CN107894465A - The detection method of food additives in a kind of moon cake - Google Patents
The detection method of food additives in a kind of moon cake Download PDFInfo
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Abstract
The detection method of food additives, comprises the following steps in a kind of moon cake：(1) mixed standard solution preparation：By dehydroactic acid, benzoic acid, sorbic acid, four kinds of P-hydroxybenzoic acid standard lipids mixed preparings in pure methanol；Acesulfame potassium, saccharin sodium, 8 kinds of food colour standard items mixed preparings are in pure water；Above-mentioned solution is mixed in proportion again, obtains hybrid standard storing solution；(2) sample extraction；(3) sample purification；(4) machine testing on：Prepare liquid and mixed standard solution are placed in the liquid chromatograph equipped with UV-detector and detected, and using quantified by external standard method, are switched by gradient elution, wavelength, upper machine surveys the content of food additives simultaneously.Pre-treatment step of the present invention is consistent, and cost is low, speed is fast, reliable results, favorable reproducibility, can disposably detect 17 kinds of food additives simultaneously.
Description
Technical field
The present invention relates to field of food detection, the detection method of food additives in more particularly to a kind of moon cake.
Background technology
Moon cake is one of Chinese tradition cake having long enjoyed a good reputation, red-letter day in Mid-autumn Festival eating habit.Moon cake circle is again round, is to divide of the whole family again
Eat, signify that reunion is harmonious.Ancient times moon cake is eaten as sacrificial offerings in the Mid-autumn Festival.It is said that the custom that the Mid-autumn Festival eats moon cake starts from Tang
Court.It is popular in imperial palace when the Northern Song Dynasty, after spread among the people, be commonly called as " little cake " and " moon group " at that time.Being developed to the Ming Dynasty then turns into
The catering customs that the whole people bought jointly.Moon cake blends with various regions catering customs, develops Cantonese, capital formula, Soviet Union's formula, tidal again,
The moon cakes such as Yunnan formula, liked by the people of Chinese north and south various regions.
Nowadays, moon cake is that seasonal is very strong, consumption phase shorter food.Manufacturer will in a short time centralized production it is big
The moon cake of amount is put on sale, and has certain difficulty, and often to produce in advance for commercial distribution.Carrying with living standard simultaneously
Height, requirement of the people to moon cake quality also more and more higher.Businessmans are in order that moon cake color is various to meet the need of people
Ask, all the food additives such as preservative, sweetener, pigment can be added when producing moon cake has certain application in moon cake.
The food additives of addition can strengthen the sensory properties of food, prevent putrid and deteriorated, extend the shelf life, if but improper use also can
Certain potential safety hazard is produced to health.
Because the kind of the preservative in moon cake, sweetener and pigment is more and more, at present detection method again it is inconsistent,
It is comparatively laborious, it there is no a performance while determine the chromatographic detection method of numerous food additive.The detection side of food additives
Method mainly has high performance liquid chromatography, gas chromatography, capillary electrophoresis, thin-layered chromatography and liquid chromatogram-matter at present
Compose combination method.But Determination of Preservatives, pigment and sweetener, and national standard can not be determined simultaneously using national standard method
In specify only assay method in the drinks such as carbonated drink, juice, drinks, this method is only adjusted to neutrality with ammoniacal liquor, after filtering
HPLC measure is directly used in, therefore this method is not suitable for being applied to high protein, the moon cake of high grease, and can determine simultaneously in moon cake
There is not been reported so far for the method for numerous food additive.
The content of the invention
It is an object of the invention to provide a kind of detection method of food additives in moon cake, this detection method can be once
Property determine numerous food additive in moon cake simultaneously, cost is low, speed is fast, reliable results, favorable reproducibility.
