CN105259288A - Method for simultaneously detecting multiple drug residues in poultry tissues - Google Patents
Method for simultaneously detecting multiple drug residues in poultry tissues Download PDFInfo
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Abstract
The invention discloses a method for detecting drug residues in poultry tissues. The method comprises the following steps of selecting a poultry sample, adding sodium chloride and acetonitrile, shaking out, adding sodium hydroxide solution, and vibrating for 30s to 5min; then, ultrasonic extracting for 10 to 40min, centrifuging, absorbing supernate for standby use, drying other solution by distillation, adding concentrated hydrochloric acid in the residues, adjusting pH to be about 1 to 3; then, adding acetonitrile, vibration extracting, centrifuging solution subjected to acetonitrile vibration extracting, absorbing supernate to merge with the above supernate, concentrating to dry to obtain sample extracting solution residues; adding a matrix solid-phase dispersion agent and 2ml ethyl acetate, swirling for 2min, taking supernatant liquid, drying through nitrogen, then adding 30 percent of methanol solution containing 0.1 percent of methanoic acid, ultrasonic dissolving, filming and injecting the sample; adopting HLPC-MS to carry out gradient elution detection. According to the method provided by the invention, the current standard one-time one-sample one-item is adjusted to one-time one-sample five-items, so that the time consumed for detection is greatly shortened.
Description
Technical field
The present invention relates to a kind of chromatographic detection method, a kind of method that particularly Ultra Performance Liquid Chromatography tandem mass spectrum detection of drugs is residual, belongs to stratographic analysis field.
Background technology
Poultry refers generally to chicken, duck, goose etc., with high protein, low fat, high nutrition and famous, plays and have very important meaning in daily life.The poultry now commercially sold is large-scale cultivation mostly, because bird in large-scale cultivation process gathers denseness of set greatly, once there be a poultry sick, can infect fast and come, occur epidemic disease epidemic disease.So general raiser can add appropriate medicine to prevent the appearance of disease epidemic disease in breeding process, is also having clear and definite drug use kind and the requirement of time accordingly in GB.
Some illegal raisers are in order to improve the cultivation density of bird, and prevent the medicine of the appearance illegal use state's laws regulation plaintext forbidding of disease epidemic disease to prevent and treat poultry diease, What is more uses forbidden drug to promote bird.After state food drug regulatory department finds, make quite high attention, the birds meat products for various places carries out strict work of making a random inspection, and the object of detection mainly includes following drug ingedient:
Diamantane amine drug, mainly contains amantadine hydrochloride, rimantadine hydrochloride, as people's antiviral agent, is usually used in the control of catching a cold, and No. 560th, Ministry of Agriculture bulletin is classified as cultivation class forbidden drugs.
Olaquindox, also known as olaquindox, have teratogenesis shape, mutagenesis, carcinogenesis to people, forbidding feed addictive is listed in by the U.S. and European Union, and " Chinese veterinary pharmacopoeia " (2005 editions) also clear stipulaties forbid using olaquindox in poultry and aquaculture.
Tilmicosin, European Union is poultry muscle, skin, lipase 37 5 μ g/kg, liver 1000 μ g/kg, kidney 250 μ g/kg to the limit standard of Tilmicosin; Japan is water 1mg/kg under chicken gizzard and edible to the limit standard of Tilmicosin, and China only requires to have formulated limit standard to the limitation of chicken meat product with reference to European Union.
Dexamethasone, a kind of sugared sebaceous glands hormone, Japan specifies that in Positive List System its limitation is for 0.003mg/kg, and uses only for part beast-like animals, and the Ministry of Agriculture of China No. 235 bulletin forbids that bird uses dexamethasone.
But the detection method of existing poultry drug residue is mostly more old, is from other detection method amendment adjustment, there is the problems such as kind is uneven, cost is high, sense cycle is long, operation easier is large.What the detection as diamantane sulfonamides residues was conventional is vapor-phase chromatography; Olaquindox and metabolin many employings high performance liquid chromatograph thereof are equipped with UV-detector; Dexamethasone and tilmicosin residue are commonly used Liquid Chromatography-Tandem Mass Spectrometry and are detected.All there is difference in various degree in instrument, detection method, reagent etc. that various detection method adopts, and all there is greatest differences in detectable scope and precision, how to propose a kind of easy, quick, economical, reliable detection method, the disposable residue detection completing multi-medicament is imperative.
