CN103245660A - Edible oil peroxide value measurement combined reagent and detection method - Google Patents
Edible oil peroxide value measurement combined reagent and detection method Download PDFInfo
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Abstract
The invention relates to measurement of peroxide values, and in particular relates to an edible oil peroxide value measurement combined reagent and a detection method. The edible oil peroxide value measurement combined reagent comprises a polarity organic solvent, a reducing agent, a color development reagent and a color development stabilizing agent, wherein the polarity organic solvent comprises ethanol with the mass concentration of 237-710 g/L in percentage by mass; the reducing agent comprises ferrous sulfate with the mass concentration of 1-20 g/L in percentage by weight; the color development reagent is a sulfosalicylic acid solution with the mass concentration of 300-1,000 g/L; and the color development stabilizing agent is a pyridoxamine dihydrochloride solution with the mass concentration of 20-800 g/L. By a colorimetric analysis method, the operation is quick and simple, and interference caused by heating on a detection result is reduced; the types of the needed detection reagents is few, and the reagents are convenient to combine and carry; and the edible oil peroxide value measurement combined reagent can develop and detect a plurality of oil samples at the same time, is uniform in developing, is low in toxicity and even non-toxic, and is safe and environmentally-friendly.
Description
Technical field
The present invention relates to the mensuration of peroxide value, relate in particular to a kind of edible oil and fat peroxide value and measure composite reagent and detection method.
Background technology
As everyone knows, edible oil and fat or oil-containing food, from producing to the consumer need certain hour, sometimes need Long-term Storage, and between storage or operating period, grease or oil-containing food are subject to the influence of airborne oxygen, illumination etc. and carry out being oxidized to the reaction of main body, not only change at sense organ, deepen, produce unhappy smell (mainly being that the aldehyde material that the superoxide that generates easily partially decomposes forming causes) as color and luster, and can make the bad change of nutrition, bad range degree is serious even can produce noxious material, threatens the healthy of consumer.
At present with the Oxidation of Fat and Oils product be the method for detected object mainly contain following these:
Iodimetric titration is the standard method that superoxide detects, its principle is in acid medium, the reaction of superoxide and iodide ion generates iodine, the iodine of separating out with the sodium thiosulfate standard solution titration again, and calculate the peroxide value of grease according to the volume of consumption sodium thiosulfate standard solution.This method complex operation, sensitivity is low, and sampling amount is big, and the testing cost height is only applicable to contain the mensuration of the above superoxide of 0.2mEq/kg, easily environment is polluted simultaneously.
The thiobarbituricacid method is the method that laboratory Oxidation of Fat and Oils commonly used detects, the existing patent (patent No. JP51151191) that detects test paper of the method.MDA is one of primary product in the many unsaturated aldehyde ketone product that produces of Oxidation of Fat and Oils, and there is absorption maximum it and the red compound that thiobarbituricacid generates at the 530nm place, and the absorbance of mensuration colored substance comes the degree of oxidation of measure oil with this.The shortcoming of this method is mainly: because heating makes testing result error is arranged.
Thiocyanate-ferric, its principle is that ferrous ion can be become ferric ion by peroxide oxidation under acid condition, when adding thiocyanate ion, then form orange-red complex compound with ferric ion, in 480-515 nm, obtained the maximum absorption is arranged, can measure the content of superoxide by colorimetric.But in this method, methyl alcohol mix reagent dissolving 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 is to take by weighing 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 the vibration, add a certain amount of distilled water diluting certain proportion immediately, vibration back time of repose is measured the conductivity value that mixes water with conductivity meter.These class methods require height to operating personnel, and simultaneously equipment complexity, cost are too high, are unfavorable for promoting the use of.
In sum, all there is larger difference in aspects such as the sensitivity of various detection methods, degree of accuracy, the simplified and traditional degree of detection and cost, thereby set up new simple and easy to do, highly sensitive, accuracy is good, the testing conditions gentleness, 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 purpose is to provide a kind of edible oil and fat peroxide value to measure composite reagent, and the use of composite reagent is simple, highly sensitive, reduces cost greatly, and is 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 the color method principle, detect by the peroxide value of composite reagent to testing sample, simple and efficient to handle, accuracy rate height, Environmental Safety again.
