CN109632446A - The Sample pretreatment method of acid value and the detection method using it in a kind of measurement food - Google Patents
The Sample pretreatment method of acid value and the detection method using it in a kind of measurement food Download PDFInfo
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- CN109632446A CN109632446A CN201910138204.4A CN201910138204A CN109632446A CN 109632446 A CN109632446 A CN 109632446A CN 201910138204 A CN201910138204 A CN 201910138204A CN 109632446 A CN109632446 A CN 109632446A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/3103—Atomic absorption analysis
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Abstract
The present invention provides a kind of Sample pretreatment methods of acid value in measurement food, including with n-hexane dissolution food samples, after standing, take supernatant, dehydrated alcohol is added, add sodium hydroxide and phenolphthalein, mix to be checked.The Sample pretreatment method that the present invention measures acid value in food uses non-toxic or low-toxic reagent, and Sample pretreatment process is simple, smaller to operator and environmental hazard.The present invention also provides a kind of methods of food samples acid value with the above-mentioned Sample pretreatment method processing of spectrophotometric determination.Detection method is simple, and quickly, and detection sensitivity and accuracy are higher.
Description
Technical field
The present invention relates to a kind of Sample pretreatment method of acid value in measurement food and using its detection method, belong to point
Analyse detection field.
Background technique
Acid value of lipids is also known as " acid value of oil and fat ".The standard of free fatty acid content in grease.Under conditions of fat production,
Acid value can be used as the index of hydrolysis degree, under conditions of its preservation, then can be used as rancid index.Free fatty acid in grease
Content it is more, then acid value is high, and content is low, then acid value is low, and acid value is lower, illustrates that oil quality is better, freshness and refinement
Better.Therefore acid value of lipids is the important indicator of edible oil quality evaluation, it reflects oil quality, oil and fat refining degree and storage
Hide quality comparison.Currently, China's rice bran crude oil acid value requires to be not more than 25 mg/g, palm, corn, olive, cottonseed, coconut are former
Oil requires to be not more than 10 mg/g, and other plant crude oil is not more than 4 mg/g, and edible vegetable oil (including ready-mixed oil) is not more than 3
Mg/g, the edible vegetable oil during frying are not more than 5 mg/g, and edible animal fatty acid valence is not more than 2.5 mg/g.
The method of the acid value of lipids of national standard detection at present is three kinds of titrations: the first is cold solvent indicator titration method, should
Grease sample is dissolved into sample solution with organic solvent by method, then in potassium hydroxide or Standard Volumetric Solutions for Sodium Hydroxide and dripping
Determine the free fatty acid in sample solution, titration end-point is determined with indicator corresponding color change, eventually eventually by titration
The standard titration solution of consumption is put to determine acid value.Second is hot ethanol indicator titration method, the method and first method class
Seemingly, the difference is that being heated to 70 DEG C with ethyl alcohol makes lipid solubilization.The first, second method can all use petroleum ether or second
The toxic reagents such as ether not only can generate harm to operator's body, also pollute the environment, and the more difficult judgement of titration end-point,
The especially deeper oil of color, such as red chilli oil, chafing dish red oil, titration end-point is blush, and titration error is larger.Third
It kind is potentiometric titration, which is first dissolved into sample solution for grease sample with organic solvent, then with potassium hydroxide or hydroxide
Sodium standard titration solution carries out constant-current titration, occurs caused by neutralization reaction " pH jumping " to be according to judgement with free fatty acid
Titration end-point calculates the acid value of grease sample finally by the volume of the standard solution of titration end-point consumption, but there is still a need for having
Malicious petroleum ether, entire detection is time-consuming also very long, and national standard method and existing detection method predominantly detect vegetable oil, test sample
Range is smaller.
Summary of the invention
The object of the present invention is to provide a kind of Sample pretreatment methods of acid value in measurement food, avoid conventional titration mistake
Reagent ether, isopropanol for being more toxic used in journey etc., agents useful for same of the present invention is non-toxic or low-toxic reagent, and it can
To be suitable for the quick detection of large amounts of food oil product or oil product seasoning.
The Sample pretreatment method of acid value in a kind of measurement food provided by the invention, including with n-hexane dissolution food sample
This, after standing, takes supernatant, dehydrated alcohol is added, add sodium hydroxide and phenolphthalein, mix to be checked.
A kind of Sample pretreatment method of acid value in measurement food, comprising the following steps:
1) edible oil sample: weighing 1 g of liquid edible oil and be placed in 2 mL polystyrene centrifuge tubes, and it is molten that 1 mL n-hexane is added
Solution stands 1 ~ 2 h after mixing, take supernatant after the clarification of sample liquid.If edible oil sample is in solidification state, then heating is needed to melt it
Change, then weighs.