Summary of the invention
The object of the invention is to overcome that prior art drug residue detection method is old, the skimble-scamble defect of standard, the detection method that in a kind of poultry tissues, multi-medicament is residual is provided.The inventive method can detect the medicament residue compositions such as amantadine hydrochloride, rimantadine hydrochloride, Tilmicosin, dexamethasone, olaquindox metabolite, both can detect wherein a kind of composition, also can disposable detection multiple.Method of the present invention is QuEChERS/ Ultra Performance Liquid Chromatography tandem mass spectrometry (ultrapressureliquidchromatography-tandemmassspectrometry, UPLC-MS/MS), is the method for quick of a kind of advanced person.
In order to realize foregoing invention object, the invention provides following technical scheme:
A detection method for poultry tissues drug residue, comprises the following steps:
(1) poultry sample is got, add sodium chloride and acetonitrile, (acutely) jolting 1 ~ 5 minute, preferably 2 minutes, add the sodium hydroxide solution of 1 ~ 4mol/L of 2 ~ 5 times of volumes, preferably 2mol/L sodium hydroxide solution 3 times (volume mass is than ml/g), then vibrate 30 seconds to 5 minutes, preferably 1 ~ 2 minute.Then, 30 ~ 50 DEG C of ultrasonic extractions 10 ~ 40 minutes, best ultrasonic extraction 20 ~ 30 minutes.Centrifugal, Aspirate supernatant (this supernatant can be stored in evaporative flask for subsequent use) for subsequent use, all the other solution evaporates to dryness.In centrifugal process, rotating speed is greater than 3000r/min, 5min as centrifugal in 4000r/min, draws upper strata acetonitrile 10ml in evaporative flask, all the other solution evaporates to dryness.
(2) add concentrated hydrochloric acid in the residue obtained to step (1) evaporate to dryness, regulate pH about 1 ~ 3.Preferably, 1 ~ 6mol/L hydrochloric acid is used to carry out pH adjustment.Through overregulating PH=1 ~ 3, preferably pH approximates 2.0.Then, add acetonitrile mechanical shaking extraction, preferably add 10.0ml acetonitrile vibration 2min.Through the solution of acetonitrile mechanical shaking extraction, after centrifugal, Aspirate supernatant and the middle supernatant drawn of step (1) merge (be incorporated in evaporative flask, facilitate concentration), and 30 ~ 50 DEG C are concentrated into dry, obtain sample extracting solution residue.
Preferably, in centrifugal process, rotating speed is greater than 3000r/min, 5min as centrifugal in 4000r/min, draws upper strata acetonitrile 10ml and is incorporated in first time extract, is preferably concentrated at 40 DEG C near dry, obtains sample extracting solution residue, to be clean.
(3) in sample extracting solution residue, matrix solid phase dispersion agent is added, ethyl acetate 2ml, vortex 2min, get supernatant liquid, nitrogen dries up, and then adds 1.0mL30vol% methanol solution (containing 0.1vol% formic acid), ultrasonic dissolution, crosses film, treats sample introduction.
Specifically, the solution that ultrasonic dissolution uses is that surplus is the mixed solution of water containing 30% methyl alcohol and 0.1% formic acid.
Described matrix solid phase dispersion agent is consumption is 0.6g.Described matrix solid phase dispersion agent is C
18and anhydrous sodium sulfate.Wherein, C
18use amount is 50mg, and anhydrous sodium sulfate use amount is 550mg.
(4) high performance liquid chromatography-tandem mass is adopted to detect.With containing 5mmol/L ammonium acetate 0.1% formic acid water and methyl alcohol for eluent gradient wash-out.
Further, gradient elution degree is: mobile phase: A is the 0.1% formic acid water containing 5mmol/L ammonium acetate, and B is methyl alcohol; Gradient elution program, gradient condition is: 10%B:0 ~ 30 second; 10% ~ 90%B:30 second ~ 2 minutes; 90%B:2 ~ 4 minute; 90% ~ 10%B:4 ~ 4.5 minute; 10%B:4.5 ~ 6.5 minute.
