CN106383190A - Method for detecting malonaldehyde in edible vegetable oil through efficient liquid chromatography - Google Patents
Method for detecting malonaldehyde in edible vegetable oil through efficient liquid chromatography Download PDFInfo
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Abstract
The invention discloses a method for detecting malonaldehyde in edible vegetable oil through efficient liquid chromatography. The method comprises the steps of (1) preparing a standard product; (2) preparing a sample; (3) taking supernatant of a malonaldehyde standard product solution and a malonaldehyde sample solution after centrifugation; respectively adding a thiobarbituric acid water solution for taking a derivatization reaction; (4) manufacturing a standard curve; (5) performing efficient liquid chromatography determination; (6) calculating the content of malonaldehyde in the edible vegetable oil through the determination result in the fifth step according to the standard curve in the step (4). The method has the advantages that green and economical mobile phases are used; a widely used ultraviolet detector with low cost is used; the malonaldehyde in various kinds of common use edible vegetable oil in daily life can be fast, simply and accurately detected at low cost.
Description
Technical field
The present invention relates to the method for malonaldehyde is and in particular to a kind of high performance liquid chromatography detection is eaten in detection edible vegetable oil
Method with malonaldehyde in vegetable oil.
Background technology
Oils and fatss are one of seven major nutrients needed by human, are not only the main source of body energy, more have promotion liposoluble
Property vitamin absorption and adjust some hormone metabolisms effect.However, when oils and fatss heat treatment or storage improperly in the case of,
Often there is a series of oxidation, hydrolysis, polyreaction, produce primary oxidation product and secondary oxidative product, primary oxidation
Product hydroperoxide easily decomposes generation secondary oxidative product, including small-molecule substances such as aldehyde, ketone or acid, wherein the third two
Aldehyde (Malondialdehyde, MDA) be in aldehydes important a kind of it is considered to be the end-product of Oxidation of Fat and Oils.Malonaldehyde is in state
Only it is classified as third level carcinogen, it is in biological internal lipid peroxy in the carcinogen list of border DKFZ (IARC)
Change in reaction and can produce, and the cross-linked polymeric of the life macromolecules such as protein, nucleic acid can be caused, therefore its cytotoxicity is subject to
The concern of researcher.Have many researchs to think, malonaldehyde can cause a disease carcinogenic, relevant with tremulous pulse medicated porridge hardening.
1980, Porter research thought that malonaldehyde is the peroxide by having the double bond fatty acid of more than 3 or 4
Derivative (Porter NA.Prostaglandin endoperoxides [A] the .Free radicals in biolog obtaining
[C].Pryor W A,Ed；Academic Press:London,1980.).1992, research is had to think that malonaldehyde is likely to spread out
It is born in the linolenic acid in vegetable oil.In the same year, there is 0.2 μM of FeCl of researcher2Solution and 0.1 μM of H2O2Aoxidize when 37 DEG C and lure
Lead fatty acid 16 hours, find that arachidonic acid produces maximum malonaldehyde (60.5nmol/mg fatty acid), be secondly linolenic acid
(33.4nmol/mg fatty acid) and linoleic acid (33.9nmol/mg fatty acid), and Oleic acid does not produce malonaldehyde.It follows that containing
The unsaturated fatty acid oxidable generation malonaldehyde of more than 2 double bonds, and its growing amount is relevant with fatty acid degree of unsaturation.Oil
Fat can produce hydroperoxides in oxidative rancidity process, and further hydroperoxides can resolve into secondary oxidative product, therefore
Peroxide value can not definitely as Oxidation of Fat and Oils index, and as oxidation end-product malonaldehyde have stable, sensitive
Feature, can as evaluation grease oxidation rancid index.At present, regulation has been made to mda content in Adeps Sus domestica by China, no
Can exceed that 0.25mg/100g, but the Mda method for edible vegetable oil and content no clear stipulaties.
