CN103760291A - Method for measuring benzo (a) pyrene content in peanut oil by using reversed-phase high-performance liquid chromatography - Google Patents
Method for measuring benzo (a) pyrene content in peanut oil by using reversed-phase high-performance liquid chromatography Download PDFInfo
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Abstract
The invention discloses a method for measuring a benzo (a) pyrene content in a peanut oil by using reversed-phase high-performance liquid chromatography. The method comprises the following steps: (1) preparing a sample treatment liquid; (2) carrying out reversed-phase high-performance liquid chromatography analysis on the sample treatment liquid to obtain chromatography analyze data; (3) taking a benzo (a) pyrene standard substance to prepare a standard operation liquid by using an acetonitrile water mixing solution, measuring on a machine, and drawing a standard curve to obtain a benzo (a) pyrene correction factor; (4) comparing the obtained chromatography analyze data and the benzo (a) pyrene correction factor to obtain the benzo (a) pyrene content in the peanut oil. The method provided by the invention has the advantages that the operation is simple, convenient and is practical, an operation error is effectively reduced, and the result of the measuring benzo (a) pyrene content in peanut oil is exact and reliable.
Description
Technical field
The present invention relates to the improvement of a kind of method of benzo (a) pyrene content in detection peanut oil, the method for benzo (a) pyrene content in especially a kind of reversed-phase high-performance liquid chromatography external standard method peanut oil.
Background technology
Peanut oil is flavor type edible oil, the strong fragrance that fries peanut because having is subject to liking of part consumer, owing to need heating a period of time at higher temperature in the process at frying peanut and extraction peanut oil, so can cause may containing a small amount of benzo (a) pyrene in finished product peanut oil, benzo (a) pyrene in the peanut oil that different technology conditions is produced is had nothing in common with each other.Benzo (a) pyrene is a kind of generally acknowledged carcinogen, the edible vegetable oil hygienic standard of China requires benzo (a) pyrene content must not surpass 10 μ g/kg, therefore, peanut oil manufacturer and quality supervision organization at different levels can be monitored the benzo in peanut oil product (a) pyrene content, guarantee to comply with the national standard requirements.The mensuration > > of benzo (a) pyrene in the mensuration reversed-phased high performace liquid chromatographic > > that current relevant examination criteria is standard GB/T/T 22509-2008 < < animal and plant fat benzo (a) pyrene and standard GB/T/T 5009.27-2003 < < food, in detecting, reality adopts the methods in GB/T 22509-2008 more, the detection method of GB/T 5009.27-2003 is because of complex operation, measurement result is unstable, in reality, seldom use, also revised on the internet at present the collection that suggestion is solicited original text, it intends revised method of operating and first standard is basic identical.
But, the method for standard GB/T 22509-2008 is measured and in peanut oil, in benzo (a) pyrene content, also still to be had following problem:
(1) take toluene as solvent preparing standard solution, the chromatographic peak that sample introduction while measuring there will be forward position deformity, also there will be forward position deformity when the eluting power of solvent is better than mobile phase greatly.
(2) take acetonitrile-tetrahydrofuran mixed solution is solvent, and the retention time of chromatographic peak that the sample preparation liquid of preparing with it goes out and the retention time of standard solution exist larger departing from.
(3) accuracy in detection is low: when filling chromatographic column, be prone to empty situation, the absorption of each component and the efficiency of wash-out that comprise fat are unstable, the components such as fat are few when many during by amount out of wash-out, few when many while causing the level of residue of final acquisition, thereby recovery of standard addition is unstable, be difficult to benzo in Accurate Determining peanut oil (a) pyrene content.
(4) reagent utilization factor is low: toluene belongs to precursor chemicals, buying formality is complicated, and the specification of buying back is 4L even more than 500ml at least, yet the utilization factor of toluene is not high, conventionally 100ml toluene just can be used the several years, and unnecessary reagent is by leaving unused waste state increased the government pressure of relevant precursor chemicals.
(5) testing cost is high: the tetrahydrofuran price of 2.5L specification dress needs 800 ~ 900 yuan/bottle, and the chromatographically pure tetrahydrofuran price of 2.5L specification dress, by nearly 1,000 yuan, makes testing cost higher.
Summary of the invention
The invention provides the method for benzo (a) pyrene content in a kind of reversed-phase high-performance liquid chromatography external standard method peanut oil, the method is easy to operation, can effectively reduce operate miss, measure the result of benzo (a) the pyrene content in peanut oil accurately and reliably.