Detection method of the present invention, applies up-to-date QuEChERS technology and Ultra Performance Liquid Chromatography tandem mass spectrometry detection, easily sample pretreatment technique simple and direct by QuEChERS technology, achieves the efficient fast processing of poultry sample.Meanwhile, by making sample preparation speed to the improvement of continuing to optimize of sample handling processes, the target detection thing recovery improve, make multi-medicament composition can in disposable detection analytic process Rapid Detection, testing result favorable reproducibility, is applicable to applying.
Compared with prior art, beneficial effect of the present invention:
1. detection method of the present invention saves detection time: by being originally reduced to 4 hours/mono-sample five project by GB and provincial standard or 4 hours industry standard used times/mono-sample individual event, the used time was only less than original 1/3.
2. detection method of the present invention saves material (consumable material) cost: reagent consumption, by original 50 yuan/sample, is reduced to 10 yuan/sample.If detected with kit, only single Project Cost is at about 50 yuan in test box detection, and new method is larger compared to the RNA isolation kit cost range of decrease, and advantage is more obvious.
3. detection method of the present invention reduces equipment investment: because the used time analyzed simultaneously original individual event detects 5 times shortens to 1 times of used time, the frequency of utilization of instrument reduces to 1/5, and maintenance cost and failure cost reduce greatly.
4. detection method environmental protection of the present invention: pre-treatment is simple, and reagent dosage greatly reduces, and also decreases, achieve low-carbon environment-friendly to the pollution of environment.
Accompanying drawing illustrates:
Fig. 1 is that the Q1 of seven kinds of compounds scans mass spectrogram.
Embodiment
Below to the further optimum choice of the partial parameters in testing process, through the experimental study that inventor is a large amount of for a long time, when adopting following Chromatography/Mass Spectrometry detected parameters, testing result more accurately and reliably.
Chromatography-mass spectroscopy condition:
Chromatographic column: PhenomenexKinetex (100mm × 4.6mm, 2.6 μm), column temperature: 35 DEG C, flow velocity: 0.3mL/min, sample size: 5 μ L, mobile phase: A are the 0.1% formic acid water containing 5mmol/L ammonium acetate, and B is methyl alcohol; Gradient elution program is in table 1.
Table 1 gradient program
Mass spectrum running parameter:
Electric spray ion source (electrosprayionizationmassspectrometryconditions, ESI); Positive and negative ion scan pattern; Multiple-reaction monitoring (MRM) acquisition mode; Desolventizing temperature: 350 DEG C; Dry gas flow velocity: 10.0L/min; Atomization gas pressure: 275.8kPa (40.0psi); Capillary voltage: 5500V; Resolution: MS1, MS2 are Unit; The mass spectrum acquisition parameter of each compound is in table 2.
The mass spectrum acquisition parameter of table 27 kind of compound
Below in conjunction with test example and embodiment, the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on content of the present invention all belong to scope of the present invention.
First, the instrument used in embodiment of the present invention process of the test, reagent and early-stage preparations process are described.
1. instrument
ABSCIEX
4500LC/MS/MS system (American AB sciex company); Solid-phase extracting instrument (Agilent company of the U.S.); YP402N electronic balance (Shanghai Precision Scientific Apparatus Co., Ltd); RV10 rotary evaporator (German IKA); Constant temperature oscillator (German IKA); Ultrasonic oscillator KQ-600E (Fauna of Kunshan, Jiangsu).
2. reagent
Acetonitrile, methyl alcohol (chromatographically pure, Merck Chemical Engineering Technology (Shanghai) Co., Ltd., German Merck company); Formic acid, ammonium acetate (analyzing pure, Ke Miou chemical reagent, Tianjin); Pure water (Wahaha company); NaOH (analyzing pure, traditional Chinese medicines group, Shanghai); Sodium chloride (analyzing pure, traditional Chinese medicines group, Shanghai); Anhydrous sodium sulfate (analyzing pure, traditional Chinese medicines group, Shanghai); Florisil silica pillar (6CC, 100mg, Guangzhou Féraud door); Activated charcoal pillar (6CC, 100mg, Guangzhou Féraud door); C
18solid phase extraction column (6CC, 60mg, Waters, US); Dispersive solid phase extraction adsorbent (C18, graphitized carbon, florisil silica, Tianjin Bonaaijieer Technology Co., Ltd).Amantadine hydrochloride, rimantadine hydrochloride standard items (purity 99%, Dr.Ehrenstorfer company of Germany), amantadine hydrochloride internal standard compound (purity 99%, Dr.Ehrenstorfer company of Germany), Tilmicosin standard items (content 95.0% ± 2.0%, Dr.Ehrenstorfer company), dexamethasone standard items (99.0% ± 1.0%, Dr company of Germany), olaquindox metabolite (3-methyl-quinoline-2-carboxylic acid, MQCA, 97%, Dr company of Germany), mark (3-methyl-quinoline-2-carboxylic acid-D4 in olaquindox metabolite-D4, MQCA-D4, 94%, Dr company of Germany).
