CN108802209A - A kind of cod-liver oil Adulteration identification method - Google Patents
A kind of cod-liver oil Adulteration identification method Download PDFInfo
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Abstract
The present invention relates to cod-liver oil Adulteration identification methods, using column front derivation capillary gas chromatography, peak area measurement is carried out to cod-liver oil and the adulterated aliphatic acid in fish oil, percentage composition of the contained Typical fatty acid with respect to total fatty acids is calculated by area normalization method, limits are established, vegetable fatty acids limits refer to national standard；By comparing cod-liver oil and the adulterated aliphatic acid limits with oil, the aliphatic acid with characteristic index is found out, adulterated fish oil draws adulterated discriminance analysis figure, realize the Rapid identification of the adulterated fish oil of cod-liver oil by establishing the adulterated model of characteristic fatty acid two dimension；Adulterated vegetable oil, as characteristic index, will be measured linoleic acid content and carry out analysis comparison with cod-liver oil limit, realize the Rapid identification to adulterated vegetable oil in cod-liver oil using vegetable oil Linoleic acid in sample.The present invention is a kind of Adulteration identification method based on cod-liver oil, fish oil and vegetable fatty acids characteristic index, has the characteristics that science, accurate, simplicity.
Description
Technical field
The present invention relates to adulterated unrest existing for cod-liver oil Quality Identification more particularly to a kind of effective specification cod-liver oil market
As the cod-liver oil Adulteration identification method that can quick and precisely differentiate.
Background technology
Cod-liver oil is the grease extracted from the deep-sea cod liver of the states such as the Norway of high latitude, and gadus is not omnivorous, dry
Only, pollution-free, the cod-liver oil safety best in quality of extraction.Cod-liver oil is rich in natural vitamin A, vitamin D3And a variety of lands
No omega-3 unsaturated fatty acid (DHA, EPA, DPA) on the ground, wherein DHA (docosahexaenoic acid) and EPA (20 light dydrocarbons
Olefin(e) acid) content is very high, be human body itself cannot synthesize but indispensable important nutrient, especially DHA, be human nerve
The essential element that system grows and maintains, is the important composition ingredient of brain and retina, the intelligence to infant and eyesight hair
Educate it is most important, have the effects that protect retina, improve eyesight, promote infant intelligent development, improve memory.Norway's political affairs
Mansion is annual《Health-nutrition guide》In mention：It is recommended that baby be born start within the 4th week it is edible in the form of cod-liver oil existing for tie up
Raw element A, vitamin D3And DHA.DHA, EPA and vitamin A, D contained in cod-liver oil3Meet world health organisation recommendations mark
Standard, therefore cod-liver oil has the laudatory title of " liquid golden " and " docosapentaenoic acid ".As people are to the day of infant's intelligence and body development
Gradually pay attention to, the market demand of cod-liver oil is also increasing, and the cod-liver oil brand of current whole world pharmaceutical grade and health care grade is up to
Hundred kinds, account for 95% or more of entire cod-liver oil market.Pure cod-liver oil natural resources shortage, expensive, illegal manufacturer is to try to gain
Profit mixes a certain proportion of cheap fish oil or vegetable oil to reduce cost in cod-liver oil, seriously damages the interests of consumer.
It is only relied only in supervision and measures significant ingredients Vitamin A and D3Content and DHA and EPA content (ratio) be it is inadequate, no
Method manufacturer can synthesize vitamin A and D by addition3, ethyl ester type DHA so that every significant ingredient is met index
It is required that but be adulterated cod-liver oil actually, it is true and false to be difficult to identify.
Invention content
It is an object of the invention in order to establish a kind of method quick and precisely differentiating pure cod-liver oil, effective specification fish liver
Adulterated confusion existing for oil market.The present invention is based on the aliphatic acid in natural animal-plant oil, type, quantity and content are presented
Stronger characteristic, the present invention use column front derivation capillary gas chromatography technology, to cod-liver oil and adulterated in fish oil
Aliphatic acid carries out peak area measurement, and percentage composition of the contained Typical fatty acid with respect to total fatty acids is calculated by area normalization method,
And limits are established, vegetable fatty acids limits refer to national standard.By analysis compare cod-liver oil and it is adulterated with oil
Aliphatic acid limits find out the aliphatic acid with characteristic index, and adulterated fish oil is by establishing the adulterated mould of characteristic fatty acid two dimension
Type draws adulterated discriminance analysis figure, realizes the Rapid identification of the adulterated fish oil of cod-liver oil；Adulterated vegetable oil can be used in vegetable oil
The linoleic acid of high-content will measure linoleic acid content and carry out analysis comparison with cod-liver oil limit as characteristic index in sample,
Rapidly adulterated vegetable oil in cod-liver oil can be identified, it is this special based on cod-liver oil, fish oil and vegetable fatty acids
The Adulteration identification method of sign index has the characteristics that science, accurate, simplicity.