The object of the invention is to realize by following technical measures: a kind of method of benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil, it comprises the following steps:
(1) take peanut oil sample, by sherwood oil I, sample dissolution is diluted, then proceed in pre-filled alumina chromatographic column, by sherwood oil I, carry out drip washing and wash-out, collect eluent, eluent through nitrogen flushing be concentrated into dry after, residue adopts acetonitrile water mixed solution vortex to dissolve, and obtains sample preparation liquid;
(2) sample thief treating fluid carries out rp-hplc analysis, obtains chromatogram analysis data;
(3) get benzo (a) pyrene standard items and make solvent preparation standard operation liquid with acetonitrile water mixed solution, upper machine is measured, and drawing standard curve, tries to achieve benzo (a) pyrene correction factor;
(4), by the correction factor comparison of the chromatogram analysis data of the acquisition of step (2) and benzo (a) pyrene, obtain the content of benzo in peanut oil (a) pyrene.
In step of the present invention (1), the sample weighting amount of peanut oil sample is 4 ~ 5g, and larger sample size has more representativeness relatively, can reduce the accidental error because benzo (a) pyrene skewness causes in large oil sample.
As one embodiment of the present invention, pre-filled alumina chromatographic column in described step (1) adopts following methods to make: 22g neutral alumina is packed in the glass chromatography column that internal diameter is 14 ~ 16mm, rap post jamb, make alumina filled closely knit, add anhydrous sodium sulfate, the packed height of anhydrous sodium sulfate is 27 ~ 33mm, close post cock, at capital end, cover one deck Parafilm, be filled with after the nitrogen of dried and clean, with the sealed membrane capital end of obturaging, make pre-filled alumina chromatographic column, lucifuge is standby.In this operation, inserts first fills post, raps post jamb, fill material is filled closely knit, even, is difficult for forming cavity, then fills the airtight preservation of nitrogen, guarantees that the performance of fill material is not influenced by ambient temperature, can shift to an earlier date batch making standby, easy to use.
In step of the present invention (1), the preparation of sample preparation liquid is specially: in alumina chromatographic column, add sherwood oil I to activate, then weigh peanut oil sample, with moving in alumina chromatographic column after sherwood oil I dissolved dilution, with the drip washing of sherwood oil I and wash-out, collect eluent, eluent is placed in nitrogen flushing in flask and in 40 ~ 50 ℃ of water-baths and is concentrated into 0.1 ~ 0.5ml, concentrate is transferred in sample bottle, with sherwood oil I washing flask inwall, cleansing solution moves in sample bottle, nitrogen flushing is to dry, repeat again once washing nitrogen flushing operation, residue dissolves with acetonitrile water mixed solution, obtain sample preparation liquid.
In described acetonitrile water mixed solution, the volume ratio of acetonitrile and water is 9:1 ~ 19:1.
The chromatographic condition that step of the present invention (2) is recommended is: capillary chromatographic column adopts palycyclic aromatic analytical column; Column temperature is 25 ~ 35 ℃; Flow velocity is 1.0 ~ 1.2ml/min; Auto injection 10 ~ 20 μ l, mobile phase is the acetonitrile water mixed solution that acetonitrile and water volume ratio are 9 ~ 19:1, yield value 12 ~ 16, excitation wavelength 380nm, emission wavelength 410nm.
The analytical column of palycyclic aromatic described in the present invention requires residual interfering material and target components to be realized to good separation, can not use guard column, recommend adoption Ultimate PAH, Supelcosil LC-PAH, Eclipse PAH etc.
In step of the present invention (4), the correction factor of benzo (a) pyrene is by use, to contain benzo (a) the pyrene series standard solution of 0 ~ 12 μ g/L, under the reversed-phase high-performance liquid chromatography condition of identical setting, records.
The solvent of benzo of the present invention (a) pyrene series standard working fluid adopts acetonitrile water mixed solution, and the solvent of standard reserving solution preparation adopts acetonitrile.
compared with prior art, the invention has the advantages that:
(1) error is few, and precision is high, and testing result accuracy is highly reliable:
Sample weighting amount of the present invention is 4 ~ 5g, is 10 times of 0.4g sample weighting amount in GB/T 22509-2008, and larger sample size obviously has more representativeness relatively, can reduce the accidental error because benzo (a) pyrene uneven distribution causes in large oil sample.The elution efficiency of object is high, chaff interference residual little, and level of residue can stably be controlled at below 0.0050g.From chromatogram, only have a small amount of impurity peaks, and within 2 minutes, go out above peak in advance than object, can not form interference.