At present, both at home and abroad the research of malonaldehyde is concentrated mainly in biomedicine, the method for detection is mainly test kit
Method and enzyme linked immunosorbent assay etc..On food, the detection method of studies in China is both referred to greatly《GB/T 5009.181-
The mensure of malonaldehyde in 2003 Adeps Sus domesticas》In thiobarbituricacidα- method, or improve on its basis, the method is to be referred from plant
The mensure of mda content in plant tissue in thing, its principle is malonaldehyde through trichloroacetic acid to be extracted and to remove Deproteinization etc. miscellaneous
Matter, malonaldehyde and thiobarbituricacidα- react generation pink material under high mild acidic conditions, and this material is in 532-538nm
There is absorption peak under wavelength.But, the reaction of thiobarbituricacidα- and malonaldehyde belongs to nonspecific reaction, except malonaldehyde it
Outer others aldehyde, ketone secondary oxidative product all can lead to measurement result higher with thiobarbituric acid reaction.Meanwhile, sulfur
Complicated for barbital acid system detection program, affected larger during absorbance measurement by personal error.Therefore, in order to overcome above lacking
Point, in domestic and foreign literature report detection food, the novel detection method of malonaldehyde mainly has high performance liquid chromatography, gas chromatogram
The application of method, combined gas chromatography mass spectrometry, near-infrared spectrum technique etc., wherein high performance liquid chromatography is relatively many.Wei Zuojun adopts
Do not derive under acid condition, directly in 250nm about carry out high performance liquid chromatography detection kitchen degras and olive oil, principle be
Most malonaldehyde are made not exist with keto-acid so that enol form exists under acid condition, indirect with double bond content under 250nm
Quantitative malonaldehyde, but measurement result is easily affected by the secondary oxidative product with double bond such as other olefine aldehydrs, ketenes, light alkene
(Wei ZJ,Li XH,Dilantha Thushara,et al.Determination and removal of
malondialdehyde and other 2-thiobarbituric acid reactive substances in waste
cooking oil[J].Journal of Food Engineering,2011,(107):379-384.)、Fuad Al-
Rimawi(Fuad Al-Rimawi.Development and Validation of a Simple Reversed-Phase
HPLC-UV Method for Determination of Malondialdehyde[J].J Am Oil Chem Soc,
2015,(92):933-937.).With methyl malonaldehyde as internal standard substance, 2,4 dinitrophenyl hydrazine is derivating agent to Caroline Douny
Set up the detection method that LC-MS/MS measures malonaldehyde in Semen Lini oil, this method is relatively costly, and derivating agent toxicity is stronger
(Caroline Douny,Angelique Tihon,Pierre Bayonnet,et al.Validation of the
Analytical Procedure for the Determination of Malondialdehyde and Three Other
Aldehydes in Vegetable Oil Using Liquid Chromatography Coupled to Tandem Mass
Spectrometry(LC-MS/MS)and Application to Linseed Oil[J].Food Anal Methods,
2015,(8):1425-1435.).Yang Haifeng, Zhu Hongliang are respectively using malonaldehyde in high performance liquid chromatography detection feedstuff, edible oil
Content, concrete operations be by sample malonaldehyde solution of trichloroacetic acid extract after perform the derivatization with thiobarbituricacidα-
Reaction, through liquid chromatograph separate after fluoroscopic examination (Yang Haifeng, Liu Dan, Yang Junhua, etc. in Feeds by HPLC
Malonaldehyde [J]. Shanghai Agricultural journal, 2013,29 (4):14-17.) (Zhu Hongliang, Ge Fangfang, pipe beauty. high performance liquid chromatography
Malonaldehyde [J] in detection edible oil. food industry science and technology, 2015,1 (36):309-315.).Yu Wei uses high-efficient liquid phase color
Spectrum detection numerous food in malonaldehyde content, concrete operations be by the malonaldehyde in sample with solution of trichloroacetic acid extraction after with
Thiobarbituricacidα- performs the derivatization reaction, through liquid chromatograph separate after two aurora detection (Yu Wei, Zhao Yongfeng, Wu Yujie, etc.
Malonaldehyde [J] in food by HPLC. assay laboratory, 2014,33 (4):445-447.).Although these sides
Method can preferably detect the malonaldehyde in food, but makees mobile phase using organic reagent and buffer, exist high cost,
It is easily damaged the problem of pillar.Therefore, existing invention optimizes and set up the high performance liquid chromatography of green economy and be used for measuring
Malonaldehyde in multiple daily visible edible vegetable oils has great importance.
Content of the invention
The invention aims to overcoming the defect of above-mentioned prior art presence and providing a kind of green economy, can be fast
Speed, accurately, in easy detection edible vegetable oil malonaldehyde method.
The purpose of the present invention is achieved through the following technical solutions.
