CN103278594A - Universal rapid detection method for micromolecule poisonous and harmful materials in powdery food - Google Patents
Universal rapid detection method for micromolecule poisonous and harmful materials in powdery food Download PDFInfo
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Abstract
The invention discloses a universal rapid detection method for micromolecule poisonous and harmful materials in powdery food. The method is characterized by comprising the following steps of performing simple and universal extraction and purification on a liquid milk sample to obtain a loading solution; preparing a mixed standard solution and making a matrix-adding standard curve; then measuring the micromolecule poisonous and harmful materials in the liquid milk by high performance liquid chromatography-triple quadrupole tandem mass spectrometry combination; and finally performing concentration calculation. The method has the advantages that the pretreatment and instrumental analysis processes have strong compatibility for compounds with different physicochemical properties; detection efficiency is high; operability is strong; detection limit can meet all subjected target objects; and detection cost is reduced.
Description
Technical field
The present invention relates to the detection method of the medium and small molecule poisonous and harmful substance of a kind of powdered food, especially the general method for quick that relates to correlation detection technologies such as the liquid chromatography-tandem mass spectrometry of the little molecule poisonous and harmful substance of ultramicron in the dry powder shape food such as a kind of baby formula milk powder, milk powder, ground rice, oatmeal (as veterinary drug, environmental hormone, biotoxin, pesticide, adjuvant, illegal additive etc., molecular weight is less than 1500) or liquid chromatography-high resolution mass spectrum.
Background technology
Dry powder shape food such as baby formula milk powder, milk powder, ground rice, oatmeal are the huge nutriment of consumption figure.It is a lot of to influence its quality safety factor, wherein of a great variety, the different small organic molecule hazardous material (molecular weight is less than 2000) of character constitutes in various degree hidden danger to the consumer health, comprise veterinary drug (microbiotic, antibiotic, hormone, growth accelerator, antiparasitic agent, antiphlogistic class etc.), agricultural chemical insecticide (as organic Phosphorus), biotoxin (as aflatoxin), illegal additive etc., these materials may produced, processing (bringing into auxiliary material), transportation or storage link enter in the product, these substance classes quantity are quite huge, physicochemical property differs greatly, add the albumen that contains high level in the powdered food, fat and matrix interference composition such as carbohydrates bring great challenge to the quality control of the little molecule poisonous and harmful substance of ultra micro content.
Along with mass spectrum especially LC-Tof Ms, the appearance of LC-MS/MS and constantly development, multi-class many method for detecting residue reports have been arranged successively, Yamada delivered first piece of LC-MS/MS method (document 1:Yamada R that screens 130 kinds of veterinary drugs in beef, pork and the chicken simultaneously in 2006, Kozono M, Ohmori T, Morimatsu F, Kitayama M (2006) Biosci Biotechnol Biochem70:54 – 65), owing to use normal hexane layering degrease, be not suitable for the low pole compound test, versatility is not strong; Ortelli etc. adopt acetonitrile to extract, UPLC-Tof Ms detected 150 kinds of veterinary drugs (document 2:Ortelli D., Cognard E., Jan P.and Edder P. in the milk after supernatant carried out ultrafiltration, J.Chrom.B, 2009,877 (23): 2363-2374.), macrolides is lower than 10%, quinolones recovery scope 98-807%, Tetracyclines 141-258%, part benzene forces imidazoles to be higher than 436%, and the recovery is undesirable; Hans(2008) put forward universal extracting method (Generic Extraction Method) concept, adopt UPLC-MS/MS to analyze 172 all agricultural chemicals, biotoxin, phytotoxin and veterinary drug in the matrix such as feed, milk, honey, meat simultaneously, be considered to universal detection method (document 3:Mol HG the earliest
P, Zomer P, de Rijk TC, Stolker AA, Mulder PP.Toward a generic extraction method for simultaneous determination of pesticides mycotoxins, plant toxins, and veterinary drugs in feed and food matrixes.Anal.Chem.2008,80 (24), 9450-9459.), but sample does not pass through purified treatment basically in this method, and detectability can not satisfy all objects that tried, and pollutes instrument easily yet.Stolker etc. have analyzed 101 kinds of veterinary drugs in the milk, and sample adopts acetonitrile to extract the little column purification of Strata-X (document 4:Stolker A.A.M., Rutgers P., Oosterink E., Lasaroms J.J.P., Peters R.J.B., van Rhijn J.A., Nielen M.W.F., Anal.Bioanal.Chem., 2008,391 (6): 2309-2322.), by purification styles such as Solid-Phase Extraction, part of compounds may be lost in this process, and the pre-treatment time is longer again simultaneously.The many method for detecting residue of above-mentioned bibliographical information multiclass are all having problems aspect versatility and the clean-up effect.