Sample preparation liquid in the present invention, standard operation liquid, solvent in mobile phase adopts the acetonitrile water mixed solution of unified proportioning, with acetonitrile water mixed solution, replace toluene to carry out the preparation of benzo (a) pyrene series standard solution on the one hand, effectively improved the peak shape of object, improved separating effect, on the other hand, in the time of can avoiding difference because of solvent to cause sample preparation liquid and standard operation liquid to go up respectively machine measuring there is fluctuation in object retention time, thereby improve the stability of object appearance time, further reduce operate miss, improve the accuracy of benzo (a) the pyrene qualitative and quantitative detection in peanut oil.
(2) detection sensitivity of the present invention is high, and its detection limit is low to moderate 0.5 μ g/kg.
(3) the present invention adopts 40 ~ 50 ℃ of temperature between sherwood oil I boiling range to be rotated evaporation, can accelerate to concentrate, and can guarantee again the completely reservation of target components in concentrated residue, and recovery of standard addition is high, result favorable reproducibility.
(4) simple to operate: the invention provides the preparation method of pre-filled alumina chromatographic column, column filling quality is good, can preserve one month, ready access upon use, easy to operate.
(5) loss of chromatographic column is low: elution efficiency of the present invention is high, and interfering component is residual little, cleans to dry rear reusablely, reduces the loss of chromatographic column.In the situation that not using guard column, separating effect and sensitivity after the column length time is used are still very good, and long service life is saved the consumption costs of guard column and chromatographic column.
(6) reduce costs, reduce pollution, be beneficial to environmental protection:
The present invention does not need to use expensive disposable commodities solid phase extraction column, and alumina chromatographic column is reused after can cleaning and drying, and saves the expense of alumina chromatographic column, reduces testing cost.
The present invention does not need to use toluene and tetrahydrofuran, thereby reduced the environmental pollution causing because of use toluene, tetrahydrofuran reagent and operating personnel's health has been endangered, and saved the buying expenses of expensive chromatographically pure toluene and tetrahydrofuran, avoid unnecessary reagent in idle waste state, and reduce the government pressure of relevant precursor chemicals.
(7) be easy to promote, social benefit is good: condition required for the present invention is uncomplicated, not expensive, utilize detecting instrument and the reagent of conventional configuration, can reach and measure accurately effect, this is conducive to this test item popularization and application Yu Ge grease production producer, thereby raw material, semi-manufacture and end product quality are strictly controlled, reduced the generation of correlated quality accident, increase the degree of belief of the public to oils Product quality and safety.The present invention is not limited in the detection to peanut oil, also can be applied to the detection of benzo (a) pyrene content in other category oil that sesame oil, camellia seed wet goods may contain benzo (a) pyrene.
accompanying drawing explanation:
Fig. 1 is the liquid chromatogram of the embodiment of the present invention one.
Fig. 2 is the liquid chromatogram of the embodiment of the present invention two.
Fig. 3 is the liquid chromatogram of the embodiment of the present invention three.
Fig. 4 is the liquid chromatogram of the embodiment of the present invention four.
Embodiment
Pre-filled alumina chromatographic column in following examples adopts following methods to make: 22g neutral alumina is packed in the glass chromatography column that internal diameter is 14 ~ 16mm, rap post jamb, make alumina filled closely knitly, add anhydrous sodium sulfate, the packed height of anhydrous sodium sulfate is 27 ~ 33mm, close post cock, at capital end, cover one deck Parafilm, be filled with after the nitrogen of dried and clean, with the sealed membrane capital end of obturaging, make pre-filled alumina chromatographic column, lucifuge is standby.In this operation, inserts first fills post, raps post jamb, fill material is filled closely knit, even, is difficult for forming cavity, then fills the airtight preservation of nitrogen, guarantees that the performance of fill material is not influenced by ambient temperature, can shift to an earlier date batch making standby, easy to use.
