CN1308685C - Kit for detecting chloromycetin - Google Patents
Kit for detecting chloromycetin Download PDFInfo
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- CN1308685C CN1308685C CNB2004100372547A CN200410037254A CN1308685C CN 1308685 C CN1308685 C CN 1308685C CN B2004100372547 A CNB2004100372547 A CN B2004100372547A CN 200410037254 A CN200410037254 A CN 200410037254A CN 1308685 C CN1308685 C CN 1308685C
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Abstract
The present invention discloses a reagent box for detecting chloramphenicol. The reagent box for detecting chloramphenicol provided by the present invention comprises enzyme marked chloramphenicol and covered chloramphenicol specific antibodies. The reagent box for detecting chloramphenicol of the present invention can simultaneously and rapidly detect numerous samples, and main reagents are arranged in the form of working fluid and are convenient for use. The present invention has the characteristics of high specificity, high sensitivity, high precision, high accuracy, etc., and is capable of having important functions on the chloramphenicol residue detection of animal food.
Description
Technical field
The present invention relates to the kit of a kind of chlorine detection mycin in enzyme linked immunological and the detection of veterinary drugs in food analysis technical field.
Background technology
(chloramphenicol CAP) is widely used antibiotic to chloromycetin, has played vital role in livestock and poultry control and treatment.But because there is serious adverse in chloromycetin, can suppress mitochondrial protein synthesis in the bone marrow cell, cause bone marrow cell and hepatocellular toxicity, thereby cause people's reproducibility aplastic anemia, agranulocytosis, diseases such as neonate, premature's ash baby syndrome, developed countries such as America and Europe forbid in succession or strictness bans use of.In Dec, 2002, the China Ministry of Agriculture announced the literary composition regulation No. 235, and chloromycetin and salt thereof, ester (comprising Chloramphenicol Succinate) must not detect in all edible tissues of all food animals.But because chloromycetin low price and be broad-spectrum antibiotic, illegal use is still very general.Therefore strengthen the residue detection of chloromycetin in the animal food is very important.
At present, be usually used in the method that residual chloromycetin detects and mainly contain microbial method and instrumental method.Though the microorganism detection method is economical, easy and simple to handle, when having other microbial inhibitors to exist in sample, its sensitivity and specificity are restricted; Simple instrument analytical methods such as high efficiency liquid phase chromatographic analysis method, gas spectrum, GC-MS method, though highly sensitive, sample pre-treatment and measurement operation are loaded down with trivial details, the expense height is unwell to the great amount of samples examination, can be used as residual conclusive evidence analysis.Its remolding sensitivity microbial method height of immuno-chemical method, especially enzyme-linked immunosorbent assay (ELISA), sample pre-treatments is simpler than instrumental method again, is particularly suitable for on-site supervision and great amount of samples examination.
The innovation and creation content
The kit that the purpose of this invention is to provide a kind of chlorine detection mycin.
The kit of chlorine detection mycin provided by the present invention comprises the chloromycetin specific antibody of enzyme mark chloromycetin and bag quilt.
Wherein, described chloromycetin specific antibody can be chloromycetin monoclonal antibody or chloromycetin polyclonal antibody, described chloromycetin monoclonal antibody or polyclonal antibody can be mouse source, Ma Yuan, Yang Yuan, rabbit source and cavy source antibody, described chloromycetin monoclonal antibody is preferably the chloromycetin mouse monoclonal antibody, and described chloromycetin polyclonal antibody is preferably the chloromycetin rabbit polyclonal antibody.Above antibody all can prepare as immunogene according to a conventional method with the conjugate of chloromycetin and carrier protein.Described carrier protein can be bovine serum albumin(BSA) (BSA), human serum albumins (HSA), ovalbumin (OVA) or hemocyanin common carrier albumen such as (KLH).Described marker enzyme can be horseradish peroxidase or alkaline phosphatase, is preferably horseradish peroxidase.
The material that can be used as the carrier of fixing described chloromycetin specific antibody is a lot, as polystyrene, cellulose, polyacrylamide, tygon, polypropylene, cross-link dextran, glass, silicon rubber, Ago-Gel etc.The form of this carrier can be test tube, micro-reaction plate shrinkage pool, globule, sequin etc.
For more convenient on-site supervision and great amount of samples examination, described kit also comprises chloromycetin standard solution, developer, stop buffer, concentrated cleaning solution and concentrates redissolution liquid.