Solves above-mentioned technical problem, the technical solution adopted in the present invention is：The detection of food additives in a kind of moon cake
Method, comprise the following steps：
(1) mixed standard solution preparation：
By dehydroactic acid, benzoic acid, sorbic acid, methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, para hydroxybenzene first
Sour third fat, parahydroxybenzoate butyl standard items mixed preparing are in pure methanol；
Acesulfame potassium, saccharin sodium, lemon yellow, newly red, amaranth, saccharin sodium, famille rose, sunset yellow, red, the erythrosine mark of temptation
Quasi- product mixed preparing is in pure water；
Above-mentioned solution is mixed in proportion, obtains 17 kinds of additive hybrid standard storing solutions；
Again with methanol-aqueous solution progressively dilutes, with obtained 17 kinds of additive hybrid standard working solutions；
(2) sample extraction：The moon cake after grinding is weighed in centrifuge tube pipe, first adds methanol-water solution extraction, Ran Houjia
Enter sodium hydroxide solution, be well mixed；
Solution of zinc sulfate is added, is mixed, is added sodium hydroxide solution, shake up, finally with pure water constant volume, obtain sample extraction
Liquid；
(3) sample purification：Sample extracting solution is centrifuged, filtered with filter paper, removes upper strata grease；Filtrate is through filter membrane mistake again
Filter, obtains prepare liquid；
(4) machine testing on：Mixed standard solution in the prepare liquid obtained in step (3) and step (1) is placed in and is furnished with
The liquid chromatograph of UV-detector is detected, and using quantified by external standard method, is switched by gradient elution, wavelength, upper machine is simultaneously
Measure the content of 17 kinds of food additives.
Preferably, hybrid standard storing solution is 100mg/L hybrid standard storing solutions in step (1).
Preferably, methanol-water solution is that volume ratio is 1 in step (1), step (2)：1 methanol-water solution.
Preferably, dehydroactic acid, benzoic acid, sorbic acid and methyl p-hydroxybenzoate, P-hydroxybenzoic acid in step (1)
Second fat, the fat of P-hydroxybenzoic acid third, parahydroxybenzoate butyl standard items are configured to after being completely dissolved with chromatographically pure level methanol
200-400mg/L mixed liquor；
Acesulfame potassium, saccharin sodium and lemon yellow, newly red, amaranth, saccharin sodium, famille rose, sunset yellow, the red, erythrosine of temptation
Standard items are configured to 200-400mg/L mixed liquor after being completely dissolved with laboratory one-level water, then mixed in equal amounts obtains
100mg/L hybrid standard storing solutions；It is again 1 with volume ratio:1 methanol aqueous solution progressively dilutes preparation, and it is dense to respectively obtain quality
Spend for 0.1,0.5,1.0,5.0,10.0,20.0mg/L 17 kinds of additive hybrid standard working solutions.
Preferably, step (2) first adds accurately to weigh the moon cake after 5.0g is ground in 50.0ml centrifuge tube pipes
30.0ml volume ratios are 1：1 methanol aqueous solution extracts, and then adds 0.5mol/L sodium hydroxide solution 1.0ml, is mixed in whirlpool
On instrument after vortex 1min, 0.42mol/L solution of zinc sulfate 1.5ml are added, mixes, adds 0.5mol/L sodium hydroxide solutions
1.4~1.8ml, shake up, be finally settled to 50.0ml with pure water, obtain sample extracting solution.
Preferably, it by sample extracting solution in temperature is 4.0~6.0 DEG C that step (3), which is, and rotating speed is 4000~5000r/min
Centrifuge in centrifuge 3~5min, with middling speed quantitative filter paper filter, remove upper strata grease, filtrate obtains through 0.22 μm of membrane filtration
Prepare liquid；
Preferably, the condition of liquid chromatograph is in step (4)：Chromatographic column：Pre-column is C18 posts (4.6mm × 4.5cm)；
Analytical column is Eclipse Plus C18 reverse chromatograms post (250mm × 4.6mm, 5.0 μm)；Mobile phase is 0.02mol/L acetic acid
Aqueous ammonium (A)+methanol (B)；Detector wavelength is 223~629nm；Sample size is 5.0~10.0uL, flow velocity is 0.8~
1.0mL/min, column oven are 40 DEG C.
It is further preferred that the gradient elution program of step (4) mobile phase is：0~12min, 5%B；12~20min,
5%~30%B；20~35min, 30%~68%B；35~44min, 68%B.