Grease-contained solid food sample: taking solid sample to be smashed to pieces with tissue mashing machine, weighs the sample after 1 g is smashed to pieces and sets
In the polystyrene centrifuge tube of 5 mL, 1 mL of n-hexane is added, be vortexed 0.5 ~ 1 min, and 3000 r/min are centrifuged 3 min, take
Supernatant；
2) it pipettes 2.5 mL of dehydrated alcohol to have in plug polystyrene centrifuge tube in 5 mL, adds 50 μ of supernatant obtained by step 1)
L adds 50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent, closes the lid, and mixes, to be checked.
It is a further object to provide a kind of methods for measuring edible oil acid value, are with spectrophotometer method pair
With above-mentioned Sample pretreatment method, treated that sample is detected comprising following steps:
1) acid value determination Specification Curve of Increasing: the standard of National Standard Method of learning from else's experience measurement uses solution, is added to anhydrous with 2.5 mL
In the 5 mL tool plug polystyrene centrifuge tube of ethyl alcohol, 50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent are added, is closed the lid
Son shakes up, and 0.5 mL of the solution after shaking up is taken to be placed in 1 cm cuvette, and absorbance value is measured at 490 ~ 580 nm wavelength, with
The acid value of series standard solution is ordinate, using the absorbance value of measurement as abscissa, draws curve；
2) sample to be tested alternative steps 1 measurement of sample to be tested acid value: are taken) in standard using solution, remaining detecting step phase
Together, the absorbance value for measuring sample to be tested, by absorbance value and above-mentioned steps 1) draw standard curve be compared, calculate to
The acid value of test sample sheet.
Further, it is preferable that acid value determination Specification Curve of Increasing specifically includes the following steps:
It takes the series standard that National Standard Method measurement acid value is 0.4,0.8,1.0,1.5,2.0,3.0,3.5 mg/g using solution, is added
Into the 5 mL tool plug polystyrene centrifuge tube with 2.5 mL dehydrated alcohols, 50 μ L of sodium hydroxide reagent, phenolphthalein examination are added
50 μ L of agent, closes the lid, shakes up, and 0.5 mL of the solution after shaking up is taken to be placed in 1 cm cuvette, measures and inhales at 560 nm wavelength
Shading value, using the absorbance value of measurement as abscissa, is drawn curve, is obtained using the acid value of series standard solution as ordinate
Calibration curve equation: y=- 6.22x+4.67, y are acid value, and x is sample absorbance value, r2=0.997。
Beneficial effect
The present invention measures the Sample pretreatment method of edible oil acid value, and agents useful for same is nontoxic and hypotoxicity, operating method
Simply, to operator and environmental hazard very little.The present invention measures edible oil acid value using spectrophotometer method, in 560 nm
There is maximum absorbance, detection method is simple, and the detection used time is short, has highly sensitive and high accuracy, detects sensitive under wavelength
Degree is up to 0.07 mg/g, solves the problems, such as that conventional titration method dark oil titration end-point error in judgement is big.And present invention detection side
Method is suitable for any edible oil product or oil product seasoning, including but not limited to vegetable oil, animal oil, miscella, red oil, chafing dish bottom
The detection of material, oil product seasoning etc..
Detailed description of the invention
Fig. 1 canonical plotting of the present invention；
Influence of Fig. 2 medium wavelength of the present invention to chromogenic reaction absorbance.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described below with reference to embodiment, still
It should be appreciated that these descriptions are only further explanation the features and advantages of the present invention, rather than to the claims in the present invention
Limitation, reagent of the present invention are that analysis is pure unless otherwise instructed.
One Sample pretreatment of embodiment
Instrument, consumptive material: pipettor: 200 μ L, 50 μ L；Pipette: 10 mL, 1 mL；Electronic balance: 0.01 g of induction amount；It is double
Steam water；Polystyrene centrifuge tube: 2 mL, 5 mL；Tissue mashing machine；Visible spectrophotometer；Vortex instrument；Centrifuge.
Reagent: n-hexane is stored in Brown Glass Brown glass bottles and jars only.
Dehydrated alcohol is stored in Brown Glass Brown glass bottles and jars only.
Sodium hydroxide reagent: weighing 0.2 g sodium hydroxide and be dissolved in 100 mL distilled water, is stored in white plastic examination
Agent bottle.
Phenolphthalein reagent: it weighs 0.5 g phenolphthalein and is dissolved in 100 mL dehydrated alcohols, be stored in white plastic reagent bottle.
Sample pretreatment process
(1) edible oil sample: weighing 1 g(of edible oil if solid animal grease, then needing heating to melt it is liquid, then is claimed
Take) in 2 mL polystyrene centrifuge tubes, 1 mL n-hexane dissolution is added, 1 ~ 2 h is stood after shaking up, is taken after the clarification of sample liquid
Supernatant is for detecting.