Described concentrated cleaning solution is the phosphate buffer that contains the 0.8-1.2% tween; Described developer is made up of colour developing liquid A liquid and colour developing liquid B liquid, and described colour developing liquid A liquid is hydrogen peroxide or urea peroxide, and described colour developing liquid B liquid is o-phenylenediamine (OPD) or tetramethyl benzidine (TMB); Described concentrated redissolution liquid is the citrate buffer that contains the 0.8-1.2% tween.
Detection principle of the present invention is for to be adsorbed in the chloromycetin specific antibody on the solid phase carrier as coating antigen, add sample and enzyme mark chloromycetin, chloromycetin and enzyme mark chloromycetin residual in the testing sample are competed the chloromycetin specific antibody that wraps quilt on the solid phase carrier, the colour developing back stops, the working sample light absorption value, chloramphenicol residue is negative correlation in this value and the sample, relatively can draw the content of chloromycetin with typical curve.Simultaneously according to the depth of the color sample on the ELISA Plate, but with the concentration range of the comparison judgement sample of the chloromycetin standard solution color of series concentration.
The kit of chlorine detection mycin of the present invention mainly adopts the residual quantity of chloromycetin in the samples such as the qualitative or detection by quantitative animal tissue (musculature of pig, chicken, ox, sheep and liver, fishes and shrimps etc.) of direct competitive ELISA method, urine sample, serum blood plasma, feed, honey, milk; Pre-treatment requirement to sample is low, and sample pretreatment process is simple, simultaneously the fast detecting gross sample; This kit adopts the chloromycetin monoclonal antibody or the polyclonal antibody of high specific, main agents all provides with the working fluid form, easy to use, have characteristics such as high specific, high sensitivity, pinpoint accuracy, pin-point accuracy, can in the animal food residual chloromycetin detects, play a significant role.
Description of drawings
Fig. 1 is the structural representation of the kit of chlorine detection mycin
Embodiment
The preparation of embodiment 1, antigen and antibody
(1) enzyme-labelled antigen is synthetic
Chloromycetin and horseradish peroxidase are carried out coupling, obtain the chloromycetin of horseradish peroxidase-labeled.
(2) immunogenic synthetic
Adopt active ester method to carry out coupling chloromycetin and bovine serum albumin(BSA) (BSA) and obtain immunogene.
(3) chloromycetin mouse monoclonal antibody preparation
The animal immune program adopts the Balb/c mouse as immune animal, with chloromycetin and bovine serum albumin(BSA) conjugate is immunogene, immunizing dose is 80-100 μ g/, Freund's complete adjuvant with immunogene and equivalent when head exempts from is mixed and made into emulsifying agent, the subcutaneous multi-point injection of nape portion, At intervals of two to three weeks are got the same dose immunogene and are added equivalent incomplete Freund mixing and emulsifying, and booster immunization once, four exempt from the pneumoretroperitoneum booster immunization once, extracting spleen cell after 3 days.
Immune BALB/c mouse splenocyte is got in Fusion of Cells and cloning, and in 5-10: 1 ratio and SP2/0 myeloma cell are merged, and adopts indirect competitive ELISA to measure cell conditioned medium liquid, screens positive hole.Utilize limiting dilution assay that cloning is carried out in positive hole, obtain the hybridoma cell strain of stably excreting monoclonal antibody.
Cell cryopreservation and recovery are got the hybridoma that is in exponential phase and are made 1-5 * 10 with cryopreserving liquid
6The cell suspension of individual/ml is sub-packed in frozen pipe, in the medium-term and long-term preservation of liquid nitrogen.Take out frozen pipe during recovery, put into 37 ℃ of water-bath middling speeds immediately and melt, behind the centrifugal removal cryopreserving liquid, move in the culture flask and cultivate.
MONOCLONAL ANTIBODIES SPECIFIC FOR and purifying adopt in the body and induce method, and Balb/c mouse (8 age in week) abdominal cavity is only injected sterilization paraffin oil 0.5ml/, 7-14 days pneumoretroperitoneum injection hybridomas 5 * 10
5-10
6Individual/as only, to gather ascites after 7-10 days.Carry out the ascites purifying through sad-saturated ammonium sulfate method, bottle packing ,-20 ℃ of preservations.