At present, though China at powdered food in existing national standard and the industry standard detection method quantity of these venomous injurants more, but these methods are divided into a plurality of sample preparation methods, operation steps is too much, testing staff's energy disperses, not only waste time and energy, detection efficiency is low, operability is not strong, and the project coverage rate is still not comprehensive, brings the risk of omission easily, and the cycle is long, experimental cost is high, is difficult to finish to the how residual efficient detection of venomous injurant in the powdered food and supervision.Trace it to its cause, the pre-treatment of these methods is difficult to compatibility at the compound of different physicochemical properties nothing more than, lacks versatility, therefore is difficult in and significantly improves monitoring efficient on the conventional method.Therefore, in the face of the complicated various and quantity of kind in the problem that ever-increasing organic contaminant and current detection standard method disperse, improve detection efficiency and flux, reducing omission and reducing analysis cost has become pressing for of industry development.
Summary of the invention
Technical matters to be solved by this invention provides the general method for quick of the medium and small molecule poisonous and harmful substance of a kind of powdered food, the pre-treatment of this method is compatible strong at the compound of different physicochemical properties with the instrumental analysis process, the detection efficiency height, workable, detectability can satisfy all objects that tried and reduce the detection cost.
The present invention solves the problems of the technologies described above the technical scheme that adopts: the general method for quick of the medium and small molecule poisonous and harmful substance of a kind of powdered food specifically may further comprise the steps:
(1) sample extraction
5.00g milk powder or ground rice are added in the plastic centrifuge tube, and in test tube, add 400mg solid EDTA-Na protective agent, 25mL extract acetonitrile, 20000rmp homogenate 1min, the centrifugal 5min of 4500rmp obtains first supernatant then; After the solid residue of centrifugal gained being joined in 60% ethanol water of 12mL fully mixing, adding the 15mL acetonitrile repeats to extract 1 time, the centrifugal 5min of 4500rmp obtains second supernatant then, first supernatant and second supernatant is merged be settled to the 50mL scale mark with acetonitrile and obtain sample extracting solution;
(2) purification method
A. high levels of organic solvents precipitated carbohydrate
Sample extracting solution is placed centrifuge tube, in the centrifugal 15min of 4500rmp, precipitated carbohydrate;
B. superfreeze purifies
Place metal test tube cover to put into-40 ℃ of refrigerator 2h centrifuge tube, then behind the centrifugal 5min of 4500rmp, to get 40mL(immediately and to bring bottom settlings in order reducing, only get top 40mL part), supernatant is filtered in the heart bottle fast, and in the heart bottle, adding the 30mL isopropyl alcohol, 40 ℃ of steamings are done near;
C. the solid phase dispersion extraction purifies (d-SPE)
Add 5mL by acetonitrile, the composite mixed extract of second alcohol and water and 2.5mL normal hexane at the heart bottle, then the liquid in the heart bottle is transferred in the centrifuge tube, 40 ℃ of nitrogen blows after evaporation removes normal hexane, adds the amino filler (NH of 150mg
2) and 75mg N-propyl group ethylenediamine (PSA) filler in centrifuge tube, fully behind the mixing, behind the centrifugal 5min of 13000rmp, get the 4mL supernatant to another centrifuge tube, 40 ℃ of nitrogen blow evaporation and blow to 0.8-1mL, add the dimethyl sulfoxide (DMSO) of 0.6mL then, fully behind the mixing, water is settled to 1.5mL, measures with acquisition method LC Run1 pattern and LC Run2 pattern;
(3) mixed standard solution is formed and the preparation of matrix typical curve
