production is exceeded.Since in Septembers, 2010, the product of domestic Duo Jia Camellia oil manufacturing enterprise
It is detected high carcinogen benzopyrene exceeded, causes and extremely shake in the industry, causes consumer to lack confidence to Flos Camelliae Japonicae oil product, make mountain
Oleum Camelliae market is greatly hit, and is unfavorable for the healthy and rapid development of Camellia oil industry.According to Chinese GB2716-2005《Food
Use vegetable oil sanitary standard》Require, in edible vegetable oils type product, the threshold limit valuess of benzopyrene are less than 10 microgram/thousand
Gram.
The method of conventional detection benzopyrene has thin layer chromatography, fluorescence spectrophotometry, gas chromatography, gas phase color
Spectrum-mass spectrography, HPLC UV method, stationary phases for HPLC.Wherein fluorescence method has sensitivity high, favorable reproducibility
The advantages of, it is more common method.However, in current benzopyrene detection method, the pre-treating method of sample is complicated, and step is numerous
Trivial, it is easy to which that, as careless manipulation is sucked or skin touches benzopyrene in front process sample, this is to experimenter's
Safety brings greatly threat.
Content of the invention
It is an object of the invention to provide a kind of easy to operate and benzo in the accurate edible oil of safety, low cost, result
The method for quick of pyrene.
For achieving the above object, the present invention provides following technical scheme：
A kind of method for quick of benzopyrene from edible oil, comprises the following steps：
（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, is added equivalent to edible oil
The organic solvent of 16～18 times of amounts of sample, shakes after jumping a queue；15～22min of supersound extraction, collected after centrifugation supernatant；Residual food
With the organic solvents added in oil samples equivalent to residual 8～12 times of edible oil sample amount, 12～18min of supersound extraction, from
The heart, merges supernatant；Taking organic solvent adds solid-phase extraction column to be activated, and after the completion of activation is transferred to the supernatant of merging
In solid-phase extraction column after activation, organic solvent eluting is used, eluent is collected to measuring bottle, constant volume, cross 0.22 m microporous filter membrane extremely
Sample injection bottle, standby；
（2）The preparation of standard solution：Compound concentration be 1 g/kg, 2.5 g/kg, 5 g/kg, 10 g/kg, 20 g/kg, 30
The benzopyrene standard serial solution of g/kg, constant volume solution are organic solvent, cross 0.22 m microporous filter membrane to sample injection bottle after shaking up, standby
With；
（3）Detection：Successively by benzopyrene standard serial solution and need testing solution injection HPLC, HPLC testing conditions are：ODS
Post；Column temperature：30℃；Gradient elution and the corresponding relation of mobile phase：0～3min, acetonitrile 5～10%, formic acid 10～15%, water 75～
85%；3 → 10min, acetonitrile 15～25%, formic acid 20～25%, water 50～65%；10 → 13min, acetonitrile 35～50%, formic acid 15～
20%th, water 30～50%；13 → 20min, acetonitrile 70～80%, formic acid 10～15%, water 5～20%；Fluorescence detector：Excitation wavelength
350nm, launch wavelength 420nm.
As further scheme of the invention：Described organic solvent be chloroform, acetone, benzene,toluene,xylene, ether,
Any one in petroleum ether and hexamethylene.
As further scheme of the invention：Described step（1）In, ultrasonic extraction process parameter is：Ultrasonic frequency
It is 400～450W for 35～40kHz, ultrasonic power；The bottleneck of conical flask with cover need to be sealed with sealed membrane during supersound extraction.
As further scheme of the invention：Described step（1）In, the ultrasonic extraction process parameter of edible oil sample
For：Ultrasonic frequency is 40kHz, ultrasonic power is 400W, and the time is 20min；The supersound extraction work of residual edible oil sample
Skill parameter is：Ultrasonic frequency is 35kHz, ultrasonic power is 450W, and the time is 15min.
As further scheme of the invention：Described step（1）In, centrifugation rate is 12000～18000r/min, when
Between be 5～10min.
As further scheme of the invention：Described step（1）In, centrifugation rate is 15000r/min, and the time is
8min.
As further scheme of the invention：Described step（1）In, the activity of solid-phase extraction column completes standard and is：Take
Organic solvent adds solid-phase extraction column to be activated, and drips 18～22ml to organic solvent is naturally lower, and activation is completed.
As further scheme of the invention：Described step（1）In, the activity of solid-phase extraction column completes standard and is：Take
Organic solvent adds solid-phase extraction column to be activated, and to the naturally lower drop 20ml of organic solvent, activation is completed.