To achieve the goals above, the present invention uses following technical scheme：
A kind of cod-liver oil Adulteration identification method, the identification method are using column front derivation capillary gas chromatography, to cod
Cod-liver oil and the adulterated aliphatic acid in fish oil carry out peak area measurement；It is opposite that contained each aliphatic acid is calculated by area normalization method
The percentage composition of total fatty acids, establishes each aliphatic acid limits, and vegetable fatty acids limits refer to national standard；Pass through analysis
Compare cod-liver oil and the adulterated aliphatic acid limits with oil, finds out the aliphatic acid with characteristic index, adulterated fish oil passes through
The adulterated model of characteristic fatty acid two dimension is established, adulterated discriminance analysis figure is drawn, realizes the Rapid identification of the adulterated fish oil of cod-liver oil；
The linoleic acid of high-content in vegetable oil can be used as characteristic index in adulterated vegetable oil, and linoleic acid content and cod will be measured in sample
Cod-liver oil limit carries out analysis comparison, realizes the Rapid identification to adulterated vegetable oil in cod-liver oil；In addition the present invention passes through survey
The rate of recovery for determining 5 kinds of representative fatty acids in cod-liver oil carries out Accuracy Verification to column front derivation method, by measuring each fat
Fat acid relative mass correction factor (f) carries out applicability verification to computational methods.
In the technical scheme, institute's fatty acids are existed with glyceride form in cod-liver oil, and boiling point is high, cannot be direct
Gas Chromatographic Determination is carried out after being extracted using organic solvent, need to carry out column front derivation processing, i.e., by first after the saponification of cod-liver oil elder generation
Esterification, is converted into low-boiling fatty acid methyl ester.
Aliphatic acid in cod-liver oil includes saturated fatty acid, monounsaturated fatty acids and polyunsaturated fatty acid, type
Up to more than 20 kind, wherein content is higher 13 kinds.The present invention measures each relatively total fat of aliphatic acid using area normalization method
The percentage composition of fat acid, the key factor of measurement are：(1) efficiently separating between each aliphatic acid chromatographic peak, (2) column front derivation side
The accuracy of method.Factor (1) is preferred by being carried out to experimental condition, using the separating degree of fatty acid isomer chromatographic peak as
Inspection target；Factor (2) to representative five kinds in cod-liver oil fatty acid recovering rates by carrying out analysis and investigation.
Positioning confirmation is carried out using methyl esters reference substance to 15 kinds of Typical fatty acid in cod-liver oil, by the priority of peak sequence
Arrangement is myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, vaccenic acid, linoleic acid, parinaric acid (SDA), suitable
9- eicosenoic acids, suitable 11- eicosenoic acids, EPA, cetoleic acid, erucic acid, clupanodonic acid (DPA), DHA.Gadus liver
Include 3 pairs of isomers in fatty acid oil, is oleic acid (along 9- octadecenoic acids)/vaccenic acid (suitable 11- octadecenes respectively
Acid), cis-9-20 carbon acid/along 11- eicosenoic acids, cetoleic acid (along 11- docosenoic acids)/erucic acid is (along 13- 20
Two carbon enoic acids), difference lies in the differences of position of double bond for the structure of this three pairs of isomers.
The opposite total fatty acids of each aliphatic acid that the cod-liver oil sample of multisample is measured through column front derivation gas chromatography
Percentage composition analyzed, the percentage composition limits of aliphatic acid are obtained, with the fish oil percentage measured using same method
Content limit range is compared, and the two characteristic aliphatic acid index is found out, first, cod-liver oil has with DHA/EPA values in fish oil
Significant difference, in view of the two characteristic indexs, establishes gadus second is that cetoleic acid content is apparently higher than fish oil in cod-liver oil
The adulterated model of adulterated fish oil in liver oil, and using cetoleic acid percentage composition as abscissa, DHA/EPA values are painted as ordinate
Adulterated discriminance analysis figure is made, the whether adulterated fish oil of cod-liver oil and adulterated degree can quickly be identified by the analysis chart.
The aliphatic acid limit of common 6 vegetable oil refers to national standard, by comparing each aliphatic acid of cod-liver oil and vegetable oil
Limit index, content of the display linoleic acid in vegetable oil are far above cod-liver oil, when mixing 10% vegetable oil in cod-liver oil
When, linoleic acid content can be more than the upper limit, and feature can carry out Rapid identification to the adulterated vegetable oil of cod-liver oil accordingly.