(4) preparation of chloromycetin rabbit polyclonal antibody
Adopt new zealand white rabbit as immune animal, with chloromycetin and bovine serum albumin(BSA) conjugate is immunogene, immunizing dose is 1mg/kgb.w., Freund's complete adjuvant with immunogene and equivalent when head exempts from is mixed and made into emulsifying agent, the subcutaneous multi-point injection of nape portion, getting the same dose immunogene 3-4 week adds equivalent incomplete Freund mixing and emulsifying at interval, and booster immunization is once, immunity is 5 times altogether, does not add adjuvant for the last time.Take a blood sample behind the last immune 7-10d, measure serum antibody titer, heart is taken a blood sample, and obtains the polyclonal antibody of purifying through ammonium sulfate precipitation.
The kit of embodiment 2, chlorine detection mycin
(1) structure of the kit of chlorine detection mycin
The structure of kit as shown in Figure 1, mainly by box body 1, vacuum-packed 96 holes of aluminium film/2,6 bottles of chloromycetin series concentration standard items 3 of 40 hole ELISA Plate, stop buffer 4, enzyme labeling thing working fluid 5, substrate colour developing liquid A liquid 6, substrate colour developing liquid B liquid 7, concentrated cleaning solution 8, concentrated liquid 9 and the foam carriage 10 of redissolving, be shaped on shrinkage pool on the foam carriage 10, mentioned reagent bottle 3-9 is placed in the shrinkage pool of foam carriage, and foam carriage 10 and ELISA Plate 2 are placed in the box body.Wherein ELISA Plate 2 is made up of plastic stent and detachable plastic strip.
(2) preparation of agents useful for same
A. chloromycetin standard solution: 6 bottles of chloromycetin series standard solution, 0,0.01,0.25,2.5,5.0,10.0ng/ml, 1-3ml/ bottle.
B. bag is cushioned liquid: pH9.6, the carbonate buffer solution of 0.05mol/L.
C. confining liquid: 3-10% calf serum.
D. concentrated cleaning solution: contain the phosphate buffer (0.01M pH7.4) of 0.8-1.2% tween, for the 15-25 of normal working concentration doubly, 30-50ml/ bottle, 1 bottle.
E. enzyme labeling thing working fluid: enzyme mark chloromycetin working fluid, 5-8ml/ bottle, 1 bottle.
F. substrate colour developing liquid A liquid: hydrogen peroxide or urea peroxide, 5-8ml/ bottle, 1 bottle.
G. substrate colour developing liquid B liquid: o-phenylenediamine (OPD) or tetramethyl benzidine (TMB), 5-8ml/ bottle, 1 bottle.
H. stop buffer: 1-2mol/L sulfuric acid or hydrochloric acid, 5-8ml/ bottle, 1 bottle.
I. concentrate to redissolve liquid: contain the citrate buffer of 0.8-1.2% tween, for the 5-10 of normal working concentration doubly, 30-50ml/ bottle, 1 bottle.
(3) preparation of ELISA Plate
Be cushioned liquid with bag the chloromycetin specific antibody is diluted to 0.1-1 μ g/ml, every hole adds 100 μ l, 37 ℃ of incubation 2h, and 4 ℃ are spent the night, coating buffer inclines, with cleansing solution washing 3 times, each 1min pats dry, in every hole, add 150-200 μ l confining liquid then, 37 ℃ of incubation 1-2h, liquid in the hole of inclining, preserve with the vacuum seal of aluminium film dry back.
The pre-treatment of embodiment 3, sample
(1) animal tissue takes by weighing the sample after the 5g homogenate, adds 20ml ethyl acetate, whirling motion, and centrifugal 5 minutes of 3000g gets organic phase in the heart bottle.Add 20ml ethyl acetate again and repeat to extract once in sample, 3000g is centrifugal, gets supernatant, and twice supernatant is incorporated in the heart bottle.50 ℃ of evaporated under reduced pressure, the 4.5ml dissolved in distilled water adds normal hexane, and mixing leaves standstill, and abandons upper organic phase.Add 0.5ml and concentrate redissolution liquid, obtain testing sample solution, get 50 μ l and analyze.
(2) urine sample is got the 1ml urine, add 0.5ml 100mM sodium-acetate buffer, pH4.8 adds 8 μ l GRD beta-glucuronidase, hatches 3h for 37 ℃, add the mixing of 2ml ethyl acetate after returning to room temperature, centrifugal 10 minutes of 3000g takes out the 1ml supernatant liquid and dries up with nitrogen, with concentrating redissolution liquid dilute sample, obtain testing sample solution, get 50 μ l and analyze.