A. mixed standard solution is formed: hybrid standard working solution concentration sees Table 2, table 3.
B. the matrix typical curve is quantitative: add the above-mentioned standard solution of 0,50,100,200,400,800 μ L in the 5g sample, handle according to above-mentioned 1-2 pre-treatment step;
(4) Ultra Performance Liquid Chromatography-triple quadrupole bar tandem mass spectrum coupling is measured
A. efficient liquid phase separation
Liquid-phase condition is:
A) chromatographic column: Acquity Waters HSS-T
3, 2.1mm * 100mm, 1.8 μ m;
B) if mass spectrum adopts the Run1 positive ion mode to measure, adopt mobile phase A so
1: 0.1% aqueous formic acid, Mobile phase B
1: the methyl alcohol that contains 0.1% formic acid; Gradient elution program (in percent by volume) is: 0-7min, 100%A
17-10.5min, 0%A
110.5-12min, 100%A
1Each elution time Duan Zhongjun is with Mobile phase B
1Supply remaining percentage;
If mass spectrum adopts Run2 to measure at negative ions switch mode (waters instrument positive ion mode and negative ion mode can be done together, switching time 20ms), adopt mobile phase A so
2: 2mmol/L ammonium acetate solution, Mobile phase B
2: methyl alcohol; Gradient elution program (in percent by volume) is: 0-7min, 100%A
27-10.5min, 0%A
210.5-12min, 100%A
2Each elution time Duan Zhongjun is with Mobile phase B
2Supply remaining percentage;
C) flow velocity: 0.4mL/min;
D) chromatogram column temperature: 45 ℃;
E) sample size: 10 μ L;
F) stream switches: 0～1.5min transfer valve is opened, and chromatographic column flows out liquid and enters waste liquid, beginning mass spectrum collection behind the 1.5min, treat the whole wash-outs of target analytes after, flow switches to waste liquid;
B. Mass Spectrometer Method
The mass spectrum condition is:
A) adopt the electro-spray ionization ionization source, capillary voltage: positive ion, negative ion voltage are respectively 3.5kV and 3.0kV;
B) desolventizing temperature degree: 500 ℃, ion source temperature: 150 ℃;
C) desolventizing flow velocity: 900L/h, taper hole flow velocity: 50L/h;
D) collision gas: argon gas 3.3 * 10
-3Millibar;
E) grouping scheme: the medium and small molecule poisonous and harmful substance of powdered food detects by mode shown in table 2 and 3;
(4) density calculating method
The content of determinand obtains according to following computing formula (1) in the sample:
In the formula:
Determinand content in the X-sample, unit are every liter of microgram (μ g/L);
Testing concentration in the C-sample treatment solution calculates according to the matrix typical curve, and unit is every milliliter of nanogram (ng/mL);
V-constant volume, unit are milliliter (mL);
M-volume of sample, unit are milliliter (mL).
The mixed volume of acetonitrile, second alcohol and water is than being 67:10:33 in the mixed extract described in the c step of step (2).
Compared with prior art, the invention has the advantages that:
1, adopt two kinds of extraction solvents successively to extract: first pass adopts acetonitrile homogenate to extract; After adopting 60% ethanol water mixing earlier second time, add acetonitrile solution again and extract.Ensured that simultaneously polarity determinand and low pole compound all obtain higher recovery, and can not cause dry powder shape food chance water caking phenomenon; In addition, EDTA-Na
2As protective agent, prevent that Tetracyclines, macrolides from being caused that by metal ion-chelant the recovery reduces;
2, the high levels of organic solvents precipitated carbohydrate is effective: organic solvent concentration reaches about 92% in the extract, and the water in the extract can add behind the isopropyl alcohol rotary evaporation and do near, realizes concentrating;
3, superfreeze method: being reduced in when not influencing the target compound recovery of temperature, make the solubleness of the higher fat of content in the extract, carbohydrates impurity reduce and part is separated out, separate by centrifugal reaching with filtration, otherwise can't carry out follow-up purified treatment;
4, disperse Solid-Phase Extraction (DSPE) purified treatment to utilize amino filler (NH
2) and PSA filler (150mg+75mg), solvent environment is 5mL acetonitrile+ethanol+water (67+10+33, volume ratio), the purification of simple and efficient realization sample, and the method repeatability is good.Compare with solid phase extraction, improve treatment effeciency greatly, with low cost.Utilize stream to cut, namely preceding 1.5min switches to waste liquid, removes salinity, sugar, has remedied the defective that does not adopt solid phase extraction column to purify.