embodiment mono-
1. chromatographic condition:
The liquid chromatograph of configuration fluorescence detector;
Chromatographic column is Eclipse PAH(4.6 * 250mm), column temperature: 30 ℃
Mobile phase is acetonitrile water mixed solution, wherein, and V
acetonitrile﹕ V
water=9:1
Flow velocity 1.0ml/min
Excitation wavelength 380nm, emission wavelength 410nm, slit 10nm, PMT gain 16
2. correction factor is measured
Preparation standard reserving solution: accurately take benzo (a) pyrene g standard items 0.0125g, dissolve and be settled to 100ml with acetonitrile;
Preparation series standard working fluid: pipette standard reserving solution 1 ml, with acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1) dilute and be settled to 25ml, obtaining the benzo that concentration is 5 μ g/ml (a) pyrene standard solution, therefrom drawing respectively 0,2,4,8,16,24 μ l, then using acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1) dilute and be settled to 10ml, making serial benzo (a) pyrene standard solution, concentration is respectively 0,1,2,4,8,12 μ g/L.
Under above-mentioned chromatographic condition, with series standard solution sample introduction, with peak area and concentration drawing standard curve, obtain the correction factor of benzo (a) pyrene, set up corresponding external standard analytical approach.
3. sample preparation
1. take out pre-filled alumina chromatographic column, add sherwood oil I, open post cock, make sherwood oil I naturally flow out to the top of sodium sulphate layer.
2. measure 80mL sherwood oil I for drip washing and the wash-out of sample.Take 4g peanut oil sample A and be placed in 100ml volumetric flask, add sherwood oil I to dissolve and constant volume, make sample diluting liquid, therefrom pipette 10ml sample diluting liquid and put into alumina chromatographic column, after adding 10 mL sherwood oil I, open post cock, regulate flow velocity be every 2 ~ 3 seconds 1, under post, consumption socket joint is got leacheate, during to 20ml, change ground round-bottomed flask, collect eluent.
3. in 45 ℃ of water-baths, nitrogen flushing is concentrated into about 0.2ml, and concentrate is transferred in 2ml sample bottle, and with 1.5ml sherwood oil I washing flask inwall, cleansing solution moves in 2ml sample bottle, and nitrogen flushing is extremely dry, then repeats once washing nitrogen flushing operation.In sample bottle, add 400 μ L acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1), vortex dissolved residue, obtains sample solution, and the machine of keeping supplying is used.
4. sample determination
Under above-mentioned chromatographic condition, choose corresponding external standard analytical approach, sample introduction is measured, and from chromatographic work station, reads measurement result, and chromatogram is as shown in Figure 1.
5. pair series standard working fluid sample introduction is measured, and the concentration value of benzo (a) pyrene of take is horizontal ordinate, take corresponding peak area as ordinate, and drawing standard curve, obtains equation of linear regression and related coefficient.
The blank mark-on sample of the benzo of low concentration (a) pyrene is carried out to 20 times and detect, by C
l=3S
b/ b computing method detection limit.
The peanut oil sample that mark-on concentration is respectively to 0.5,5,10 μ g/kg carries out respectively replicate determination, calculate recovery rate and the coefficient of variation 6 times.
From table result, detection method of the present invention meets correlation technique requirement, highly sensitive, and detection limit is lower, and accuracy is good, and typical curve good linearity, recovery of standard addition are high, and precision is good, and the coefficient of variation is less.
embodiment bis-
1. chromatographic condition:
The liquid chromatograph of configuration fluorescence detector;
Chromatographic column is Supelcosil LC-PAH(4.6 * 250mm), column temperature: 35 ℃
Mobile phase is acetonitrile water mixed solution, wherein, and V
acetonitrile﹕ V
water=10:1
Flow velocity 1.0ml/min
Excitation wavelength 380nm, emission wavelength 410nm, slit 10nm, PMT gain 16
2. correction factor is measured
Preparation standard reserving solution: accurately take benzo (a) pyrene g standard items 0.0125g, dissolve and be settled to 100ml with acetonitrile;
Preparation series standard working fluid: pipette standard reserving solution 1ml, with acetonitrile water mixed solution (V
acetonitrile﹕ V
water=10:1) dilute and be settled to 25ml, obtaining the benzo that concentration is 5 μ g/ml (a) pyrene standard solution, therefrom drawing respectively 0,2,4,8,16,24 μ l, then using acetonitrile water mixed solution (V
acetonitrile﹕ V
water=10:1) dilute and be settled to 10ml, making serial benzo (a) pyrene standard solution, concentration is respectively 0,1,2,4,8,12 μ g/L.