Preferably, step (4) wavelength changeover program is：Initial detecting wavelength 254nm, 2.00min：226nm6.00min：
223nm, 11.00min：267nm, 14.50min：293nm, 20.00min：254nm, 34.50min：629nm, 35.50min：
256nm。
Compared with prior art, the invention has the advantages that：
(1) present invention disposably detects the 17 kinds of food additives commonly used in moon cake using liquid chromatography simultaneously, wherein
Including 2 kinds of sweeteners, 7 kinds of preservatives and 8 kinds of synthetic food colors, cost is low, speed is fast, reliable results, favorable reproducibility.
(2) according to similar compatibility principle, corresponding standard mother liquor first is respectively configured using pure methanol and pure water, then mixes
Be configured to mix storing solution, then with volume ratio be 1：1 methanol-water is configured to hybrid standard working solution, solve 8 kinds of pigments with
And saccharin sodium, acesulfame potassium are soluble in water is not readily dissolved in organic solvent, and 4 kinds of parabenses and benzoic acid, sorb
Acid and dehydroactic acid are soluble in the problem of organic solvent is not readily dissolved in water.
(3) sodium hydroxide-zinc sulfate is selected extraction sample impurity to be made few, the rate of recovery is high as precipitating reagent.
(4) mobile phase selects methanol+ammonium acetate solution, makees with acetonitrile+ammonium acetate solution, acetonitrile+ammonium citrate solution
For mobile phase ratio, the separating degree of all each materials and analysis time are all ideal, as ammonium acetate 0.02mol/L, each group
Point peak shape, separating degree is preferable, the serious conditions that solve that erythrosine is overlapping with butyl p-hydroxybenzoate, dehydroactic acid trails.
The effect of ammonium acetate is exactly mainly to adjust PH, prevents sample from dissociating, and is easy to separate, and improves peak shape, phenomena such as suppressing to trail, is improved
The reappearance of sample retention.
(5) gradient elution program is used, peak type is improved, improve sensitivity and stability, shortens minute, make
Whole analysis process can meet the needs of 17 kinds of food additives detections simultaneously.
(6) according to 17 food additives ultra-violet absorption spectrum features, most suitable wavelength is selected；And according to its peak sequence
And the time difference, by wavelength switching mode, realize the single-time measurement simultaneously of 17 food additives.
(7) 17 kinds of food additives detection method minimum detectabilities of the invention are 0.013~0.055mg/kg, and mark-on returns
Yield is 82.3%~109.2%, and precision is 0.75~8.73%, has good sensitivity, the degree of accuracy and reappearance.
(8) the invention provides a kind of disposable detection method of a variety of mixed food additives in moon cake, the present invention are logical
Cross and standard liquid collocation method and pre-treatment precipitating reagent are effectively changed, improve mobile phase and use gradient elution journey
Sequence and Detection wavelength handoff technique reach disposable quick, 17 kinds of food additives purposes of Accurate Determining.
Brief description of the drawings
Fig. 1 is 17 kinds of food additives liquid chromatogram peak sequence figures in moon cake of the present invention.
In figure：1- acesulfame potassiums (Acesulfame Potassium), 2- benzoic acid (Benzoic Acid), 3- sorbic acids
(Sorbic Acid), 4- dehydroactic acids (Dehydroacetic), 5- lemon yellows (Tartrazine), 6- newly reds (New red),
7- amaranths (Amaranthus Red), 8- saccharin sodiums (Sodium Saccharin), 9- famille roses (Ponceau 4R), 10-
Sunset yellow (Sunset Yellow), 11- lure red (Allura Red), 12- methyl p-hydroxybenzoates (Methyl
), paraben 13- light blues (Brilliant Blue), 14- ethyl p-hydroxybenzoates (Ethyl paraben), 15- is to hydroxyl
The fat of benzoic acid third (Propyl paraben), 16- erythrosines (Erythrosine), 17- parahydroxybenzoate butyls (Butyl
paraben)。
Embodiment
The invention will be further described for the following examples, so that those skilled in the art can be better understood from this hair
It is bright and can be practiced, but illustrated embodiment is not as a limitation of the invention.
In embodiment, 4 kinds of P-hydroxybenzoic acid lipids refer to methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate,
The fat of P-hydroxybenzoic acid third, parahydroxybenzoate butyl.8 clock pigments refer to lemon yellow, newly red, amaranth, saccharin sodium, kermes
Red, sunset yellow, lure red, erythrosine.
