CN105021737B - It is a kind of at the same detect milk product in dicyandiamide and content of melamine method - Google Patents
It is a kind of at the same detect milk product in dicyandiamide and content of melamine method Download PDFInfo
- Publication number
- CN105021737B CN105021737B CN201510485741.8A CN201510485741A CN105021737B CN 105021737 B CN105021737 B CN 105021737B CN 201510485741 A CN201510485741 A CN 201510485741A CN 105021737 B CN105021737 B CN 105021737B
- Authority
- CN
- China
- Prior art keywords
- dicyandiamide
- standard
- solution
- content
- milk product
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Dairy Products (AREA)
Abstract
The invention discloses a kind of while the method for detecting dicyandiamide and content of melamine in milk product, methods described comprises the steps：（1）It is prepared by standard solution；（2）Testing sample process：Testing sample is dissolved in the water, Jing isolating protein → except obtaining prepare liquid after lipid → purification → filtration treatment；（3）Detected using high performance liquid chromatography tandem mass spectrum method.Method of the present invention simultaneously can carry out quantitative and qualitative analysis to the dicyandiamide and tripolycyanamide in milk product, with it is accurate, sensitive, the response rate is high the features such as.
Description
Technical field
The invention belongs to chemical analysis technology field, further belongs to field of detection of food safety, and in particular to Yi Zhongtong
When detect milk product in dicyandiamide and content of melamine method.
Background technology
At present, in food, the detection of feed additives not yet causes enough attention, especially the non-egg in milk product
The residual of white nitrogen, it is adulterated the problems such as had a strong impact on the survival and development of dairy enterprises or even whole industry, be detrimental to health,
Affect the orthobiosiss of people.On January 25th, 2013, the U.S.《Wall Street Journal》Report, the permanent natural collection of New Zealand milk giant
In part milk and milk powder that group produces, a small amount of toxic chemical substance dicyandiamide (dicyandiamide, abbreviation is detected
DCD).On the one hand, New Zealand claims that the food containing a small amount of dicyandiamide residue is harmless；Separately-aspect, orders to prohibit again
Sell the milk fish containing dicyandiamide.The milk product enterprise and milk supplier in the whole world 60% adopts New Zealand's milk supply, New Zealand's breast
Product accounts for the 40% or so of Chinese full import infant food.After " Sanlu milk powder case " occurs at home, compatriots are to domestic baby
In the case that Follow Up Formula confidence is lost entirely, the hypotoxicity even avirulence how thing each side explain dicyandiamide no matter is related to, this
Secondary event has undoubtedly caused the Chinese common people to the panic of a milk product new round and has caused people again to milk product safety problem
Concern.
Impact to health can not be with " poison be dead " of parties concerned's explanation as standard.Can carcinogenic, cause as dicyandiamide
Abnormal, mutagenesis, allergy, and infringement human organ, seldom have experimental data.For health, " there is no experimental data " when
Wait, just should more take conservative careful attitude.And milk product industry prestige is low, food safety extreme sensitivity society
In face of reality, the residue detection safety criterion for formulating dicyandiamide is the thing being compelled to do.
Dicyandiamide is a kind of important chemical products intermediate, another name dicyanodiamine, dicyandiamide（It is the dimerization of cyanamide
Body, and the cyano derivative of guanidine）, its Pathological experiment data display：Its mouse oral LD50>4000 mg/kg, the oral LD of rabbit50
>3000 mg/kg, if may result in half mice death by poisoning more than 4000 mg to the mice nursing dicyandiamide of a collection of 1kg
Die.Although though the toxicity of dicyandiamide is little, a large amount of or harm of the Long Term Contact to human body is very important.Especially for baby children
For youngster, each internal organs is not yet developed completely, and the metabolic function of liver, renal excretion system be not perfect, internal material generation
Thank different from adult, toxic reaction is also variant, the long-term accumulated of noxious substance there certainly will be certain infringement to its internal organs gesture, and
And the determination of three cause " teratogenesis, carcinogenic, mutagenesis " properties of poisonous substance is a long-term physiological disposition, then infant is being grown up
During must just prevent the intake of poisonous and harmful substance.
