CN103245660B - A kind of edible oil and fat determination of POV composite reagent and detection method - Google Patents
A kind of edible oil and fat determination of POV composite reagent and detection method Download PDFInfo
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Abstract
The present invention relates to the mensuration of peroxide value, particularly relate to a kind of edible oil and fat determination of POV composite reagent and detection method, comprising: polar organic solvent, reductive agent, chromogenic reagent, color stability agent; Described polar organic solvent is remembered by mass percentage, and the mass concentration comprising ethanol is 237 ~ 710g/L; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1 ~ 20g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 300 ~ 1000g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 20 ~ 800g/L.The present invention adopts the method for colorimetric analysis, operates easy rapidly, decreases the interference of heating to testing result; Detection reagent type used is less, is convenient to assembling and carries; Can develop the color to multiple grease sample and detect simultaneously, colour developing evenly; The equal low toxicity of detection reagent used is even nontoxic, safety and environmental protection.
Description
Technical field
The present invention relates to the mensuration of peroxide value, particularly relate to a kind of edible oil and fat determination of POV composite reagent and detection method.
Background technology
As everyone knows, edible oil and fat or oil-containing food, from production to consumer need certain hour, sometimes Long-term Storage is needed, and between storage or operating period, grease or oil-containing food are subject to the impact of oxygen, illumination etc. in air and carry out being oxidized to the reaction of main body, not only change on sense organ, as color and luster is deepened, produced unhappy smell (aldehyde material that the superoxide mainly generated easily partially decomposes forming causes), and nutrition deterioration can be made, deterioration degree seriously even can produce noxious material, threatens the healthy of consumer.
The method being at present detected object with Oxidation of Fat and Oils product mainly contain following these:
Iodimetric titration is the standard method that superoxide detects, its principle is in acid medium, superoxide and iodide ion react and generate iodine, then with the iodine that sodium thiosulfate standard solution titration is separated out, and according to consuming the volume of sodium thiosulfate standard solution and calculate the peroxide value of grease.This method complex operation, sensitivity is low, and sampling amount is large, and testing cost is high, is only applicable to the mensuration containing more than 0.2mEq/kg superoxide, simultaneously easy in environment.
Thiobarbituricacidα-method is the method for the Oxidation of Fat and Oils detection that laboratory is commonly used, and the method has the patent (patent No. JP51151191) of Test paper.MDA is one of primary product in the many unsaturated aldehyde ketone product of Oxidation of Fat and Oils generation, and the red compound that it and thiobarbituricacidα-generate, has absorption maximum at 530nm place, measures the absorbance of colored substance, carrys out the degree of oxidation of measure oil with this.The shortcoming of the method is mainly: because heating makes testing result have error.
Thiocyanate-ferric, its principle is that ferrous ion can be become ferric ion by peroxide oxidation in acid condition, when adding thiocyanate ion, then form orange-red complex compound with ferric ion, in 480-515nm, there is obtained the maximum absorption, the content of superoxide can be measured by colorimetric.But in the method, methyl alcohol mix reagent dissolves oil sample, is harmful to human body and contaminated environment.
A kind of edible oil determination method of peroxide value based on conductivity (number of patent application CN201210324117.6), its principle takes a certain amount of edible oil sample, add a certain amount of isooctane and concentrated hydrochloric acid, mix, add saturated solution of potassium iodide, after vibration, add a certain amount of distilled water diluting certain proportion immediately, time of repose after vibration, measures the conductivity value of mixing aqueous phase with conductivity meter.These class methods require high to operating personnel, equipment complexity, high cost, be unfavorable for promoting the use of simultaneously.
In sum, all there is larger difference in the aspects such as the sensitivity of various detection method, degree of accuracy, the simplified and traditional degree of detection and cost, thus set up new simple and easy to do, highly sensitive, accuracy is good, testing conditions is gentle, with low cost, few green test method consuming time is still an urgent demand of society and important goal in research.
Summary of the invention
The present invention overcomes above-mentioned weak point, and object is to provide a kind of edible oil and fat determination of POV composite reagent, and the use of composite reagent is simple, highly sensitive, greatly reduces costs, nuisanceless to human body.
