CN106546731A - A kind of chloromycetin detection method and detection card - Google Patents
A kind of chloromycetin detection method and detection card Download PDFInfo
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- CN106546731A CN106546731A CN201611114230.6A CN201611114230A CN106546731A CN 106546731 A CN106546731 A CN 106546731A CN 201611114230 A CN201611114230 A CN 201611114230A CN 106546731 A CN106546731 A CN 106546731A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9446—Antibacterials
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Abstract
The invention discloses a kind of chloromycetin detection method and detection card, take 5 10g samples and be put in pulverizer, add extracting solution and 10 15ml distilled water, take out after crushing 2 3min；Sample after crushing completely is added in centrifuge tube, 2 5ml ethyl acetate are added, 3 7min are centrifuged with 2000 2300r/min, upper solution is removed after standing 10 15min standby；Remove in the sample addition centrifuge tube of upper solution, add 0.5ml buffer, 5 10min are centrifuged with 3000 4000r/min, it is to be checked using filtrate is taken after 0.22um filtering with microporous membranes.The chloromycetin detection method and detection card, the extraction ratio of chloromycetin can be effectively ensured, the accuracy of testing result is effectively ensured, step is simple, easy to operate, sensitivity is high, operator can grasp rapidly detection method, be suitable to promote, intuitively clearly can express experimental result, facilitate the detection of testing staff, it is practical.
Description
Technical field
The present invention relates to detect card technique field, specially a kind of chloromycetin detection method and detection block.
Background technology
Chloromycetin is the antibiotic produced by Venezuela Streptothrix.Chloromycetin be in the world first kind completely by synthetic method
Many different types of microorganisms are played works use by the broad ectrum antibiotic of a large amount of manufactures.Because of the cheap relation of price, chloromycetin
Still prevailing but to use other western countries Jing is very few in some low income countries, this is the pass of the side effect due to it
System.Chloromycetin can cause fatal hypoplastic anemia.Chloromycetin is mainly used in the collyrium for curing bacterial conjunctivitis
Or on ointment.
ELISA kit, mainly uses the base of antigen and the specific immuno-chemical reaction of antibody
Present principles are carrying out, in other words, as being carried out using the antibody in the chloromycetin in enzyme marker and sample and micropore competing
Strive reaction.In the middle of whole reaction, if the residual chloromycetin in sample is more, relatively will the more enzymes of competitive reaction
Label so that can relatively reduce with reference to the enzyme marker of upper antibody, is developed the color with tmb substrate, the chloromycetin content in sample
Absorbance with sample is in inverse ratio, and chloromycetin content is drawn by comparing with standard curve.
Although chloromycetin be a kind of effective antibiotic, but toxic and side effects are larger, can not only bring serious food to pacify
Full problem, can also bring serious threat to the health of the mankind.Therefore there is important meaning to the accurate quick detection of chloromycetin
Justice.
The content of the invention
（One）The technical problem of solution
For the deficiencies in the prior art, the invention provides a kind of chloromycetin detection method and detection card, can be to the chlorine in food
Mycin is detected, solves chloromycetin and can bring serious food-safety problem, can also bring serious to the health of the mankind
The problem of threat.
（Two）Technical scheme
To realize object above, the present invention is achieved by the following technical programs：
A kind of chloromycetin detection method, comprises the following steps：
S1, take 5-10g samples and be put in pulverizer, add extracting solution and 10-15ml distilled water, take out after crushing 2-3min.
S2, will crush completely after sample add centrifuge tube, add 2-5ml ethyl acetate, with 2000-2300r/min
Centrifugation 3-7min, removes upper solution after standing 10-15min standby.
S3, remove upper solution sample add centrifuge tube in, add 0.5ml buffer, with 3000-4000r/min
Centrifugation 5-10min, it is to be checked using filtrate is taken after 0.22um filtering with microporous membranes.
Preferably, the sample can be muscle, fat, liver and the kidney of the animals such as chicken, duck, fish, cattle, pig, sheep, rabbit.
Preferably, 8-10h is stood in the environment of the liquid to be detected being placed on 28 DEG C.
Preferably, containing one or two in ethanol and glycerol, the ethanol and glycerol in the extracting solution
Concentration is 60%-70%.
Preferably, the buffer is 0.05-0.08M PBS.
A kind of chloromycetin detection card, including detection card body, one end of the detection card body is provided with handle, the inspection
One end that card is surveyed away from handle is provided with standard color comparison card, and the detection card body includes base plate, is provided with the base plate
Lid, offers well and detection zone, is provided with sample pad, protecting film, reagent paper between the upper lid and base plate on the upper lid
Bar and adsorptive pads, are provided with detection line and control line in the sample pad.
Preferably, the sample pad, test strips, adsorptive pads and protecting film are pasted onto on base plate successively.