At present, China there is now 22388-2008 raw milks of GB/T and melamine in dairy products detection method standard,
But not yet set up dicyandiamide residues detection standard in baby milk powder, for simultaneously detect in baby milk powder tripolycyanamide and
The document report of dicyandiamide is also few.Therefore, set up it is a kind of quick and precisely and can detect simultaneously melamine in dairy products and pair
The method of cyanamide residual quantity is extremely urgent, can be that dicyandiamide in accurate objective evaluation milk product and content of melamine provide technology
Means；Evaluation to milk product safety and protection people's is healthy significant.
The content of the invention
Present invention aim at providing a kind of while the method for detecting dicyandiamide and content of melamine in milk product.The mesh
Be realized in：Described is a kind of while detecting that the method for dicyandiamide and content of melamine in milk product includes following step
Suddenly：
（1）It is prepared by standard solution：Dicyandiamide and tripolycyanamide standard reserving solution are prepared respectively first；Again two kinds of storing solutions are mixed
Conjunction is diluted to standard mixed solution；Standard mixed solution is carried out into gradient dilution finally, standard hybrid working solution series is obtained；
（2）Testing sample process：Testing sample is dissolved in the water, adds protein precipitant, vortex, centrifugation to obtain
Clear liquid, is vortexed after adding grease removal agent, is centrifuged, reject fat oxidant layer, adds purification bag, be vortexed, supernatant be centrifuged to obtain, most in clear liquid
By filtering with microporous membrane, prepare liquid is obtained；
（3）Detected using high performance liquid chromatography-tandem mass method, by step（1）Described standard hybrid working is molten
Liquid series and step（2）Described prepare liquid difference sample introduction, first makes standard curve, according to standard curve and the prepare liquid
Sample introduction result is calculating the content of dicyandiamide and tripolycyanamide；Wherein：
Chromatographic condition is：
Chromatographic column：C18, 150mm × 2.1mm × 5 μm；
Flow velocity：0.2 mL/min；
Column temperature：25℃；
Sample size：5 μL；
Mobile phase A：10mmol/L ammonium acetate solutions second acid for adjusting pH to 3.0, Mobile phase B：Acetonitrile；Gradient elution,
Elution program is that elution program is as shown in table 1.
1 Gradient Elution condition of table
Mass Spectrometry Conditions are：
Electron spray ionisation source（ESI）；
Scan mode：Cation is scanned；
Detection mode：Multiple-reaction monitoring（MRM）；
Electron spray voltage：500 V；
It is dried temperature degree：325℃；
Dry gas stream amount：6L/min；
Sheath temperature degree：380℃；
Sheath throughput：10 L/min；
Capillary voltage：3500 V；
Mass spectrometry parameters are as shown in table 2.
2 mass spectrometry parameters of table
| Title | [M+H]+Quasi-molecular ions | Taper hole voltage（V） | Quota ion（Collision energy） | Qualitative ion（Collision energy） |
| Dicyandiamide | 85.2 | 23 | 68.1（17） | 43.1（13） |
| Tripolycyanamide | 127.1 | 97 | 85.1（18） | 68.1（34） |
As the content of dicyandiamide and tripolycyanamide in milk product is relatively low, particularly dicyandiamide is only trace sometimes, traditional
Method is difficult to detect, and the method for the high performance liquid chromatography-tandem mass that the present invention is adopted overcomes traditional detection method
Deficiency, meets the detection requirement of micro dicyandiamide and tripolycyanamide in food, and with the spy that specificity is strong, sensitivity is high
Point.
Description of the drawings
Fig. 1 is Agela Venusil AS-T C18Total ion current figure；
In figure, 2.323 is dicyandiamide quasi-molecular ions, and 8.458 is tripolycyanamide quasi-molecular ions.
Ion figures of the Fig. 2 for melamine quantitative ion 85.1；
Fig. 3 is the ion figure of the qualitative ion of tripolycyanamide 68.1；
Ion figures of the Fig. 4 for dicyandiamide quota ion 68.1；
Fig. 5 is the ion figure of the qualitative ion of dicyandiamide 43.1；
Fig. 6 is the total ion current figure of certain brand baby formulas milk powder.
Specific embodiment
The invention will be further described below in conjunction with the accompanying drawings, but never in any form the present invention is any limitation as, and is based on
Present invention teach that any conversion made, each falls within the scope of the present invention.
