CN108931600A - The detection method of content of bisphenol A in a kind of food - Google Patents
The detection method of content of bisphenol A in a kind of food Download PDFInfo
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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
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Abstract
The invention discloses a kind of detection methods of content of bisphenol A in food, its key points of the technical solution are that including the following steps: step 1: weighing 5~10g food to be measured into container, 20~40mL extractant is added；Step 2: container being placed under refiner, is homogenized and is extracted using 3000~9000rpm revolving speed, extraction time is 5~15 minutes；Step 3: above-mentioned solution is transferred to centrifugal treating in centrifuge tube, 5000~20000rpm of condition, 5~20min of centrifugation time；Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition methanol, acetonitrile, acetone mixed solvent redissolution；Step 5: using Solid Phase Extraction column purification sample, collect efflux, nitrogen, which is blown to, closely to be done, and solvent is added and redissolves；Step 6: being tested using high performance liquid chromatograph, wherein liquid-phase chromatographic column is C8 or C18 reversed-phase liquid chromatography column, and column oven temperature is 20~40 DEG C.The detection method of bisphenol-A in a kind of food of the invention, can it is simple and efficient, accurately to the carry out quantitative detection of bisphenol-A in food.
Description
[technical field]
The invention belongs to technical field of food detection, and in particular to the detection method of content of bisphenol A in a kind of food.
[background technique]
Bisphenol-A is industrially used to the materials such as polycarbonate synthesis (PC) and epoxy resin, is in the world using most wide
One of general industrial compound is widely used in packaging material for food, medical instrument, feeding bottle, water bottle, spectacle lens and other is hundreds of
The manufacturing process of kind daily necessities, after heated, BPA is likely to move in food and beverage from container or packaging material, perhaps
It is more to influence whether the function of nervous function development and immune system research shows that contact BPA, especially to infant and
Speech.BPA can also simulate female sex hormone, upset endocrine system, may cause fertility Issue, sex premature, influence breast development
And reproductive function, various cancers relevant to hormone are even resulted in, therefore more paid close attention to.
As the food-safe requirement of people is higher and higher, there is relevant food for tableware and the vessel for containing food
The test method of product contact, including test method the GB 31604.10-2016 and GB/ of the content of bisphenol A contacted to food
T 23296.16-2009, import and export food engaged test standard SN/T 2282-2009.And European Union, the U.S., China etc. are right
Bisphenol-A is disabled in food contact articles for children.GB 9685-2008 standard " food containers, packet that revision in 2008 updates
Package material with additive use sanitary standard " in regulation, bisphenol-A for food contact plastics, coating, adhesive when, it is specific
The amount of migration should be no more than 0.6mg/kg.
National standard is more also more mature for the bisphenol-A test method of food contact material but temporary for food residue bisphenol-A
When there is no relevant test method, relevant limit value requirement is not made yet to the content of bisphenol A of food residue, food is connect
The test of material bisphenol-A is touched, has and tests bisphenol-A using gas chromatography-mass spectrometry using derivatization method, before such method
Reason is complicated and is related to derivative reaction, also has and is directly tested using Liquid Chromatography-Tandem Mass Spectrometry instrument, such method is related to valuable set
It is standby to popularize, it is difficult to cope with detection demand.
[summary of the invention]
The present invention provides a kind of detection method of bisphenol-A in food, can it is simple and efficient, accurately to bisphenol-A in food
Progress total content quantitative detection.
Technical solution of the present invention is as follows:
The detection method of content of bisphenol A in a kind of food, comprising the following steps:
Step 1: weighing 5~10g food to be measured into container, 20~40mL extractant is added；
Step 2: container is placed under refiner, using 3000~9000rpm revolving speed be homogenized extract, extraction time be 5~
15 minutes；
Step 3: above-mentioned solution is transferred to centrifugal treating in centrifuge tube, 5000~20000rpm of condition, centrifugation time 5~
20min；
Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition methanol, acetonitrile, acetone mixed solvent redissolution；
Step 5: using Solid Phase Extraction column purification sample, collect efflux, nitrogen, which is blown to, closely to be done, and solvent is added and redissolves；
Step 6: being tested using high performance liquid chromatograph, wherein liquid-phase chromatographic column is C8 or C18 reversed-phase liquid chromatography column, column
Temperature of Warm Case is 20~40 DEG C.