3. standard solution preparation
Standard substance stock solution: accurately take appropriate amantadine hydrochloride, rimantadine hydrochloride, Tilmicosin, dexamethasone, olaquindox metabolite and internal standard compound standard items thereof, is 100.0mg/L standard solution with methyl alcohol compound concentration, keeps in Dark Place in 4 DEG C.
4. standard uses solution reagent preparation
Standard substance uses liquid: accurately pipette appropriate standard solution, be diluted to 2,5,10,25,50 and 100 μ g/L, now with the current.In amantadine hydrochloride, mark becomes 10 μ g/L with standard configuration in olaquindox metabolite.
2mol/L sodium hydroxide solution: take after 40.0g NaOH is dissolved in water, turn in 500mL volumetric flask, add water and be settled to scale.
The instrument more than selected, reagent, standard solution etc., will be applied in following examples, the model specification of above reagent can be thought more preferably to select, but should as the absolute restriction of the model specification selective gist of reagent.Those skilled in the art can carry out adjusting and optimizing to the preparation of the supplier of various reagent, standard solution/use solution, above-mentioned solution reagent should not thought to realize necessary condition of the present invention.
Embodiment
Adopt high performance liquid chromatography-tandem mass to carry out detection to analyze, in instrument, the conditional parameter of chromatography-mass spectroscopy arranges as follows:
Chromatogram running parameter: chromatographic column: PhenomenexKinetex (100mm × 4.6mm, 2.6 μm), column temperature: 35 DEG C, flow velocity: 0.3mL/min, sample size: 5 μ L, mobile phase: A are for containing 5mmol/L ammonium acetate 0.1% formic acid water, and B is methyl alcohol; Gradient elution program is in table 1.
Mass spectrum running parameter: electric spray ion source (electrosprayionizationmassspectrometryconditions, ESI); Positive and negative ion scan pattern; Multiple-reaction monitoring (MRM) acquisition mode; Desolventizing temperature: 350 DEG C; Dry gas flow velocity: 10.0L/min; Atomization gas pressure: 275.8kPa (40.0psi); Capillary voltage: 5500V; Resolution: MS1, MS2 are Unit.Use the analysis of standard model sample introduction to obtain the mass spectrum acquisition parameter of each compound, the results are shown in Table 2.
1. preparation of samples:
Take 2.00g chicken respectively, chicken gizzard, chicken blood sample, add 8g sodium chloride, add 20.0mL acetonitrile again, 2mol/L sodium hydroxide solution 5.0mL is added after violent jolting 2min, vibrate again after 1min and extract 30 minutes in 40 DEG C of water bath sonicator, after in the centrifugal 5min of 4000r/min, draw upper strata acetonitrile 10ml in evaporative flask, in residue, add 6mol/L hydrochloric acid regulate PH about 2.0, add 10.0ml acetonitrile vibration 2min again, in the centrifugal 5min of 4000r/min, drawing upper strata acetonitrile 10ml is incorporated in first time extract, 40 DEG C are concentrated near dry, obtain chicken meat sample extract residue, to be clean.
0.6g matrix solid phase dispersion agent (C is added in chicken meat sample extract residue
18, 50mg, anhydrous sodium sulfate 550mg), ethyl acetate 2ml vortex 2min, gets after supernatant liquid nitrogen dries up and adds 1.0mL30% methanol solution (containing 0.1% formic acid), ultrasonic dissolution, crosses film, treats sample introduction.
2. linear equation and detection limit
The hybrid standard series of working liquids of preparation and testing sample same matrix, make its concentration for difference 2,5,10,25,50 and 100 μ g/L, do typical curve with selected quota ion peak area to concentration, five kinds of medicines present good linear relationship in concentration range, and linear equation in chicken, chicken gizzard, chicken heme of five kinds of medicines and related coefficient list in table 3.Adopt the method for adding target compound in blank sample to press in row relax with 3 times of signal to noise ratio (S/N ratio) determination detectabilities, the results are shown in table 3 with 10 times of signal to noise ratio (S/N ratio) determination quantitative limit.
