CN106610403A - Method for measuring residual amount of diaveridine in chicken tissue - Google Patents

Method for measuring residual amount of diaveridine in chicken tissue Download PDF

Info

Publication number
CN106610403A
CN106610403A CN201510687285.5A CN201510687285A CN106610403A CN 106610403 A CN106610403 A CN 106610403A CN 201510687285 A CN201510687285 A CN 201510687285A CN 106610403 A CN106610403 A CN 106610403A
Authority
CN
China
Prior art keywords
nsc
carrying
eyeball
sample
detection
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.)
Pending
Application number
CN201510687285.5A
Other languages
Chinese (zh)
Inventor
洪霞
张淑雅
刘静
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Xian Chuan Bio Tech Ltd Zhenjiang
Original Assignee
Xian Chuan Bio Tech Ltd Zhenjiang
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Xian Chuan Bio Tech Ltd Zhenjiang filed Critical Xian Chuan Bio Tech Ltd Zhenjiang
Priority to CN201510687285.5A priority Critical patent/CN106610403A/en
Publication of CN106610403A publication Critical patent/CN106610403A/en
Pending legal-status Critical Current

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N2030/022Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027Liquid chromatography

Abstract

The invention belongs to the field of food examination and relates to a method for measuring the residual amount of diaveridine. The method comprises carrying out extraction through acetonitrile and acetone, removing fat through n-hexane, carrying out MCX solid phase extraction small column purification, orderly carrying out leaching by 0.1mol/L of a hydrochloric acid solution and methanol, then carrying out elution through 10% of ammoniated acetonitrile, concentrating the eluate, re-dissolving the concentrate through a mobile phase, filtering the solution, and carrying out high performance liquid chromatography through a C18 chromatographic column under conditions of 0.01mol/L of acetonitrile as a mobile phase, potassium dihydrogen phosphate aqueous solution (13/87v/v), UV detection wavelength of 270nm and external standard quantification. The method has a detection limit of 0.02mg/kg, a quantification limit of 0.05mg/kg, a recovery rate of 73.63% to 92.44% in muscle, and a recovery rate of 75.09% to 98.72% in liver. The method has an important meaning for detecting and monitoring the residual amount of diaveridine in chicken tissue.