Preferably, its preparation method is comprised the following steps：
1）Colloid gold label chloramphenicol resistance monoclonal is coated with test strips.
2）Detection line is the conjugate of chloromycetin and BSA, and nature controlling line is sheep anti-mouse igg.
（Three）Beneficial effect
The invention provides a kind of chloromycetin detection method and detection card, possess following beneficial effect：
（1）The present invention can be effectively ensured the extraction ratio of chloromycetin, and the accuracy of testing result has been effectively ensured.
（2）Step of the present invention is simple, easy to operate, and sensitivity is high, and operator can grasp rapidly detection method, be suitable to push away
Extensively.
（3）Experimental result intuitively clearly can be expressed by the present invention, facilitate the detection of testing staff, practical.
Description of the drawings
Fig. 1 is the structural representation of chloromycetin detection card of the present invention；
Fig. 2 is the section of structure of chloromycetin detection card of the present invention.
In figure：1 detection card body, 2 handles, 3 standard color comparison cards, 4 base plates, lid, 6 wells, 7 detection zones, 8 samples on 5
Pad, 9 protecting film, 10 test strips, 11 adsorptive pads, 12 detection lines, 13 control lines.
Specific embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the invention, rather than the embodiment of whole.It is based on
Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under the premise of creative work is not made
Embodiment, belongs to the scope of protection of the invention.
Embodiment 1
A kind of chloromycetin detection method, comprises the following steps：
S1, take 5g samples and be put in pulverizer, sample can be the muscle of the animals such as chicken, duck, fish, cattle, pig, sheep, rabbit, fat,
Liver and kidney, add extracting solution and 10ml distilled water, containing one or two in ethanol and glycerol in extracting solution, ethanol and third
The concentration of triol is 64%, is taken out after crushing 2.2min；
S2, will crush completely after sample add centrifuge tube, add 2.3ml ethyl acetate, 3min be centrifuged with 2060r/min,
Upper solution is removed after standing 10min standby；
S3, remove upper solution sample add centrifuge tube in, add 0.5ml buffer, buffer be 0.05M PBS, with
3000r/min is centrifuged 7min, and using taking after 0.22um filtering with microporous membranes, filtrate is to be checked, and liquid to be detected is placed on 28 DEG C of ring
8.2h is stood under border.
Embodiment 2
A kind of chloromycetin detection method, comprises the following steps：
S1, take 7g samples and be put in pulverizer, sample can be the muscle of the animals such as chicken, duck, fish, cattle, pig, sheep, rabbit, fat,
Liver and kidney, add extracting solution and 12ml distilled water, containing one or two in ethanol and glycerol in extracting solution, ethanol and third
The concentration of triol is 66%, is taken out after crushing 2.6min；
S2, will crush completely after sample add centrifuge tube, add 3.5ml ethyl acetate, be centrifuged with 2180r/min
4.2min, removes upper solution after standing 12.5min standby；
S3, remove upper solution sample add centrifuge tube in, add 0.5ml buffer, buffer be 0.066M PBS, with
3400r/min is centrifuged 8min, and using taking after 0.22um filtering with microporous membranes, filtrate is to be checked, and liquid to be detected is placed on 28 DEG C of ring
8.8h is stood under border.
Embodiment 3
A kind of chloromycetin detection method, comprises the following steps：
S1, take 9g samples and be put in pulverizer, sample can be the muscle of the animals such as chicken, duck, fish, cattle, pig, sheep, rabbit, fat,
Liver and kidney, add extracting solution and 14ml distilled water, containing one or two in ethanol and glycerol in extracting solution, ethanol and third
The concentration of triol is 70%, is taken out after crushing 3min；
S2, will crush completely after sample add centrifuge tube, add 5ml ethyl acetate, 6.6min be centrifuged with 2300r/min,
Upper solution is removed after standing 14min standby；
S3, remove upper solution sample add centrifuge tube in, add 0.5ml buffer, buffer be 0.08M PBS, with
3800r/min is centrifuged 5min, and using taking after 0.22um filtering with microporous membranes, filtrate is to be checked, and liquid to be detected is placed on 28 DEG C of ring
9.4h. is stood under border
Details see attached list：
A kind of chloromycetin detection card, refers to Fig. 1-2, including detection card body 1, detects that one end of card body 1 is provided with handle
2, detection card is provided with standard color comparison card 3 away from one end of handle 2, and detection card body 1 includes base plate 4, is provided with base plate 4
Lid 5, offers well 6 and detection zone 7 on upper lid 5, sample pad 8, protecting film 9, reagent paper are provided between upper lid 5 and base plate 4
Bar 10 and adsorptive pads 11, are provided with detection line 12 and control line 13 in sample pad 8.