The linear equation related coefficient detectability of table 3 five kinds of medicines and quantitative limit
3. recovery experimental result in different substrates
Adopt concentration to do recovery testu to different substrates for respectively 5 μ g/L, 10 μ g/L and 50 μ g/L mixed standard solutions, each concentration do 5 times parallel, the recovery experimental result of 5 kinds of medicines in different substrates lists in table 4.
Table 4 five kinds of medicines are in the recovery experimental result (n=5) of different substrates
The actual analysis result that detects shows, interpolation concentration is 0.01 ~ 0.05mg/kg, amantadine hydrochloride in poultry, rimantadine hydrochloride, dexamethasone, Tilmicosin and olaquindox metabolite TIANZHU XINGNAO Capsul are 76.9% ~ 96.1%, relative standard deviation (n=5) is 0.6% ~ 3.0%, amantadine hydrochloride in poultry, rimantadine hydrochloride, dexamethasone, the method detection limit of Tilmicosin and olaquindox metabolite is respectively 0.001, 0.001, 0.005, 0.005, 0.002mg/kg, quantitative limit is respectively 0.005, 0.005, 0.010, 0.010, 0.005mg/kg.
4 results and discussion
4.1 Mass Spectrometry Conditions optimizations
Adamantanamine hydrochloride and rimantadine hydrochloride ionize out Cl under solution state
-, Cl
-usually losing under high-temperature electric nebulization, the adduct ion peak of amantadine, Rimantadine can only be detected, carry out Q1 parent ion full scan, finding that adduct ion peak that they form is for mainly containing [M+H]
+, [M+Na]
+[M+NH
4]
+deng, under mobile phase existent condition, carry out the optimization of Mass Spectrometry Conditions, when mobile phase be acetonitrile and water, acetonitrile and 0.1% formic acid, acetonitrile and 0.1% acetic acid time, [M+H]
+clearly, [M+NH
4]
+[M+Na]
+not obvious; When containing ammonium formate in mobile phase, [M+H]
+present each compound ions abundance to strengthen; When ammonium acetate is mobile phase, [M+H]
+reach mxm., therefore in mobile phase, add certain density ammonium acetate, thus ensure that the adduction peak of compound is more stable, and sensitivity is also high.Mass spectrographic capillary voltage, taper hole voltage, desolventizing temperature, desolventizing gas and taper hole gas are optimized simultaneously, finally determine Mass Spectrometry Conditions.The mass spectrogram carrying out parent ion Q1 scanning after optimization is shown in Fig. 1, then has carried out daughter ion scanning to parent ion, thus determines daughter ion and collision energy.
4.2 optimize chromatography condition
Each compound retains on a cl 8 column all good reservation, and experiment confirms all in 5 minutes, to be eluted peak containing all target compounds of mobile phase of 30% methyl alcohol.In order to ensure that each compound can have good separation, this test adopts gradient elution, saves analysis time, by optimizing the condition of gradient elution, can ensure the sensitivity of each compound, can ensure again its good peak type.Test compares methanol system and acetonitrile system, result under showing methanol system each compound sensitivity higher, therefore select methyl alcohol and containing 0.1% aqueous formic acid of 5mmol/L ammonium acetate.
4.3 Pretreatment optimizations
Amantadine hydrochloride, Rimantadine hydrochloric acid can be extracted well by organic solvent in alkaline solution, and Tilmicosin, dexamethasone and biological tissue's affinity are stronger, must saltout in neutral salt solution and improve extraction efficiency, olaquindox metabolite is combined with animal tissue closely, be hydrolyzed by sodium hydroxide solution, it is dissociated out from tissue, make it be reduced into organic acidity compound to utilize solvent extraction by adding hydrochloric acid, so this experiment adopts first basic hydrolysis biological tissue, then by regulating PH to extract target compound.Experiment display, under alkali condition, neutral compound and acid compound olaquindox metabolite do not reclaim at all, therefore, this experiment first time extract get half as subsequent operation, be actually and be only extracted amantadine hydrochloride and the rimantadine hydrochloride that displaced half in sample.And then add the acetonitrile of 10ml, with the extraction of other three kinds of medicines, then dividing and get 10ml and be incorporated in primary extract, is also the residual drug that displaced half in sample.Therefore the carrying out of sample is settled to 1ml after purifying, to keep the dilutability of sample for 1.