Preferably, identification method includes the following steps：
A) prepared by mixed reference substance solution：
It weighs methyl hexadecanoate, Methyl palmitoleinate, methyl oleate, EPA methyl esters, DHA methyl esters to set in 20mL brown volumetric flasks, add
Isooctane dissolves and is diluted to scale, and 4.97mg/mL containing methyl hexadecanoate, Methyl palmitoleinate 2.94mg/mL, oleic acid first is made
The mixed reference substance solution of ester 6.21mg/mL, EPA methyl esters 3.84mg/mL, DHA methyl esters 6.05mg/mL；
B) sample aliphatic acid background concentration measures：
It weighs cod-liver oil sample 100mg to set in 20mL band screw socket reaction bulbs, sodium hydroxide-methanol solution of 1.5mL is added, revolves
Tight bottle cap, whirlpool mixing 30s set in 90 DEG C of thermostats and heat 20min, cooling；The methanolic (2g → 100mL) of 2mL is added
Solution screws bottle cap, and whirlpool mixing 30s sets in 100 DEG C of thermostats and heats 10min, cooling；2mL isooctane is added, screws bottle
5mL saturated nacl aqueous solutions, jog, stratification is added in lid, whirlpool mixing 1min immediately；Aspirate supernatant, which is transferred to, to be equipped with
In the tool plug test tube of anhydrous sodium sulfate, shaking dehydration is parallel to prepare three parts to get test solution；C) recovery of standard addition is tested：
The above-mentioned sample 50mg for having predicted content is weighed, totally 9 parts, is set in 20mL band screw socket reaction bulbs, precision plus hybrid standard product
Each 3 parts of solution 0.5mL, 1.0mL, 1.5mL, are added sodium hydroxide-methanol solution of 1.5mL, screw bottle cap, whirlpool mixing 30s,
It sets in 90 DEG C of thermostats and heats 20min, it is cooling；The methanolic solution of 2mL is added, screws bottle cap, whirlpool mixing 30s sets 100
10min is heated in DEG C thermostat, it is cooling；2mL isooctane is added, screws bottle cap, 5mL saturations are added in whirlpool mixing 1min immediately
Sodium chloride solution, jog, stratification；Aspirate supernatant is transferred in the tool plug test tube equipped with anhydrous sodium sulfate, shaking dehydration,
Up to mark-on testing liquid；
D) chromatographic condition is pressed, sample introduction, the peak area of 5 kinds of aliphatic acid of measurement are pressed successively for mixed reference substance solution and test solution
External standard method calculates the content of fatty acid methyl ester, then is converted into content of fatty acid.
Preferably, the mass fraction for the sodium hydroxide-methanol solution being added in step b) is 2%；The sulfuric acid first of addition
The mass fraction of alcoholic solution is 5%.
Preferably, the calculation formula of relative mass correction factor f is：
In formula, Ai：Component peak area；As：Reference substance peak area；mi：Concentration of component；ms：Reference substance concentration.
Preferably, chromatographic column：The specification of DB-23 capillary columns, capillary column is：60m×0.25mm×0.25μm；Color
Compose column temperature program：Initial temperature is 170 DEG C, is warming up to 225 DEG C with the rate of 1 DEG C/min and keeps 5min；Injector temperature：
250℃；Detector：FID；Detector temperature：280℃；Carrier gas：High-purity N2, flow velocity 1.0mL/min；Sample size：1μL；Into sample prescription
Formula：Split sampling, split ratio：150:1.
Preferably, in identification cod-liver oil when adulterated fish oil, using cetoleic acid percentage composition as abscissa, DHA%/
EPA% values are ordinate, establish the adulterated model of two dimension for mixing 5%, 15%, 25% and 50% fish oil, draw adulterated differentiation point
Analysis figure.
Preferably, adulterated vegetable oil works as cod-liver oil using linoleic acid content as characteristic index in identification cod-liver oil
In when mixing any type vegetable oil >=10%, linoleic percentage composition exceeds the upper limit, and differentiating in cod-liver oil accordingly is
No adulterated vegetable oil.
The beneficial effects of the invention are as follows：The present invention is based on the aliphatic acid in natural animal-plant oil, type, quantity and contents
It is presented stronger characteristic, the present invention uses column front derivation capillary gas chromatography technology, to cod-liver oil and adulterated uses fish
Aliphatic acid in oil is measured, and calculates contained Typical fatty acid percentage composition by area normalization method, and establish limits,
Vegetable fatty acids limits refer to national standard.Compare cod-liver oil and the adulterated aliphatic acid limits with oil by analysis,
The aliphatic acid with characteristic index is found out, adulterated fish oil draws adulterated identification by establishing the adulterated model of characteristic fatty acid two dimension
Analysis chart realizes the Rapid identification of the adulterated fish oil of cod-liver oil；The linoleic acid of high-content in vegetable oil can be used in adulterated vegetable oil
As characteristic index, linoleic acid content will be measured in sample and carries out analysis comparison with cod-liver oil limit, it can be rapidly to gadus
Adulterated vegetable oil is identified in liver oil, this adulterated mirror based on cod-liver oil, fish oil and vegetable fatty acids characteristic index
The method of determining has the characteristics that science, accurate, simplicity.
Description of the drawings
Fig. 1 is cod-liver oil feature gas chromatogram of the present invention.In figure, 1, myristic acid 2, palmitic acid 3, palmitoleic acid
4, stearic acid 5, oleic acid 6, vaccenic acid 7, linoleic acid 8, SDA 9, cis-9-20 carbon acid 10, suitable 11- eicosenoic acids
11, EPA 12, cetoleic acid 13, erucic acid 14, DPA 15, DHA.
Fig. 2 is reference substance gas chromatogram of the present invention.In figure, 1, methyl hexadecanoate 2, Methyl palmitoleinate 3, oleic acid first
Ester 4, EPA methyl esters 5, DHA methyl esters.
Fig. 3 is adulterated cod-liver oil aliphatic acid discriminant analysis figure.