5, decide that the ratio of organic solvent and water has a significant impact testing result in the solution.This method is in order to the final constant volume of 40% dimethyl sulfoxide (DMSO) aqueous solution, can ensure good peak type and the linearity of polar compound, can ensure being unlikely to from decide solution, to separate out of polar compound again and cause linear poor phenomenon, avoid because deciding the different requirements that need additionally to increase LC-MS/MS analysis number of run of solution;
6, flow with methyl alcohol as organic phase, not only have price and environment-friendly advantage, and the relative acetonitrile of sensitivity of most compounds acquisition is higher; Determinand respectively with formic acid or ammonium acetate as mobile phase adjuvant, each testing compound can be realized improving chromatographic resolution flexibly and improve Ionization Efficiency.
7, this method is through the demonstration test of the concrete medicine of table 2, table 3, and the average interpolation recovery of three levels is at 51.7-157.1%, its corresponding RSD(withinday precision) 1.1-28.8% and.For second interpolation level, carried out the test of day to day precision, the result shows that the RSD of the recovery is at 2.4-26.2%.The average recovery and the precision (in a few days and in the daytime) of adding sees form 234 for details.For the determinand more than 80%, between 70-120% and 1-15%, precision is good respectively for the average recovery rate in milk powder and milk and repeatability, all can satisfy the requirement of limiting the quantity of of present domestic and European Union.This method is compared with the method for document 2 in the background technology, this method recovery ideal, and precision is good; Compare with the method for document 3, this method not only comprises universal extracting method, also possesses the versatility purification method simultaneously, and detectability can reach 0.01～5 μ g/L; Compare with the method for document 4, rapid and simple, recovery height, precision is good.
In sum, compare with the many method for detecting residue of above-mentioned bibliographical information multiclass, this method has had bigger breakthrough at the aspects such as versatility, clean-up effect and processing speed of pre-treatment except the detection coverage rate is wider.Extractive technique and purification techniques have versatility, the detected object coverage is wide, reduce the omission risk, the poisonous and hazardous organism of little molecule is only handled by a pre-treating method, add that pre-treatment is easy fast, with the detection method of a plurality of methods in the past, detection efficiency has obtained the lifting of essence, shortens sense cycle; Simultaneously, compare with adopting solid phase extraction column, not only guarantee polarity by force and the recovery of the determinand that polarity is weak, and reduced the cost consumption of sample preparation.
The present invention is from the general general character of micromolecular compound, fully takes into account the dissolubility, thermal stability, ph stability, polarity power, coexistence stability of various materials etc., develops universal quick pretreatment technology (Generic﹠amp; Rapid sample preparation), MRM(multi-channel reaction monitoring by UPLC-ESI-MS/MS) pattern is measured, satisfying under the prerequisite that detects the requirement of limiting the quantity of, can bring the effect of raising the efficiency significantly and reducing the detection cost to production run control and food inspection work.
Description of drawings
Fig. 1 is the general method for quick process flow diagram of the medium and small molecule poisonous and harmful substance of powdered food of the present invention.
Embodiment
Describe in further detail below in conjunction with the present invention of accompanying drawing embodiment.
The invention will be further described with embodiment below, but protection scope of the present invention is not limited to this, and protection domain is as the criterion with claim.
1, material
1.1 main agents and material
1.2 key instrument
Waters ultra high efficiency liquid phase systems (U.S. Waters, ACQUITY UPLC) and triple quadrupole bar tandem mass spectrum (U.S. Waters, XEVO TQ); Refrigerated centrifuge (German Sigma, 3-15k), Rotary Evaporators (Switzerland Buchi, Rota vapor R210) Nitrogen evaporator (U.S. OA, N-EVAP-24), Superpure water machine (French Mi Libo, Milliporeco).