Chloromycetin detection card result judges：
It is negative（-）：T lines develop the color（Detection line, near well one end）C lines（Control line）Colour developing, represents medicine to be checked in sample
Concentration is less than test limit or does not contain medicine to be checked.
It is positive（+）：C lines develop the color, and T lines do not develop the color, and in representing sample, drug level to be checked is higher than test limit.
It is invalid：There are not C lines, it may be possible to which misoperation or agent plate fail.
It should be noted that herein, term " including ", "comprising" or its any other variant are intended to non-row
His property is included, so that a series of process, method, article or equipment including key elements not only include those key elements, and
And also include other key elements being not expressly set out, or also include for this process, method, article or equipment institute inherently
Key element.In the absence of more restrictions.By sentence " include one ... the key element for limiting, it is not excluded that including
Also there is other identical element in the process of the key element, method, article or equipment ".
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding can carry out various changes, modification, replacement to these embodiments without departing from the principles and spirit of the present invention
And modification, the scope of the present invention be defined by the appended.
Claims (8)
1. a kind of chloromycetin detection method, it is characterised in that：Comprise the following steps：
S1, take 5-10g samples and be put in pulverizer, add extracting solution and 10-15ml distilled water, take out after crushing 2-3min；
S2, will crush completely after sample add centrifuge tube, add 2-5ml ethyl acetate, be centrifuged with 2000-2300r/min
3-7min, removes upper solution after standing 10-15min standby；
S3, remove upper solution sample add centrifuge tube in, add 0.5ml buffer, with 3000-4000r/min be centrifuged
5-10min, it is to be checked using filtrate is taken after 0.22um filtering with microporous membranes.
2. a kind of chloromycetin detection method according to claim 1, it is characterised in that：The sample can for chicken, duck,
The muscle of the animals such as fish, cattle, pig, sheep, rabbit, fat, liver and kidney.
3. a kind of chloromycetin detection method according to claim 1, it is characterised in that：The liquid to be detected is placed on into 28
8-10h is stood in the environment of DEG C.
4. a kind of chloromycetin detection method according to claim 1, it is characterised in that：In the extracting solution containing ethanol and
One or two in glycerol, the concentration of the ethanol and glycerol is 60%-70%.
5. a kind of chloromycetin detection method according to claim 1, it is characterised in that：The buffer is 0.05-0.08M
PBS。
6. a kind of chloromycetin detection blocks, including detection card body（1）, it is characterised in that：The detection card body（1）One end set
It is equipped with handle（2）, the detection card is away from handle（2）One end be provided with standard color comparison card（3）, the detection card body（1）
Including base plate（4）, the base plate（4）On be provided with lid（5）, the upper lid（5）On offer well（6）And detection zone
（7）, the upper lid（5）With base plate（4）Between be provided with sample pad（8）, protecting film（9）, test strips（10）And adsorptive pads（11）,
The sample pad（8）On be provided with detection line（12）And control line（13）.
7. a kind of chloromycetin detection according to claim 6 blocks, it is characterised in that：The sample pad（8）, test strips
（10）, adsorptive pads（11）And protecting film（9）Base plate is pasted onto successively（4）On.
8. a kind of chloromycetin detection according to claim 6 blocks, it is characterised in that：Detection card preparation method, including with
Lower step：
A, test strips（10）On be coated with colloid gold label chloramphenicol resistance monoclonal；
B, detection line（12）For chloromycetin and the conjugate of BSA, nature controlling line is sheep anti-mouse igg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611114230.6A CN106546731A (en) | 2016-12-07 | 2016-12-07 | A kind of chloromycetin detection method and detection card |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611114230.6A CN106546731A (en) | 2016-12-07 | 2016-12-07 | A kind of chloromycetin detection method and detection card |
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| Publication Number | Publication Date |
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| CN106546731A true CN106546731A (en) | 2017-03-29 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201611114230.6A Pending CN106546731A (en) | 2016-12-07 | 2016-12-07 | A kind of chloromycetin detection method and detection card |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107727850A (en) * | 2017-10-10 | 2018-02-23 | 北京康华源科技发展有限公司 | A kind of lateral flow chromatography detection reaction starts control method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104515834A (en) * | 2014-09-20 | 2015-04-15 | 中山鼎晟生物科技有限公司 | Detection kit for chloramphenicol in cosmetics and detection method thereof |
-
2016
- 2016-12-07 CN CN201611114230.6A patent/CN106546731A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104515834A (en) * | 2014-09-20 | 2015-04-15 | 中山鼎晟生物科技有限公司 | Detection kit for chloramphenicol in cosmetics and detection method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107727850A (en) * | 2017-10-10 | 2018-02-23 | 北京康华源科技发展有限公司 | A kind of lateral flow chromatography detection reaction starts control method |
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Application publication date: 20170329 |
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