Another object of the present invention is to provide a kind of edible oil and fat determination method of peroxide value, based on to color method principle, detected by the peroxide value of composite reagent to testing sample, simple and efficient to handle, accuracy rate is high, again Environmental Safety.
The present invention achieves the above object by the following technical programs: a kind of edible oil and fat determination of POV composite reagent, comprising: polar organic solvent, reductive agent, chromogenic reagent, color stability agent;
Described polar organic solvent is remembered by mass percentage, and the mass concentration comprising ethanol is 237 ~ 710g/L;
Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1 ~ 20g/L;
Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 300 ~ 1000g/L;
Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 20 ~ 800g/L.
A kind of edible oil and fat determination method of peroxide value, comprises the following steps:
(1) liquid testing sample is pipetted with liquid-transfering gun, the sample volume of described testing sample is 20 ~ 40 μ L, record quality simultaneously, add the mixed liquor of polar organic solvent and color stability agent, the volume ratio of described testing sample and polar organic solvent is 1:15 ~ 1:25, the volume ratio of color stability agent and polar organic solvent is 1:30 ~ 1:40, and eddy current shakes;
(2) in the mixed liquor of step (1), add reductive agent, the reductive agent volume of every gram of testing sample dropping is 5 ~ 12mL, and eddy current shakes, left at room temperature;
(3) in the solution of step (2), add developer, the developer volume that every gram of testing sample drips is 0.01 ~ 2mL, and solution is with the rotating speed centrifugal rotation 10 minutes of more than 10000r/min;
(4) get step (3) and obtain the supernatant fraction of mixed solution at 500 ~ 550nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from ferric ion canonical plotting, and the iron concentration being converted into unit mass is as peroxide value.
As preferably, described testing sample is liquid.
The making of described ferric ion canonical plotting comprises the following steps:
A reduced iron powder is added hydrochloric acid and hydrogen peroxide dissolving by () after, heating removing hydrogen peroxide, after cooling, dilute with water mixes, the iron standard reserving solution of obtained 10mg/mL;
B solution that () utilizes step (a) obtained with the method for distilled water diluting obtain iron concentration be 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5, a series of gradient solutions of 4.0mg/mL, to add after described polar solvent and color stability agent concussion respectively 30 seconds;
Concussion reaction in 30 seconds 3 minutes after adding reductive agent in c each solution that () obtains in step (b), then add developer;
D each solution that step (c) obtains by () centrifugal 10 minutes with the rotating speed of more than 10000r/min is that the reaction tube of 0 is for measuring absorbance with reference at 500 ~ 550nm place with iron concentration;
E () for horizontal, ordinate curve plotting, and obtains the straight-line equation of matching with absorbance, iron concentration respectively by least square method.
Beneficial effect of the present invention is: the present invention adopts the method for colorimetric analysis, and grease sample and composite reagent react at normal temperatures and develop the color, and operates easy rapidly, and decrease the interference of heating to testing result, testing result intuitive is good; Detection reagent type used is less, is convenient to assembling and carries; Can develop the color to multiple grease sample and detect simultaneously, colour developing be evenly, convenient and swift, is easy to Site Detection; The equal low toxicity of detection reagent used is even nontoxic, safety and environmental protection.
Embodiment
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:
Embodiment 1: the composite reagent of the present embodiment is polar organic solvent, reductive agent, chromogenic reagent, color stability agent; Described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
First, need to make ferric ion canonical plotting.The making of described ferric ion canonical plotting comprises the following steps:
A reduced iron powder is added hydrochloric acid and hydrogen peroxide dissolving by () after, heating removing hydrogen peroxide, after cooling, dilute with water mixes, the iron standard reserving solution of obtained 10mg/mL;
B solution that () utilizes step (a) obtained with the method for distilled water diluting obtain iron concentration be 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5, a series of gradient solutions of 4.0mg/mL, add the polar solvent of 350 μ L and the color stability agent of 10 μ L respectively, shake 30 seconds;
Concussion reaction in 30 seconds 3 minutes after adding reductive agent 150 μ L in c each solution that () obtains in step (b), then add chromogenic reagent 10 μ L;
D each solution that step (c) obtains by () centrifugal 10 minutes with the rotating speed of more than 10000r/min is that the reaction tube of 0 is for measuring absorbance with reference at 510nm place with iron concentration;
E () for horizontal, ordinate curve plotting, and obtains the straight-line equation of matching with absorbance, iron concentration respectively by least square method.