Fig. 4 is the flow chart of the present invention.
Specific implementation mode
Below in conjunction with specific embodiment and attached drawing, the present invention is further explained：
Instrument and material
Agilent 7890A gas chromatographs (Agilent companies of the U.S.)；Fatty acid methyl ester reference substance is purchased from U.S. NU-
CHEK companies and SIGMA companies of the U.S., the equal > of purity 99%；Isooctane, methanol (chromatographically pure, J.T.Baker companies), sample is equal
From commercially available and net purchase.
With reference to Fig. 4, a kind of adulterated fish oil identification method of cod-liver oil, the identification method is using column front derivation capillary
Gas chromatography is measured cod-liver oil and the adulterated aliphatic acid in fish oil；Contained allusion quotation is calculated by area normalization method
Type aliphatic acid percentage composition, and establish limits；Compare cod-liver oil and the adulterated aliphatic acid limit model with oil by analysis
It encloses, finds out the aliphatic acid with characteristic index, adulterated fish oil realizes gadus liver by drawing the adulterated model of characteristic fatty acid two dimension
The Rapid identification of the adulterated fish oil of oil.
The linoleic acid of high-content in vegetable oil can be used as characteristic index in the adulterated vegetable oil of cod-liver oil, will be surveyed in sample
It obtains linoleic acid content and carries out analysis comparison with cod-liver oil limit, realize the Rapid identification to adulterated vegetable oil in cod-liver oil.
The optimization of chromatographic condition
Isomer structure is close, and the separating difficulty in chromatography is very big, it is desirable to obtain satisfied separating degree, it is necessary to chromatography
Condition optimizes.The most important factor that compound separation is influenced in gas-chromatography is chromatographic column and column temperature, and determination of fatty acid is logical
Frequently with polarity chromatographic column, the chromatographic column difference stationary phase of same type, the different trades mark and specification have the separation of aliphatic acid larger
Influence.It is 30m × 0.25mm × 0.25 μm, the different trades mark (DB-23, DB-WAX, Rtx-WAX) that specification is used in the present invention
Capillary column, as a result show DB-23 chromatographic columns to Separation of Fatty Acids best results, oleic acid/vaccenic acid isomery physical efficiency reaches base
Line detaches, but the separating effect of another two pairs of isomers is bad, and separating degree is less than 1.3.Reprogramming Elevated Temperature Conditions and flow velocity also without
Method reaches baseline separation, therefore considers to increase the length of chromatographic column, uses specification for the DB-23 of 60m × 0.25mm × 0.25 μm hairs
Capillary column, the cis-9-20 carbon acid/separating effect of isomers is obviously changed along 11- eicosenoic acids, cetoleic acid/erucic acid two
It is kind.By the analysis and investigation influenced on separating degree on different temperature programs, finally it is determined that optimum temperature rise program is：Initial temperature
It is 170 DEG C, is warming up to 225 DEG C with the rate of 1 DEG C/min and keeps 5min.To injector temperature, detector temperature, load in experiment
Gas velocity, split ratio are investigated, and as a result show influence unobvious of the above-mentioned several factors to Separation of Fatty Acids effect.
The optimum chromatogram condition that the present invention establishes is：
Chromatographic column：DB-23 capillary columns (60m × 0.25mm × 0.25 μm)；Chromatographic column temperature program：Initial temperature is 170 DEG C,
It is warming up to 225 DEG C with the rate of 1 DEG C/min and keeps 5min；Injector temperature：250℃；Detector：FID；Detector temperature：
280℃；Carrier gas：High-purity N2, flow velocity 1.0mL/min；Sample size：1μL；Input mode：Split sampling, split ratio：150:1.
The separating degree of the three pairs of isomers measured by above-mentioned chromatographic condition is 2.0,1.4,1.6 respectively, meets area normalization
Change method, which measures, to be required, and cod-liver oil Typical gas chromatograph figure is shown in Fig. 1, which can be used as the characteristic spectrum of cod-liver oil, be used for
Each aliphatic acid fast qualitative confirms.
Aliphatic acid Derivatization Method accuracy is investigated
Derivatization Method
It saltouts and is dehydrated including saponification, esterification reaction of organic acid, extraction, step is：
A. saponification：It weighs cod-liver oil to set in band screw socket reaction bulb, 1.5mL2% sodium hydrate methanol solutions is added, whirlpool is mixed
It closes 30 seconds, 90 DEG C of heating 20min take out cooling, obtain saponification solution；
B. esterification reaction of organic acid：In the saponification solution that step a is obtained, 2mL5% methanolic solutions, whirlpool mixing is added
30 seconds, 100 DEG C of heating 10min took out cooling, obtain esterification reaction of organic acid solution；
C. extraction is saltoutd：2mL isooctane solutions are added in the esterification reaction of organic acid solution that step b is obtained, whirlpool mixing 1min is stood
5mL saturated nacl aqueous solutions, jog, stratification is added；
D. dehydration：The supernatant that aspiration step c is obtained is transferred in the test tube equipped with a small amount of anhydrous sodium sulfate, and shaking makes
Dehydration, solution is fatty acid methyl ester solution after dehydration.