Under above-mentioned chromatographic condition, with series standard solution sample introduction, with peak area and concentration drawing standard curve, obtain the correction factor of benzo (a) pyrene, set up corresponding external standard analytical approach.
3. sample preparation
1. take out pre-filled alumina chromatographic column, add sherwood oil I, open post cock, make sherwood oil I naturally flow out to the top of sodium sulphate layer.
2. measure 80mL sherwood oil I for drip washing and the wash-out of sample.Take 4g peanut oil sample B and be placed in 100ml volumetric flask, add sherwood oil I to dissolve and constant volume, make sample diluting liquid, therefrom pipette 10ml sample diluting liquid and put into alumina chromatographic column, after adding 10 mL sherwood oil I, open post cock, regulate flow velocity be every 2 ~ 3 seconds 1, under post, consumption socket joint is got leacheate, during to 20ml, change ground round-bottomed flask, collect eluent.
3. in 45 ℃ of water-baths, nitrogen flushing is concentrated into about 0.5ml, and concentrate is transferred in 2ml sample bottle, and with 1.5ml sherwood oil I washing flask inwall, cleansing solution moves in 2ml sample bottle, and nitrogen flushing is extremely dry, then repeats once washing nitrogen flushing operation.In sample bottle, add 400 μ L acetonitrile water mixed solution (V
acetonitrile﹕ V
water=10:1), vortex dissolved residue, sample solution, the machine of keeping supplying is used.
4. sample determination
Under above-mentioned chromatographic condition, choose corresponding external standard analytical approach, sample introduction is measured, and from chromatographic work station, reads measurement result, and chromatogram is as shown in Figure 2.
embodiment tri-
1. chromatographic condition:
The liquid chromatograph of configuration fluorescence detector;
Chromatographic column is Ultimate PAH(4.6 * 250mm), column temperature: 25 ℃
Mobile phase is acetonitrile water mixed solution, wherein, and V
acetonitrile﹕ V
water=19:1
Flow velocity 1.0ml/min
Excitation wavelength 380nm, emission wavelength 410nm, slit 10nm, PMT gain 16
2. correction factor is measured
Preparation standard reserving solution: accurately take benzo (a) pyrene g standard items 0.0125g, dissolve and be settled to 100ml with acetonitrile;
Preparation series standard working fluid: pipette standard reserving solution 1 ml, with acetonitrile water mixed solution (V
acetonitrile﹕ V
water=19:1) dilute and be settled to 25ml, obtaining the benzo that concentration is 5 μ g/ml (a) pyrene standard solution, therefrom drawing respectively 0,2,4,8,16,24 μ l, then using acetonitrile water mixed solution (V
acetonitrile﹕ V
water=19:1) dilute and be settled to 10ml, making serial benzo (a) pyrene standard solution, concentration is respectively 0,1,2,4,8,12 μ g/L.
Under above-mentioned chromatographic condition, with series standard solution sample introduction, with peak area and concentration drawing standard curve, obtain the correction factor of benzo (a) pyrene, set up corresponding external standard analytical approach.
3. sample preparation
1. take out pre-filled alumina chromatographic column, add sherwood oil I, open post cock, make sherwood oil naturally flow out to the top of sodium sulphate layer.
2. measure 80mL sherwood oil I for drip washing and the wash-out of sample.Take 4g peanut oil sample C and be placed in 100ml volumetric flask, add sherwood oil I to dissolve and constant volume, make sample diluting liquid, therefrom pipette 10ml sample diluting liquid and put into alumina chromatographic column, after adding 10 mL sherwood oil I, open post cock, regulate flow velocity be every 2 ~ 3 seconds 1, under post, consumption socket joint is got leacheate, during to 20ml, change ground round-bottomed flask, collect eluent.
3. in 45 ℃ of water-baths, nitrogen flushing is concentrated into about 0.3ml, and concentrate is transferred in 2ml sample bottle, and with 1.5ml sherwood oil I washing flask inwall, cleansing solution moves in 2ml sample bottle, and nitrogen flushing is extremely dry, then repeats once washing nitrogen flushing operation.In sample bottle, add 400 μ L acetonitrile water mixed solution (V
acetonitrile﹕ V
water=19:1), vortex dissolved residue, sample solution, the machine of keeping supplying is used.