The present invention achieves the above object by the following technical programs: a kind of edible oil and fat peroxide value is measured composite reagent, comprising: polar organic solvent, reductive agent, chromogenic reagent, color stability agent;
Described polar organic solvent remembers that by mass percentage the mass concentration that comprises ethanol is 237～710g/L;
Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 1～20g/L;
Described chromogenic reagent is sulfosalicylic acid solution, and mass concentration is 300～1000 g/L;
Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 20～800 g/L.
A kind of edible oil and fat determination method of peroxide value may further comprise the steps:
(1) pipettes liquid testing sample with liquid-transfering gun, the sample volume of described testing sample is 20～40 μ L, note quality simultaneously, the mixed liquor that adds 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 polar organic solvent and color stability agent is 1:30～1:40, the eddy current concussion;
(2) add reductive agent in the mixed liquor of step (1), the reductive agent volume that every gram testing sample drips is 5～12mL, and the eddy current concussion is left standstill under the room temperature;
(3) add developer in the solution of step (2), the developer volume that every gram testing sample drips is 0.1～2mL, and solution was with the centrifugal rotation of the rotating speed more than the 10000r/min 10 minutes;
(4) get supernatant part that step (3) obtains mixed solution at 500～550nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from the ferric ion canonical plotting, and the iron concentration that is converted into unit mass is as peroxide value.
As preferably, described testing sample is liquid.
The making of described ferric ion canonical plotting may further comprise the steps:
(a) reduced iron powder is added the dissolving of hydrochloric acid and hydrogen peroxide after, hydrogen peroxide is removed in heating, cooling back dilute with water mixing makes the iron standard reserving solution of 10mg/mL;
(b) utilize solution that step (a) makes to obtain a series of gradient solutions that iron concentration is 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5,4.0 mg/mL with the method for distilled water diluting, added after described polar solvent and the color stability agent concussion respectively 30 seconds;
(c) in each solution that step (b) obtains, shake reaction in 30 seconds 3 minutes behind the adding reductive agent, add developer again;
(d) each solution that step (c) is obtained is with the rotating speed more than the 10000r/min centrifugal 10 minutes, is that 0 reaction tube serves as with reference at 500～550nm place mensuration absorbance with iron concentration;
(e) be horizontal stroke, ordinate curve plotting with absorbance, iron concentration respectively, and obtain the straight-line equation of match with 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 easyly rapidly, has reduced heating to the interference of testing result, and the testing result intuitive is good; Used detection reagent type is less, is convenient to assembling and carries; Can simultaneously a plurality of grease samples be developed the color and detect, colour developing evenly, and is convenient and swift, is easy to on-the-spot the detection; The equal low toxicity of used detection reagent even nontoxic, safety and environmental protection.
Embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1: the composite reagent of present embodiment is polar organic solvent, reductive agent, chromogenic reagent, color stability agent; Described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic acid solution, and mass concentration is 528 g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225 g/L.
At first, need to make the ferric ion canonical plotting.The making of described ferric ion canonical plotting may further comprise the steps:
(a) reduced iron powder is added the dissolving of hydrochloric acid and hydrogen peroxide after, hydrogen peroxide is removed in heating, cooling back dilute with water mixing makes the iron standard reserving solution of 10mg/mL;
(b) utilize solution that step (a) makes to obtain a series of gradient solutions that iron concentration is 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5,4.0 mg/mL with the method for distilled water diluting, add the color stability agent of polar solvent and the 10 μ L of 350 μ L respectively, shook 30 seconds;
(c) in each solution that step (b) obtains, shake reaction in 30 seconds 3 minutes behind the adding reductive agent 150 μ L, add chromogenic reagent 10 μ L again;
(d) each solution that step (c) is obtained is with the rotating speed more than the 10000r/min centrifugal 10 minutes, is that 0 reaction tube serves as with reference at 510nm place mensuration absorbance with iron concentration;
(e) be horizontal stroke, ordinate curve plotting with absorbance, iron concentration respectively, and obtain the straight-line equation of match with least square method.