Embodiment one
(1) prepared by mixed standard solution：
By dehydroactic acid, benzoic acid, sorbic acid and 4 kinds of P-hydroxybenzoic acid standard lipids mixed preparings in methyl alcohol,
Above-mentioned solution is mixed in pure water, obtains 100mg/L by acesulfame potassium, saccharin sodium and 8 kinds of pigment standard items mixed preparings in proportion
The storing solution of standard liquid.It is again 1 with volume ratio:1 methanol-water solution progressively dilutes preparation, respectively obtains mass concentration and is
0.1st, 0.5,1.0,5.0,10.0,20.0mg/L hybrid standard working solution.
(2) sample extraction：The experiment material of the present embodiment is lotus seed paste mooncake, and 5.0g samples are weighed after lotus seed paste mooncake is ground
In 50.0ml centrifuge tube pipes, it is 1 first to add 30.0ml volume ratios：1 methanol aqueous solution extracts, and then adds 0.5mol/LNaOH
Solution 1.0ml, in the 1min that is vortexed on whirlpool blending instrument, 0.42mol/L solution of zinc sulfate 1.5ml mixings are added, are added
0.5mol/L sodium hydroxide solution 1.5ml, shake up, are finally settled to 50.0ml with pure water.
(3) sample purification：It it is 4.0 DEG C in temperature, rotating speed is 5000r/min centrifuge after said extracted liquid is mixed
Middle centrifugation 4min, middling speed quantitative filter paper filtering, removes upper strata grease, filtrate supplies liquid chromatogram through 0.22 μm of membrane filtration, filtrate
It is to be measured.
(4) liquid chromatogram measuring：Mixed standard solution in the prepare liquid obtained in step (3) and step (1) is placed in
Liquid chromatograph equipped with UV-detector is detected, using quantified by external standard method, the wherein liquid chromatogram of standard liquid such as
Shown in Fig. 1.
The condition of liquid chromatograph is：
1. chromatographic column：Pre-column is C18 posts (4.6mm × 4.5cm)；Analytical column is Agilent Eclipse Plus C18 posts
(250mm × 4.6mm, 5.0 μm).
2. mobile phase：0.02mol/L ammonium acetate solutions (A)+methanol (B), refer to table 1.
The chromatography gradient elution program of table 1
| Time/min | A：Ammonium acetate solution (%) | B：Methanol (%) |
| 0~12min | 95% | 5% |
| 12~20min | 95%~70% | 5%~30% |
| 20~35min | 70%~32% | 30%~68% |
| 35~44min | 32% | 68% |
3. Detection wavelength and wavelength switching times table, refer to table 2.
The Detection wavelength and retention time table of 2 17 kinds of food additives of table
4. flow velocity is 0.8mL/min, column oven is 40 DEG C；Sample size is 10uL.
(5) rate of recovery：Detected by above-mentioned steps, food additives sample recovery rate data are shown in Table 3.
The rate of recovery of 3 17 kinds of food additives samples of table
(6) precision：Detected by above-mentioned steps, RSD (relative standard deviation) data of 17 kinds of food additives are shown in
Table 4.
The RSD of 4 17 kinds of food additives samples of table
(7) test material is the lotus seed paste mooncake that 5 parts of different companys' products are up-sampled from market, is examined by above-mentioned steps
Survey, measure peak area result and settle accounts its food additives content, testing result is as shown in table 5.
The residue detection result of 17 kinds of food additives in 5 commercially available lotus seed paste mooncake of table
Testing result content unit is mg/kg in table 5, and ND represents not detect.The detection limit of this method, acesulfame potassium ﹤
0.026mg/kg, benzoic acid ﹤ 0.032mg/kg, sorbic acid ﹤ 0.025mg/kg, dehydroactic acid ﹤ 0.055mg/kg, lemon yellow ﹤
0.036mg/kg, newly red ﹤ 0.023mg/kg, amaranth ﹤ 0.039mg/kg, saccharin sodium ﹤ 0.049mg/kg, carmine ﹤
0.037mg/kg, sunset yellow ﹤ 0.013mg/kg, lure red ﹤ 0.027mg/kg, methyl p-hydroxybenzoate ﹤ 0.015mg/kg, bright
Blue ﹤ 0.017mg/kg, ethyl p-hydroxybenzoate ﹤ 0.015mg/kg, the third fat of P-hydroxybenzoic acid ﹤ 0.016mg/kg, erythrosine ﹤
0.021mg/kg, parahydroxybenzoate butyl ﹤ 0.019mg/kg.