It is of the present invention a kind of while detecting that the method for dicyandiamide and content of melamine in milk product includes following step
Suddenly：
（1）It is prepared by standard solution：Dicyandiamide and tripolycyanamide standard reserving solution are prepared respectively first；Again two kinds of storing solutions are mixed
Conjunction is diluted to standard mixed solution；Standard mixed solution is carried out into gradient dilution finally, standard hybrid working solution series is obtained.
Preferred as one kind of the present invention, the preparation method of the standard solution is：Dicyandiamide and melamine are weighed accurately
The each 25.0mg of amine standard substance, is dissolved with methanol respectively and is settled to 25.0mL, obtains the standard reserving solution of 1000 μ g/mL；Pipette institute
The each 1.0mL mixing of standard reserving solution is stated, and 100mL is settled to deionization, is made the standard mixed solution of 10 μ g/mL；Finally use
Negative dairy base solution carries out gradient dilution, obtains standard hybrid working solution series.Standard hybrid working solution series
The mass concentration of dicyandiamide is 1～209 ng/mL, and the mass concentration of tripolycyanamide is 2～420 ng/mL.
（2）Testing sample process：Testing sample is dissolved in the water, adds protein precipitant, vortex, centrifugation to obtain
Clear liquid, is vortexed after adding grease removal agent, is centrifuged, reject fat oxidant layer, adds purification bag, be vortexed, supernatant be centrifuged to obtain, most in clear liquid
By filtering with microporous membrane, prepare liquid is obtained；
Due to milk product, particularly dry milk component is complicated, and protein and lipopoteins matter and fats content are higher, therefore
In the pretreatment process of milk powder, extremely important is exactly to remove protein and lipid.Now conventional removing method of protein is
Select certain protein precipitant to make protein precipitation, be then centrifuged for, be filtered to remove.Conventional protein precipitant includes trichlorine
Acetic acid solution, ammonia acetonitrile and its aqueous solution, acetonitrile.By test, compare the sedimentation effect of three kinds of precipitant, as a result show：
To be purified with solid phase extraction column and be concentrated after extracting using trichloroacetic acid, solid phase extraction column is to dicyandiamide and trimerization
Extraction effect is bad simultaneously for cyanamide；It is simply square that dicyandiamide and tripolycyanamide in milk powder are extracted simultaneously with ammonification acetonitrile solution and acetonitrile
Just, extracting solution can be without carrying out Solid-Phase Extraction.Wherein, the effect extracted by 5% ammonification acetonitrile and acetonitrile does not have significant difference,
Acetonitrile is slightly better, but because preparing solution, the use of acetonitrile is more convenient.
Remove solution in lipid material method be using grease removal agent extract lipid material, then be vortexed, centrifugation.It is conventional
Grease removal agent includes normal hexane and petroleum ether etc., general grease removal 1 time, but removes complete and matrix interference for guarantee lipid material
It is less, present invention preferably uses normal hexane grease removal 2 times.
After removing isolating protein and lipid material, in order to further eliminate the interference of substrate, carried out only using purification bag
Change is processed.Conventional purification bag is Agilent QuEChERs purifications bag and the dicyandiamide purification of moon rising sun Welchrom QuChERS-
Pipe.But, the latter is preferable to the clean-up effect of dicyandiamide, poor to the clean-up effect of tripolycyanamide, therefore the present invention is preferably
Agilent QuEChERs purification bags（25mg C18With 150mg MgSO4）Extracting solution is purified.
Preferred as one kind of the present invention, the aperture of the microporous filter membrane is 0.2 μm.
（3）Detected using high performance liquid chromatography-tandem mass method, by step（1）Described standard hybrid working is molten
Liquid series and step（2）Described prepare liquid difference sample introduction, first makes standard curve, according to standard curve and the prepare liquid
Sample introduction result is calculating the content of dicyandiamide and tripolycyanamide；Wherein：
Chromatographic condition is：
Chromatographic column：C18, 150mm × 2.1mm × 5 μm；
Flow velocity：0.2 mL/min；
Column temperature：25℃；
Sample size：5 μL；
Mobile phase A：10mmol/L ammonium acetate solutions second acid for adjusting pH to 3.0, Mobile phase B：Acetonitrile；Gradient elution,
Elution program is as shown in table 1.