2, the detection method (as shown in Figure 1) of the medium and small molecule poisonous and harmful substance of powdered food
(1) sample extraction
5.00g milk powder or ground rice are added in the plastic centrifuge tube, and in test tube, add 400mg solid EDTA-Na protective agent, 25mL extract acetonitrile, 20000RMP homogenate 1min, the centrifugal 5min of 4500RMP obtains first supernatant then; After the solid residue of centrifugal gained being joined in 60% ethanol water of 12mL fully mixing, adding the 15mL acetonitrile repeats to extract 1 time, centrifugal second supernatant that obtains of the centrifugal 5min of 4500RMP then merges first supernatant and second supernatant and is settled to 50mL with acetonitrile and obtains sample extracting solution;
(2) purification method
A. high levels of organic solvents precipitated carbohydrate
Sample extracting solution is placed centrifuge tube, in the centrifugal 15min of 4500RMP, precipitated carbohydrate;
B. superfreeze purifies
Place metal test tube cover to put into the 2h of-40 ℃ of refrigerators centrifuge tube, behind the centrifugal 5min of 4500RMP, get the 40mL supernatant liquid filtering immediately to the heart bottle then, and add the 30mL isopropyl alcohol in the heart bottle, 40 ℃ of steamings are done near;
D. the solid phase dispersion extraction purifies (d-SPE)
Add 5mL by acetonitrile, the composite mixed extract of second alcohol and water (three's volume ratio is followed successively by 67:10:33) and 2.5mL normal hexane at the heart bottle, then the liquid in the heart bottle is transferred in the centrifuge tube, 40 ℃ of nitrogen blows after evaporation removes normal hexane, adds the amino filler (NH of 150mg
2) and 75mg PSA(N-propyl group ethylenediamine) filler is in centrifuge tube, fully behind the mixing, behind the centrifugal 5min of 13000RMP, get the 3mL supernatant to another centrifuge tube, 40 ℃ of nitrogen blow evaporation and blow to 0.8-1mL, add the dimethyl sulfoxide (DMSO) of 0.6mL then, fully behind the mixing, water is settled to 1.5mL, measures with acquisition method LC Run1 and LC Run2;
(3) mixed standard solution is formed and the preparation of matrix typical curve
A. mixed standard solution is formed: hybrid standard working solution concentration sees Table 2, table 3.
B. the matrix typical curve is quantitative: add the above-mentioned standard solution of 0,50,100,200,400,800 μ L in the 5g sample, handle according to above-mentioned 1-2 pre-treatment step;
(4) Ultra Performance Liquid Chromatography-triple quadrupole bar tandem mass spectrum coupling is measured
A. efficient liquid phase separation
Liquid-phase condition is:
A) chromatographic column: Acquity Waters HSS-T
3, 2.1mm * 100mm, 1.8 μ m;
B) if mass spectrum adopts the Run1 pattern to measure, adopt mobile phase A so
1: 0.1% aqueous formic acid, Mobile phase B
1: the methyl alcohol that contains 0.1% formic acid; Gradient elution program (in percent by volume) is: 0-7min, 100%A
17-10.5min, 0%A
110.5-12min, 100%A
1Each elution time Duan Zhongjun is with Mobile phase B
1Supply remaining percentage;
If mass spectrum adopts the Run2 pattern to measure, adopt mobile phase A so
2: 2mmol/L ammonium acetate solution, Mobile phase B
2: methyl alcohol; Gradient elution program (in percent by volume) is: 0-7min, 100%A
27-10.5min, 0%A
210.5-12min, 100%A
2Each elution time Duan Zhongjun is with Mobile phase B
2Supply remaining percentage (as shown in table 1);
The UPLC eluent gradient parameter of table 1. object arranges
C) flow velocity: 0.4mL/min;
D) chromatogram column temperature: 45 ℃;
E) sample size: 10 μ L;
F) stream switches: 0～1.5min transfer valve is opened,, chromatographic column flows out liquid and enters waste liquid, beginning mass spectrum collection behind the 1.5min;
B. Mass Spectrometer Method
The mass spectrum condition is:
A) adopt the electro-spray ionization ionization source, capillary voltage: positive ion, negative ion voltage are respectively 3.5kV and 3.0kV;
B) desolventizing temperature degree: 500 ℃, ion source temperature: 150 ℃;
C) desolventizing flow velocity: 900L/h, taper hole flow velocity: 50L/h;
D) collision gas: argon gas 3.3 * 10
-3Millibar;
E) grouping scheme: the medium and small molecule poisonous and harmful substance of powdered food detects by mode shown in table 2 and 3;
(4) density calculating method
The content of determinand obtains according to following computing formula (1) in the sample:
In the formula:
Determinand content in the X-sample, unit are every liter of microgram (μ g/L);
Testing concentration in the C-sample treatment solution calculates according to the matrix typical curve, and unit is every milliliter of nanogram (ng/mL);
V-constant volume, unit are milliliter (mL);
M-volume of sample, unit are milliliter (mL).