A kind of method of malonaldehyde in high performance liquid chromatography detection edible vegetable oil, the method concretely comprises the following steps：
(1) preparation of standard substance：The malonaldehyde standard substance preparing 10 μ g/mL using 1,1,3,3- tetraethoxypropane are molten
Liquid；
(2) preparation of sample：Accurately weigh food plant oil samples, add the trichloroacetic acid that concentration is 7.5wt%, shaking table
Shaking, filters, and filtrate is placed in centrifuge tube and is centrifuged, and Aspirate supernatant is stand-by；
(3) derivative reaction：Take the supernatant after isopyknic malonaldehyde standard solution and step (2) centrifugation, respectively
Add and supernatant isopyknic thiobarbituricacidα- aqueous solution, whirlpool mixes, be incubated in water-bath, take out, rapid with ice-water bath
It is cooled to room temperature, by solution filter membrane, stand-by；
(4) make standard curve：Compound concentration is the malonaldehyde standard solution of 0.02～2.5 μ g/mL respectively, and through same
The derivative reaction of step (3), respectively after high effective liquid chromatography for measuring, with standard concentration as x-axis, corresponding chromatographic peak
Area makes linear standard curve for y-axis；
(5) high-performance liquid chromatogram determination：The solution that step (3) is obtained carries out high-performance liquid chromatogram determination；
(6) measurement result of step (5) is calculated containing of malonaldehyde in edible vegetable oil according to the standard curve of step (4)
Amount.
Further, described edible vegetable oil includes Petiolus Trachycarpi oil, Semen Maydis oil, Camellia oil, oleum lini or Oleum Camelliae.
Further, in step (2), the concentration of the supernatant obtaining is 0.038～0.042g/ml.
Further, in step (2), concentration contains 0.1wt%EDTA in the trichloroacetic acid for 7.5wt%.
Further, in step (2), described shaking table shakes and shakes 30～60min for 140～180r/min.
Further, in step (2), described centrifugation is centrifugation 5～10min under rotating speed 5000～8000r/min.
Further, in step (3), described water bath heat preservation is in 80～100 DEG C of water bath heat preservation 20～40min.
Further, in step (3), the concentration of described thiobarbituricacidα- aqueous solution is 0.02mol/L.
Further, step (4), in (5), described high-performance liquid chromatogram determination is with anti-phase C18Post is chromatographic column；Mobile phase
With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, volume ratio is 10～15 to composition:85～90；Flow velocity is 1.0mL/min
Carry out isocratic elution；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is 532nm；Column temperature is
30℃；Sample size is 10 μ L.
Further, in step (4), the concentration of described malonaldehyde standard solution is 0.02,0.1,0.2,0.3,0.4,
0.5、0.6、0.7、0.8、0.9、1.0、2.5μg/mL.
Further, the method is limited to 0.035mg/kg to the detection of malonaldehyde, is quantitatively limited to 0.14mg/kg.
Further, add malonaldehyde in edible vegetable oil, then be measured with the detection method of the present invention, obtain third
The content of dialdehyde, is compared with the content of MDA of actual interpolation, to verify the accuracy of detection method.
Further, the response rate obtaining malonaldehyde in edible vegetable oil is between 88.75～106.99%.
Compared with prior art, the present invention has advantages below and technique effect：
(1) mobile phase that high performance liquid chromatography detection of the present invention adopts is acetonitrile and ultra-pure water, and ethane nitrile content is minimum only to be accounted for
10%, compared to the organic phase flows such as the ammonium acetate used by other technologies, potassium dihydrogen phosphate, more green economy, more environment
Friendly；
(2) detector that high performance liquid chromatography detection of the present invention adopts is UV-detector, compared to other technologies institute
Fluorescence detector, UV-detector has the characteristics that universal, inexpensive, is more suitable for colleges and universities' research worker, industrial applications.
Brief description
Fig. 1 is the chromatogram of malonaldehyde standard substance；
Fig. 2 is the linear standard curve of malonaldehyde；
Fig. 3 is the chromatogram of Flos Camelliae Japonicae oil samples in embodiment 5.