The sampling grease of present embodiment is ground sesameseed oil, and a kind of edible oil and fat determination method of peroxide value specifically may further comprise the steps:
(1) pipettes ground sesameseed oil 20 μ L with liquid-transfering gun, note quality, add the mixed liquor of polar organic solvent 350 μ L and color stability agent 10 μ L, eddy current concussion 30 seconds;
(2) add reductive agent 150 μ L in the mixed liquor of step (1), eddy current concussion 30 seconds was left standstill under the room temperature 3 minutes;
(3) in the solution of step (2), add chromogenic reagent 10 μ L after, with the centrifugal rotation of the rotating speed more than the 10000r/min 10 minutes;
(4) get supernatant part that step (3) obtains mixed solution at 510nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from the ferric ion canonical plotting, and the iron concentration that is converted into unit mass is as peroxide value.
Measure the POV value that obtains ground sesameseed oil at last and be 4.820mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.087mmol/L.
Embodiment 2: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 30% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 20g/L; Described chromogenic reagent is sulfosalicylic acid solution, and mass concentration is 528 g/L; Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 225 g/L.
The sampling grease of present embodiment is corn oil, pipettes corn oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains corn oil at last and be 1.051mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.094mmol/L.
Embodiment 3: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 90% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is rapeseed oil, pipettes rapeseed oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains rapeseed oil at last and be 1.119mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.121mmol/L.
Embodiment 4: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is soybean oil, pipettes soybean oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains soybean oil at last and be 0.960mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.134mmol/L.
Embodiment 5: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 20g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is peanut oil, pipettes peanut oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains peanut oil at last and be 1.910mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.100mmol/L.
Embodiment 6: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is the cereal blending stock, pipettes cereal blending stock 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains the cereal blending stock at last and be 0.920mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.115mmol/L.
Embodiment 7: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is olive oil, pipettes olive oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains olive oil at last and be 5.47mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.114mmol/L.
Embodiment 8: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 1g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is oil residues in the dace with black bean can, pipettes oil residues 20 μ L in the dace with black bean can with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains oil residues in the dace with black bean can at last and be 2.960mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.099mmol/L.
Embodiment 9: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is the miscella that peanut oil, rapeseed oil, olive oil, ground sesameseed oil, cereal blending stock, soybean oil, corn oil mix by equal proportion, pipettes miscella 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains miscella at last and be 7.730mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.087mmol/L.
Embodiment 10: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is fish oil, pipettes fish oil 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains fish oil at last and be 8.920mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.087mmol/L.
Embodiment 11: in the present embodiment, described polar organic solvent is the number percent note by volume, comprises 70% ethanol; Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 3.6g/L; Described chromogenic reagent is sulfosalicylic 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 present embodiment is lard, pipettes lard 20 μ L with liquid-transfering gun, and other embodiments are identical with embodiment 1.
Measure the POV value that obtains lard at last and be 3.810mmol/Kg.
With ethanol stepwise dilution oil sample, the sensitivity that obtains described method is 0.087mmol/L.
Above described be specific embodiments of the invention and the know-why used, if the change of doing according to conception of the present invention, when its function that produces does not exceed spiritual that instructions contains yet, must belong to protection scope of the present invention.
Claims (4)
1. an edible oil and fat peroxide value is measured composite reagent, it is characterized in that comprising: polar organic solvent, reductive agent, chromogenic reagent, color stability agent;
Described polar organic solvent remembers that by mass percentage the mass concentration that comprises ethanol is 237～710g/L;
Described reductive agent remembers that by weight percentage the mass concentration that comprises ferrous sulphate is 1～20g/L;
Described chromogenic reagent is sulfosalicylic acid solution, and mass concentration is 300～1000 g/L;
Described color stability agent is hydrochloric acid pyridoxamine solution, and mass concentration is 20～800 g/L.