Embodiment two
The experiment material of embodiment two is fruit moon cake, and remaining step is identical with embodiment one.
The food additives sample recovery rate data of the present embodiment are shown in Table 6, and the RSD data of 17 kinds of food additives samples are shown in
Table 7,5 parts of samples are adopted with commercially available fruit moon cake, peak area result is measured and calculates food additives content, testing result is shown in Table 8.
The rate of recovery of 17 kinds of food additives samples in the fruit moon cake of table 6
The RSD of 17 kinds of food additives samples in the fruit moon cake of table 7
The residue detection result of 17 kinds of food additives in 8 commercially available fruit moon cake of table
Testing result content unit is mg/kg, and ND represents not detect.The detection limit of this table 8 is the same as table 5.
Embodiment three
The experiment material of embodiment three is kernel moon cake, and remaining step is identical with embodiment one.
The food additives sample recovery rate data of the present embodiment are shown in Table 9, and the RSD data of 17 kinds of food additives samples are shown in
Table 10,5 parts of samples are adopted with commercially available kernel moon cake, peak area result is measured and calculates food additives content, testing result is shown in Table
11。
The rate of recovery of 17 kinds of food additives samples in the kernel moon cake of table 9
The RSD of 17 kinds of food additives samples in the kernel moon cake of table 10
The residue detection result of 17 kinds of food additives in 11 commercially available kernel moon cake of table
Testing result content unit is mg/kg, and ND represents not detect.The detection limit of table 11 is the same as table 5.
Example IV
The experiment material of example IV is other moon cakes in addition to lotus-seed paste, fruit, nut stuffing, remaining step and embodiment
One is identical.
The present embodiment adopts 5 parts of samples with other commercially available filling moon cakes, measures peak area result and calculates food additives content,
Testing result is shown in Table 12.
The residue detection result of 17 kinds of food additives in other the commercially available filling moon cakes of table 12
Testing result content unit is mg/kg, and ND represents not detect.The detection limit of table 12 is the same as table 5.
The above embodiment of the present invention is not limiting the scope of the present invention, and embodiments of the present invention are not limited to
This, all this kind, according to the ordinary technical knowledge and customary means of this area, is not departing from this according to the above of the present invention
Under the premise of inventing above-mentioned basic fundamental thought, modification, replacement or the change of the other diversified forms made to said structure of the present invention
More, all should fall within the scope and spirit of the invention.
Claims (9)
1. the detection method of food additives in a kind of moon cake, it is characterised in that comprise the following steps：
(1) mixed standard solution preparation：
By dehydroactic acid, benzoic acid, sorbic acid, methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, P-hydroxybenzoic acid third
Fat, parahydroxybenzoate butyl standard items mixed preparing are in pure methanol；
Acesulfame potassium, saccharin sodium, lemon yellow, newly red, amaranth, saccharin sodium, famille rose, sunset yellow, red, the erythrosine standard items of temptation
Mixed preparing is in pure water；
Above-mentioned solution is mixed in proportion, obtains 17 kinds of additive hybrid standard storing solutions；
Again with methanol-aqueous solution progressively dilutes, with obtained 17 kinds of additive hybrid standard working solutions；
(2) sample extraction：The moon cake after grinding is weighed in centrifuge tube pipe, methanol-water solution extraction is first added, then adds hydrogen
Sodium hydroxide solution, it is well mixed；
Solution of zinc sulfate is added, is mixed, is added sodium hydroxide solution, shake up, finally with pure water constant volume, obtain sample extracting solution；
(3) sample purification：Sample extracting solution is centrifuged, filtered with filter paper, removes upper strata grease；Filtrate is filtered again through filter membrane,
Obtain prepare liquid；
(4) machine testing on：Mixed standard solution in the prepare liquid obtained in step (3) and step (1) is placed in equipped with ultraviolet
The liquid chromatograph of detector is detected, and using quantified by external standard method, is switched by gradient elution, wavelength, upper machine measures simultaneously
The content of 17 kinds of food additives.
2. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Mixed in step (1)
Standard reserving solution is 100mg/L hybrid standard storing solutions.
3. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Step (1), step
(2) methanol-water solution is that volume ratio is 1 in：1 methanol-water solution.
4. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Dehydrogenation in step (1)
Acetic acid, benzoic acid, sorbic acid and methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, the fat of P-hydroxybenzoic acid third, to hydroxyl
Benzoic acid fourth fat standard items are configured to 200-400mg/L mixed liquor after being completely dissolved with chromatographically pure level methanol；
Acesulfame potassium, saccharin sodium and lemon yellow, newly red, amaranth, saccharin sodium, famille rose, sunset yellow, red, the erythrosine standard of temptation
Product are configured to 200-400mg/L mixed liquor after being completely dissolved with laboratory one-level water, then mixed in equal amounts obtains 100mg/L and mixed
Standardization storing solution；It is again 1 with volume ratio:1 methanol aqueous solution progressively dilutes preparation, respectively obtain mass concentration for 0.1,
0.5th, 1.0,5.0,10.0,20.0mg/L 17 kinds of additive hybrid standard working solutions.
5. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Step (2) is accurate
The moon cake after 5.0g is ground is weighed in 50.0ml centrifuge tube pipes, it is 1 first to add 30.0ml volume ratios：1 methanol aqueous solution extracts,
Then 0.5mol/L sodium hydroxide solution 1.0ml are added, after vortex 1min on whirlpool blending instrument, add 0.42mol/L sulfuric acid
Zinc solution 1.5ml, mix, add 0.5mol/L 1.4~1.8ml of sodium hydroxide solution, shake up, be finally settled to pure water
50.0ml, obtain sample extracting solution.
6. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Step (3) is by sample
Product extract solution is 4.0~6.0 DEG C in temperature, centrifuges 3~5min in the centrifuge that rotating speed is 4000~5000r/min, uses middling speed
Quantitative filter paper filters, and removes upper strata grease, filtrate obtains prepare liquid through 0.22 μm of membrane filtration.
7. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Liquid phase in step (4)
Chromatographic condition is：Chromatographic column：Pre-column is C18 posts (4.6mm × 4.5cm)；Analytical column is the reverse colors of EclipsePlusC18
Compose post (250mm × 4.6mm, 5.0 μm)；Mobile phase is 0.02mol/L ammonium acetate solutions (A)+methanol (B)；Detector wavelength
For 223~629nm；Sample size is 5.0~10.0uL, and flow velocity is 0.8~1.0mL/min, and column oven is 40 DEG C.
8. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Step (4) mobile phase
Gradient elution program be：0~12min, 5%B；12~20min, 5%~30%B；20~35min, 30%~68%B；35
~44min, 68%B.
9. the detection method of food additives in moon cake according to claim 1, it is characterised in that：Step (4) wavelength is cut
Changing program is：Initial detecting wavelength 254nm, 2.00min：226nm6.00min：223nm, 11.00min：267nm,
14.50min：293nm, 20.00min：254nm, 34.50min：629nm, 35.50min：256nm.
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| CN109580847A (en) * | 2019-01-22 | 2019-04-05 | 梧州市食品药品检验所 | A kind of method of acesulfame potassium in measurement cake |
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| CN113341004A (en) * | 2021-05-12 | 2021-09-03 | 河南恒晟检测技术有限公司 | Method for determining benzoic acid, sorbic acid, saccharin sodium, acesulfame and dehydroacetic acid in food |
| CN115452963A (en) * | 2021-10-14 | 2022-12-09 | 王丽蓉 | Method for detecting mixed food additive in pickled vegetables |
| CN115452963B (en) * | 2021-10-14 | 2023-07-25 | 王丽蓉 | Method for detecting mixed food additive in pickled vegetables |
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