Preferably, extractant is methanol, n-hexane, acetonitrile, isopropanol, methylene chloride, one in acetone in the step 1
Kind or a variety of mixing.
Preferably, solid-phase extraction column is one of HLB, silica gel, C18 in the step 5.
Preferably, when liquid chromatograph is tested in the step 6, mobile phase is methanol and aqueous solution mixed solution, ratio
It is 50~90%, flow velocity is 0.8~1.5mL/min, and sampling volume is 10~40uL.
Preferably, column oven temperature setting described in step 6 is 10 DEG C higher than room temperature, and the fluorescence detector temperature compares column temperature
Box temperature degree is 10 DEG C high.
Preferably, fluorescence detector parameter setting described in step 6,235~275nm of excitation wavelength, launch wavelength 297~
313nm, detector sensitivity are 1~8.
The present invention compared with the existing technology, has the advantage that
The detection method of bisphenol-A in a kind of food of the invention is extracted in conjunction with homogenate, using extractant, by centrifugation, only
Change, after concentration, is dissolved in bisphenol-A in suitable solvent, analysis calibration is carried out into liquid chromatograph, since bisphenol-A belongs to
Highly polar substance, based on the similar principle that mixes, bisphenol-A is suitable for being extracted using highly polar solvent, and the application selects methanol
Equal organic solvents are based on this principle as extractant, and this test method advantage is:
Firstly, the equipment of homogenate extraction is relatively cheap, more other extracting process, as ultrasonic extraction needs to buy valuableness
Ultrasonic extraction instrument, cost performance is higher；
Secondly, the time that homogenate is extracted is shorter, detection speed is fast, is suitble to quickly test, improves detection efficiency；
Again, using liquid chromatogram fluorescence detector, bisphenol-A in the food of complicated component is detected, sample is used
Amount is few, and detection sensitivity is high, and equipment cost is low, and pretreatment process is simple and fast, as a result accurately, bisphenol-A in obtained food
Total content data detection limit is lower, can accurate quantitative analysis bisphenol-A lower loading food.
[Detailed description of the invention]
Fig. 1 is the A factor trend figure that field cause for gossip is tested in the embodiment of the present invention 1；
Fig. 2 is the Factor B tendency chart that field cause for gossip is tested in the embodiment of the present invention 1；
Fig. 3 is the C factor tendency chart that field cause for gossip is tested in the embodiment of the present invention 1；
Fig. 4 is the D factor trend figure that field cause for gossip is tested in the embodiment of the present invention 1；
Fig. 5 is the A factor trend figure that field cause for gossip is tested in the embodiment of the present invention 2；
Fig. 6 is the Factor B tendency chart that field cause for gossip is tested in the embodiment of the present invention 2；
Fig. 7 is the C factor tendency chart that field cause for gossip is tested in the embodiment of the present invention 2；
Fig. 8 is the D factor trend figure that field cause for gossip is tested in the embodiment of the present invention 2；
Fig. 9 is the A factor trend figure that field cause for gossip is tested in the embodiment of the present invention 3；
Figure 10 is the Factor B tendency chart that field cause for gossip is tested in the embodiment of the present invention 3；
Figure 11 is the C factor tendency chart that field cause for gossip is tested in the embodiment of the present invention 3；
Figure 12 is the D factor trend figure that field cause for gossip is tested in the embodiment of the present invention 3.
[specific embodiment]
1~3 illustrates the specific technical solution of the present invention in conjunction with specific embodiments:
Embodiment 1:
The detection method of bisphenol-A in a kind of food:
Step 1: weighing 5g food to be measured and be added to the container, after 20mL methylene chloride is added as extractant；
Step 2: sample being placed under refiner, refiner, rotational speed regulation to 3000rpm are opened, homogenate is extracted 5 minutes；
Step 3: above-mentioned solution is transferred to centrifuge tube, using high speed freezing centrifuge, centrifugal rotational speed 5000rpm, from
Heart disengaging time is 20 minutes；
Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition acetonitrile redissolution；
Step 5: activating HLB solid-phase extraction column using water/methanol, sample to be clean is added into solid-phase extraction column, bear
Column is pressed through, efflux is collected, elutes solid-phase extraction column using ethanol/methylene, collects eluent, nitrogen, which is blown to, closely to be done, and is added
Acetonitrile redissolves；
Step 6: carrying out quantitative analysis, liquid chromatograph test condition are as follows: mobile phase using liquid chromatograph testing result
For methanol and aqueous solution mixed solution, ratio 50%, flow velocity 1.5mL/min；Sampling volume is 40uL；Liquid-phase chromatographic column is
C8 reversed-phase liquid chromatography column；Column oven temperature setting is 30 DEG C (20 DEG C of room temperature)；Fluorescence detector temperature is 40 DEG C；Fluorescence detection
Device parameter is set as excitation wavelength 275nm, launch wavelength 313nm, detector sensitivity 8.