As further scheme of the invention：Described step（3）In, the corresponding relation of gradient elution and mobile phase：0～
3min, acetonitrile 8%, formic acid 12%, water 80%；3 → 10min, acetonitrile 20%, formic acid 22%, water 58%；10 → 13min, acetonitrile 42%, first
Acid 18%, water 40%；13 → 20min, acetonitrile 75%, formic acid 12%, water 13%.
Compared with prior art, the invention has the beneficial effects as follows：
The present invention is initially with supersound extraction twice, then with the benzopyrene in solid-phase extraction column enriching and purifying edible oil sample, is somebody's turn to do
Method is easy to operate, and step is simple, and reducing artificial maloperation causes suction or skin to touch the probability of benzopyrene, greatly improves
The safety of experimenter's work, and the benzopyrene in the method extracts completely, and obtained certain purification effect, make into
Free from admixture peak interference when entering high performance liquid chromatography detection, testing result are accurately, true and reliable.Side of the present invention using gradient elution
Formula eluting benzopyrene, the method elution time are short, and can guarantee that benzopyrene eluting completely, save time and mobile phase, make detection into
This is low.
Specific embodiment
Below in conjunction with the embodiment of the present invention, to the embodiment of the present invention in technical scheme be clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the invention, rather than whole embodiment.In based on the present invention
Embodiment, the every other embodiment obtained under the premise of creative work is not made by those of ordinary skill in the art, all
Belong to the scope of protection of the invention.
Embodiment 1
In the embodiment of the present invention, a kind of method for quick of benzopyrene from edible oil is comprised the following steps：
（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, is added equivalent to edible oil
The petroleum ether of 16 times of amounts of sample, shakes after jumping a queue；After sealed by the bottleneck of conical flask with cover, supersound extraction, ultrasound
Wave frequency is 35kHz, ultrasonic power is 400W, and the time is 22min, 12000r/min centrifugation 10min, collects supernatant；Residual
The petroleum ether added in edible oil sample equivalent to residual 12 times of edible oil sample amount are stayed, with sealed membrane by conical flask with cover
After bottleneck is sealed, supersound extraction, ultrasonic frequency is 35kHz, ultrasonic power is 400W, and the time is 18min, 12000r/min
Centrifugation 10min, merges supernatant；Taking petroleum ether adds solid-phase extraction column to be activated, to the naturally lower drop 18ml of petroleum ether, activation
After the completion of the supernatant of merging is transferred in the solid-phase extraction column after activation, use petroleum ether eluting, collect eluent to measuring bottle,
Constant volume, crosses 0.22 m microporous filter membrane to sample injection bottle, standby；
（2）The preparation of standard solution：Compound concentration be 1 g/kg, 2.5 g/kg, 5 g/kg, 10 g/kg, 20 g/kg, 30
The benzopyrene standard serial solution of g/kg, constant volume solution are petroleum ether, cross 0.22 m microporous filter membrane to sample injection bottle after shaking up, standby
With；
（3）Detection：Successively by benzopyrene standard serial solution and need testing solution injection HPLC, HPLC testing conditions are：ODS
Post；Column temperature：30℃；Gradient elution and the corresponding relation of mobile phase：0～3min, acetonitrile 5%, formic acid 10%, water 85%；3→
10min, acetonitrile 15%, formic acid 20%, water 65%；10 → 13min, acetonitrile 35%, formic acid 15%, water 50%；13 → 20min, acetonitrile
70%th, formic acid 10%, water 20%；Fluorescence detector：Excitation wavelength 350nm, launch wavelength 420nm.