Specific embodiment
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1
High performance liquid chromatography detection eats mda content in oleum lini：
(1) preparation of standard substance：Accurately weigh 0.315g 1,1,3,3- tetraethoxypropane, deionized water dissolves simultaneously
It is diluted to 1000mL, accurately pipette above-mentioned solution 10mL and be diluted to 100mL further, as 10 μ g/mL malonaldehyde standard substance are molten
Liquid；
(2) preparation of sample：Weigh 2.0334g oleum lini, add the trichloroacetic acid mixed liquor of 50mL concentration 7.5wt%,
Contain 0.1wt%EDTA in trichloroacetic acid mixed liquor, 140r/min shaking table shakes 40min, filtered once with middling speed double-layer filter paper,
Take whole filtrates, be placed in centrifuge tube 5000r/min centrifugation 5min, Aspirate supernatant, stand-by；
(3) derivative reaction：After taking above-mentioned centrifugation respectively, the supernatant 5mL of solution, malonaldehyde standard solution 5mL are placed in
In test tube, it is separately added into the thiobarbituricacidα- aqueous solution that 5mL concentration is 0.02mol/L, whirlpool mixes, is placed in 90 DEG C of water-baths
Insulation 20min, takes out, is rapidly cooled to room temperature with ice-water bath, solution is crossed 0.45 μm of microporous filter membrane, stand-by；
(4) high-performance liquid chromatogram determination：Chromatographic column is anti-phase C18Post (250mm × 4.6mm, 5 μm, Agilent ZOTBAX
Eclipse XDB-C18)；With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, ratio is 15 for flowing phase composition:85；Flow velocity
Carry out isocratic elution for 1.0mL/min；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is
532nm；Column temperature is 30 DEG C；Sample size is 10 μ L；
The chromatogram of malonaldehyde standard substance as shown in figure 1, as shown in Figure 1 the appearance time of malonaldehyde be 7.681min, peak
Shape is good；The chromatogram peak type of obtained oleum lini is good, good with the separating degree of impurity；
(5) make linear standard curve：Respectively compound concentration be 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8th, the malonaldehyde standard solution of 0.9,1.0,2.5 μ g/mL, through high effective liquid chromatography for measuring (derivative reaction and chromatostrip
Part is identical with Caulis et Folium Lini oil samples) after, with standard concentration as x-axis, it is bent that corresponding chromatographic peak area makes linear criterion for y-axis
Line, as shown in Figure 2；The linear standard curve equation of malonaldehyde：Y=3.8051x+0.0582, R2=0.9979；
(6) detection of detection method is limited to 0.035mg/kg, is quantitatively limited to 0.14mg/kg；
(7) it is the accuracy verifying detection method, add quantitative malonaldehyde in oleum lini, then use the present invention
Detection method be measured, obtain the content of malonaldehyde, be compared with the content of MDA of actual interpolation, obtain in oleum lini
The response rate of malonaldehyde, as shown in table 1.
The response rate of malonaldehyde in table 1 oleum lini
Known by table 1, in oleum lini, the response rate of malonaldehyde, between 97.84～104.94%, shows present invention detection side
The method response rate is good, can meet the actually detected needs of oleum lini.
Embodiment 2
Mda content in high performance liquid chromatography detection edible corn oil：
(1) preparation of standard substance：Accurately weigh 0.315g 1,1,3,3- tetraethoxypropane, deionized water dissolves simultaneously
It is diluted to 1000mL, accurately pipette above-mentioned solution 10mL and be diluted to 100mL further, as 10 μ g/mL malonaldehyde standard substance are molten
Liquid；
(2) preparation of sample：Weigh 2.0093g Semen Maydis oil, add the trichloroacetic acid mixed liquor of 50mL concentration 7.5wt%,
Contain 0.1wt%EDTA in trichloroacetic acid mixed liquor, 150r/min shaking table shakes 60min, filtered once with middling speed double-layer filter paper,
Take whole filtrates, be placed in centrifuge tube 6000r/min centrifugation 5min, Aspirate supernatant, stand-by；
(3) derivative reaction：After taking above-mentioned centrifugation respectively, the supernatant 5mL of solution, malonaldehyde standard solution 5mL are placed in
In test tube, it is separately added into the thiobarbituricacidα- aqueous solution that 5mL concentration is 0.02mol/L, whirlpool mixes, is placed in 80 DEG C of water-baths
Insulation 30min, takes out, is rapidly cooled to room temperature with ice-water bath, solution is crossed 0.45 μm of microporous filter membrane, stand-by；
(4) high-performance liquid chromatogram determination：Chromatographic column is anti-phase C18Post (250mm × 4.6mm, 5 μm, Agilent ZOTBAX