3. demonstration test
Following form is the grouping scheme that is used for 300 kinds of medicines of this method of checking
The compound Chinese and English title that table 2.LC Run1 gathers, chemical classification number, molecular weight, retention time, adduct ion, quota ion are to, qualitative ion pair, ion ratio and standard operation solution concentration
The compound Chinese and English title that table 3.LC Run2 gathers, chemical classification number, molecular weight, retention time, adduct ion, quota ion are to, qualitative ion pair, ion ratio and standard operation solution concentration
Certainly, above-mentioned explanation is not limitation of the present invention, and the present invention also is not limited to above-mentioned giving an example.Those skilled in the art are in essential scope of the present invention, and the variation of making, remodeling, interpolation or replacement also should belong to protection scope of the present invention.
Claims (2)
1. the general method for quick of the medium and small molecule poisonous and harmful substance of powdered food is characterized in that specifically may further comprise the steps:
(1) sample extraction
5.00g milk powder or ground rice are added in the plastic centrifuge tube, and in test tube, add 400mg solid EDTA-Na protective agent, 25mL extract acetonitrile, 20000rmp homogenate 1min, the centrifugal 5min of 4500rmp obtains first supernatant then; After the solid residue of centrifugal gained being joined in 60% ethanol water of 12mL fully mixing, adding the 15mL acetonitrile repeats to extract 1 time, the centrifugal 5min of 4500rmp obtains second supernatant then, first supernatant and second supernatant is merged be settled to the 50mL scale mark with acetonitrile and obtain sample extracting solution;
(2) purification method
A. high levels of organic solvents precipitated carbohydrate
Sample extracting solution is placed centrifuge tube, in the centrifugal 15min of 4500rmp, precipitated carbohydrate;
B. superfreeze purifies
Place metal test tube cover to put into-40 ℃ of refrigerator 2h centrifuge tube, behind the centrifugal 5min of 4500rmp, get 40mL immediately then, supernatant is filtered in the heart bottle fast, and adds the 30mL isopropyl alcohol in the heart bottle, and 40 ℃ of steamings are done near;
C. the solid phase dispersion extraction purifies
Add 5mL by acetonitrile at the heart bottle, the composite mixed extract of second alcohol and water and 2.5mL normal hexane, then the liquid in the heart bottle is transferred in the centrifuge tube, 40 ℃ of nitrogen blows after evaporation removes normal hexane, add the amino filler of 150mg and 75mg N-propyl group ethylenediamine filler in centrifuge tube, fully behind the mixing, behind the centrifugal 5min of 13000rmp, get the 4mL supernatant to another centrifuge tube, 40 ℃ of nitrogen blow evaporation and blow to 0.8-1mL, the dimethyl sulfoxide that adds 0.6mL then, fully behind the mixing, water is settled to 1.5mL, measures with acquisition method LC Run1 pattern and LC Run2 pattern;
(3) mixed standard solution is formed and the preparation of matrix typical curve
A. mixed standard solution is formed: hybrid standard working solution concentration sees Table 1, table 2;
B. the matrix typical curve is quantitative: add 0 μ L, 50 μ L, 100 μ L, 200 μ L, 400 μ L, the above-mentioned standard solution of 800 μ L in the 5g sample, handle according to above-mentioned (1)-(2) pre-treatment step;
(4) Ultra Performance Liquid Chromatography-triple quadrupole bar tandem mass spectrum coupling is measured
A. efficient liquid phase separation
Liquid-phase condition is:
A) chromatographic column: Acquity Waters HSS-T
3, 2.1mm * 100mm, 1.8 μ m;
B) if mass spectrum adopts the Run1 positive ion mode to measure, adopt mobile phase A so
1: 0.1% aqueous formic acid, Mobile phase B
1: the methyl alcohol that contains 0.1% formic acid; Gradient elution program (in percent by volume) is: 0-7min, 100%A
17-10.5min, 0%A
110.5-12min, 100%A
1Each elution time Duan Zhongjun is with Mobile phase B
1Supply remaining percentage;