The sampling grease of the present embodiment is ground sesameseed oil, and a kind of edible oil and fat determination method of peroxide value, specifically comprises the following steps:
(1) pipette ground sesameseed oil 20 μ L with liquid-transfering gun, record quality, add the mixed liquor of polar organic solvent 350 μ L and color stability agent 10 μ L, eddy current shakes 30 seconds;
(2) in the mixed liquor of step (1), add reductive agent 150 μ L, eddy current shakes 30 seconds, left at room temperature 3 minutes;
(3) add chromogenic reagent 10 μ L in the solution of step (2) after, with the rotating speed centrifugal rotation 10 minutes of more than 10000r/min;
(4) get step (3) and obtain the supernatant fraction of mixed solution at 510nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from ferric ion canonical plotting, and the iron concentration being converted into unit mass is as peroxide value.
Finally measuring the POV value obtaining ground sesameseed oil is 4.820mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.087mmol/L.
Embodiment 2: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 30%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 20g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is corn oil, and pipette corn oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining corn oil is 1.051mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.094mmol/L.
Embodiment 3: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 90%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is rapeseed oil, and pipette rapeseed oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining rapeseed oil is 1.119mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.121mmol/L.
Embodiment 4: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 1000g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is soybean oil, and pipette soybean oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining soybean oil is 0.960mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.134mmol/L.
Embodiment 5: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 20g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 300g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is peanut oil, and pipette peanut oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining peanut oil is 1.910mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.100mmol/L.
Embodiment 6: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 1000g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 200g/L.
The sampling grease of the present embodiment is cereal blending stock, and pipette cereal blending stock 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining cereal blending stock is 0.920mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.115mmol/L.
Embodiment 7: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 300g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 800g/L.
The sampling grease of the present embodiment is olive oil, and pipette olive oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining olive oil is 5.47mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.114mmol/L.
Embodiment 8: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 800g/L.
The sampling grease of the present embodiment is oil residues in dace with black bean can, and pipette oil residues 20 μ L in dace with black bean can with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining oil residues in dace with black bean can is 2.960mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.099mmol/L.
Embodiment 9: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The miscella that the sampling grease of the present embodiment is peanut oil, rapeseed oil, olive oil, ground sesameseed oil, cereal blending stock, soybean oil, corn oil mix by equal proportion, pipette miscella 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining miscella is 7.730mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.087mmol/L.
Embodiment 10: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is fish oil, and pipette fish oil 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining fish oil is 8.920mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.087mmol/L.
Embodiment 11: in the present embodiment, described polar organic solvent is number percent note by volume, comprises the ethanol of 70%; Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 528g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225g/L.
The sampling grease of the present embodiment is lard, and pipette lard 20 μ L with liquid-transfering gun, other embodiments are identical with embodiment 1.
Finally measuring the POV value obtaining lard is 3.810mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity obtaining described method is 0.087mmol/L.
The know-why being specific embodiments of the invention and using described in above, if the change done according to conception of the present invention, its function produced do not exceed that instructions contains yet spiritual time, must protection scope of the present invention be belonged to.
Claims (3)
1. an edible oil and fat determination of POV composite reagent, is characterized in that comprising: polar organic solvent, reductive agent, chromogenic reagent, color stability agent;
Described polar organic solvent is remembered by mass percentage, and the mass concentration comprising ethanol is 237 ~ 710g/L;
Described reductive agent is remembered by weight percentage, and the mass concentration comprising ferrous sulphate is 1 ~ 20g/L;
Described chromogenic reagent is sulfosalisylic acid solution, and mass concentration is 300 ~ 1000g/L;
Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 20 ~ 800g/L.