(3) serum blood plasma is got 1ml serum blood plasma, adds 2ml ethyl acetate, mixes vibration, and centrifugal 10 minutes of 3000g gets 1ml upper strata liquid and dries up with nitrogen, with the concentrated liquid dilute sample that redissolves, obtains testing sample solution, gets 50 μ l and analyzes.
(4) feed is got 5g feed adding 20ml ethyl acetate, mixes vibration, centrifugal 10 minutes of 3000g.Getting 1ml upper strata liquid dries up with nitrogen.With 1ml isooctane+chloroform (40+60) dissolution residual substance.With concentrating redissolution liquid dilute sample, centrifugal 10 minutes of 3000g obtains testing sample solution, gets 50 μ l and analyzes.
(5) honey is got 1g honey, adds 4ml distilled water, adds 4ml ethyl acetate again, mixes vibration, centrifugal 10 minutes of 3000g.Get 2ml upper strata liquid and dry up,, obtain testing sample solution, get 50 μ l and analyze with concentrating redissolution liquid dilute sample with nitrogen.
(6) milk centrifugal 10 minutes of the sample 6000g that will suckle absorbs the fat on upper strata, gets 0.5ml and removes fat milk with redissolving the liquid dilution, obtains testing sample solution, gets 50 μ l and analyzes.
Residual chloromycetin in embodiment 4, the detection chicken gizzard sample
(1) sample pre-treatments
Take by weighing the sample after the 5g homogenate, add 20ml ethyl acetate, whirling motion was left standstill 20 minutes, and centrifugal 5 minutes of 3000g gets organic phase in the heart bottle.Add 20ml ethyl acetate again and repeat to extract once in sample, centrifugal 10 minutes of 3000g gets supernatant, and twice supernatant is incorporated in the heart bottle.50 ℃ of evaporated under reduced pressure add the 4.5ml dissolved in distilled water, add normal hexane 5ml, and mixing leaves standstill, and abandons upper organic phase.Add 0.5ml and concentrate redissolution liquid, obtain testing sample solution, get 50 μ l and analyze.
(2) sample detection
In 96 hole ELISA Plate micropores, add series standard solution or sample solution (each 2 hole) 50 μ l, add enzyme mark chloromycetin working fluid 50 μ l then,, react 1h in 37 ℃ of constant temperature ovens with cover plate film shrouding with chloromycetin specific antibody bag quilt.Pour out liquid in the hole, every hole adds 250 μ l concentrated cleaning solutions (phosphate buffer (0.01M pH7.4) that contains 1.2% tween), pours out liquid in the hole after 1 minute, pats dry with thieving paper, and so repetitive operation is washed plate 5 times altogether.Every hole adds substrate colour developing liquid A liquid (urea peroxide) 50 μ l, adds B liquid (o-phenylenediamine) 50 μ l again, the mixing that vibrates gently, 37 ℃ of constant temperature oven lucifuge colour developing 5-10min.Every hole adds stop buffer (1.5mol/L sulfuric acid) 50 μ l, and the mixing that vibrates is gently measured every hole absorbance (OD value) with microplate reader.
The used reagent of present embodiment is all prepared according to the reagent compound method among the embodiment 2.
Each the concentration standard solution that is obtained and the mean value (B) of sample absorbance are divided by the absorbance (B of first standard (0 standard)
0) multiply by 100 again, i.e. percentage absorbance.
B is the mean light absorbency value of standard solution or sample solution in the formula, B
0It is the mean light absorbency value of 0 μ g/L standard solution.Natural logarithm value with chloramphenicol concentration (μ g/L) is an X-axis, and the percentage absorbance is a Y-axis, the drawing standard curve map, and the concentration of corresponding each sample (μ g/L) can be read from typical curve.Also can calculate sample solution concentration with regression equation method.Utilize computer professional software, the express-analysis of a large amount of samples of being more convenient for.Whole testing process only needed just can finish in 1.5 hours.