Eclipse XDB-C18)；With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, ratio is 15 for flowing phase composition:85；Flow velocity
Carry out isocratic elution for 1.0mL/min；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is
532nm；Column temperature is 30 DEG C；Sample size is 10 μ L；
The chromatogram of malonaldehyde standard substance is referring to Fig. 1；The chromatogram peak type of obtained Semen Maydis oil is good, with dividing of impurity
Good from degree
(5) make linear standard curve：Respectively compound concentration be 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8th, the malonaldehyde standard solution of 0.9,1.0,2.5 μ g/mL, through high effective liquid chromatography for measuring (derivative reaction and chromatostrip
Part is identical with Semen Maydiss oil samples) after, with standard concentration as x-axis, it is bent that corresponding chromatographic peak area makes linear criterion for y-axis
Line, as shown in Figure 2；The linear standard curve equation of malonaldehyde：Y=3.8051x+0.0582, R2=0.9979；
(6) detection of detection method is limited to 0.035mg/kg, is quantitatively limited to 0.14mg/kg；
(7) it is the accuracy verifying detection method, add quantitative malonaldehyde in Semen Maydis oil, then use the present invention
Detection method be measured, obtain the content of malonaldehyde, be compared with the content of MDA of actual interpolation, obtain in Semen Maydis oil
The response rate of malonaldehyde, as shown in table 2.
The response rate of malonaldehyde in table 2 Semen Maydis oil
Known by table 2, in Semen Maydis oil, the response rate of malonaldehyde, between 89.98～102.96%, shows present invention detection side
The method response rate is good, can meet the actually detected needs of Semen Maydis oil.
Embodiment 3
Mda content in high performance liquid chromatography detection edible rapeseed oil：
(1) preparation of standard substance：Accurately weigh 0.315g 1,1,3,3- tetraethoxypropane, deionized water dissolves simultaneously
It is diluted to 1000mL, accurately pipette above-mentioned solution 10mL and be diluted to 100mL further, as 10 μ g/mL malonaldehyde standard substance are molten
Liquid；
(2) preparation of sample：Weigh 2.0164g Oleum Brassicae campestriss, add the trichloroacetic acid mixed liquor of 50mL concentration 7.5wt%,
Contain 0.1wt%EDTA in trichloroacetic acid mixed liquor, 180r/min shaking table shakes 30min, filtered once with middling speed double-layer filter paper,
Take whole filtrates, be placed in centrifuge tube 8000r/min centrifugation 10min, Aspirate supernatant, stand-by；
(3) derivative reaction：After taking above-mentioned centrifugation respectively, the supernatant 5mL of solution, malonaldehyde standard solution 5mL are placed in
In test tube, it is separately added into the thiobarbituricacidα- aqueous solution that 5mL concentration is 0.02mol/L, whirlpool mixes, is placed in 85 DEG C of water-baths
Insulation 40min, takes out, is rapidly cooled to room temperature with ice-water bath, solution is crossed 0.45 μm of microporous filter membrane, stand-by；
(4) high-performance liquid chromatogram determination：Chromatographic column is anti-phase C18Post (250mm × 4.6mm, 5 μm, Agilent ZOTBAX
Eclipse XDB-C18)；With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, ratio is 14 for flowing phase composition:86；Flow velocity
Carry out isocratic elution for 1.0mL/min；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is
532nm；Column temperature is 30 DEG C；Sample size is 10 μ L；
The chromatogram of malonaldehyde standard substance is referring to Fig. 1；The chromatogram peak type of obtained Oleum Brassicae campestriss is good, with dividing of impurity
Good from degree.
(5) make linear standard curve：Respectively compound concentration be 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8th, the malonaldehyde standard solution of 0.9,1.0,2.5 μ g/mL, through high effective liquid chromatography for measuring (derivative reaction and chromatostrip
Part is identical with Semen Allii Tuberosi oil samples) after, with standard concentration as x-axis, it is bent that corresponding chromatographic peak area makes linear criterion for y-axis
Line, as shown in Figure 2；The linear standard curve equation of malonaldehyde：Y=3.8051x+0.0582, R2=0.9979；
(6) detection of detection method is limited to 0.035mg/kg, is quantitatively limited to 0.14mg/kg；
(7) it is the accuracy verifying detection method, add quantitative malonaldehyde in Oleum Brassicae campestriss, then use the present invention
Detection method be measured, obtain the content of malonaldehyde, be compared with the content of MDA of actual interpolation, obtain in Oleum Brassicae campestriss
The response rate of malonaldehyde, as shown in table 3.