The present inventor has found in the course of the research, and when using the test method, influencing maximum parameter to quantitative result is A
(example weight), B (extraction solution volume), C (Homogenization time) and D (homogenate speed) are tested using field cause for gossip to above-mentioned each parameter
It is verified, four controlling elements, three levels of each correspondence, each controlling elements use three levels, used three levels
It is that data of the invention can be achieved, as shown in table 1:
Table 1: controlling elements and its corresponding three levels
Food for detection is Chinese cabbage (Brassica rapa L), and straight friendship table L9 (34) the experiment side of three levels is used to this food
Case is tested, totally 9 groups of tests, this 9 groups of experiments are all made of high performance liquid chromatography detection, the sample preparation in each experiment
Parameter etc., other than changing four conditions in table 1, other conditions are consistent.
Table 2: each sample carries out 2 Duplicate Samples tests simultaneously, obtains following tests result
In above-mentioned table 2, rate of recovery calculation formula:
Table 3: the straight rate of recovery average value for handing over table L9 (34) experimental program and test result in field mouthful
Above-mentioned test result and calculated result are obtained into trend Fig. 1 to Fig. 4 using the Tian Kou mapping software of mintab.
Embodiment 2:
The detection method of bisphenol-A in a kind of food:
Step 1: weighing 8g food to be measured and be added in centrifuge tube, after 35mL acetonitrile is added as extractant；
Step 2: sample being placed under refiner, refiner, rotational speed regulation to 7500rpm are opened, homogenate is extracted 10 minutes；
Step 3: above-mentioned solution is transferred to centrifuge tube, using high speed freezing centrifuge, centrifugal rotational speed 20000rpm, from
Heart disengaging time is 10 minutes；
Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition methanol redissolution；
Step 5: using water/methanol activated silica gel solid-phase extraction column, sample to be clean is added into solid-phase extraction column, bear
Column is pressed through, efflux is collected, elutes solid-phase extraction column using methanol, collects eluent, nitrogen, which is blown to, closely to be done, and methanol is added and redissolves；
Step 6: carrying out quantitative analysis, liquid chromatograph test condition are as follows: mobile phase using liquid chromatograph testing result
For methanol and aqueous solution mixed solution, ratio 72%, flow velocity 1.2mL/min；Sampling volume is 40uL；Liquid-phase chromatographic column is
C8 reversed-phase liquid chromatography column；Column oven temperature setting is 32 DEG C (20 DEG C of room temperature)；Fluorescence detector temperature is 42 DEG C；Fluorescence detection
Device parameter is set as excitation wavelength 255nm, launch wavelength 305nm, detector sensitivity 7.
The present inventor has found in the course of the research, and when using the test method, influencing maximum parameter to quantitative result is A
(example weight), B (extraction solution volume), C (Homogenization time) and D (homogenate speed) are tested using field cause for gossip to above-mentioned each parameter
It is verified, four controlling elements, three levels of each correspondence, each controlling elements use three levels, used three levels
It is that data of the invention can be achieved, as shown in table 4:
Table 4: controlling elements and its corresponding three levels
| Controlling elements | Explanation | Lever 1 | Lever 2 | Lever 3 |
| A | Example weight (g) | 4 | 8 | 12 |
| B | Extraction solution volume (mL) | 25 | 35 | 45 |
| C | Homogenization time (min) | 5 | 10 | 15 |
| D | It is homogenized speed (rpm) | 5000 | 7500 | 10000 |
Food for detection is steamed bun, is tried using straight friendship table L9 (34) experimental program of three levels this food
It tests, totally 9 groups of tests, this 9 groups of experiments are all made of high performance liquid chromatography detection, the parameter etc. of the sample preparation in each experiment,
Other than changing four conditions in table 4, other conditions are consistent.