Because extract coextraction material composition is complicated, matrix interference can cause the sensitivity decrease that the recovery declines, reappearance is bad and detect usually, therefore, takes suitable purification to be conducive to improving accuracy and the precision of detection.This experiment investigation C
18tripoli (Florisil) post in pillar, ketjenblack EC (GCB) and fluorine sieve, has been C simultaneously
18, GCB, Florisil adsorbent adopts Dispersive solid phase extraction technology to test: GCB has good adsorpting pigment effect, but also adsorbed evaluating objects compound simultaneously, experiment confirms that active carbon purifying needs a large amount of eluting solvent, otherwise the recovery cannot reach requirement, but strengthen eluting solvent consumption, pigment is entered sample by wash-out again, does not reach the object of rationally purification; C
18pillar and Florisil post well can retain compound and by wash-out.Confirm C by experiment
18the eluent of little column purification, ammoniated methanol and formic acid methyl alcohol two kinds of different PH carries out elute effect better, but complicated operation, and also cost is higher.Adopt C
18for adsorbent carries out the purification of QuEChERS dispersive solid-phase extraction, while obtaining good clean-up effect and the satisfied recovery, operating process is simplified significantly.Therefore, finally C is adopted
18for adsorbent carries out the purification of QuEChERS dispersive solid-phase extraction.
4.4 matrix effects are investigated
2 have been prepared with methyl alcohol, 5, the target compound solvent standard specimen of the series of 10,25,50 and 100 μ g/L, take blank sample, chicken, chicken gizzard, chicken blood simultaneously, according to pre-treatment step extraction and cleaning, and blank sample sample introduction is analyzed, determine it not containing after trace compound, again with the matrix standard specimen of a series of same solvent standard specimen concentration of blank sample liquid dilution, respectively with standard specimen concentration be horizontal ordinate, peak area carries out linear regression for ordinate.The curve of matrix standard specimen lists in table 3.Extraction standard curve linear is good, correlation coefficient r
2all be greater than 0.995.
The ratio (theratioofslope) of matrix and solvent slope of standard curve is the characteristic parameter of the power examining or check matrix effect, and in chicken, chicken gizzard, chicken blood, the slope ratio of target compound matrix standard specimen and solvent standard specimen is respectively: 0.972,0.843 and 0.876.In this test matrix, the matrix effect of chicken gizzard and chicken blood is comparatively obvious, and target compound has substrate inhibition effect in these two kinds of matrix.
5 actual sample analyses
Adopt 112 samples (comprise chicken, duck and blood and liver etc.) of this method to sampling observation to detect, the positives rate of sample is 12.45%; Main containing amantadine hydrochloride, olaquindox metabolite, Tilmicosin.Do not detect dexamethasone and rimantadine hydrochloride.The chicken positive accounts for 86.7% of positive sample total amount, and what content was the highest is present in liver, reaches 115 μ g/kg.
Innovative approach contrast test situation
Innovative approach of the present invention is adopted to test the content of amantadine and olaquindox metabolite in Fresh Grade Breast.Environment temperature 20 DEG C, temperature 40%, ion source temperature: 550 DEG C, chromatographic column: kinetexC18 post.Adopt the gradient elution mode in embodiment to carry out wash-out, test record and result are as following table 5, table 6.
New method
Table 5 innovative approach amantadine test result calculations table
Olaquindox metabolite test result calculations table in table 6 innovative approach Fresh Grade Breast
From the test result showing 5-6 above, the result of innovative approach test has higher accuracy, meets the requirement for accuracy in testing process, can be applied to the analytical test of five kinds of medicament residues.For the analytical test accelerating medicament residue, there is outstanding meaning.