Mass Spectrometry Conditions are：
Electron spray ionisation source（ESI）；
Scan mode：Cation is scanned；
Detection mode：Multiple-reaction monitoring（MRM）；
Electron spray voltage：500 V；
It is dried temperature degree：325℃；
Dry gas stream amount：6L/min；
Sheath temperature degree：380℃；
Sheath throughput：10 L/min；
Capillary voltage：3500 V；
Mass spectrometry parameters are as shown in table 2.
Preferred as one kind of the present invention, chromatographic column adopts Agela Venusil AS-T C18Chromatographic column.
After detection, in milk product, the qualitative method of dicyandiamide and tripolycyanamide is：
Under same experimental conditions, if certain in the retention time of the chromatographic peak detected in prepare liquid and standard solution
The retention time of component peaks is consistent（Table 3, Fig. 1）, and the mass-to-charge ratio of selected two pairs of daughter ions is consistent, prepare liquid it is qualitative
When ion relative abundance is compared with standard working solution, relative deviation then can determine that sample less than the scope of the regulation of table 4
Middle presence component.
The retention time and qualitative ion relative abundance of 3 dicyandiamide of table and tripolycyanamide
| Title | Retention time（min） | Abundance（%） |
| Dicyandiamide | 2.323 | 56.5 |
| Tripolycyanamide | 8.458 | 50.3 |
During 4 qualitative determination of table with respect to abundance of ions maximum allowable offset
| Relative ion abundance/% | ＞ 50 | ＞ 20 ~ 50 | ＞ 10 ~ 20 | ≤10 |
| Relative deviation/the % of permission | ±20 | ±25 | ±30 | ±50 |
After detection, in milk product, the quantitative approach of dicyandiamide and tripolycyanamide is：
According to the testing result of the standard hybrid working solution series of standard solution, standard curve is made, double cyanogen in sample
The mass fraction quantified by external standard method of amine and tripolycyanamide, by formula（1）Calculated：
（1）
In formula：XFor the mass fraction of certain component in sample, ng/g；c i Be in the sample drawn by standard curve certain
The mass concentration of component, ng/mL；c 0For the mass concentration of certain component of blank solution, ng/mL；VFor sample constant volume, mL；mFor sample quality, g.
Meansigma methodss with parallel assay twice are accurate to 0.01 ng/g as measurement result, as a result；Parallel determinations twice
Between relative deviation not should be greater than 10.0%.
In addition, general milk product include liquid milk, milk powder and other milk product all can with method of the present invention while
Detection dicyandiamide and content of melamine.
1 testing sample of embodiment is detected
（1）Instrument
Liquid Chromatography-Tandem Mass Spectrometry instrument：1290 high performance liquid chromatographs（Agilent companies of the U.S.）, Agilent 6,460 3
Weight level Four bar liquid chromatography-tandem mass spectrometry instrument, is furnished with electric spray ion source（Agilent companies of the U.S.）；Analytical balance（Switzerland prunus mume (sieb.) sieb.et zucc.
Teller；Model：AB204-S）；Vortex vortex mixer；Centrifuge；Tool screw lid politef centrifuge tube（50mL）.
（2）Reagent and material
Methanol（Chromatographically pure, Dikma）；Acetonitrile（Chromatographically pure, Dikma）；Acetic acid（Analysis is pure）；Formic acid（Analysis is pure）；It is ultrapure
Water (resistivity >=18.2M Ω cm)；Trichloroacetic acid（Analysis is pure）；Ammonia（Analysis is pure）.
Chromatographic column：Agela Venusil AS-T C18（150 × 2.1mm, 5 μm）；Agilent QuEChERs substrate point
Powder purification pipe；Syringe and filter；High pure nitrogen (purity is more than 99.999%).
Testing sample：Certain brand baby formulas milk powder.
（3）It is prepared by standard solution
The preparation of dicyandiamide and tripolycyanamide standard reserving solution：The each 25.0mg of standard substance is accurately weighed, distinguishes molten with methanol
Solution is settled to 25.0mL, obtains the standard reserving solution of about 1000 μ g/mL, the freezen protective at -18 DEG C.