Table 5: each sample carries out 2 Duplicate Samples tests simultaneously, obtains following tests result
In above-mentioned table 5, rate of recovery calculation formula:
Table 6: the straight rate of recovery average value for handing over table L9 (34) experimental program and test result in field mouthful
Above-mentioned test result and calculated result are obtained into tendency chart 5 to Fig. 8 using the Tian Kou mapping software of mintab.
Embodiment 3:
The detection method of bisphenol-A in a kind of food:
Step 1: weighing 6g food to be measured and be added in centrifuge tube, after 40mL acetone is added as extractant；
Step 2: sample being placed under refiner, refiner, rotational speed regulation to 10000rpm are opened, homogenate extracts 20 points
Clock；
Step 3: above-mentioned solution is transferred to centrifuge tube, using high speed freezing centrifuge, centrifugal rotational speed 10000rpm, from
Heart disengaging time is 10 minutes；
Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition methanol redissolution；
Step 5: activating amino solid-phase extraction column using water, sample to be clean is added into solid-phase extraction column, negative pressure mistake
Column collects efflux, elutes solid-phase extraction column using ammonia/methanol, collects eluent, and nitrogen, which is blown to, closely to be done, and acetonitrile is added and redissolves；
Step 6: carrying out quantitative analysis, liquid chromatograph test condition are as follows: mobile phase using liquid chromatograph testing result
For methanol and aqueous solution mixed solution, ratio 90%, flow velocity 0.8mL/min；Sampling volume is 40uL；Liquid-phase chromatographic column is
Reversed-phase liquid chromatography column；Column oven temperature setting is 38 DEG C (20 DEG C of room temperature)；Fluorescence detector temperature is 48 DEG C；Fluorescence detector
Parameter is set as excitation wavelength 275nm, launch wavelength 313nm, detector sensitivity 6.
The present inventor has found in the course of the research, and when using this method, influencing maximum parameter to quantitative result is A (sample
Product weight), B (extraction solution volume), C (Homogenization time) and D (homogenate speed) tested using field cause for gossip and carried out to above-mentioned each parameter
Verifying, four controlling elements, three levels of each correspondence, each controlling elements use three levels, and used three levels are
Data of the invention can be achieved, as shown in table 7:
Table 7: controlling elements and its corresponding three levels
| Controlling elements | Explanation | Lever 1 | Lever 2 | Lever 3 |
| A | Example weight (g) | 6 | 12 | 24 |
| B | Extraction solution volume (mL) | 30 | 45 | 55 |
| C | Homogenization time (min) | 10 | 15 | 20 |
| D | It is homogenized speed (rpm) | 7500 | 9000 | 10500 |
Food for detection is milk powder, is tried using straight friendship table L9 (34) experimental program of three levels this food
It tests, totally 9 groups of tests, this 9 groups of experiments are all made of high performance liquid chromatography detection, the parameter etc. of the sample preparation in each experiment,
Other than changing four conditions in table 7, other conditions are consistent.
Table 8: each sample carries out 5 Duplicate Samples tests simultaneously, obtains following tests result
In above-mentioned table 8, rate of recovery calculation formula:
Table 9: the straight rate of recovery average value for handing over table L9 (34) experimental program and test result in field mouthful
Above-mentioned test result and calculated result are obtained into tendency chart 9 to Figure 12 using the Tian Kou mapping software of mintab.
Embodiment 1-3 uses high performance liquid chromatography, and combines four Optimal Parameters, contains to the bisphenol-A in numerous food
Amount is detected, including Chinese cabbage (Brassica rapa L), steamed bun and milk powder, and the test knot of food samples made of bisphenol-A standard substance is added
Fruit analysis, is tested, the results are shown in Table 10 by liquid chromatography.
Table 10: liquid-phase chromatographic analysis result
| Embodiment | 1 | 2 | 3 |
| Sample ID | Chinese cabbage (Brassica rapa L) | Steamed bun | Milk powder |
| The rate of recovery | 86.73% | 86.23% | 87.58% |
As can be seen from Table 10, this method obtain as a result, the rate of recovery in 80% to 120% range, thus illustrates this
The detection method of invention can actually accomplish the total content for accurately detecting bisphenol-A in food, and detection method of the invention, behaviour
Make simple and fast and low in cost.