Claims (10)
1. a detection method for poultry tissues drug residue, comprises the following steps:
(1) get poultry sample, add sodium chloride and acetonitrile, jolting 1 ~ 5 minute, adds the sodium hydroxide solution of 1 ~ 4mol/L of 2 ~ 5 times of volumes, vibrates 30 seconds to 5 minutes; Then, 30 ~ 50 DEG C of ultrasonic extractions 10 ~ 40 minutes, centrifugal, Aspirate supernatant is for subsequent use, all the other solution evaporates to dryness;
(2) add concentrated hydrochloric acid in the residue obtained to step (1) evaporate to dryness, regulate pH about 1 ~ 3; Then, add acetonitrile mechanical shaking extraction, preferably add 10.0ml acetonitrile vibration 2min; Through the solution of acetonitrile mechanical shaking extraction, after centrifugal, Aspirate supernatant and the middle supernatant drawn of step (1) merge, and 30 ~ 50 DEG C are concentrated into dry, obtain sample extracting solution residue;
(3) in sample extracting solution residue, add matrix solid phase dispersion agent, ethyl acetate 2ml, vortex 2min, gets supernatant liquid, and nitrogen dries up, and then adds 30% methanol solution of 1.0mL containing 0.1% formic acid, ultrasonic dissolution, crosses film, treat sample introduction;
(4) high performance liquid chromatography-tandem mass is adopted to detect; With containing 5mmol/L ammonium acetate-0.1% formic acid water and methyl alcohol for eluent gradient wash-out.
2. the detection method of poultry tissues drug residue as claimed in claim 1, is characterized in that, after step (1) adds sodium chloride and acetonitrile, and jolting 2 minutes.
3. the detection method of poultry tissues drug residue as claimed in claim 1, is characterized in that, ultrasonic extraction 20 ~ 30 minutes in step (1).
4. the detection method of poultry tissues drug residue as claimed in claim 1, it is characterized in that, in step (1) centrifugal process, rotating speed is greater than 3000r/min, draws upper strata acetonitrile 10ml in evaporative flask, all the other solution evaporates to dryness.
5. the detection method of poultry tissues drug residue as claimed in claim 1, it is characterized in that, step (2) uses 1 ~ 6mol/L hydrochloric acid to carry out pH adjustment.
6. the detection method of poultry tissues drug residue as claimed in claim 1, it is characterized in that, the described matrix solid phase dispersion agent of step (3) is the agent of 0.6g matrix solid phase dispersion.
7. the detection method of poultry tissues drug residue as claimed in claim 6, it is characterized in that, described matrix solid phase dispersion agent is C
18or anhydrous sodium sulfate.
8. the detection method of poultry tissues drug residue as claimed in claim 1, is characterized in that, step (2) is concentrated near doing at 40 DEG C, obtains sample extracting solution residue.
9. the detection method of poultry tissues drug residue as claimed in claim 1, it is characterized in that, gradient elution degree is: mobile phase: A is the 0.1% formic acid water containing 5mmol/L ammonium acetate, and B is methyl alcohol.
10. the detection method of poultry tissues drug residue as claimed in claim 9, it is characterized in that, gradient elution degree is: mobile phase: A is the 0.1% formic acid water containing 5mmol/L ammonium acetate, and B is methyl alcohol; Gradient elution program, gradient condition is: 10%B:0 ~ 30 second; 10% ~ 90%B:30 second ~ 2 minutes; 90%B:2 ~ 4 minute; 90% ~ 10%B:4 ~ 4.5 minute; 10%B:4.5 ~ 6.5 minute.
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| CN107664671A (en) * | 2017-08-18 | 2018-02-06 | 广东省药品检验所（广东省药品质量研究所、广东省口岸药品检验所） | A kind of left drug detection method of animal-derived food |
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| CN109406249B (en) * | 2018-11-30 | 2022-10-14 | 江苏美正生物科技有限公司 | Pretreatment method for food safety drug residue detection |
| CN110129728A (en) * | 2019-06-14 | 2019-08-16 | 东莞市广正模具塑胶有限公司 | A kind of modified AF coating material and its preparation method and application |
| CN113960236A (en) * | 2021-10-11 | 2022-01-21 | 大连海洋大学 | Method for determining geosmin and dimethyl isoborneol in fish body based on rapid pretreatment technology |
| CN114965812A (en) * | 2022-05-25 | 2022-08-30 | 山东省食品药品检验研究院 | Method for simultaneously determining residues of 15 quaternary ammonium salt disinfectants in livestock and poultry meat |
| CN114965812B (en) * | 2022-05-25 | 2024-04-05 | 山东省食品药品检验研究院 | Method for simultaneously measuring 15 quaternary ammonium salt disinfectant residues in livestock and poultry meat |
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