Embodiment 5, kit sensitivity, specificity, precision, accuracy and storage life test
(1) kit sensitivity test
50% inhibition concentration (both IC
50, referring to the drug concentration of 50% place correspondence of the absorbance of 0 standard solution) and Chang Zuowei estimates the index of competitive ELISA kit sensitivity.Measure 50% inhibition concentration of 20 typical curves respectively, determine this kit standard curve I C
50Should be within the scope of 1.5-4.0 μ g/L.Measure blank sample and 20 parts of chloromycetin 0 standard items of 20 portions of chicken, chicken gizzard, pork, pork liver, fishes and shrimps, serum blood plasma, urine sample, honey, milk and feed respectively.The result judges that this kit is limited to 0.05 μ g/L to chloromycetin 0 standard items lowest detection.To the lowest detectable limit of muscle, liver, fishes and shrimps, serum blood plasma, urine sample, honey and feed all less than 0.1 μ g/L; Lowest detection to milk sample is limited to 0.15 μ g/L.
(2) kit specificity test
Select 4 kinds of chloromycetin drug monitoring cross reacting rates with chloromycetin similar structures and similar functions.Typical curve by various medicines obtains its 50% inhibition concentration respectively.Calculate the cross reacting rate of kit with following formula to other medicines.Cross reacting rate is littler, shows that the specificity of kit is higher.
The specificity of table 1. kit
| Medicine name | Cross reacting rate (%) |
| Chloromycetin (CAP) | 100.0 |
| Chloramphenicol Succinate | ＞100.0 |
| Chloramphenicol Base | ＜25.0 |
| Thiamphenicol (TAP) | ＜0.5 |
| Florfenicol (FF) | ＜1.0 |
| Penicillin | ＜0.01 |
| Streptomysin | ＜0.01 |
| Sulfadimidine (SM 2) | ＜0.01 |
Kit specificity test findings is as shown in table 1, the result shows that (annotate: Chloramphenicol Succinate enters in the body to clear up through enzyme becomes chloromycetin performance pharmacological action to this kit to the cross reacting rate height of chloromycetin and Chloramphenicol Succinate, so in animal-derived food, all exist with the chloromycetin form), can guarantee reliability to residual chloromycetin testing result in animal-derived food and the feed.
(3) kit precision test
The standard repeatability
From three batches of elisa plates, every plate is extracted 20 micropores out, measures same concentration standard solution absorbency value (0D value), and replication 10 times calculates the coefficient of variation.Coefficient of variation scope is at 3.5-9.7%.
The sample repeatability
Get the chloromycetin standard specimen of variable concentrations, add in the sample, get each three of the kits of three different batches respectively, each concentration repeats 5 times.Calculate plate interior, batch interior, interassay coefficient of variation respectively.The Variation Lines number average of animal muscle, animal's liver, fishes and shrimps, serum blood plasma, urine sample, milk and honey is lower than 20.0%, and the Variation Lines number average of feed is lower than 15.0%.
(4) accuracy of kit
Get the chloromycetin standard specimen of 4 concentration, sample added recovery test, each concentration establish 4 parallel, calculate recovery rate respectively.The interpolation recovery of animal's liver, animal muscle, fishes and shrimps, serum blood plasma, urine sample, honey, milk and feed is all in the 60.0-120.0% scope.
(5) kit storage life test
The kit preservation condition is 2-8 ℃, and through 6 months mensuration, the maximum absorbance value (zero standard) of kit, 50% inhibition concentration, chloromycetin added the practical measurement value all within normal range.Consideration has improper preservation condition and occurs in transportation and use, and kit was placed 6 days under the condition of 37 ℃ of preservations, carries out the accelerated deterioration experiment, and the result shows that the every index of this kit meets the requirements fully.Consider that the freezing situation of kit takes place, kit was put into-20 ℃ of refrigerators freezing 5 days, measurement result shows that also the every index of kit is normal fully.Can draw kit from above result can preserve more than 6 months at least at 2-8 ℃.