2. an edible oil and fat determination method of peroxide value, is characterized in that comprising the following steps:
(1) liquid testing sample is pipetted with liquid-transfering gun, the sample volume of described testing sample is 20 ~ 40 μ L, record quality simultaneously, add the mixed liquor of polar organic solvent and color stability agent, the volume ratio of described testing sample and polar organic solvent is 1:15 ~ 1:25, the volume ratio of color stability agent and polar organic solvent is 1:30 ~ 1:40, and eddy current shakes;
(2) in the mixed liquor of step (1), add reductive agent, the reductive agent volume of every gram of testing sample dropping is 5 ~ 12mL, and eddy current shakes, left at room temperature;
(3) in the solution of step (2), add developer, the developer volume that every gram of testing sample drips is 0.01 ~ 2mL, and solution is with the rotating speed centrifugal rotation 10 minutes of more than 10000r/min;
(4) get step (3) and obtain the supernatant fraction of mixed solution at 500 ~ 550nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from ferric ion canonical plotting, and the iron concentration being converted into unit mass is as peroxide value.
3. a kind of edible oil and fat determination method of peroxide value according to claim 2, is characterized in that, the making of described ferric ion canonical plotting comprises the following steps:
A reduced iron powder is added hydrochloric acid and hydrogen peroxide dissolving by () after, heating removing hydrogen peroxide, after cooling, dilute with water mixes, the iron standard reserving solution of obtained 10mg/mL;
B solution that () utilizes step (a) obtained with the method for distilled water diluting obtain iron concentration be 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5, a series of gradient solutions of 4.0mg/mL, to add after described polar solvent and color stability agent concussion respectively 30 seconds;
Concussion reaction in 30 seconds 3 minutes after adding reductive agent in c each solution that () obtains in step (b), then add developer;
D each solution that step (c) obtains by () centrifugal 10 minutes with the rotating speed of more than 10000r/min is that the reaction tube of 0 is for measuring absorbance with reference at 500 ~ 550nm place with iron concentration;
E () for horizontal, ordinate curve plotting, and obtains the straight-line equation of matching with absorbance, iron concentration respectively by least square method.
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| CN103776976B (en) * | 2014-01-17 | 2015-08-05 | 广东美味鲜调味食品有限公司 | A kind of peanut oil stabilization of peroxide values quick judgment method |
| CN104515770A (en) * | 2014-12-09 | 2015-04-15 | 苏州东辰林达检测技术有限公司 | Waste oil rapid detection kit and use method thereof |
| CN106198419B (en) * | 2016-08-03 | 2018-12-07 | 哈尔滨普凡农牧有限公司 | Detect the reagent and method of alpha, beta-unsaturated aldehyde content in oil plant or grease |
| CN106383119B (en) * | 2016-11-08 | 2019-01-01 | 自贡勃生表面技术推广有限公司 | Quickly detect the photometry of peroxide concentrations in edible oil |
| CN107831126A (en) * | 2017-12-14 | 2018-03-23 | 广州傲农生物科技有限公司 | A kind of assay method of oil peroxidation value and its application |
| CN109283161B (en) * | 2018-09-19 | 2021-04-02 | 青岛大学 | Method for detecting peroxide value in grease by using fluorescence quenching method |
| CN110687108B (en) * | 2019-10-17 | 2022-05-06 | 山东瑞思康生物科技有限公司 | Multilayer film dry chemical reagent strip for quantitatively detecting urine malondialdehyde |
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| CN102706872A (en) * | 2012-06-13 | 2012-10-03 | 安徽省农业科学院农产品加工研究所 | Combined reagent for detecting peroxide value of edible oil and fat and detection method thereof |
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| CN1540319A (en) * | 2003-04-27 | 2004-10-27 | 莫庆奎 | Method for testing peroxide value of liquid edible oil |
| CN102706872A (en) * | 2012-06-13 | 2012-10-03 | 安徽省农业科学院农产品加工研究所 | Combined reagent for detecting peroxide value of edible oil and fat and detection method thereof |
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