Aliphatic acid recovery of standard addition measures
The present invention has chosen 5 kinds, and content is higher and representative aliphatic acid in cod-liver oil, is palmitic acid, palm oil respectively
Acid, oleic acid, EPA and DHA, wherein palmitic acid represent saturated fatty acid, and palmitoleic acid, oleic acid represent monounsaturated fatty acids,
EPA, DHA represent polyunsaturated fatty acid.
Measurement takes following steps：
A. prepared by mixed reference substance solution
Precision weighs methyl hexadecanoate, Methyl palmitoleinate, methyl oleate, EPA methyl esters, DHA methyl esters and sets 20mL browns appearance in right amount
In measuring bottle, add isooctane to dissolve and be diluted to scale, 4.97mg/mL containing methyl hexadecanoate, Methyl palmitoleinate 2.94mg/ is made
The mixed reference substance solution of mL, methyl oleate 6.21mg/mL, EPA methyl esters 3.84mg/mL, DHA methyl esters 6.05mg/mL.
B. sample aliphatic acid background concentration measures
Precision weighs cod-liver oil sample about 100mg and sets in 20mL band screw socket reaction bulbs, adds 1.5mL2% sodium hydroxide methanol molten
Liquid screws bottle cap, and whirlpool mixing 30s sets in 90 DEG C of thermostats and heats 20min, cooling.Add 2mL5% methanolic solutions, revolves
Tight bottle cap, whirlpool mixing 30s set in 100 DEG C of thermostats and heat 10min, cooling.2mL isooctane is added in precision, screws bottle cap,
Whirlpool mixing 1min adds 5mL saturated nacl aqueous solutions, gently shakes, stratification immediately.Aspirate supernatant is transferred to equipped with few
In the tool plug test tube for measuring anhydrous sodium sulfate, shaking dehydration is parallel to prepare three parts to get test solution.By above-mentioned chromatographic condition,
Sample introduction, the peak area of 5 kinds of aliphatic acid of measurement calculate fatty acid methyl by external standard method successively for mixed reference substance solution and test solution
The content of ester, then it is converted into content of fatty acid, the conversion coefficient that fatty acid methyl ester is converted into aliphatic acid is palmitic acid：0.948；Palm fibre
Palmitic acid oleic acid：0.948；Oleic acid：0.953；EPA：0.956；DHA：0.959, measurement result is shown in Table 1.Mix reference substance gas-chromatography
Figure is shown in Fig. 2.
15 kinds of content of fatty acid measurement results of table
C. aliphatic acid recovery of standard addition measures
Take the sample for having predicted content, totally 9 parts, respectively precision weigh about 50mg to 20mL band screw socket reaction bulb in, precision adds
Each 3 parts of standard liquid 0.5mL, 1.0mL, 1.5mL are stated, 1.5mL2% sodium hydrate methanol solutions are added, screws bottle cap, whirlpool mixing
30s sets and heats 20min in 90 DEG C of dry-type thermostats, takes out cooling.Add 2mL5% methanolic solutions, screw bottle cap, whirlpool is mixed
30s is closed, sets and heats 10min in 100 DEG C of dry-type thermostats, takes out cooling.The accurate addition isooctane 1.5mL, 1.0mL of difference,
Each 3 parts, whirlpool mixing 1min of 0.5mL adds 5mL saturated nacl aqueous solutions, gently shakes, stratification immediately.Aspirate supernatant
It is transferred in the test tube equipped with a small amount of anhydrous sodium sulfate, shaking takes supernatant to distinguish sample introduction, the sample recovery rate of 5 kinds of aliphatic acid
Measurement result is shown in Table 2.By table as it can be seen that the recovery of standard addition of 5 kinds of aliphatic acid is between 98.30%~99.54%, RSD exists
Between 1.09%~1.42%, show at the conditions of the experiments described above, aliphatic acid column front derivation method is accurate and reliable in cod-liver oil.
2 determination of recovery rates result of table
Aliphatic acid relative mass correction factor (f) measures
Quantitative accuracy is carried out to each component using area normalization method and detects mass-basis response value depending on the FID of each component
(mi/Ai), the higher aliphatic acid mi/Ai values of 13 kinds of contents are measured, as a result show DHA measured value highests, therefore selection is done with DHA
For reference substance, f value of each aliphatic acid relative to DHA, result of calculation are shown in Table 3 according to the following formula, by table as it can be seen that cod-liver oil
The f values of middle Typical fatty acid methyl esters are all higher than 0.95, therefore directly can carry out quantitative analysis with peak area normalization.