Claims (3)
1, a kind of kit of chlorine detection mycin, the chloromycetin that comprises horseradish peroxidase or alkaline phosphate ester enzyme labeling, bag is by the ELISA Plate of chloromycetin monoclonal antibody or chloromycetin polyclonal antibody, concentration is respectively 0,0.01,0.25,2.5,5.0 and 6 bottles of chloromycetin series standard solution of 10.0ng/ml, the developer of forming by colour developing liquid A liquid and colour developing liquid B liquid, concentrated cleaning solution, stop buffer and the concentrated liquid that redissolves;
Wherein, described colour developing liquid A liquid is hydrogen peroxide or urea peroxide, and described colour developing liquid B liquid is o-phenylenediamine or tetramethyl benzidine; Described concentrated cleaning solution is the phosphate buffer that contains the 0.8-1.2% tween; The citrate buffer that described concentrated redissolution liquid is the 0.8-1.2% tween;
Described ELISA Plate prepares with following method: be cushioned liquid with bag chloromycetin monoclonal antibody or polyclonal antibody are diluted to 0.1-1 μ g/ml, every hole adds 100 μ l, 37 ℃ of incubation 2h, and 4 ℃ are spent the night, coating buffer inclines, with cleansing solution washing 3 times, each 1min pats dry, in every hole, add 150-200 μ l confining liquid then, 37 ℃ of incubation 1-2h, liquid in the hole of inclining, preserve with the vacuum seal of aluminium film dry back; It is pH9.6 that described bag is cushioned liquid, the carbonate buffer solution of 0.05mol/L; Described confining liquid is the 3-10% calf serum.
2, kit according to claim 1 is characterized in that: described chloromycetin monoclonal antibody or chloromycetin polyclonal antibody are that the conjugate with chloromycetin and carrier protein obtains as immunogene.
3, kit according to claim 2 is characterized in that: described carrier protein is bovine serum albumin(BSA), human serum albumins, ovalbumin or hemocyanin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100372547A CN1308685C (en) | 2004-04-30 | 2004-04-30 | Kit for detecting chloromycetin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100372547A CN1308685C (en) | 2004-04-30 | 2004-04-30 | Kit for detecting chloromycetin |
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| Publication Number | Publication Date |
|---|---|
| CN1690707A CN1690707A (en) | 2005-11-02 |
| CN1308685C true CN1308685C (en) | 2007-04-04 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB2004100372547A Expired - Fee Related CN1308685C (en) | 2004-04-30 | 2004-04-30 | Kit for detecting chloromycetin |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105759040A (en) * | 2015-12-31 | 2016-07-13 | 贵州勤邦食品安全科学技术有限公司 | Enzyme-linked immunosorbent reagent kit for detecting chloramphenicol residues |
| CN112239751B (en) * | 2020-10-13 | 2023-02-03 | 苏州大学 | Florfenicol/thiamphenicol monoclonal antibody hybridoma cell strain, monoclonal antibody secreted by same and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RO82727A2 (en) * | 1981-10-01 | 1983-11-01 | Institutul Pentru Controlul De Stat Al Medicamentului Si Cercetari Farmaceutice,Ro | CHROMATOGRAPHIC METHOD FOR DETERMINING FREE CLORAMPHENICOL IN CLORAMPHENIC HEMISUCCINATE |
| DE4013004A1 (en) * | 1990-04-24 | 1991-10-31 | Elvira Schecklies | ELISA determn. of low mol. wt. pesticides, antibiotics or toxins - using enzyme labelled antibody reactant, providing lower detection limit |
| CN1389729A (en) * | 2002-07-12 | 2003-01-08 | 江南大学 | Chloromycetin enzyme immunoassay kit for animal food and its immunoassay method |
| CN1429845A (en) * | 2001-12-30 | 2003-07-16 | 中国科学院上海生物工程研究中心 | Mouse antichloromycetin monoclonal antibody and its use |
-
2004
- 2004-04-30 CN CNB2004100372547A patent/CN1308685C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RO82727A2 (en) * | 1981-10-01 | 1983-11-01 | Institutul Pentru Controlul De Stat Al Medicamentului Si Cercetari Farmaceutice,Ro | CHROMATOGRAPHIC METHOD FOR DETERMINING FREE CLORAMPHENICOL IN CLORAMPHENIC HEMISUCCINATE |
| DE4013004A1 (en) * | 1990-04-24 | 1991-10-31 | Elvira Schecklies | ELISA determn. of low mol. wt. pesticides, antibiotics or toxins - using enzyme labelled antibody reactant, providing lower detection limit |
| CN1429845A (en) * | 2001-12-30 | 2003-07-16 | 中国科学院上海生物工程研究中心 | Mouse antichloromycetin monoclonal antibody and its use |
| CN1389729A (en) * | 2002-07-12 | 2003-01-08 | 江南大学 | Chloromycetin enzyme immunoassay kit for animal food and its immunoassay method |
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