The preparation of standard hybrid working solution：1000 μ g/mL standard reserving solutions 1.0mL mixing is pipetted respectively, uses deionization
Constant volume makes the standard mixed solution that concentration is 10 μ g/mL in 100mL volumetric flasks, preserves at 4 DEG C.Finally with negative breast
Article substrate solution carries out gradient dilution, obtains standard hybrid working solution series.Double cyanogen in standard hybrid working solution series
The mass concentration of amine is respectively 1.05,4.19,20.9,41.9,105,209ng/mL, the mass concentration of tripolycyanamide is 2.10,
8.41、42.0、84.1、210、420ng/mL。
（4）Sample treatment
1.0g milk powder is weighed, is placed in 50 mL centrifuge tubes, be initially charged 2mL deionized waters, vortex 1min, ultrasonic 5min make
Milk powder fully dissolves, and adds 2mL acetonitriles, and then vortex 1min divides 3 addition 2mL acetonitriles again, be vortexed after adding every time
1min, in making milk powder, protein is fully precipitated, and 10000 r/min are centrifuged 10 min, the supernatant of 3 times are transferred completely into another
Prop up in 50 mL centrifuge tubes, add normal hexane 10mL, vortex 1min, 10000 r/min centrifugation 5min, discard normal hexane layer, again
Add 10mL normal hexane to repeat above-mentioned grease removal process, then take out clear liquid 2mL.
2mL clear liquids are transferred to into 10mL centrifuge tubes, QuEChRS dispersed solid phase reagent bags are added（25mg C18And 150mg
MgSO4）, vortex 1min, 10000 r/min are centrifuged 5 min, take 0.2 μm of membrane filtration of supernatant Jing into sample injection bottle, to be measured
It is fixed.
（4）High performance liquid chromatography-Tandem Mass Spectrometry Analysis
High performance liquid chromatography reference conditions：Chromatographic column：Agela Venusil AS-T C18（150 × 2.1mm, 5 μm）；Stream
Speed：0.2 mL/min；Column temperature：25℃；Sample size：5 μL；Mobile phase A：10mmol ammonium acetate solutions second acid for adjusting pH is extremely
3.0, Mobile phase B：Acetonitrile solution；Condition of gradient elution is shown in Table 1.
Mass Spectrometry Conditions：Electron spray ionisation source（ESI）；Scan mode：Cation is scanned；Detection mode：Multiple-reaction monitoring
（MRM）；Electron spray voltage：500 V；It is dried temperature degree：325℃；Dry gas stream amount：6 L/min；Sheath temperature degree：380℃；Sheath
Throughput：10 L/min；Capillary voltage：3500 V；Mass spectrometry parameters are shown in Table 2.
（5）Testing result
According to the measurement result of standard mixed solution series（Fig. 1～5）, with peak area as vertical coordinate, mass concentration is horizontal stroke
Coordinate, makes standard curve, linear regression is carried out to correction data, and regression equation and correlation coefficient are shown in Table 5.According to linear pass
System, calculates through formula, and the testing result for obtaining certain brand baby formulas milk powder is as shown in table 6.
The linear relationship of 5 dicyandiamide of table and tripolycyanamide
| Title | Linear equation | The range of linearity（ng/mL） | Correlation coefficient |
| Dicyandiamide | A=909.235C+1339.07 | 1~209 | 0.9984 |
| Tripolycyanamide | A=244.175C+1035.82 | 2~420 | 0.9992 |
Note：In linear equationAFor peak area；CFor the mass concentration of each component.
6 sample detection result of table
| Sample ID | Dicyandiamide (ng/g) | Tripolycyanamide (ng/g) |
| Certain brand baby formulas milk powder | - | 20.75 |
According to table 6：Detect containing tripolycyanamide in certain brand baby formulas milk powder, its mass fraction is
20.75 ng/g, do not detect dicyandiamide.
The reappearance test of 2 method of embodiment
Reappearance test is carried out using mark-on method.1.0 g of negative powdered milk sample is taken, is carried out as described in Example 1 to be measured
Sample treatment, is separately added into standard substance so that dicyandiamide concentration is 10.48 ng/mL, and melamine concentration is to use 21.01 ng/
ML, arranges 6 repetitions.In the same terms（Chromatograph and Mass Spectrometry Conditions are with embodiment 1）Lower measure dicyandiamide and content of melamine,
METHOD FOR CONTINUOUS DETERMINATION 3 days, and its RSD% in a few days and in the daytime is calculated, as a result as shown in table 6 and table 7.