The response rate of malonaldehyde in table 3 Oleum Brassicae campestriss
Known by table 3, in Oleum Brassicae campestriss, the response rate of malonaldehyde, between 89.71～106.79%, shows present invention detection side
The method response rate is good, can meet the actually detected needs of Oleum Brassicae campestriss.
Embodiment 4
Mda content in high performance liquid chromatography detection edible palm oil：
(1) preparation of standard substance：Accurately weigh 0.315g 1,1,3,3- tetraethoxypropane, deionized water dissolves simultaneously
It is diluted to 1000mL, accurately pipette above-mentioned solution 10mL and be diluted to 100mL further, as 10 μ g/mL malonaldehyde standard substance are molten
Liquid；
(2) preparation of sample：Weigh 2.0078g Petiolus Trachycarpi oil, add the trichloroacetic acid mixed liquor of 50mL concentration 7.5wt%,
Contain 0.1wt%EDTA in trichloroacetic acid mixed liquor, 160r/min shaking table shakes 40min, filtered once with middling speed double-layer filter paper,
Take whole filtrates, be placed in centrifuge tube 7000r/min centrifugation 8min, Aspirate supernatant, stand-by；
(3) derivative reaction：After taking above-mentioned centrifugation respectively, the supernatant 5mL of solution, malonaldehyde standard solution 5mL are placed in
In test tube, it is separately added into the thiobarbituricacidα- aqueous solution that 5mL concentration is 0.02mol/L, whirlpool mixes, is placed in 95 DEG C of water-baths
Insulation 30min, takes out, is rapidly cooled to room temperature with ice-water bath, solution is crossed 0.45 μm of microporous filter membrane, stand-by；
(4) high-performance liquid chromatogram determination：Chromatographic column is anti-phase C18Post (250mm × 4.6mm, 5 μm, Agilent ZOTBAX
Eclipse XDB-C18)；With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, ratio is 12 for flowing phase composition:88；Flow velocity
Carry out isocratic elution for 1.0mL/min；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is
532nm；Column temperature is 30 DEG C；Sample size is 10 μ L；
The chromatogram of malonaldehyde standard substance is referring to Fig. 1；The chromatogram peak type of obtained Petiolus Trachycarpi oil is good, with dividing of impurity
Good from degree
(5) make linear standard curve：Respectively compound concentration be 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8th, the malonaldehyde standard solution of 0.9,1.0,2.5 μ g/mL, through high effective liquid chromatography for measuring (derivative reaction and chromatostrip
Part is identical with Petiolus Trachycarpi oil samples) after, with standard concentration as x-axis, it is bent that corresponding chromatographic peak area makes linear criterion for y-axis
Line, as shown in Figure 2；The linear standard curve equation of malonaldehyde：Y=3.8051x+0.0582, R2=0.9979；
(6) detection of detection method is limited to 0.035mg/kg, is quantitatively limited to 0.14mg/kg；
(7) it is the accuracy verifying detection method, add quantitative malonaldehyde in Petiolus Trachycarpi oil, then use the present invention
Detection method be measured, obtain the content of malonaldehyde, be compared with the content of MDA of actual interpolation, obtain in Petiolus Trachycarpi oil
The response rate of malonaldehyde, as shown in table 4.
The response rate of malonaldehyde in table 4 Petiolus Trachycarpi oil
Known by table 4, in Petiolus Trachycarpi oil, the response rate of malonaldehyde, between 88.75～98.98%, shows detection method
The response rate is good, can meet the actually detected needs of Petiolus Trachycarpi oil.