2. edible oil and fat determination method of peroxide value is characterized in that may further comprise the steps:
(1) pipettes liquid testing sample with liquid-transfering gun, the sample volume of described testing sample is 20～40 μ L, note quality simultaneously, the mixed liquor that adds 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 polar organic solvent and color stability agent is 1:30～1:40, the eddy current concussion;
(2) add reductive agent in the mixed liquor of step (1), the reductive agent volume that every gram testing sample drips is 5～12mL,
The eddy current concussion is left standstill under the room temperature;
(3) add developer in the solution of step (2), the developer volume that every gram testing sample drips is 0.1～2mL, and solution was with the centrifugal rotation of the rotating speed more than the 10000r/min 10 minutes;
(4) get supernatant part that step (3) obtains mixed solution at 500～550nm place mensuration absorbance;
(5) obtain corresponding iron concentration according to absorbance from the ferric ion canonical plotting, and the iron concentration that is 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, described testing sample is liquid.
4. 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 may further comprise the steps:
(a) reduced iron powder is added the dissolving of hydrochloric acid and hydrogen peroxide after, hydrogen peroxide is removed in heating, cooling back dilute with water mixing makes the iron standard reserving solution of 10mg/mL;
(b) utilize solution that step (a) makes to obtain a series of gradient solutions that iron concentration is 0,0.2,0.5,1,1.5,2.0,2.5,3.0,3.5,4.0 mg/mL with the method for distilled water diluting, added after described polar solvent and the color stability agent concussion respectively 30 seconds;
(c) in each solution that step (b) obtains, shake reaction in 30 seconds 3 minutes behind the adding reductive agent, add developer again;
(d) each solution that step (c) is obtained is with the rotating speed more than the 10000r/min centrifugal 10 minutes, is that 0 reaction tube serves as with reference at 500～550nm place mensuration absorbance with iron concentration;
(e) be horizontal stroke, ordinate curve plotting with absorbance, iron concentration respectively, and obtain the straight-line equation of match with least square method.
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| CN103776976A (en) * | 2014-01-17 | 2014-05-07 | 广东美味鲜调味食品有限公司 | Fast judging method for stability of peanut oil peroxide value |
| 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 |
| CN106198419A (en) * | 2016-08-03 | 2016-12-07 | 哈尔滨普凡农牧有限公司 | Detect reagent and the method for α, β unsaturated aldehyde content in oil plant or oils and fats |
| CN106383119A (en) * | 2016-11-08 | 2017-02-08 | 自贡勃生表面技术推广有限公司 | Spectrophotometry of rapidly detecting concentration of peroxide 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 |
| CN109283161A (en) * | 2018-09-19 | 2019-01-29 | 青岛大学 | A kind of method using peroxide value in fluorescent quenching method detection grease |
| CN110687108A (en) * | 2019-10-17 | 2020-01-14 | 山东瑞思康生物科技有限公司 | Multilayer film dry chemical reagent strip for quantitatively detecting urine malondialdehyde |
| WO2024130740A1 (en) * | 2022-12-23 | 2024-06-27 | 江南大学 | Color-developing hydrogel for testing freshness of vegetable oil, and preparation method therefor and use thereof |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776976A (en) * | 2014-01-17 | 2014-05-07 | 广东美味鲜调味食品有限公司 | Fast judging method for stability of peanut oil peroxide value |
| 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 |
| CN106198419A (en) * | 2016-08-03 | 2016-12-07 | 哈尔滨普凡农牧有限公司 | Detect reagent and the method for α, β unsaturated aldehyde content in oil plant or oils and fats |
| 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 |
| CN106383119A (en) * | 2016-11-08 | 2017-02-08 | 自贡勃生表面技术推广有限公司 | Spectrophotometry of rapidly detecting concentration of peroxide in edible oil |
| 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 |
| CN109283161A (en) * | 2018-09-19 | 2019-01-29 | 青岛大学 | A kind of method using peroxide value in fluorescent quenching method detection grease |
| CN110687108A (en) * | 2019-10-17 | 2020-01-14 | 山东瑞思康生物科技有限公司 | Multilayer film dry chemical reagent strip for quantitatively detecting urine malondialdehyde |
| WO2024130740A1 (en) * | 2022-12-23 | 2024-06-27 | 江南大学 | Color-developing hydrogel for testing freshness of vegetable oil, and preparation method therefor and use thereof |
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