Formula：
In formula, Ai：Component peak area；As：Reference substance peak area；mi：Constituent mass；ms：Reference substance quality
3 fatty acid methyl ester relative mass correction factor of table
| Component | F values | Component | F values |
| Methyl myristate | 0.96 | Parinaric acid methyl esters | 0.97 |
| Methyl hexadecanoate | 0.97 | Along 11- eicosenoic acid methyl esters | 1.00 |
| Methyl palmitoleinate | 0.96 | EPA methyl esters | 0.98 |
| Methyl stearate | 0.97 | Cetoleic acid methyl esters | 0.98 |
| Methyl oleate | 0.98 | DPA methyl esters | 0.96 |
| Along vaccenic acid methyl esters | 0.98 | DHA methyl esters | 1.00 |
| Methyl linoleate | 0.97 | / | / |
Cod-liver oil and fish oil fatty acid signature analysis
Cod-liver oil is the fat oil extracted from gadus liver, and fish oil is the fat oil extracted from fatty,fiss, contained by the two
Fatty acid species and quantity are essentially identical, but content difference is apparent, respectively has feature.The present invention is from the two characteristic content of fatty acid
Identification and analysis is carried out with content ratio changing rule.Inventor collect altogether domestic and imported cod-liver oil raw material, oral solution and
Totally 46 batches, soft capsule sample, fish oil raw material and totally 30 batches, soft capsule sample, by above-mentioned sample by area after column front derivation is handled
Normalization method calculates aliphatic acid percentage composition.Aliphatic acid has because of the difference in the gadus place of production and grease extraction process in cod-liver oil
Institute's difference, there are one relatively narrow waving intervals (limits) for the content measured.Fish oil then by fingerling, season, the place of production, plus
The influence of the factors such as Ministry of worker position and extraction process, content waving interval are more than cod-liver oil.In cod-liver oil and fish oil sample
The Typical fatty acid content range measured is shown in Table 4, by table as it can be seen that two big spies are presented compared to fish oil in cod-liver oil institute fatty acids
Sign：1. DHA content is apparently higher than EPA content, in 46 batches of samples in addition to 2 batches of doubtful adulterated samples, DHA, EPA percentage composition ratio
(DHA/EPA) in 1.40~1.68 ranges, mean value 1.55, and the DHA content in fish oil sample contains significantly lower than EPA
Amount, DHA/EPA are 0.64~0.75, mean value 0.68.2. along 11- eicosenoic acids and cetoleic acid (along bis- dodecylenes of 11-
Acid) content is apparently higher than fish oil.Mix a certain amount of fish oil in cod-liver oil and will lead to DHA/EPA, along 11- eicosenoic acids and
The decline of the content of cetoleic acid, especially cetoleic acid, in other cod-liver oil in (dogfish oil, tuna liver oil) almost
No, content is very low in fish oil, only has high level in cod-liver oil, and average content is about the 5 of fish oil in cod-liver oil
Times, there is very strong characteristic, therefore can be to adulterated in cod-liver oil from DHA/EPA and the changing rule of cetoleic acid content
Fish oil is differentiated.
4 cod-liver oil of table and fish oil fatty acid percentage composition limit
Aliphatic acid changing rule in adulterated model
According to the characteristic index and its changing rule of cod-liver oil aliphatic acid, the present invention establishes a kind of comparison intuitively adulterated mould
Type carries out Analysis and Screening to modeling with sample, selectes wherein 28 batches of cod-liver oil samples, 17 batches of fish oil sample (determination of fatty acid knots
The almost the same different batches sample of fruit, only selects a wherein batch) adulterated data be used for mode input, with cetoleic acid percentage
Content is abscissa, and DHA/EPA values are ordinate, establishes the adulterated model of two dimension for mixing 5%, 15%, 25% and 50% fish oil,
Adulterated discriminant analysis figure is drawn, sees Fig. 3.As seen from the figure, when mixing the fish oil of different proportion in cod-liver oil, different adulterated water
Flat that certain regularity of distribution is presented, fatty acid profile has more apparent regional.
The differentiation of adulterated sample
Each aliphatic acid peak area in sample to be identified is measured, percentage composition of each aliphatic acid with respect to total fatty acids is calculated, works as spermaceti
Olefin(e) acid content and (or) DHA/EPA values go beyond the limit range when (DHA/EPA is with 1.40~1.68 for limits), can tentatively sentence
The fixed sample is doubtful adulterated, then corresponding coordinate position is found from discriminant analysis figure, you can is made intuitively to adulterated degree
Differentiate and identifies.
Adulterated analysis of cases
When analyzing cod-liver oil sample, discovery has DHA/EPA values and cetoleic acid content in 2 batches of samples to deviate limit
Degree, a collection of DHA/EPA are 1.16, and cetoleic acid percentage composition is 6.74, and another crowd of DHA/EPA is 1.13, cetoleic acid percentage
Content is 5.32, and coordinate points (figure orbicular spot position), sees Fig. 2 where 2 batches of samples are drawn in adulterated model, as seen from the figure,
The position of dot is fallen in 25% adulterated region, thus can determine that mixed in 2 batches of cod-liver oils about 25% fish oil.