The repeatability result of 6 dicyandiamide of table（n=6）
The repeatability result of 7 tripolycyanamide of table（n=6）
As shown in Table 6, the wherein dicyandiamide in a few days RSD of 3 days is respectively 1.31%, 2.74% and 5.10%, and RSD is in the daytime
4.33%；The tripolycyanamide in a few days RSD of 3 days is respectively 2.94%, 3.69% and 2.23%, and RSD is 3.07% in the daytime, contrasts existing skill
Art, the method have preferable repeatability to two materials.
3 test limit of embodiment is tested
Detection limit is defined as concentration amount corresponding when the response signal of detector generation is 3 times of noise figure；It is quantitative to limit
Justice concentration amount corresponding when being 10 times of noise figure for the response signal that detector is produced, is calculated and is somebody's turn to do with 3 times and 10 times of levels of noise
The concentrations and quantitative concentrations of method, and pass through computing formula（1）, bring concentration into formula, constant volume 10mL and sample
Amount 1g, conversion obtain the detection limit and quantitative limit of sample, are shown in Table 8.
The concentrations of 8 dicyandiamide of table and tripolycyanamide, quantitative concentrations, detection limit and quantitative limit
As shown in Table 8, dicyandiamide is quantitatively limited to 25.7ng/g, and tripolycyanamide is quantitatively limited to 7.10ng/g, the method inspection
Rising limit and quantitative limit are less than 10 ng/g of quantitative limit of tripolycyanamide in GB GB/T 23888-2008, less than lot of documents report
50 ng/g of dicyandiamide quantitative limit in road, illustrates that the method has preferable sensitivity, relatively low test limit, preferably can meet
Trace detection is required.
4 response rate of embodiment
Dicyandiamide is added in negative powdered milk sample so that low quality concentration is 4.19ng/mL, mean quality concentration is
10.48g/mL, high concentration are 20.98g/mL；Add tripolycyanamide so that low quality concentration is 8.41ng/mL, medium
Mass concentration is 21.02g/mL, high concentration is 42.03g/mL, carries out response rate experiment, and each mass concentration is parallel to be carried out
3 times, the response rate of dicyandiamide and tripolycyanamide is obtained, 9 are shown in Table.
The response rate of 9 dicyandiamide of table and tripolycyanamide
As shown in Table 9, the response rate of dicyandiamide is 84.75%～95.49%, better than 80.1%～88.2% He of document report
79%～83%, the tripolycyanamide response rate is 96.13%%～100.63%, better than 223888-2008 standard recovery rates 80% of GB/T
~ 110%, illustrate that the response rate of the method is higher.
5 stability test of embodiment
In negative sample, dicyandiamide standard substance is added to cause concentration for 10.48g/mL, addition tripolycyanamide reference material
It is 21.02 ng/mL that matter causes concentration, then will be detected after the sample handled well 0,6,12,24,48 h of placement, and investigate sample
The stability of product, each time period are tested 3 times and are averaged, and have obtained the stability experiment result of dicyandiamide and tripolycyanamide,
It is shown in Table 10.
10 dicyandiamide of table and tripolycyanamide stability experiment result
In sum, method of the present invention can determine dicyandiamide and tripolycyanamide simultaneously, the method have it is sensitive,
Accurately, favorable reproducibility, the features such as the response rate is high, disclosure satisfy that dicyandiamide and content of melamine detection in baby milk powder will
Ask.