Embodiment 2
In the embodiment of the present invention, a kind of method for quick of benzopyrene from edible oil is comprised the following steps：
（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, is added equivalent to edible oil
The dimethylbenzene of 18 times of amounts of sample, shakes after jumping a queue；After sealed by the bottleneck of conical flask with cover, supersound extraction, ultrasound
Wave frequency is 40kHz, ultrasonic power is 450W, and the time is 15min, 18000r/min centrifugation 5min, collects supernatant；Residual
The dimethylbenzene equivalent to residual 8 times of edible oil sample amount are added in edible oil sample, with sealed membrane by the bottle of conical flask with cover
After mouth is sealed, supersound extraction, it is 450W that ultrasonic frequency is 40kHz, ultrasonic power, and the time is 12min, 18000r/min from
Heart 5min, merges supernatant；Taking dimethylbenzene adds solid-phase extraction column to be activated, and to the naturally lower drop 22ml of dimethylbenzene, has activated
The supernatant of merging is transferred in the solid-phase extraction column after activation into after, uses dimethylbenzene eluting, eluent is collected to measuring bottle, fixed
Hold, 0.22 m microporous filter membrane is crossed to sample injection bottle, standby；
（2）The preparation of standard solution：Compound concentration be 1 g/kg, 2.5 g/kg, 5 g/kg, 10 g/kg, 20 g/kg, 30
The benzopyrene standard serial solution of g/kg, constant volume solution are dimethylbenzene, cross 0.22 m microporous filter membrane to sample injection bottle after shaking up, standby
With；
（3）Detection：Successively by benzopyrene standard serial solution and need testing solution injection HPLC, HPLC testing conditions are：ODS
Post；Column temperature：30℃；Gradient elution and the corresponding relation of mobile phase：0～3min, acetonitrile 10%, formic acid 15%, water 75%；3→
10min, acetonitrile 25%, formic acid 25%, water 50%；10 → 13min, acetonitrile 50%, formic acid 20%, water 30%；13 → 20min, acetonitrile
80%th, formic acid 15%, water 5%；Fluorescence detector：Excitation wavelength 350nm, launch wavelength 420nm.
Embodiment 3
In the embodiment of the present invention, a kind of method for quick of benzopyrene from edible oil is comprised the following steps：
（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, is added equivalent to edible oil
The chloroform of 17 times of amounts of sample, shakes after jumping a queue；After sealed by the bottleneck of conical flask with cover, supersound extraction, ultrasound wave
Frequency is 40kHz, ultrasonic power is 400W, and the time is 20min, 15000r/min centrifugation 8min, collects supernatant；Residual food
With the chloroform added in oil samples equivalent to residual 10 times of amounts of edible oil sample, the bottleneck of conical flask with cover is sealed with sealed membrane
After firmly, supersound extraction, it is 450W that ultrasonic frequency is 35kHz, ultrasonic power, and the time is 15min, and 15000r/min is centrifuged
8min, merges supernatant；Taking chloroform adds solid-phase extraction column to be activated, to the naturally lower drop 20ml of chloroform, will after the completion of activation
The supernatant of merging is transferred in the solid-phase extraction column after activation, uses chloroform eluting, collects eluent to measuring bottle, constant volume, mistake
0.22 m microporous filter membrane is to sample injection bottle, standby；
（2）The preparation of standard solution：Compound concentration be 1 g/kg, 2.5 g/kg, 5 g/kg, 10 g/kg, 20 g/kg, 30
The benzopyrene standard serial solution of g/kg, constant volume solution are chloroform, cross 0.22 m microporous filter membrane to sample injection bottle after shaking up, standby；
（3）Detection：Successively by benzopyrene standard serial solution and need testing solution injection HPLC, HPLC testing conditions are：ODS
Post；Column temperature：30℃；Gradient elution and the corresponding relation of mobile phase：0～3min, acetonitrile 8%, formic acid 12%, water 80%；3→
10min, acetonitrile 20%, formic acid 22%, water 58%；10 → 13min, acetonitrile 42%, formic acid 18%, water 40%；13 → 20min, acetonitrile
75%th, formic acid 12%, water 13%；Fluorescence detector：Excitation wavelength 350nm, launch wavelength 420nm.
The present invention initially with supersound extraction twice, then with the benzo in solid-phase extraction column enriching and purifying edible oil sample
Pyrene, the method are easy to operate, and step is simple, and reducing artificial maloperation causes suction or skin to touch the probability of benzopyrene, greatly
The safety of experimenter's work is improve greatly, and the benzopyrene in the method is extracted completely, and obtained certain purification effect
Really, when making entrance high performance liquid chromatography detection, free from admixture peak is disturbed, and testing result is accurately, true and reliable.The present invention uses gradient
The mode eluting benzopyrene of eluting, the method elution time are short, and can guarantee that benzopyrene eluting completely, saves time and mobile phase,
Make testing cost low.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of spirit or essential attributes without departing substantially from the present invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit is required rather than described above is limited, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.
Moreover, it will be appreciated that although this specification is been described by according to embodiment, not each embodiment is only wrapped
Contain an independent technical scheme, this narrating mode of description is only that those skilled in the art should for clarity
Using description as an entirety, the technical scheme in each embodiment can also form those skilled in the art through appropriately combined
Understandable other embodiment.