Grease-contained solid food sample: sampling this and smashed to pieces with tissue mashing machine, weighs the sample after 1 g is smashed to pieces in 5 mL
Polystyrene centrifuge tube in, be added 1 mL of n-hexane, be vortexed 0.5 ~ 1 min, 3000 r/min be centrifuged 3 min, take supernatant
For detecting.
(2) pipette pipettes 2.5 mL of dehydrated alcohol in 5 mL polystyrene centrifuge tubes, adds on step (1) gained
50 μ L of clear liquid adds 50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent, closes the lid, shake up, to be checked.
Two edible oil acid value determination of embodiment
Edible oil acid value determination includes the following steps:
National Standard Method of learning from else's experience GB 5009.229-2016 measures acid value as 0.4,0.8,1.0,1.5,2.0,3.0,3.5 mg/g
Column standard uses solution, is added in the 5 mL tool plug polystyrene centrifuge tube with 2.5 mL dehydrated alcohols, adds hydrogen-oxygen
Change 50 μ L of sodium reagent, 50 μ L of phenolphthalein reagent, closes the lid, shake up.It is returned to zero with air, 0.5 mL of the solution after shaking up is taken to be placed in
1 cm cuvette measures absorbance value, using the acid value of series standard solution as ordinate, with the suction of measurement at 560 nm wavelength
Shading value is abscissa, draws curve, obtained calibration curve equation: y=- 6.22x+4.67, y are acid value, and x is sample extinction
Angle value, r2=0.997, as shown in Figure 1.
When measuring the acid value of edible oil sample, according to method described in embodiment one, by the solution after Sample pretreatment,
It is placed in 1 cm cuvette, absorbance value is measured at 560 nm wavelength, absorbance value and the standard curve of drafting are compared,
Calculate the acid value of sample to be tested.
When detection, if sample acid value is more than 3.5 mg/g, needs to increase extension rate and detected again later.
Influence of three wavelength of embodiment to chromogenic reaction absorbance
According to the method for embodiment two, influence of the different wave length to absorbance value is measured, as shown in Fig. 2, being at 560nm in wavelength
With maximum absorbance value.
Example IV difference sodium hydroxide, influence of the phenolphthalein additional amount to chromogenic reaction absorbance
According to the method for embodiment two, the influence of different sodium hydroxides, phenolphthalein sample-adding amount to absorbance value is measured, measures hydrogen respectively
200 μ L of sodium oxide molybdena sample-adding amount, 50 μ L of phenolphthalein sample-adding amount；Sodium hydroxide, phenolphthalein sample-adding amount are 200 μ L；Sodium hydroxide sample-adding amount
50 μ L, 200 μ L of phenolphthalein sample-adding amount, sodium hydroxide, phenolphthalein sample-adding amount are 50 μ L, and sodium hydroxide, phenolphthalein sample-adding amount are 100 μ
L is 0.778 the results show that having maximum absorbance value when sodium hydroxide, phenolphthalein sample-adding amount are 50 μ L.
Five edible oil acid value determination of embodiment
Learn from else's experience measure acid value be 0,0.07,0.15,0.3,0.5,0.8,1.0,1.5,2.0,3.0,4.0 mg/g series standard
Using solution, it is added in the 5 mL tool plug polystyrene centrifuge tube with 2.5 mL dehydrated alcohols, adds sodium hydroxide examination
50 μ L of agent, 50 μ L of phenolphthalein reagent, close the lid, shake up.It is returned to zero with air, 0.5 mL of the solution after shaking up is taken to be placed in 1 cm ratio
Color ware measures absorbance value, using the absorbance value of measurement as ordinate, with the acid for series standard solution at 560 nm wavelength
Valence is abscissa, draws curve, obtained calibration curve equation: y=- 0.15x+0.74, X are acid value, and Y is sample absorbance value,
r2=0.994。
When measuring the acid value of edible oil sample, according to method described in embodiment five, by the solution after Sample pretreatment,
It is placed in 1 cm cuvette, absorbance value is measured at 560 nm wavelength, absorbance value and the standard curve of drafting are compared,
Calculate the acid value of sample to be tested.
Experimental example
Accuracy experiment
Using the Sample pretreatment method of embodiment one, the acid value determination method of embodiment two, to 5 parts of random random samples into
Row processing measurement, while being surveyed with the measuring method of acid value in GB 5009.229-2016 national food safety standard food
It is fixed, it the results are shown in Table 1, sample of the present invention Detection accuracy range is 86%-117%.
The experiment of 1 sample of the present invention accuracy in detection of table
| Sample names | National Standard Method measures concentration (mg/g) | The present invention measures concentration (mg/g) | Accuracy rate |
| Chafing dish bottom flavorings | 0.91 | 0.78 | 86% |
| Rapeseed oil | 0.4 | 0.47 | 117% |
| Vinegar-pepper flavoring | 2.1 | 1.97 | 94% |
| Garlic aromatic material | 1.36 | 1.27 | 93% |
| Paste flavor condiment | 3.86 | 3.41 | 88% |
Precision Experiment
5 measurements, present invention detection side are carried out respectively to the rapeseed oil sample that acid value is 1.0,2.0 mg/g with the method for the present invention
The standard deviation of method is respectively 2.4%, 2.6%, and detection method precision is preferable.
The precision verifying of 2 sample of the present invention of table detection
| Measurement result mg/g | 1 | 2 | 3 | 4 | 5 | Average value | Standard deviation % |
| 1.0 | 1.01 | 0.98 | 1.00 | 1.02 | 0.96 | 0.99 | 2.4 |
| 2.0 | 2.02 | 2.04 | 1.97 | 2.01 | 2.02 | 2.01 | 2.6 |
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.To this
A variety of modifications of a little embodiments will be readily apparent to those skilled in the art, as defined herein general
Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.
Claims (5)
1. a kind of Sample pretreatment method of acid value in measurement food, including with n-hexane dissolution food samples, after standing, take
Dehydrated alcohol is added in clear liquid, adds sodium hydroxide and phenolphthalein, mixes to be checked.
2. measuring the Sample pretreatment method of acid value in food as described in claim 1, which is characterized in that including following step
It is rapid:
Edible oil sample: weighing 1 g of liquid edible oil sample sheet and be placed in 2 mL polystyrene centrifuge tubes, and it is molten that 1 mL n-hexane is added
Solution stands 1 ~ 2 h after mixing, take supernatant after the clarification of sample liquid, if edible oil sample is in solidification state, then heating is needed to melt it
Change, then weighs；
Grease-contained solid food sample: taking solid sample to be smashed to pieces with tissue mashing machine, weighs the sample after 1 g is smashed to pieces and is placed in 5
In the polystyrene centrifuge tube of mL, 1 mL of n-hexane is added, be vortexed 0.5 ~ 1 min, and 3000 r/min are centrifuged 3 min, take supernatant
Liquid；
2) it pipettes 2.5 mL of dehydrated alcohol to have in plug polystyrene centrifuge tube in 5 mL, adds 50 μ of supernatant obtained by step 1)
L adds 50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent, closes the lid, and mixes, to be checked.
3. a kind of method of acid value in measurement food, it is characterised in that: pass through the described in any item measurement food of claim 1 ~ 2
The Sample pretreatment method of middle acid value carries out pre-treatment to sample and to treated, sample is detected with spectrophotometer.
4. the method for measuring acid value in food as claimed in claim 3, includes the following steps:
1) acid value determination Specification Curve of Increasing: the standard of National Standard Method of learning from else's experience measurement uses solution, is added to anhydrous with 2.5 mL
In the 5 mL tool plug polystyrene centrifuge tube of ethyl alcohol, 50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent are added, is closed the lid
Son shakes up, and 0.5 mL of the solution after shaking up is taken to be placed in 1 cm cuvette, and absorbance value is measured at 490 ~ 580 nm wavelength, with
The acid value of series standard solution is ordinate, using the absorbance value of measurement as abscissa, draws curve；
2) sample to be tested alternative steps 1 measurement of sample to be tested acid value: are taken) in standard using solution, remaining detecting step phase
Together, the absorbance value for measuring sample to be tested, by absorbance value and above-mentioned steps 1) draw standard curve be compared, calculate to
The acid value of test sample sheet.
5. the method for measuring acid value in food as claimed in claim 4, includes the following steps:
Acid value determination Specification Curve of Increasing: it is 0.4,0.8,1.0,1.5,2.0,3.0,3.5 mg/g that National Standard Method of learning from else's experience, which measures acid value,
Series standard use solution, be added to 2.5 mL dehydrated alcohols 5 mL tool plug polystyrene centrifuge tube in, add
50 μ L of sodium hydroxide reagent, 50 μ L of phenolphthalein reagent, close the lid, shake up, and 0.5 mL of the solution after shaking up is taken to be placed in 1 cm ratio
Color ware measures absorbance value, using the acid value of series standard solution as ordinate, with the absorbance value of measurement at 560 nm wavelength
For abscissa, draw curve, obtained calibration curve equation: y=- 6.22x+4.67, y are acid value, and x is sample absorbance value；
The measurement of sample to be tested acid value: sample to be tested alternative steps 1 are taken) in standard using solution, remaining detecting step is identical,
The absorbance value for measuring sample to be tested, by absorbance value and above-mentioned steps 1) draw standard curve be compared, calculate it is to be measured
The acid value of sample.
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