If mass spectrum adopts Run2 to measure at the negative ions switch mode, adopt mobile phase A so
2: 2mmol/L ammonium acetate solution, Mobile phase B
2: methyl alcohol; Gradient elution program (in percent by volume) is: 0-7min, 100%A
27-10.5min, 0%A
210.5-12min, 100%A
2Each elution time Duan Zhongjun is with Mobile phase B
2Supply remaining percentage;
C) flow velocity: 0.4mL/min;
D) chromatogram column temperature: 45 ℃;
E) sample size: 10 μ L;
F) stream switches: 0～1.5min transfer valve is opened, and chromatographic column flows out liquid and enters waste liquid, beginning mass spectrum collection behind the 1.5min, treat the whole wash-outs of target analytes after, flow switches to waste liquid;
B. Mass Spectrometer Method
The mass spectrum condition is:
A) adopt the electro-spray ionization ionization source, capillary voltage: positive ion, negative ion voltage are respectively 3.5kV and 3.0kV;
B) desolventizing temperature degree: 500 ℃, ion source temperature: 150 ℃;
C) desolventizing flow velocity: 900L/h, taper hole flow velocity: 50L/h;
D) collision gas: argon gas 3.3 * 10
-3Millibar;
E) grouping scheme: the medium and small molecule poisonous and harmful substance of powdered food detects by mode shown in table 1 and 2;
Table 1.LC Run1 pattern
Table 2.LC Run2 pattern
(4) density calculating method
The content of determinand obtains according to following computing formula (1) in the sample:
In the formula:
Determinand content in the X-sample, unit are every liter of microgram (μ g/L);
Testing concentration in the C-sample treatment solution calculates according to the matrix typical curve, and unit is every milliliter of nanogram (ng/mL);
V-constant volume, unit are milliliter (mL);
M-volume of sample, unit are milliliter (mL).
2. the general method for quick of the medium and small molecule poisonous and harmful substance of a kind of powdered food according to claim 1 is characterized in that: the mixed volume of acetonitrile, second alcohol and water is than being 67:10:33 in the mixed extract described in the c step of step (2).
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| CN109490451A (en) * | 2018-10-16 | 2019-03-19 | 宁波中盛产品检测有限公司 | The universal precipitation and purification agent of strong water-soluble object and its pre-treating method of chromatography, Mass Spectrometer Method |
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| CN106770863A (en) * | 2015-11-19 | 2017-05-31 | 内蒙古蒙牛乳业（集团）股份有限公司 | The method of remains of pesticide content in detection liquid diary product |
| CN106770863B (en) * | 2015-11-19 | 2018-07-13 | 内蒙古蒙牛乳业（集团）股份有限公司 | The method for detecting remains of pesticide content in liquid diary product |
| CN106404949A (en) * | 2016-08-31 | 2017-02-15 | 天津市中升挑战生物科技有限公司 | Detection method for content of carprofen |
| CN106840836A (en) * | 2016-12-31 | 2017-06-13 | 湖北省理化分析测试中心 | Plastic cement race track poisons detection pre-treating method |
| CN106841430A (en) * | 2017-01-04 | 2017-06-13 | 华中农业大学 | A kind of method that Liquid Chromatography-Tandem Mass Spectrometry determines anti-microbial type and forbidden drug in feed |
| CN109490451A (en) * | 2018-10-16 | 2019-03-19 | 宁波中盛产品检测有限公司 | The universal precipitation and purification agent of strong water-soluble object and its pre-treating method of chromatography, Mass Spectrometer Method |
| CN109490451B (en) * | 2018-10-16 | 2021-08-31 | 宁波中盛产品检测有限公司 | Universal precipitation purifying agent for strong water-soluble target and pretreatment method for chromatographic and mass spectrometric detection of universal precipitation purifying agent |
| CN112198264A (en) * | 2019-07-08 | 2021-01-08 | 上海凯宝药业股份有限公司 | Method for detecting veterinary drug residues in bear gall powder |
| CN114778737A (en) * | 2022-04-27 | 2022-07-22 | 天津国科医工科技发展有限公司 | Liquid chromatography detection sample pretreatment method capable of shortening time |
| CN114778737B (en) * | 2022-04-27 | 2024-05-10 | 天津国科医疗科技发展有限公司 | Liquid chromatography detection sample pretreatment method capable of shortening time |
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