Claims (9)
1. a kind of method for quick of benzopyrene from edible oil, it is characterised in that comprise the following steps：
（1）The preparation of need testing solution：Edible oil sample is accurately weighed in right amount in conical flask with cover, is added equivalent to edible oil
The organic solvent of 16～18 times of amounts of sample, shakes after jumping a queue；15～22min of supersound extraction, collected after centrifugation supernatant；Residual food
With the organic solvents added in oil samples equivalent to residual 8～12 times of edible oil sample amount, 12～18min of supersound extraction, from
The heart, merges supernatant；Taking organic solvent adds solid-phase extraction column to be activated, and after the completion of activation is transferred to the supernatant of merging
In solid-phase extraction column after activation, organic solvent eluting is used, eluent is collected to measuring bottle, constant volume, cross 0.22 m microporous filter membrane extremely
Sample injection bottle, standby；
（2）The preparation of standard solution：Compound concentration be 1 g/kg, 2.5 g/kg, 5 g/kg, 10 g/kg, 20 g/kg, 30
The benzopyrene standard serial solution of g/kg, constant volume solution are organic solvent, cross 0.22 m microporous filter membrane to sample injection bottle after shaking up, standby
With；
（3）Detection：Successively by benzopyrene standard serial solution and need testing solution injection HPLC, HPLC testing conditions are：ODS
Post；Column temperature：30℃；Gradient elution and the corresponding relation of mobile phase：0～3min, acetonitrile 5～10%, formic acid 10～15%, water 75～
85%；3 → 10min, acetonitrile 15～25%, formic acid 20～25%, water 50～65%；10 → 13min, acetonitrile 35～50%, formic acid 15～
20%th, water 30～50%；13 → 20min, acetonitrile 70～80%, formic acid 10～15%, water 5～20%；Fluorescence detector：Excitation wavelength
350nm, launch wavelength 420nm.
2. the method for quick of benzopyrene from edible oil according to claim 1, it is characterised in that described is organic molten
Agent is any one in chloroform, acetone, benzene,toluene,xylene, ether, petroleum ether and hexamethylene.
3. the method for quick of benzopyrene from edible oil according to claim 1, it is characterised in that described step
（1）In, ultrasonic extraction process parameter is：Ultrasonic frequency is 35～40kHz, ultrasonic power is 400～450W；Supersound extraction
When the bottleneck of conical flask with cover need to be sealed with sealed membrane.
4. the method for quick of benzopyrene from edible oil according to claim 3, it is characterised in that described step
（1）In, the ultrasonic extraction process parameter of edible oil sample is：Ultrasonic frequency be 40kHz, ultrasonic power be 400W, the time
For 20min；Residual edible oil sample ultrasonic extraction process parameter be：Ultrasonic frequency is 35kHz, ultrasonic power is
450W, time are 15min.
5. the method for quick of benzopyrene from edible oil according to claim 1, it is characterised in that described step
（1）In, centrifugation rate is 12000～18000r/min, and the time is 5～10min.
6. the method for quick of benzopyrene from edible oil according to claim 5, it is characterised in that described step
（1）In, centrifugation rate is 15000r/min, and the time is 8min.
7. the method for quick of benzopyrene from edible oil according to claim 1, it is characterised in that described step
（1）In, the activity of solid-phase extraction column completes standard and is：Taking organic solvent adds solid-phase extraction column to be activated, to organic solvent
Naturally 18～22ml of drop under, activation are completed.
8. the method for quick of benzopyrene from edible oil according to claim 7, it is characterised in that described step
（1）In, the activity of solid-phase extraction column completes standard and is：Taking organic solvent adds solid-phase extraction column to be activated, to organic solvent
Naturally lower drop 20ml, activation are completed.
9. the method for quick of benzopyrene from edible oil according to claim 1, it is characterised in that described step
（3）In, the corresponding relation of gradient elution and mobile phase：0～3min, acetonitrile 8%, formic acid 12%, water 80%；3 → 10min, acetonitrile
20%th, formic acid 22%, water 58%；10 → 13min, acetonitrile 42%, formic acid 18%, water 40%；13 → 20min, acetonitrile 75%, formic acid 12%,
Water 13%.
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| CN109430685A (en) * | 2018-12-03 | 2019-03-08 | 中南林业科技大学 | A method of BaP in removal betel nut cigarette fruit |
| CN109580318A (en) * | 2018-12-11 | 2019-04-05 | 江南大学 | A kind of method of distributing Solid Phase Extraction processing edible oil sample detection benzo [a] pyrene |
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Application publication date: 20170315 |