Embodiment 5
High performance liquid chromatography detection eats mda content in Camellia oil：
(1) preparation of standard substance：Accurately weigh 0.315g 1,1,3,3- tetraethoxypropane, deionized water dissolves simultaneously
It is diluted to 1000mL, accurately pipette above-mentioned solution 10mL and be diluted to 100mL further, as 10 μ g/mL malonaldehyde standard substance are molten
Liquid；
(2) preparation of sample：Weigh 2.0212g Camellia oil, add the trichloroacetic acid mixed liquor of 50mL concentration 7.5wt%,
Contain 0.1wt%EDTA in trichloroacetic acid mixed liquor, 170r/min shaking table shakes 50min, filtered once with middling speed double-layer filter paper,
Take whole filtrates, be placed in centrifuge tube 8000r/min centrifugation 8min, Aspirate supernatant, stand-by；
(3) derivative reaction：After taking above-mentioned centrifugation respectively, the supernatant 5mL of solution, malonaldehyde standard solution 5mL are placed in
In test tube, it is separately added into the thiobarbituricacidα- aqueous solution that 5mL concentration is 0.02mol/L, whirlpool mixes, and is placed in 100 DEG C of water-baths
Middle insulation 20min, takes out, is rapidly cooled to room temperature with ice-water bath, solution is crossed 0.45 μm of microporous filter membrane, stand-by；
(4) high-performance liquid chromatogram determination：Chromatographic column is anti-phase C18Post (250mm × 4.6mm, 5 μm, Agilent ZOTBAX
Eclipse XDB-C18)；With acetonitrile and ultra-pure water as mobile phase A with Mobile phase B, ratio is 10 for flowing phase composition:90；Flow velocity
Carry out isocratic elution for 1.0mL/min；The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is
532nm；Column temperature is 30 DEG C；Sample size is 10 μ L；
The chromatogram of malonaldehyde standard substance is referring to Fig. 1；The chromatogram of the Camellia oil obtaining is as shown in figure 3, as shown in Figure 3 third
The appearance time of dialdehyde is 7.431min, and peak type is good, good with the separating degree of impurity.
(5) make linear standard curve：Respectively compound concentration be 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8th, the malonaldehyde standard solution of 0.9,1.0,2.5 μ g/mL, through high effective liquid chromatography for measuring (derivative reaction and chromatostrip
Part is identical with Flos Camelliae Japonicae oil samples) after, with standard concentration as x-axis, it is bent that corresponding chromatographic peak area makes linear criterion for y-axis
Line, as shown in Figure 2；The linear standard curve equation of malonaldehyde：Y=3.8051x+0.0582, R2=0.9979；
(6) detection of detection method is limited to 0.035mg/kg, is quantitatively limited to 0.14mg/kg；
(7) it is the accuracy verifying detection method, add quantitative malonaldehyde in Camellia oil, then use the present invention
Detection method be measured, obtain the content of malonaldehyde, be compared with the content of MDA of actual interpolation, obtain in Camellia oil
The response rate of malonaldehyde, as shown in table 5.
The response rate of malonaldehyde in table 5 Camellia oil
Known by table 5, in Camellia oil, the response rate of malonaldehyde, between 90.81～100.83%, shows that the inventive method is returned
Yield is good, can meet the actually detected needs of Camellia oil.
Claims (10)
1. in a kind of high performance liquid chromatography detection edible vegetable oil the method for malonaldehyde it is characterised in that comprising the following steps：
（1）The preparation of standard substance：Malonaldehyde standard solution is prepared using 1,1,3,3- tetraethoxypropane；
（2）The preparation of sample：Accurately weigh food plant oil samples, add trichloroacetic acid, shaking table shakes, filter, filtrate is placed in
It is centrifuged in centrifuge tube, Aspirate supernatant, stand-by；
（3）Derivative reaction：Take isopyknic malonaldehyde standard solution and step（2）Supernatant after centrifugation, is separately added into
With supernatant isopyknic thiobarbituricacidα- aqueous solution, whirlpool mixes, and is incubated in water-bath, takes out, is cooled down rapidly with ice-water bath
To room temperature, by solution filter membrane, stand-by；
（4）Make standard curve：Compound concentration is the malonaldehyde standard solution of 0.02 ~ 2.5 μ g/mL respectively, and through same step
（3）Derivative reaction, respectively after high effective liquid chromatography for measuring, with standard concentration as x-axis, corresponding chromatographic peak area
Make linear standard curve for y-axis；
（5）High-performance liquid chromatogram determination：By step（3）The solution obtaining carries out high-performance liquid chromatogram determination；
（6）By step（5）Measurement result according to step（4）Standard curve calculate edible vegetable oil in malonaldehyde content.
2. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（2）In, the concentration of the supernatant obtaining is 0.038 ~ 0.042g/ml；Described edible vegetable oil include Petiolus Trachycarpi oil,
Semen Maydis oil, Camellia oil, oleum lini or Oleum Camelliae.
3. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（1）The concentration of described malonaldehyde standard solution is 10 μ g/mL；Step（2）In, the concentration of described trichloroacetic acid
For 7.5wt%, and contain 0.1wt%EDTA.
4. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（2）In, described shaking table shakes and shakes 30 ~ 60 min for 140 ~ 180 r/min；Described centrifugation be rotating speed 5000 ~
It is centrifuged 5 ~ 10 min under 8000 r/min.
5. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（3）In, described water bath heat preservation is in 80 ~ 100 DEG C of water bath heat preservation 20 ~ 40 min；Described thiobarbituricacidα- is water-soluble
The concentration of liquid is 0.02 mol/L.
6. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（4）、（5）In, described high-performance liquid chromatogram determination is with anti-phase C18Post is chromatographic column；Flowing phase composition with acetonitrile and
Ultra-pure water is mobile phase A and Mobile phase B, and volume ratio is 10 ~ 15:85~90；Flow velocity carries out isocratic elution for 1.0 mL/min；
The detector that high performance liquid chromatography detection uses is UV-detector, and Detection wavelength is 532 nm；Column temperature is 30 DEG C；Sample size is
10μL.
7. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, step（4）In, the concentration of described malonaldehyde standard solution is 0.02,0.1,0.2,0.3,0.4,0.5,0.6,0.7,
0.8、0.9、1.0、2.5 μg/mL.
8. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, the method is limited to 0.035 mg/kg to the detection of malonaldehyde, be quantitatively limited to 0.14 mg/kg.
9. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 1 malonaldehyde method, its feature
It is, add malonaldehyde in edible vegetable oil, then be measured with the detection method of the present invention, obtain the content of malonaldehyde,
It is compared with the content of MDA of actual interpolation, to verify the accuracy of detection method.
10. in a kind of high performance liquid chromatography detection edible vegetable oil according to claim 9 malonaldehyde method, its feature
It is, the response rate obtaining malonaldehyde in edible vegetable oil is between 88.75 ~ 106.99%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108572224A (en) * | 2018-04-25 | 2018-09-25 | 滨州医学院 | The assay method of mda content in a kind of biological tissue |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805669A (en) * | 2010-04-15 | 2010-08-18 | 浙江大学 | Method for removing malonaldehyde in oil |
| CN102778545A (en) * | 2012-07-10 | 2012-11-14 | 北京智云达科技有限公司 | Method for comprehensively and quickly screening multiparameters of illegal cooking oil |
| CN102778454A (en) * | 2012-07-10 | 2012-11-14 | 北京智云达科技有限公司 | Testing reagent for illegal cooking oil |
| CN103134921A (en) * | 2012-12-27 | 2013-06-05 | 北京智云达科技有限公司 | Illegal cooking oil colloidal gold fast measuring card |
-
2016
- 2016-11-07 CN CN201610971477.3A patent/CN106383190A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101805669A (en) * | 2010-04-15 | 2010-08-18 | 浙江大学 | Method for removing malonaldehyde in oil |
| CN102778545A (en) * | 2012-07-10 | 2012-11-14 | 北京智云达科技有限公司 | Method for comprehensively and quickly screening multiparameters of illegal cooking oil |
| CN102778454A (en) * | 2012-07-10 | 2012-11-14 | 北京智云达科技有限公司 | Testing reagent for illegal cooking oil |
| CN103134921A (en) * | 2012-12-27 | 2013-06-05 | 北京智云达科技有限公司 | Illegal cooking oil colloidal gold fast measuring card |
Non-Patent Citations (5)
| Title |
|---|
| 中华人民共和国国家卫生和计划生育委员会: "《食品安全国家标准 食品中丙二醛的测定GB5009.181-2016》", 31 August 2016 * |
| 余巍 等: "高效液相色谱法测定食品中丙二醛", 《分析试验室》 * |
| 刘静波: "《食品科学与工程专业实验指导》", 30 September 2010, 化学工业出版社 * |
| 叶振生 主编: "《蜂产品深加工技术》", 30 April 2003, 中国轻工业出版社 * |
| 朱洪亮 等: "高效液相色谱法检测食用油中的丙二醛", 《食品工业科技》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108572224A (en) * | 2018-04-25 | 2018-09-25 | 滨州医学院 | The assay method of mda content in a kind of biological tissue |
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