Description

A kind of method of NSC 408735 residual quantity in detection chicken tissues
Technical field
The invention belongs to bromatology inspection field.The specifically related to assay method of NSC 408735 residual quantity.
Background technology
The discovery of diaminopyrimidine antibacterial synergist so that after sulfa drugss and such Drug combination, not only antimicrobial spectrum expands, and antibacterial activity is greatly enhanced, and the advantages of in addition cheap, thus is widely used in Production of Livestock and Poultry.Irrational use on veterinary clinic with sulfa drugss, or even abuse, so that residual of the Trimethoprim class medicine in animal derived food also inevitably can occur.Animal food of people's long-term consumption containing NSC 408735 residual and its product, it would be possible to which harm is produced to human health.
It is 50 μ g/kg that the most countries such as international food committee (CAC) and America and Europe define the MRL (MRL) of methoxy section eyelash in animal food, China in the animal food residue of veterinary drug limitation that the Ministry of Agriculture in 2002 promulgates new revision, the same CAC of regulation to TMP residue limits.In addition to Japan, CAC, EU countries and China do not make stipulations in the MRL of animal food to DVD.Specify in Japanese positive list that DVD is 50 μ g/kg in the residue limits of edible chicken tissues (muscle, liver, kidney and sebum).
Application along with such medicine in animal food and caused residue problem is attracted attention.Therefore in order to the detection method for strengthening setting up the monitoring in animal food about veterinary drug NSC 408735 residual quantity in a kind of animal tissue is necessary.At present, though analysis method domestic and international about NSC 408735 residue detection it has been reported that detection method research of the China to NSC 408735 residual quantity in animal tissue there is not been reported.Recognize based on more than, the HPLC detection methods of NSC 408735 residual quantity in vertical chicken tissues are planned to build in this research, it is intended to provide technological means to the residual implementing monitoring of such medicine from now on.
The content of the invention
High line liquid chromatography has the advantages that sensitivity height, qualitative, quantitative are accurate, and this project the method sets up NSC 408735 residues detection confirmation method in chicken tissues, and the residual monitoring for such medicine provides scientific basis.
The present invention is comprised the following steps:
(1)The preparation of sample and preservation:1 age in days white meat-type chickens are raised to 40 ages in days with the perfect compound feed without any medicine and are sampled.The blank chicken tissues (muscle, liver, kidney, skin+fat) of proper amount of fresh or freezing are taken, is rubbed and homogeneous;The tissue sample for preparing be placed on one 20 DEG C it is stored below standby;
(2)Sample pre-treatments:((2 scholar 0.05) g, is placed in 5Oml polypropylene centrifuge tubes, adds 4g anhydrous sodium sulfates accurately to weigh the sample after rubbing, add 15m1 second eyeball whirling motion lmin, 15000rpm is homogenized 1~2min, supersound extraction lOmin, 5000rpm centrifugation 10min;Supernatant is moved in 100m1 Cor Gigeriae Galli bottles, refiner cutter head is then washed with 15ml acetone and is proceeded to and repeat extraction, supersound extraction 1 Omin, recentrifuge l after 3 min of whirling motion Omin, merges supernatant and adds 5ml normal propyl alcohols to do near in 45 DEG C of rotary evaporations;
4 ml second eyeball -0.017mo1/L phosphoric acid (30/70) aqueous solutions are added to steaming into nearly dry Cor Gigeriae Galli bottle, vortex oscillation 5min is moved in 1 Oml teat glasses after residue dissolving;Washed with 4m1 normal hexane after Cor Gigeriae Galli bottle, merge cleaning mixture, vortex 2min;
(3)Purification:Solid phase extraction is carried out using MCX posts, activated, balanced MCX pillars with the HCl of 5ml methanol, 0.1 mol/L of 5ml successively.Above-mentioned subnatant 2m1 upper props are drawn, is flowed out under natural gravity.Then successively with 0.1 Mol/L hydrochloric acid 2m1 and methanol 6m1 drip washing pillars;Finally with 10% ammonification second eyeball 5ml eluted substances, eluent is collected in 5OmI Cor Gigeriae Galli bottles, rotates and be evaporated at 45 DEG C.Residue lml flowing phased solns, cross 0.22 μm of organic filter membrane, collect filtrate and analyze for HPLC;
(4)Upper machine is determined
Detected with Ultra Performance Liquid Chromatography, record chromatogram, internal standard peak height(Hi)With NSC 408735 residual quantity peak height(Hs), use standard curve converted score.
The assay method of aforesaid NSC 408735 residual quantity, it is characterised in that the method is to select C18Chromatographic column carries out efficient liquid phase chromatographic analysis, and mobile phase is second eyeball/0.01 mol/L, potassium dihydrogen phosphate aqueous solution (13/87 v/v), and ultraviolet detection wavelength is 270nm, quantified by external standard method.This method detection is limited to 0.02mg/kg, 0.05mg/kg is limited to quantitatively, muscle, liver the response rate respectively between 73.63% 1 %.44% and 75.09% one 98.72%, the method is significant for the detection monitoring side mask of NSC 408735 residual quantity in chicken tissues.The invention has the beneficial effects as follows:The present invention has preferable veracity and precision using high performance liquid chromatography detection Basic Orange content.
Description of the drawings
Fig. 1 is standard solution chromatogram.
Fig. 2 is blank Carnis Gallus domesticus chromatogram.
Fig. 3 is blank Carnis Gallus domesticus addition(0.05μg/g)Chromatogram.
Fig. 4 is blank liver chromatogram.
Fig. 5 is blank liver addition(0.05μg/g)Chromatogram.
Specific embodiment
The specific embodiment of the present invention is described more detail below:
1. instrument and reagent:
Instrument:
High performance liquid chromatograph Agilent 1200 is serial, with Agilent LC and CE cycle chemistries work station and ultraviolet detection
Chromatographic column Yi Lite ODS-2 chromatographic column (250X4.6 mm, 5 μm);
Tissue refiner POLYTRON PT 1200;
Vortex mixer WH-1;
Low speed centrifuge LDS-2A;
Electronic analytical balance BP110S;
Vacuum pump using circulatory water SHB-III types;
Electric heating constant-temperature blowing drying box;
Automatic dual pure water distillator SZ-93;
Rotary Evaporators RE-52 types;
Ultraviolet/visible spectrophotometer ND-1000;
Ultrasonic cleaner KQ-250B;
Syringe-driven filter, organic system, 0.45 pm of aperture;
MCX solid phase extraction columns (60mg/3m1);
Adjustable micropipettor, 1-10 μ l, 20-200 μ l, 100-1000 μ l;
Reagent:
Second eyeball:Chromatographically pure;
Methanol:Chromatographically pure;
Phosphoric acid:Top pure grade;
Acetone:Analysis is pure;
Ammonia:Analysis is pure;
Hydrochloric acid:Analysis is pure;
Normal hexane:Analysis is pure;
Ultra-pure water;
2. the preparation of solution
The preparation of test solution
Standard reserving solution:Precision weighs appropriate dimethoxy section smack one's lips and methoxy section is steep respectively, is placed in 50ml brown volumetric flasks, first with the dissolving with hydrochloric acid of a small amount of 0.1 mol/L, scale is settled to second eyeball again, the NSC 408735 standard reserving solution that concentration is 100 μ g/ml is made into, is preserved in putting 4 DEG C of refrigerators, six months effect duration.
Hybrid standard working solution (40 μ g/ml):NSC 408735 standard reserving solution (100 μ g/ml) 4m1 is measured respectively, is collectively disposed in 10ml brown volumetric flasks, with second eyeball constant volume, become the standard mixing storing solution of the NSC 408735 of 40 μ g/ml.It is placed in 4 DEG C of Refrigerator stores.
The preparation of other solution:0.1 mol/L hydrochloric acid solutions:Take 826 river hydrochloric acid to be placed in 100m1 volumetric flasks, use ultra-pure water constant volume, mixing to obtain final product;0.017mol/L phosphate aqueous solutions:Take 0.49m1 phosphoric acid to be placed in 500mI volumetric flasks, use ultra-pure water constant volume, mixing to obtain final product;10% ammonification second eyeball:Take l Oml ammonia to be placed in 100m1 volumetric flasks, with second eyeball constant volume, mixing is obtained final product;0.01mol/L potassium dihydrogen phosphate aqueous solutions:0.68g potassium dihydrogen phosphates are taken, is placed in 500mI volumetric flasks, use ultra-pure water constant volume, mixing to obtain final product.
3rd, method and result
Chromatographic condition:Chromatographic column:Erie's spy's ODS-2 chromatographic columns (250 × 4.6 mm, 5 μm);Mobile phase:Second eyeball one contains 0.01 mol/L potassium dihydrogen phosphate aqueous solutions (13/87 v/v);Detection wavelength:270 nm flow velocitys:1 ml/min;Sample size:20 μl;Column temperature:350 DEG C, under the chromatographic condition set up, well NSC 408735 in Chicken Tissues can be kept completely separate with other matrix components in solvent peak and tissue.NSC 408735 retention time is shown in Fig. 1-5 respectively in 9min or so, NSC 408735 reference substance, blank tissue, medicine addition sample chromatogram figure.
The drafting of standard curve:Accurately draw the hybrid standard working solution prepared appropriate, be diluted to the standard working solution of 4,2,1,0.5,0.25 and 0.05 μ g/ml successively with mobile phase.Under the chromatographic condition selected, by concentration order from low to high, 2 μ l of sample introduction make HPLC analyses respectively.Each concentration is repeated 5 times.With concentration of standard solution level (X) as independent variable, the average peak area (Y) of each concentration level is dependent variable, makees linear regression, draws standard curve, and obtains its regression equation and correlation coefficient.
Under the chromatographic condition set up, standard substance mean concentration and the regression equation of its average peak area of NSC 408735 are:Y = 30.753X-0.2872。
The measure of test limit (LOD) and quantitative limit (LOQ):Prepare series concentration dimethoxy section smack one's lips and methoxy section throat standard working solution 0.1,0.2,, O.5 μ g/ml, 0.2m1 is taken respectively in (in (2 scholar 0.05) g blank homogenised tissue (muscle, liver), it is obtained 0.01,0.02,0.05 Tissues of μ g/g tri-, are repeated 5 times, carry out HPLC measure.The peak height and noise (baseline peak height) of each sample are measured, signal to noise ratio is taken>Concentration when 3 is test limit, and concentration during signal to noise ratio Feng≤10 is quantitative limit.
Accuracy is determined:Accuracy is represented with the response rate for organizing addition sample.The blank tissue samples of 4 parts of each (2 scholar 0.05) g homogenizing are weighed accurately, it is placed in 50ml polypropylene centrifuge tubes, it is separately added into mixing dimethoxy section eyelash and each 0.2m1 of 0.5,10,20,40 μ g/ml of methoxy section smack one's lips standard working solution, the concentration of dimethoxy section smack one's lips and methoxy section throat in each tissue is made to be respectively 0.05,1,2 and 4 μ g/g, Jing after processing, take 20 μ l sample introductions of filtrate and make HPLC measure, simultaneously with the dimethoxy section eyelash and methoxy section shallow lake titer (0.05,1,2,4 μ g/ml) 20 μ l of sample introduction of corresponding concentration, make HPLC measure.Each concentration is repeated 5 times respectively.The computing formula of the wherein response rate is:
Note:Actual measurement peak areas of the A for NSC 408735 in organizing after pretreatment in formula;As is the peak area of NSC 408735 in correspondence titer.
TIANZHU XINGNAO Capsul:In the pitch-based sphere of tetra- concentration of 0.05mg/kg, lmg/kg, 2mg/kg, 4mg/kg, recovery test is added to chicken tissues (muscle, liver).It is shown in Table 1-4.
Table 1
Table 2
Table 3
Table 4

Claims (4)

1. in chicken tissues NSC 408735 residual quantity assay method, it is characterised in that after by testing sample pretreatment, with ultra-performance liquid chromatography detect, comprise the following steps:
(1)The preparation of sample and preservation:1 age in days white meat-type chickens are raised to 40 ages in days with the perfect compound feed without any medicine and are sampled;The blank chicken tissues (muscle, liver, kidney, skin+fat) of proper amount of fresh or freezing are taken, is rubbed and homogeneous;The tissue sample for preparing be placed on one 20 DEG C it is stored below standby;
(2 Sample pre-treatments:((2 scholar 0.05) g, is placed in 50ml polypropylene centrifuge tubes, adds 4g anhydrous sodium sulfates accurately to weigh the sample after rubbing, add 15m1 second eyeball whirling motion lmin, 15000rpm is homogenized 1~2min, supersound extraction 10min, 5000rpm centrifugation 10min;Supernatant is moved in 100m1 Cor Gigeriae Galli bottles, then refiner cutter head is washed with 15ml acetone and proceeds to and repeat extraction, supersound extraction 10min, recentrifuge 10min after 3 min of whirling motion merge supernatant and add 5ml normal propyl alcohols to do near in 45 DEG C of rotary evaporations;4 ml second eyeball -0.017mo1/L phosphoric acid (30/70) aqueous solutions are added to steaming into nearly dry Cor Gigeriae Galli bottle, vortex oscillation 5min is moved in 1 Oml teat glasses after residue dissolving;Washed with 4m1 normal hexane after Cor Gigeriae Galli bottle, merge cleaning mixture, vortex 2min;
(3)Purification:Solid phase extraction is carried out using MCX posts, activated, balanced MCX pillars with the HCl of 5ml methanol, 0.1 mol/L of 5ml successively;Above-mentioned subnatant 2m1 upper props are drawn, is flowed out under natural gravity;Then successively with 0.1 mol/L hydrochloric acid 2m1 and methanol 6m1 drip washing pillars;Finally with 10% ammonification second eyeball 5ml eluted substances, eluent is collected in 5OmI Cor Gigeriae Galli bottles, rotates and be evaporated at 45 DEG C;Residue lml flowing phased solns, cross 0.22 μm of organic filter membrane, collect filtrate and analyze for HPLC;
(4)Upper machine is determined
Detected with Ultra Performance Liquid Chromatography, record chromatogram, internal standard peak height(Hi)With NSC 408735 residual quantity peak height(Hs), use standard curve converted score.
2. the assay method of aforesaid NSC 408735 residual quantity, it is characterised in that the method is to select C18Chromatographic column carries out efficient liquid phase chromatographic analysis, and mobile phase is second eyeball/0.01 mol/L, potassium dihydrogen phosphate aqueous solution (13/87 v/v), and ultraviolet detection wavelength is 270nm, quantified by external standard method.
3. this method detection is limited to 0.02mg/kg, 0.05mg/kg is limited to quantitatively, muscle, liver the response rate respectively between 73.63% 1 %.44% and 75.09% one 98.72%, the method is significant for the detection monitoring side mask of NSC 408735 residual quantity in chicken tissues.
4. the invention has the beneficial effects as follows:The present invention has preferable veracity and precision using high performance liquid chromatography detection Basic Orange content.
CN201510687285.5A 2015-10-22 2015-10-22 Method for measuring residual amount of diaveridine in chicken tissue Pending CN106610403A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510687285.5A CN106610403A (en) 2015-10-22 2015-10-22 Method for measuring residual amount of diaveridine in chicken tissue

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510687285.5A CN106610403A (en) 2015-10-22 2015-10-22 Method for measuring residual amount of diaveridine in chicken tissue

Publications (1)

Publication Number Publication Date
CN106610403A true CN106610403A (en) 2017-05-03

Family

ID=58611353

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510687285.5A Pending CN106610403A (en) 2015-10-22 2015-10-22 Method for measuring residual amount of diaveridine in chicken tissue

Country Status (1)

Country Link
CN (1) CN106610403A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111024854A (en) * 2019-12-30 2020-04-17 大连理工大学 Method for rapidly and efficiently detecting contents of multiple antibiotics in vegetables simultaneously
CN112525893A (en) * 2019-09-19 2021-03-19 首都师范大学 Method for detecting copper ions in liver by colorimetric sensor based on nanogold catalytic ability

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101949898A (en) * 2010-08-10 2011-01-19 上海安谱科学仪器有限公司 Method for detecting residual quantity of multiple alkaline drugs in animal derived food
CN102636590A (en) * 2012-05-06 2012-08-15 扬州大学 Method for detecting dehydroacetic acid residue in chicken tissues

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101949898A (en) * 2010-08-10 2011-01-19 上海安谱科学仪器有限公司 Method for detecting residual quantity of multiple alkaline drugs in animal derived food
CN102636590A (en) * 2012-05-06 2012-08-15 扬州大学 Method for detecting dehydroacetic acid residue in chicken tissues

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112525893A (en) * 2019-09-19 2021-03-19 首都师范大学 Method for detecting copper ions in liver by colorimetric sensor based on nanogold catalytic ability
CN111024854A (en) * 2019-12-30 2020-04-17 大连理工大学 Method for rapidly and efficiently detecting contents of multiple antibiotics in vegetables simultaneously

Similar Documents

Publication Publication Date Title
CN103543224B (en) Detection method for residues of abamectin and ivermectin
CN103869041B (en) A kind of detection method of cough-relieving combination of oral medication
CN106770865B (en) A kind of organic acid content testing method in ginkgo biloba p.e
CN104391072B (en) A kind of detection method of the compound Chinese medicinal preparation treating osteoporosis
CN102119961A (en) Detection method of compound danshen dripping pills
CN113341004A (en) Method for determining benzoic acid, sorbic acid, saccharin sodium, acesulfame and dehydroacetic acid in food
CN106198810B (en) A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow suppression
CN105699500A (en) A method of measuring contents of seven components in a Huoxiangzhengqi dripping pill through ultra-high performance liquid chromatography
CN106610403A (en) Method for measuring residual amount of diaveridine in chicken tissue
CN107449846B (en) Method for measuring effective components in infantile nerve-soothing and brain-nourishing granules by HPLC-MS (high Performance liquid chromatography-Mass Spectrometry)
CN104678004B (en) A kind of method of quality control of Ge Shan blood-fat-lowering granule
CN104155383B (en) The detection method of blue or green Pu granule
CN106324167A (en) Method for determining flavone components in radix astragali extract by UPLC (ultra performance liquid chromatography)
CN106370736A (en) Method for simultaneous SMART column on-line purification and HPLC/UVE fluorescent detection of four aflatoxins in peanut product
CN105675755A (en) Method for detecting contents of flavonoid compounds in cyclocarya paliurus on basis of HPLC (high-performance liquid chromatography)
CN105758977A (en) Method for detecting quality of Lycium ruthenicum Murr fruit or Lycium ruthenicum Murr fruit extracts
CN106483260A (en) A kind of pill of Eight Treasures capsule quality standard control method
CN105616946A (en) Preparation for treating cough, preparation method and quality control method thereof
CN115389666A (en) Method for efficiently and simultaneously detecting ergothioneine and ectoine in cosmetics
CN102008541B (en) Method for simultaneously detecting three main active ingredients in sugar-free type compound wintercreeper preparation
CN115032310A (en) Method for detecting 6 dianthrone compounds in polygonum multiflorum
CN103091408B (en) Method for determining content of hesperidin in Citrus Bioflavonoids
CN103454374A (en) Quality control method of bone rehabilitation medicine
CN105823830B (en) One surveys the methods for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more
CN103364501B (en) The detection method of the adulterated pigment gold orange II in safflower

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20170503