Claims (6)
1. a kind of at the same detect milk product in dicyandiamide and content of melamine method, it is characterised in that comprise the steps：
（1）It is prepared by standard solution：Dicyandiamide and tripolycyanamide standard reserving solution are prepared respectively first；Two kinds of storing solutions are mixed into dilute again
It is interpreted as standard mixed solution；Standard mixed solution is carried out into gradient dilution finally, standard hybrid working solution series is obtained；
（2）Testing sample process：Testing sample is dissolved in the water, protein precipitant acetonitrile or 5% ammonification acetonitrile, whirlpool is added
Rotation, centrifugation obtain supernatant, are vortexed, are centrifuged, reject fat oxidant layer, add containing 25mg in clear liquid after adding grease removal agent normal hexane
C18、150mg MgSO4QuEChERs purification bags, be vortexed, be centrifuged to obtain supernatant, last via hole diameter is 0.2 μm of microporous filter membrane mistake
Filter, obtains prepare liquid；
（3）Detected using high performance liquid chromatography-tandem mass method, by step（1）Described standard hybrid working solution system
Row and step（2）Described prepare liquid difference sample introduction, first makes standard curve, according to standard curve and the sample introduction of the prepare liquid
As a result calculating the content of dicyandiamide and tripolycyanamide；Wherein：
Chromatographic condition is：
Chromatographic column：C18, 150mm × 2.1mm × 5 μm；
Flow velocity：0.2mL/min；
Column temperature：25℃；
Sample size：5μL；
Mobile phase A：10mmol/L ammonium acetate solutions second acid for adjusting pH to 3.0, Mobile phase B：Acetonitrile；Gradient elution, eluting
Program is：
Mass Spectrometry Conditions are：
Electron spray ionisation source ESI；
Scan mode：Cation is scanned；
Detection mode：Multiple-reaction monitoring MRM；
Electron spray voltage：500V；
It is dried temperature degree：325℃；
Dry gas stream amount：6L/min；
Sheath temperature degree：380℃；
Sheath throughput：10L/min；
Capillary voltage：3500V；
Mass spectrometry parameters are：
。
2. according to claim 1 a kind of while the method for detecting dicyandiamide and content of melamine in milk product, which is special
It is step to levy（1）Method prepared by the standard solution is：Dicyandiamide and each 25.0mg of tripolycyanamide standard substance are weighed accurately,
Dissolved with methanol respectively and be settled to 25.0mL, obtain the standard reserving solution of 1000 μ g/mL；Pipette the standard reserving solution each
1.0mL mixes, and is settled to 100mL with deionization, makes the standard mixed solution of 10 μ g/mL；Finally with negative dairy base
Solution carries out gradient dilution, obtains standard hybrid working solution series.
3. according to claim 1 and 2 a kind of at the same detect milk product in dicyandiamide and content of melamine method, its
The mass concentration for being characterised by dicyandiamide in described standard hybrid working solution series is 1 ~ 209ng/mL, the matter of tripolycyanamide
Amount concentration is 2 ~ 420ng/mL.
4. according to claim 1 a kind of while the method for detecting dicyandiamide and content of melamine in milk product, which is special
It is 1 ~ 4 addition of the protein precipitant point to levy.
5. according to claim 1 a kind of while the method for detecting dicyandiamide and content of melamine in milk product, which is special
It is step to levy（2）Described grease removal agent number of processes is 1 ~ 2 time.
6. according to claim 1 a kind of while the method for detecting dicyandiamide and content of melamine in milk product, which is special
It is step to levy（3）The chromatographic column is Venusil AS-T C18Chromatographic column.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510485741.8A CN105021737B (en) | 2015-08-10 | 2015-08-10 | It is a kind of at the same detect milk product in dicyandiamide and content of melamine method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510485741.8A CN105021737B (en) | 2015-08-10 | 2015-08-10 | It is a kind of at the same detect milk product in dicyandiamide and content of melamine method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105021737A CN105021737A (en) | 2015-11-04 |
| CN105021737B true CN105021737B (en) | 2017-04-05 |
Family
ID=54411862
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510485741.8A Expired - Fee Related CN105021737B (en) | 2015-08-10 | 2015-08-10 | It is a kind of at the same detect milk product in dicyandiamide and content of melamine method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105021737B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548433B (en) * | 2015-12-17 | 2017-06-06 | 浙江出入境检验检疫局检验检疫技术中心 | The method that liquid chromatography tandem mass spectrometry detects various non-protein nitrogen-containing compound contents in milk simultaneously |
| CN107271576A (en) * | 2016-04-08 | 2017-10-20 | 内蒙古蒙牛乳业（集团）股份有限公司 | The method for determining dicyandiamide residual quantity in dairy products |