The identification method of adulterated vegetable oil in cod-liver oil
Containing there are many saturations and unsaturated fatty acid, price to be typically well below cod-liver oil, there is fish in the market in vegetable oil
The phenomenon that vegetable oil is to reduce cost is admixed in liver oil.Table 5 is the percentage composition limit of 13 kinds of aliphatic acid in Vegetable Oils, will
Cod-liver oil and each vegetable oil institute fatty acids carry out analysis comparison, as a result show that prodigious difference is presented in the content of fatty acid of the two
The opposite sex, wherein vegetable oil Linoleic acid content are far above cod-liver oil, can be using linoleic acid content as characteristic index, when gadus liver
When mixing any type vegetable oil 10% in oil, linoleic percentage composition exceeds the upper limit, can differentiate in cod-liver oil accordingly
Adulterated vegetable oil.Simulation of the present invention prepares the adulterated sample of adulterated 10% vegetable oil in cod-liver oil, selects 4 Linoleic acid of table
The minimum rapeseed oil of content is analyzed as puppet is mixed with oil, and simulation cod-liver oil sample linoleic acid content is 1.6%, adulterated
It is 12.5% with rapeseed oil linoleic acid content, when adulterated 10% rapeseed oil in cod-liver oil, simulates the Asia oil in adulterated sample
Acid content rises to 2.7% by 1.6%, has exceeded the upper limit, therefore whether the content by analyzing sample Linoleic acid to be identified surpasses
Can quickly adulterated vegetable oil in cod-liver oil be identified by going out the limit upper limit.
5 vegetable fatty acids percentage composition limit of table
Note：ND is to be not detected (content is less than 0.05%)
It should be understood that those skilled in the art, it can be modified or changed according to the above description, and
All these modifications and variations should all belong to the protection domain of appended claims of the present invention.
Claims (7)
1. a kind of cod-liver oil Adulteration identification method, which is characterized in that the identification method is using column front derivation Capillary Gas
Phase chromatography carries out peak area measurement to cod-liver oil and the adulterated aliphatic acid in fish oil；Institute is calculated by area normalization method
Percentage composition containing each aliphatic acid with respect to total fatty acids establishes each aliphatic acid limits, vegetable fatty acids limits ginseng
Examine national standard；Compare cod-liver oil and the adulterated aliphatic acid limits with oil by analysis, finds out the fat with characteristic index
Acid；Adulterated fish oil draws adulterated discriminance analysis figure, realizes that cod-liver oil is adulterated by establishing the adulterated model of characteristic fatty acid two dimension
The Rapid identification of fish oil；The linoleic acid of high-content in vegetable oil can be used as characteristic index in adulterated vegetable oil, will be surveyed in sample
It obtains linoleic acid content and carries out analysis comparison with cod-liver oil limit, realize the Rapid identification to adulterated vegetable oil in cod-liver oil；
The rate of recovery by measuring 5 kinds of representative fatty acids in cod-liver oil carries out Accuracy Verification to column front derivation method, passes through survey
Fixed each aliphatic acid relative mass correction factor f carries out applicability verification to area normalization computational methods.
2. a kind of cod-liver oil Adulteration identification method according to claim 1, which is characterized in that identification method includes following
Step：
A) prepared by mixed reference substance solution：
It weighs methyl hexadecanoate, Methyl palmitoleinate, methyl oleate, EPA methyl esters, DHA methyl esters to set in 20mL brown volumetric flasks, add
Isooctane dissolves and is diluted to scale, and 4.97mg/mL containing methyl hexadecanoate, Methyl palmitoleinate 2.94mg/mL, oleic acid first is made
The mixed reference substance solution of ester 6.21mg/mL, EPA methyl esters 3.84mg/mL and DHA methyl esters 6.05mg/mL；
B) sample aliphatic acid background concentration measures：
It weighs cod-liver oil sample 100mg to set in 20mL band screw socket reaction bulbs, sodium hydroxide-methanol solution of 1.5mL is added, revolves
Tight bottle cap, whirlpool mixing 30s set in 90 DEG C of thermostats and heat 20min, cooling；The methanolic (2g → 100mL) of 2mL is added
Solution screws bottle cap, and whirlpool mixing 30s sets in 100 DEG C of thermostats and heats 10min, cooling；2mL isooctane is added, screws bottle
5mL saturated nacl aqueous solutions, jog, stratification is added in lid, whirlpool mixing 1min immediately；Aspirate supernatant, which is transferred to, to be equipped with
In the tool plug test tube of anhydrous sodium sulfate, shaking dehydration is parallel to prepare three parts to get test solution；
C) recovery of standard addition is tested
The above-mentioned sample 50mg for having predicted content is weighed, totally 9 parts, is set in 20mL band screw socket reaction bulbs, precision plus hybrid standard product
Each 3 parts of solution 0.5mL, 1.0mL, 1.5mL, are added sodium hydroxide-methanol solution of 1.5mL, screw bottle cap, whirlpool mixing 30s,
It sets in 90 DEG C of thermostats and heats 20min, it is cooling；The methanolic solution of 2mL is added, screws bottle cap, whirlpool mixing 30s sets 100
10min is heated in DEG C thermostat, it is cooling；2mL isooctane is added, screws bottle cap, 5mL saturations are added in whirlpool mixing 1min immediately
Sodium chloride solution, jog, stratification；Aspirate supernatant is transferred in the test tube equipped with anhydrous sodium sulfate, shaking dehydration to get
Mark-on testing liquid；
D) chromatographic condition is pressed, sample introduction, the peak area of 5 kinds of aliphatic acid of measurement are pressed successively for mixed reference substance solution and test solution
External standard method calculates the content of fatty acid methyl ester, then is converted into content of fatty acid.