And in order to improve the accuracy of detection method, Taguchi's method is selected from numerous experimental data optimization method and is come to shadow
The important parameter for ringing testing result optimizes, and Taguchi's method is a kind of inexpensive, high benefit quality engineering method, it is emphasized
The raising of product quality is not instead of by examining, and by design and optimal conditions, from field, oral examination is tested, can be obtained in statistical method
Optimal example weight, the volume of extractant, extraction time, Extracting temperature are obtained, bisphenol-A test experience end value in food is made
It is more accurate.
Claims (6)
1. the detection method of content of bisphenol A in a kind of food, it is characterised in that the following steps are included:
Step 1: weighing 5~10g food to be measured into container, 20~40mL extractant is added；
Step 2: container being placed under refiner, is homogenized and is extracted using 3000~9000rpm revolving speed, extraction time is 5~15 points
Clock；
Step 3: above-mentioned solution is transferred to centrifugal treating in centrifuge tube, 5000~20000rpm of condition, centrifugation time 5~
20min；
Step 4: taking out all supernatant liquors, nitrogen is blown to close dry, addition methanol, acetonitrile, acetone mixed solvent redissolution；
Step 5: using Solid Phase Extraction column purification sample, collect efflux, nitrogen, which is blown to, closely to be done, and solvent is added and redissolves；
Step 6: being tested using high performance liquid chromatograph, liquid-phase chromatographic column is C8 or C18 reversed-phase liquid chromatography column, column oven temperature
It is 20~40 DEG C.
2. the detection method of content of bisphenol A in a kind of food according to claim 1, it is characterised in that: in the step 1
Extractant is one of methanol, n-hexane, acetonitrile, isopropanol, methylene chloride, acetone or a variety of mixing.
3. the detection method of content of bisphenol A in a kind of food according to claim 1, it is characterised in that: in the step 5
Solid-phase extraction column is one of HLB, silica gel, C18.
4. the detection method of content of bisphenol A in a kind of food according to claim 1, it is characterised in that: in the step 6
When liquid chromatograph is tested, mobile phase is methanol and aqueous solution mixed solution, and ratio is 50~90%, flow velocity is 0.8~
1.5mL/min, sampling volume are 10~40uL.
5. the detection method of content of bisphenol A in a kind of food according to claim 1, it is characterised in that: described in step 6
Column oven temperature setting is 10 DEG C higher than room temperature, and the fluorescence detector temperature is 10 DEG C higher than column oven temperature.
6. the detection method of content of bisphenol A in a kind of food according to claim 1, it is characterised in that: described in step 6
Fluorescence detector parameter setting, 235~275nm of excitation wavelength, 297~313nm of launch wavelength, detector sensitivity are 1~8.
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Cited By (3)
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| CN111380984A (en) * | 2020-03-27 | 2020-07-07 | 深圳大学 | Method for pretreating vegetable sample and simultaneously detecting various bisphenol compounds in vegetable sample |
| CN111707737A (en) * | 2020-05-06 | 2020-09-25 | 深圳市农产品质量安全检验检测中心(深圳市动物疫病预防控制中心) | Method for improving detection precision of bisphenol A content in animal derived food and application thereof |
| CN113514312A (en) * | 2021-09-09 | 2021-10-19 | 佛山职业技术学院 | Intelligence food detection extraction workstation |
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| CN111380984A (en) * | 2020-03-27 | 2020-07-07 | 深圳大学 | Method for pretreating vegetable sample and simultaneously detecting various bisphenol compounds in vegetable sample |
| CN111707737A (en) * | 2020-05-06 | 2020-09-25 | 深圳市农产品质量安全检验检测中心(深圳市动物疫病预防控制中心) | Method for improving detection precision of bisphenol A content in animal derived food and application thereof |
| CN111707737B (en) * | 2020-05-06 | 2022-12-20 | 深圳市质量安全检验检测研究院(深圳市动物疫病预防控制中心) | Method for improving detection precision of bisphenol A content in animal derived food and application thereof |
| CN113514312A (en) * | 2021-09-09 | 2021-10-19 | 佛山职业技术学院 | Intelligence food detection extraction workstation |
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