4. sample determination
Under above-mentioned chromatographic condition, choose corresponding external standard analytical approach, sample introduction is measured, and from chromatographic work station, reads measurement result, and chromatogram is as shown in Figure 3.
embodiment tetra-
1. chromatographic condition:
The liquid chromatograph of configuration fluorescence detector;
Chromatographic column is Eclipse PAH(4.6 * 250mm), column temperature: 30 ℃
Mobile phase is acetonitrile water mixed solution, wherein, and V
acetonitrile﹕ V
water=9:1
Flow velocity 1.0ml/min
Excitation wavelength 380nm, emission wavelength 410nm, slit 10nm, PMT gain 16
2. correction factor is measured
Preparation standard reserving solution: accurately take benzo (a) pyrene g standard items 0.0125g, dissolve and be settled to 100ml with acetonitrile;
Preparation series standard working fluid: pipette standard reserving solution 1 ml, with acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1) dilute and be settled to 25ml, obtaining the benzo that concentration is 5 μ g/ml (a) pyrene standard solution, therefrom drawing respectively 0,2,4,8,16,24 μ l, then using acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1) dilute and be settled to 10ml, making serial benzo (a) pyrene standard solution, concentration is respectively 0,1,2,4,8,12 μ g/L.
Under above-mentioned chromatographic condition, with series standard solution sample introduction, with peak area and concentration drawing standard curve, obtain the correction factor of benzo (a) pyrene, set up corresponding external standard analytical approach.
3. sample preparation
1. take out pre-filled alumina chromatographic column, add sherwood oil I, open post cock, make sherwood oil I naturally flow out to the top of sodium sulphate layer.
2. measure 80mL sherwood oil I for drip washing and the wash-out of sample.Take 3g sesame oil sample D and be placed in 100ml volumetric flask, add sherwood oil I to dissolve and constant volume, make sample diluting liquid, therefrom pipette 10ml sample diluting liquid and put into alumina chromatographic column, after adding 10 mL sherwood oil I, open post cock, regulate flow velocity be every 2 ~ 3 seconds 1, under post, consumption socket joint is got leacheate, during to 20ml, change ground round-bottomed flask, collect eluent.
3. in 45 ℃ of water-baths, nitrogen flushing is concentrated into about 0.2ml, and concentrate is transferred in 2ml sample bottle, and with 1.5ml sherwood oil I washing flask inwall, cleansing solution moves in 2ml sample bottle, and nitrogen flushing is extremely dry, then repeats once washing nitrogen flushing operation.In sample bottle, add 400 μ L acetonitrile water mixed solution (V
acetonitrile﹕ V
water=9:1), vortex dissolved residue, obtains sample solution, and the machine of keeping supplying is used.
4. sample determination
Under above-mentioned chromatographic condition, choose corresponding external standard analytical approach, sample introduction is measured, and from chromatographic work station, reads measurement result, and chromatogram is as shown in Figure 4.
The present invention can summarize with other the concrete form without prejudice to spirit of the present invention or principal character.Above-mentioned embodiment of the present invention all can only be thought explanation of the present invention rather than restriction, therefore in the implication suitable with claims of the present invention and any change in scope, all should think to be included in the scope of claims.
Claims (8)
1. a method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil, is characterized in that, comprises the following steps:
(1) take peanut oil sample, by sherwood oil I, sample dissolution is diluted, then proceed in pre-filled alumina chromatographic column, by sherwood oil I, carry out drip washing and wash-out, collect eluent, eluent through nitrogen flushing be concentrated into dry after, residue adopts acetonitrile water mixed solution vortex to dissolve, and obtains sample preparation liquid;
(2) sample thief treating fluid carries out rp-hplc analysis, obtains chromatogram analysis data;
(3) get benzo (a) pyrene standard items and make solvent preparation standard operation liquid with acetonitrile water mixed solution, upper machine is measured, and drawing standard curve, tries to achieve benzo (a) pyrene correction factor;
(4), by the correction factor comparison of the chromatogram analysis data of the acquisition of step (2) and benzo (a) pyrene, obtain the content of benzo in peanut oil (a) pyrene.
2. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 1, it is characterized in that, pre-filled alumina chromatographic column in described step (1) adopts following methods to make: 22g neutral alumina is packed in the glass chromatography column that internal diameter is 14 ~ 16mm, rap post jamb, make alumina filled closely knit, add anhydrous sodium sulfate, the packed height of anhydrous sodium sulfate is 27 ~ 33mm, close post cock, at capital end, cover one deck Parafilm, be filled with after the nitrogen of dried and clean, with the sealed membrane capital end of obturaging, make pre-filled alumina chromatographic column, lucifuge is standby.
3. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 1, it is characterized in that, in described step (1), the preparation of sample preparation liquid is specially: in alumina chromatographic column, adding sherwood oil I to activate, then weigh peanut oil sample, with moving in alumina chromatographic column after sherwood oil I dissolved dilution, with the drip washing of sherwood oil I and wash-out, collect eluent, eluent is placed in nitrogen flushing in flask and in 40 ~ 50 ℃ of water-baths and is concentrated into 0.1 ~ 0.5ml, concentrate is transferred in sample bottle, with sherwood oil I washing flask inwall, cleansing solution moves in sample bottle, nitrogen flushing is to dry, repeat again once washing nitrogen flushing operation, residue dissolves with acetonitrile water mixed solution, obtain sample preparation liquid.
4. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 3, is characterized in that, in described acetonitrile water mixed solution, the volume ratio of acetonitrile and water is 9:1 ~ 19:1.
5. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 1, is characterized in that, the chromatographic condition of described step (2) is: capillary chromatographic column adopts palycyclic aromatic analytical column; Column temperature is 25 ~ 35 ℃; Flow velocity is 1.0 ~ 1.2ml/min; Auto injection 10 ~ 20 μ l, mobile phase is the acetonitrile water mixed solution that acetonitrile and water volume ratio are 9 ~ 19:1, yield value 12 ~ 16, excitation wavelength 380nm, emission wavelength 410nm.
6. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 5, is characterized in that, described palycyclic aromatic analytical column is Ultimate PAH, Supelcosil LC-PAH or Eclipse PAH.
7. according to the method for benzo (a) pyrene content in the reversed-phase high-performance liquid chromatography external standard method peanut oil described in claim 1 to 6 any one claim, it is characterized in that, the sample weighting amount in described step (1) is 4 ~ 5g.
8. the method for benzo (a) pyrene content in reversed-phase high-performance liquid chromatography external standard method peanut oil according to claim 7, it is characterized in that, in described step (4), the correction factor of benzo (a) pyrene is by use, to contain benzo (a) the pyrene series standard solution of 0 ~ 12 μ g/L, under identical reversed-phase high-performance liquid chromatography condition, records.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569193A (en) * | 2014-12-29 | 2015-04-29 | 山东龙大粮油有限公司 | Detection method of polycyclic aromatic hydrocarbon in edible oil |
| CN105136537A (en) * | 2015-08-14 | 2015-12-09 | 中国石油化工股份有限公司 | Pretreatment method of benzo (a) pyrene collected by glass fiber filter paper |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569193A (en) * | 2014-12-29 | 2015-04-29 | 山东龙大粮油有限公司 | Detection method of polycyclic aromatic hydrocarbon in edible oil |
| CN105136537A (en) * | 2015-08-14 | 2015-12-09 | 中国石油化工股份有限公司 | Pretreatment method of benzo (a) pyrene collected by glass fiber filter paper |
| CN105203679A (en) * | 2015-08-14 | 2015-12-30 | 中国石油化工股份有限公司 | Method for detecting content of benzo a pyrene in polycyclic aromatic hydrocarbon solution |
| CN105136537B (en) * | 2015-08-14 | 2018-02-16 | 中国石油化工股份有限公司青岛安全工程研究院 | The pre-treating method of benzo (a) pyrene gathered in glass fiber filter paper |
| CN105203694A (en) * | 2015-09-23 | 2015-12-30 | 成都艾比科生物科技有限公司 | Method for measuring 3, 4-benzopyrene in edible oil |
| CN105223301A (en) * | 2015-09-23 | 2016-01-06 | 成都艾比科生物科技有限公司 | A kind of method for measuring benzopyrene content in vegetable oil |
| CN106501411A (en) * | 2016-11-04 | 2017-03-15 | 无锡艾科瑞思产品设计与研究有限公司 | A kind of method for quick of benzopyrene from edible oil |
| CN112881539A (en) * | 2019-11-29 | 2021-06-01 | 岛津企业管理（中国）有限公司 | Method for rapidly detecting benzopyrene |
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