3. a kind of cod-liver oil Adulteration identification method according to claim 2, which is characterized in that the hydrogen being added in step b)
The mass fraction of sodium oxide molybdena-methanol solution is 2%；The mass fraction of the methanolic solution of addition is 5%.
4. a kind of cod-liver oil Adulteration identification method according to claim 1 or 2, which is characterized in that relative mass corrects
The calculation formula of factor f is：
In formula, Ai：Component peak area；As：Reference substance peak area；mi：Concentration of component；ms：Reference substance concentration.
5. a kind of cod-liver oil Adulteration identification method according to claim 2, which is characterized in that chromatographic column：DB-23 capillarys
The specification of tubing string, capillary column is：60m×0.25mm×0.25μm；Chromatographic column temperature program：Initial temperature is 170 DEG C, with 1
DEG C/rate of min is warming up to 225 DEG C and keeps 5min；Injector temperature：250℃；Detector：FID；Detector temperature：280
℃；Carrier gas：High-purity N2, flow velocity 1.0mL/min；Sample size：1μL；Input mode：Split sampling, split ratio：150:1.
6. a kind of cod-liver oil Adulteration identification method according to claim 1 or 2, which is characterized in that identification cod-liver oil
In adulterated fish oil when, using cetoleic acid percentage composition as abscissa, DHA%/EPA% values are ordinate, foundation mixes 5%,
15%, the adulterated model of two dimension of 25% and 50% fish oil, draws adulterated discriminant analysis figure.
7. a kind of cod-liver oil Adulteration identification method according to claim 1 or 2, which is characterized in that identification cod-liver oil
In adulterated vegetable oil, using linoleic acid content as characteristic index, when mixing any type vegetable oil >=10% in cod-liver oil,
Linoleic percentage composition exceeds the upper limit, differentiates whether adulterated vegetable oil in cod-liver oil accordingly.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109609599A (en) * | 2019-01-30 | 2019-04-12 | 中国水产科学研究院黄海水产研究所 | A kind of fish oil and its DNA extraction and application method with palm oil mixture |
| CN112557494A (en) * | 2020-11-05 | 2021-03-26 | 广州海关技术中心 | Pollack species identification method based on multiple elements and chemometrics |
| CN113278710A (en) * | 2021-07-13 | 2021-08-20 | 南宁海关技术中心 | Primer probe, kit and method for detecting heterogeneous oil doped in deep sea fish oil |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569277A (en) * | 2014-12-31 | 2015-04-29 | 中国农业科学院油料作物研究所 | Method for rapidly identifying deep sea fish oil |
| US20160139091A1 (en) * | 2014-10-08 | 2016-05-19 | Perkinelmer Health Sciences, Inc. | Traps for use with chromatography systems |
| CN106053677A (en) * | 2016-08-12 | 2016-10-26 | 江南大学 | Determination method for location distribution of aliphatic acid in triglyceride of fish oil |
-
2018
- 2018-03-23 CN CN201810250446.8A patent/CN108802209A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160139091A1 (en) * | 2014-10-08 | 2016-05-19 | Perkinelmer Health Sciences, Inc. | Traps for use with chromatography systems |
| CN104569277A (en) * | 2014-12-31 | 2015-04-29 | 中国农业科学院油料作物研究所 | Method for rapidly identifying deep sea fish oil |
| CN106053677A (en) * | 2016-08-12 | 2016-10-26 | 江南大学 | Determination method for location distribution of aliphatic acid in triglyceride of fish oil |
Non-Patent Citations (4)
| Title |
|---|
| PEDRO ARAUJO等: "Discrimination of n-3 Rich Oils by Gas Chromatography", 《LIPIDS》 * |
| 刘帅等: "脂肪酸甲酯化方法的研究进展", 《中国药房》 * |
| 周文斌等: "脂肪酸检测方法的研究进展", 《中国现代应用药学》 * |
| 李彩霞等: "鱼肝油中脂肪酸的气相色谱及气相色谱-质谱分析", 《中国现代应用药学》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109609599A (en) * | 2019-01-30 | 2019-04-12 | 中国水产科学研究院黄海水产研究所 | A kind of fish oil and its DNA extraction and application method with palm oil mixture |
| CN109609599B (en) * | 2019-01-30 | 2019-11-08 | 中国水产科学研究院黄海水产研究所 | A kind of fish oil and its DNA extraction and application method with palm oil mixture |
| CN112557494A (en) * | 2020-11-05 | 2021-03-26 | 广州海关技术中心 | Pollack species identification method based on multiple elements and chemometrics |
| CN113278710A (en) * | 2021-07-13 | 2021-08-20 | 南宁海关技术中心 | Primer probe, kit and method for detecting heterogeneous oil doped in deep sea fish oil |
| CN113278710B (en) * | 2021-07-13 | 2022-02-22 | 南宁海关技术中心 | Primer probe, kit and method for detecting heterogeneous oil doped in deep sea fish oil |
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