| CN105929061A (en) * | 2016-04-25 | 2016-09-07 | 广西壮族自治区梧州食品药品检验所 | Method for determining melamine in milk powder by using ASE-UPLC |
| CN105954399A (en) * | 2016-04-25 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Method for rapidly detecting content of melamine in milk powder |
| CN109557230A (en) * | 2018-12-07 | 2019-04-02 | 云南省烟草质量监督检测站 | A method of three classes dithiocarbamate residual quantity in measurement tobacco |
| CN110618227B (en) * | 2019-10-17 | 2021-07-23 | 北京君德同创生物技术股份有限公司 | Method for detecting dicyandiamide content in cyanamide solution by using HPLC method |
| CN111521458B (en) * | 2020-04-21 | 2023-02-03 | 深圳市深大检测有限公司 | Method for detecting liquid milk melamine |
| CN112505165A (en) * | 2020-07-10 | 2021-03-16 | 中检科（北京）测试技术有限公司 | Method for measuring residual quantity of dicyandiamide in dairy product |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2648641A (en) * | 1950-08-03 | 1953-08-11 | Fir Tex Insulating Board Co | Fire retardant coating containing a carbohydrate, urea or melamineformaldehyde and dicyandiamide or melamine |
| CN102419354B (en) * | 2011-09-08 | 2013-07-10 | 宁波检验检疫科学技术研究院 | General rapid detection method for small molecule poisonous and harmful substances in liquid milk |
| CN103344726B (en) * | 2013-06-14 | 2014-11-05 | 中华人民共和国温州出入境检验检疫局 | Method for extraction of dicyandiamide component in dairy product |
-
2015
- 2015-08-10 CN CN201510485741.8A patent/CN105021737B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105021737A (en) | 2015-11-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105021737B (en) | It is a kind of at the same detect milk product in dicyandiamide and content of melamine method | |
| CN104535664B (en) | A kind of method simultaneously detecting multiple mycotoxin in sesame paste | |
| CN104614479B (en) | A kind of detection method of food vitamins | |
| CN105319292B (en) | A kind of UPLC-MS/MS methods that 29 kinds of four class limits the use of residue of veterinary drug in analyzing animal food | |
| CN102692469B (en) | The method of mycotoxin levels in LC-MS/MS ginseng | |
| Vinogradova et al. | Rapid surface plasmon resonance immunobiosensor assay for microcystin toxins in blue-green algae food supplements | |
| CN106324144A (en) | Method for detecting chlorate, perchlorate and bromate from milk powder and infant formula milk powder by hydrophilic interaction chromatography-tandem mass spectrometry | |
| CN103543221A (en) | Method for simultaneously detecting multiple mycotoxins in milk | |
| CN112684030A (en) | Method for detecting perfluoroalkanoic acid compound in aquatic product by enrichment purification-liquid chromatography tandem mass spectrometry and application | |
| CN108982691A (en) | Arsenic mercury shape simultaneous quantitative determination in aquatic products | |
| CN108445133A (en) | A kind of method of glufosinate-ammonium and glyphosate drug residue in detection tealeaves | |
| CN106706829A (en) | Method for measuring diarrhetic shellfish poisons in shellfishes by use of immunoaffinity purification-liquid chromatography-tandem mass spectrometry | |
| CN105784858B (en) | Method for measuring PPCPs in environmental soil | |
| Guo et al. | Highly sensitive fluorescence methods for the determination of alfuzosin, doxazosin, terazosin and prazosin in pharmaceutical formulations, plasma and urine | |
| CN110501438B (en) | Detection method of sodium picosulfate in weight-reducing tea | |
| Sun et al. | Speciation analysis of trace inorganic arsenic in dietary supplements by slurry sampling hydride generation atomic absorption spectrometry | |
| Chen et al. | Development of a high-performance liquid chromatography method for the simultaneous quantification of four organoarsenic compounds in the feeds of swine and chicken | |
| CN105424865A (en) | Illegal cooking oil detection method | |
| CN102998305A (en) | Method for detecting melamine in raw milk by utilizing chromogenic reaction | |
| CN103217498B (en) | Method for detecting dicyandiamide in milk powder with LC-MS (liquid chromatography/mass spectrometry) and sample preparation method | |
| CN110702828A (en) | Method for determining four arsenic morphological concentrations in whole blood or red blood cells by HPLC-HG-AFS method | |
| CN115856153A (en) | Method for detecting 8 estrogen residues in edible part of crab | |
| Krata et al. | Reference values of methyl mercury mass fractions in new type of environmental matrix-matching materials for speciation analysis assigned by species-specific isotope dilution inductively coupled plasma mass spectrometry and high-performance liquid chromatography | |
| Lv et al. | A simple and high-throughput method of ultrasonic extraction-capillary electrophoresis for determination of melamine in milk | |
| CN112285243B (en) | Treatment method for detecting drug residues in animal tissue sample, confirmation detection method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170405 Termination date: 20190810 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |