CN105319369A - Enzyme linked immunosorbent assay kit used for detecting chloramphenicol, and detection method thereof - Google Patents
Enzyme linked immunosorbent assay kit used for detecting chloramphenicol, and detection method thereof Download PDFInfo
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- CN105319369A CN105319369A CN201410353404.9A CN201410353404A CN105319369A CN 105319369 A CN105319369 A CN 105319369A CN 201410353404 A CN201410353404 A CN 201410353404A CN 105319369 A CN105319369 A CN 105319369A
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Abstract
The invention discloses an enzyme linked immunosorbent assay kit used for detecting chloramphenicol, and a detection method thereof. The enzyme linked immunosorbent assay kit is high in detection sensitivity, accuracy, speed, and specificity; operation is simple; and the detection method is suitable for large-scale sample detection. The enzyme linked immunosorbent assay kit comprises an enzyme labeled plate coated with chloramphenicol antigens, a chloramphenicol standard substance, a chloramphenicol antibody working solution, a chloramphenicol enzyme labeled second antibody working solution, a substrate solution A, a substrate solution B, a stopping solution, a concentrated reconstitution fluid, and a concentrated washing liquid. Solid phase indirect competitive enzyme-linked immune response is taken as the principle of the enzyme linked immunosorbent assay kit used for detecting chloramphenicol. According to the detection method, an extracted sample, and the chloramphenicol antibody working solution are added into corresponding enzyme labeled plate micropores for a certain time of incubation; after plate washing, the chloramphenicol enzyme labeled second antibody working solution is added for a certain time of incubation; after plate washing, the substrate solution A, and the substrate solution B are added, so that the color of a color developing agent is changed to be blue under enzyme action; and the stopping solution is added, and the color is changed from blue to yellow. The shade of color developing and the content of chloramphenicol in the standard substance or the sample are inversely related. The direction method can be used for directly detecting residual quantity of chloramphenicol in samples to be detected.
Description
Technical field
The present invention relates to enzyme linked immunosorbent detection technical field, particularly the enzyme linked immunological kit of chlorine detection mycin.
Background technology
Chloromycetin (chloroamphenicol) is a kind of broad-spectrum antibacterial microbiotic, all has inhibiting effect to Grain-positive, negative bacteria.Aquaculture is widely used in for a long time because of its good anti-bacterial effect.Its antifungal mechanism is by being reversibly combined with 50S subunit, blocks the effect turning peptide acyl enzyme, disturbs and be combined with 50S subunit with amino acid whose amido acyl-tRNA terminal, thus the formation of new peptide chain is obstructed, and Profilin matter is synthesized.But because chloromycetin also can be combined with the mitochondrial 70S of human body, thus the mitochondrial albumen synthesis of human body can also be suppressed, toxicity is produced to human body, severe patient can suppress human bone marrow's hematopoiesis function and cause the disease such as aplastic anemia and granulocytic leukemia, and the residual chloromycetin therefore in animal-derived food forms huge potential hazard by the health of the mankind.The World Food Programme and health organization reduce to 0.1 μ g/kg to the regulation of the residual quantity of chloromycetin in food by 10 original μ g/kg.The Ministry of Agriculture of China is also formal in " food animal forbidding veterinary drug and the compound inventory thereof " of issue in 2002 is classified as preparations such as chloromycetin and salt esters thereof as forbidden drugs.But, continue to use this microbiotic because indivedual raiser ignores national ban, make the edible safety of animal-derived food there is hidden danger.
At present, because of testing result accurately and reliably, the mensuration of chloromycetin adopts chromatography more, but the requirement of chromatography to instrument and equipment is higher, and Sample pretreatment process more complicated, in order to save cost, residual chloromycetin in detection sample rapidly and efficiently, the present invention establishes a kind of enzyme linked immunological kit and detection method thereof of chlorine detection mycin, and the method is sensitive, accurate, quick, easy and simple to handle, high specificity, is applicable to the quick detection of gross sample.
The enzyme linked immunological kit of a kind of chlorine detection mycin of the present invention, includes ELISA Plate, chloromycetin standard items, chloramphenicol antibody working fluid, chloromycetin ELIAS secondary antibody working fluid, substrate solution A, substrate solution B, stop buffer, concentrated liquid and the concentrated cleaning solution of redissolving.
The preparation of the enzyme linked immunological kit of a kind of chlorine detection mycin of the present invention, comprise the preparation of ELISA Plate, the preparation of chloromycetin standard items, the preparation of chloramphenicol antibody working fluid, the preparation of chloromycetin ELIAS secondary antibody working fluid, the preparation of substrate solution A, the preparation of substrate solution B, stop buffer preparation, concentratedly redissolve the preparation of liquid and the preparation of concentrated cleaning solution.
It is further characterized in that: described ELISA Plate is via the antigen coated preparation of chloromycetin, concrete steps are that chloromycetin haptens and the pure albumen of carrier proteins Bovine (BSA) coupling are obtained envelope antigen, with phosphate (PBS) damping fluid of 0.05mol/LpH7.2 as coating buffer, chloromycetin envelope antigen is diluted to 1:60000 ratio, 100 μ L/ holes, 37 DEG C of lucifuges hatch 2h, take out ELISA Plate and get rid of liquid in plate, add diluted concentrated cleaning solution 300 μ L/ hole, wash plate 2 times, 30s/ time, then 0.5% bovine serum albumin(BSA) (BSA) confining liquid is added, 150 μ L/ holes, place 1.5h for 37 DEG C, get rid of confining liquid directly to pat dry, ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and is dried, inspect by random samples qualified after ELISA Plate is vacuum-sealed in 4 DEG C of conditions under preserve.
Chloromycetin standard concentration is respectively 0ng/mL, 0.01ng/mL, 0.03ng/mL, 0.09ng/mL, 0.81ng/mL, 2.43ng/mL.
Described chloramphenicol antibody working fluid adopts chloromycetin artificial antigen immune mouse to obtain monoclonal antibody, is diluted to the preparation of 1:80000 ratio with antibody diluent.
Described chloromycetin ELIAS secondary antibody working fluid adds diluted by ELIAS secondary antibody and becomes 1:4000 ratio, described substrate solution A is the citrate-phosphate disodium hydrogen buffer solution of the carbamide peroxide containing 0.5mmol/L, described substrate solution B is the ethanolic solution of tetramethyl biphenyl diamines, described stop buffer is the sulfuric acid of 2mol/L, described concentrated redissolution liquid is 10 times of concentrated liquid that redissolve, for the PBS of 0.1mol/L, between pH value range 7.0-7.5, described concentrated cleaning solution is 10 times of concentrated cleaning solutions, for containing 0.5% Tween-20, the PBST of 0.1mol/L, between pH value range 7.0-7.5.
The enzyme linked immunological kit of chlorine detection mycin and detection method thereof, based on the indirect competitive enzyme-linked immunosorbent reaction principle of antigen-antibody, the method detecting step is as follows:
Get the ELISA Plate being coated with chloromycetin antigen, add standard items/sample respectively according to the order of sequence, chloromycetin ELIAS secondary antibody working fluid, the each 50 μ L/ holes of chloramphenicol antibody working fluid are in the micropore of correspondence, to vibrate gently mixing, 30min is reacted with in the rearmounted room temperature of cover plate membrane cover plate 25 DEG C of light protected environment, liquid in hole is dried, 4 ~ 5 times are fully washed with wash operating solution, every minor tick 10s, substrate solution A50 μ L/ hole is added after patting dry, substrate solution B50 μ L/ hole, to vibrate gently mixing, 15 ~ 20min is reacted with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate, add stop buffer 50 μ L/ hole, to vibrate gently mixing, setting microplate reader detects in 450nm place or dual wavelength 450/630nm, measure every hole absorbance (please running through data in 5min), with the logarithm of standard concentration (ppb) for horizontal ordinate, standard items percentage absorbance is ordinate, drawing standard curve, the content of chloromycetin in reference standard curve calculation sample.
Wherein, described testing sample following methods carries out pre-service:
(1) milk 0.25ng/ml is by milk centrifugal segregation upper-layer fat, get 2mL removed fat milk sample add 4mL ethyl acetate, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
(2) tissue 1.5ng/g accurately takes the sample after 2g average in centrifuge tube, first add 2mL deionized water fully to mix and add 4mL ethyl acetate again, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
(3) serum 0.5ng/ml get 2mL serum add 4mL ethyl acetate vortex mixing after centrifugal, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL sample redissolve liquid mixing to be measured;
(4) honey (examining soon) 1.5ng/g accurately takes 2g sample in centrifuge tube, add people 2mL deionized water, vortex mixed adds people 4mL acetic acid ethyl ester after sample dissolves completely, centrifugal after vortex mixing, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, centrifugal after abundant vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured.
A kind of enzyme linked immunological kit of chlorine detection mycin of the present invention and the measuring principle of detection method thereof: antigen-specific sexual competition antibody fixing in the chloromycetin in sample and ELISA Plate, add ELIAS secondary antibody, with antibody response, by enzymatic chromogenic reagent, carry out the content of chloromycetin in judgement sample according to the depth of colour developing.If the chloromycetin content in sample is few, colour developing is dark; Otherwise, then develop the color shallow.Kit test method of the present invention is easy and simple to handle, detects sensitive, accurate, quick, is applicable to the quick detection of batch samples.
Embodiment
Chloromycetin protein conjugate adopts succinic anhydride method chloromycetin haptens and carrier protein BSA coupling to be prepared.
The preparation of chloramphenicol antibody:
Select the purebred BALA/C mouse of healthy adult, get the immunizing antigen 50 μ g prepared with protein molecule to mix with equivalent complete Freund's adjuvant and adopt lumbar injection to carry out initial immunity, adding equivalent incomplete Freund's adjuvant every 3 weeks with same dose immunizing antigen afterwards adopts lumbar injection to carry out secondary, three immunity, after after each immune 6 days, tail vein blood measures antiserum titre to certain titre, do not add adjuvant with same dose and carry out final immunization, get spleen after 3 days and prepare Spleen cell suspensions and myeloma cell carries out Fusion of Cells, filter out required hybridoma cell line and carry out cloning, the hybridoma being in exponential phase is selected to carry out frozen, prepare for ascites, first lumbar injection 0.5ml whiteruss is in BALB/C mouse sensitization, pneumoretroperitoneum injection 1 × 10 in 2 weeks
6individual hybridoma, ascites can be produced after inoculating cell 7-10 days, ascites is extracted with syringe when ascites is many as far as possible, repeatedly collect for several times, the centrifugal 15min of 4000rpm, collect supernatant, adopt caprylic acid-ammonium purifying ascites to carry out purifying to monoclonal antibody, freeze drying saves backup in-20 DEG C after obtaining freeze-dried powder.
Preparation is coated with the ELISA Plate of chloromycetin envelope antigen:
Chloromycetin haptens and the pure albumen of carrier proteins Bovine (BSA) coupling obtain by envelope antigen, with phosphate (PBS) damping fluid of 0.05mol/LpH7.2 as coating buffer, chloromycetin antigen diluent is become 1:60000 ratio, 100 μ L/ holes, place 2h for 37 DEG C, take out ELISA Plate and get rid of liquid in plate, with the concentrated cleaning solution 300 μ L/ hole after dilution, wash plate 2 times, 30s/ time, then add 0.5% bovine serum albumin(BSA) (BSA) to close, 150 μ L/ holes, place 1.5h for 37 DEG C, discard confining liquid, ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and is dried, inspect by random samples qualified after by rearmounted for ELISA Plate vacuum seal 4 DEG C preserve.
Chloromycetin standard items compound concentration is respectively 0ng/mL, 0.01ng/mL, 0.03ng/mL, 0.09ng/mL, 0.81ng/mL, 2.43ng/mL.
The preparation of chloramphenicol antibody working fluid: adopt chloromycetin artificial antigen immune mouse to obtain monoclonal antibody, is diluted to the preparation of 1:80000 ratio with antibody diluent.
Chloromycetin ELIAS secondary antibody working fluid adds diluted by ELIAS secondary antibody and becomes 1:4000 ratio, substrate solution A is the citrate-phosphate disodium hydrogen buffer solution of the carbamide peroxide containing 0.5mmol/L, substrate solution B is the ethanolic solution of tetramethyl biphenyl diamines, stop buffer is the sulfuric acid of 2mol/L, the concentrated liquid that redissolves is 10 times of concentrated liquid that redissolve, for the PBS of 0.1mol/L, between pH value range 7.0-7.5, concentrated cleaning solution is 10 times of concentrated cleaning solutions, for containing 0.5% Tween-20, the PBST of 0.01mol/L, between pH value range 7.0-7.5.
Based on the reagent of above-mentioned preparation, the enzyme linked immunological kit that the present invention is used for chlorine detection mycin comprises following material:
(1) 96 ELISA Plate × 1 piece, hole;
(2) titer × 6 bottle: (1mL/ bottle) 0ppm, 0.01ng/mL, 0.03ng/mL, 0.09ng/mL, 0.81ng/mL, 2.43ng/mL;
(3) antibody working fluid 7mL;
(4) ELIAS secondary antibody working fluid 7mL;
(5) substrate solution A7mL;
(6) substrate solution B7mL;
(7) stop buffer 7mL;
(8) 10 × concentrated liquid 40mL that redissolves;
(9) 10 × concentrated cleaning solution 40mL;
When kit of the present invention is for detecting chloramphenicol residue in sample, implemented by following steps: sample pretreatment, with kit of the present invention carry out detecting, analysis result.
(1) sample pretreatment
Milk: by milk centrifugal segregation upper-layer fat, get 2mL removed fat milk sample add 4mL ethyl acetate, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
Tissue: accurately take the sample after 2g average in centrifuge tube, first add 2mL deionized water fully to mix and add 4mL ethyl acetate again, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
Serum: get 2mL serum add 4mL ethyl acetate vortex mixing after centrifugal, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL sample redissolve liquid mixing to be measured;
Honey: accurately take 2g sample in centrifuge tube, add people 2mL deionized water, vortex mixed adds people 4mL acetic acid ethyl ester after sample dissolves completely, centrifugal after vortex mixing, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, centrifugal after abundant vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured.
(2) chloramphenicol residue in testing sample is detected with kit of the present invention
Pre-service testing sample, get the ELISA Plate being coated with chloromycetin antigen, add standard items/sample respectively according to the order of sequence, the each 50 μ L/ holes of chloramphenicol antibody working fluid are in the micropore of correspondence, to vibrate gently mixing, 30min is reacted with in the rearmounted room temperature of cover plate membrane cover plate 25 DEG C of light protected environment, liquid in hole is dried, 4 ~ 5 times are fully washed with wash operating solution, every minor tick 10s, chloromycetin ELIAS secondary antibody working fluid 100 μ L/ hole is added after patting dry, to vibrate gently mixing, 30min is reacted with in the rearmounted room temperature of cover plate membrane cover plate 25 DEG C of light protected environment, liquid in hole is dried, same procedure is washed, substrate solution A50 μ L/ hole is added after patting dry, substrate solution B50 μ L/ hole, to vibrate gently mixing, 15min is reacted with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate, add stop buffer 50 μ L/ hole, to vibrate gently mixing, setting microplate reader detects in 450nm place or dual wavelength 450/630nm, measure every hole absorbance (please running through data in 5min), with the logarithm of standard concentration (ppb) for horizontal ordinate, standard items percentage absorbance is ordinate, drawing standard curve, the content of chloromycetin in reference standard curve calculation sample.
(3) analysis result
The calculating of percentage absorbance, the percentage absorbance of standard items or sample equals the absorbance of mean value (diplopore) divided by first standard (0 standard) of the absorbance of standard items or sample, then is multiplied by 100%, namely
Percentage absorbance (%)=B/B
0× 100%
The wherein mean absorbance values of B-standard solution or sample solution, B
0the mean absorbance values of-0ppb standard solution.
With the logarithm of the standard concentration of chloromycetin (ppb) for horizontal ordinate, standard items percentage absorbance is ordinate, and drawing standard curve, obtains straight-line equation.Substituted in typical curve by the percentage absorbance of sample, read the concentration corresponding to sample from typical curve, the extension rate being multiplied by its correspondence is the actual concentrations of chloromycetin in sample.
Claims (8)
1. the enzyme linked immunological kit of chlorine detection mycin and a detection method thereof, comprises ELISA Plate, chloromycetin standard items, chloramphenicol antibody working fluid, chloromycetin ELIAS secondary antibody working fluid, substrate solution A, substrate solution B, stop buffer, concentrated liquid and the concentrated cleaning solution of redissolving.
2. the enzyme linked immunological kit of chlorine detection mycin and a detection method thereof, comprises the following steps: the preparation of the preparation of the preparation of the preparation of ELISA Plate, the preparation of chloromycetin standard items, chloramphenicol antibody working fluid, the preparation of chloromycetin ELIAS secondary antibody working fluid, substrate solution A, the preparation of substrate solution B, stop buffer, concentratedly redissolve the preparation of liquid and the preparation of concentrated cleaning solution.
3. the enzyme linked immunological kit of a kind of chlorine detection mycin according to claim 2 and detection method thereof, it is characterized in that: described ELISA Plate preparation method is for obtain chloromycetin envelope antigen by chloromycetin haptens and the pure albumen of carrier proteins Bovine (BSA) coupling, with phosphate (PBS) damping fluid of 0.05mol/LpH7.2 as coating buffer, envelope antigen is diluted to 1:60000 ratio, 100 μ L/ holes, hatch 2h for 37 DEG C, take out ELISA Plate and get rid of liquid in plate, add the concentrated cleaning solution after dilution 300 μ L/ hole, wash plate 2 times, 30s/ time, then add 0.5% bovine serum albumin(BSA) (BSA) to close, 150 μ L/ holes, place 1.5h for 37 DEG C, discard confining liquid directly to pat dry, ELISA Plate after patting dry is placed (25 DEG C) between constant temperature and is dried, inspect by random samples qualified after ELISA Plate is vacuum-sealed in 4 DEG C of conditions under preserve.
4. the enzyme linked immunological kit of a kind of chlorine detection mycin according to claim 2 and detection method thereof, is characterized in that: the concentration of chloromycetin standard items is respectively 0ng/mL, 0.01ng/mL, 0.03ng/mL, 0.09ng/mL, 0.81ng/mL, 2.43ng/mL.
5. the enzyme linked immunological kit of a kind of chlorine detection mycin according to claim 2 and detection method thereof, it is characterized in that: described chloramphenicol antibody working fluid adopts chloromycetin artificial antigen immune mouse to obtain monoclonal antibody, is diluted to the preparation of 1:80000 ratio with antibody diluent.
6. the enzyme linked immunological kit of a kind of chlorine detection mycin according to claim 2 and detection method thereof, it is characterized in that: described chloromycetin ELIAS secondary antibody working fluid adds diluted by ELIAS secondary antibody and becomes 1:4000 ratio, described substrate solution A is the citrate-phosphate disodium hydrogen buffer solution of the carbamide peroxide containing 0.5mmol/L, described substrate solution B is the ethanolic solution of tetramethyl biphenyl diamines, described stop buffer is the sulfuric acid of 2mol/L, described concentrated redissolution liquid is 10 times of concentrated liquid that redissolve, it is the PBS of 0.1mol/L, between pH value range 7.0-7.5, described concentrated cleaning solution is 10 times of concentrated cleaning solutions, it is for containing 0.5% Tween-20, the PBST of 0.01mol/L, between pH value range 7.0-7.5.
7. the enzyme linked immunological kit of a kind of chlorine detection mycin according to claim 2 and detection method thereof, based on the indirect competitive enzyme-linked immunosorbent reaction principle of antigen-antibody, the method is characterized in that: pre-service testing sample, get the ELISA Plate being coated with chloromycetin antigen, add standard items/sample respectively according to the order of sequence, the each 50 μ L/ holes of chloramphenicol antibody working fluid are in the micropore of correspondence, to vibrate gently mixing, 30min is reacted with in the rearmounted room temperature of cover plate membrane cover plate 25 DEG C of light protected environment, liquid in hole is dried, 4 ~ 5 times are fully washed with wash operating solution, every minor tick 10s, chloromycetin ELIAS secondary antibody working fluid 100 μ L/ hole is added after patting dry, to vibrate gently mixing, 30min is reacted with in the rearmounted room temperature of cover plate membrane cover plate 25 DEG C of light protected environment, liquid in hole is dried, same procedure is washed, substrate solution A50 μ L/ hole is added after patting dry, substrate solution B50 μ L/ hole, to vibrate gently mixing, 15min is reacted with in the rearmounted 25 DEG C of light protected environment of cover plate membrane cover plate, add stop buffer 50 μ L/ hole, to vibrate gently mixing, setting microplate reader detects in 450nm place or dual wavelength 450/630nm, measure every hole absorbance (please running through data in 5min), with the logarithm of standard concentration (ppb) for horizontal ordinate, standard items percentage absorbance is ordinate, drawing standard curve, the content of chloromycetin in reference standard curve calculation sample.
8. method according to claim 7, wherein, described testing sample following methods carries out pre-service:
Milk: by milk centrifugal segregation upper-layer fat, get 2mL removed fat milk sample add 4mL ethyl acetate, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
Tissue: accurately take the sample after 2g average in centrifuge tube, first add 2mL deionized water fully to mix and add 4mL ethyl acetate again, centrifugal after vortex mixing, get upper organic phase 2mL to dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, fully centrifugal after vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured;
Serum: get 2mL serum add 4mL ethyl acetate vortex mixing after centrifugal, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL sample redissolve liquid mixing to be measured;
Honey: accurately take 2g sample in centrifuge tube, add people 2mL deionized water, vortex mixed adds people 4mL acetic acid ethyl ester after sample dissolves completely, centrifugal after vortex mixing, get upper organic phase 2mL and dry up in 50 ~ 60 DEG C of water-bath nitrogen, add 1mL normal hexane dissolution residual substance, and add 1mL sample redissolution liquid, centrifugal after abundant vibration mixing, remove the foam between upper strata normal hexane phase and two-phase or jelly, remaining layer aqueous phase is to be measured.
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| CN109813900A (en) * | 2017-11-21 | 2019-05-28 | 南京亿特生物科技有限公司 | A kind of time-resolved fluoroimmunoassay chromatography quantitative testing test paper item detecting chloramphenicol |
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Application publication date: 20160210 |