CN106770102A - A kind of streptomysin detection method - Google Patents
A kind of streptomysin detection method Download PDFInfo
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- CN106770102A CN106770102A CN201611103945.1A CN201611103945A CN106770102A CN 106770102 A CN106770102 A CN 106770102A CN 201611103945 A CN201611103945 A CN 201611103945A CN 106770102 A CN106770102 A CN 106770102A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
Abstract
The invention discloses a kind of streptomysin detection method, comprise the following steps：（1）Sample is extracted, mechanical shaking extraction, centrifugation is successively carried out in three times to sample with the acetonitrile solution of the various concentrations containing acid, merges No. three extract solutions, obtain sample extracting solution；（2）Sample extracting solution is saltoutd, grease removal, add methyl alcohol and 0.2% ~ 0.5% ammoniacal liquor to be saltoutd in the sample extracting solution, be centrifuged, n-hexane is added in gained supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic solvent obtained solution A, alkali soluble is in water obtained solution B.Streptomysin detection method of the present invention has the advantages that easy to operate, quick, need not be complicated mechanical sensing equipment.
Description
Technical field
The present invention relates to a kind of detection method, specifically a kind of streptomysin detection method.
Background technology
Streptomysin (Streptomycin) is a kind of important aminoglycoside antibiotics, is widely used in Animal diseases
Treatment, streptomysin to chicken, sheep, ox, pig all kinds of inflammation, such as infectious coryza of chicken, pig acute bronchitis, pig dysentery characterized by white mucous stool, milk
Bovine endometritis etc. have good curative effect, therefore through being used frequently as feed addictive.If incorrect use, chain can be caused
Mycin residual is exceeded.Research has shown that streptomysin can cause irreversible loss to kidney and auditory nerve, and long-term use can also make
Bacterium produces drug resistance.In order to ensure the safety of animal derived food, No. 235 file of the Ministry of Agriculture《Animal food herbal medicine
MRL》Middle regulation streptomysin is 200 μ g/L in milk, is 600 μ g/kg in muscle, fat, liver, in kidney
It is 1000 μ g/kg.The detection of current Determination of Streptomycin Residues mainly has Instrumental Analysis and immunological method.Wherein instrumental method is main
With high performance liquid chromatography (HPLC), gas chromatography (GC), these methods have that sensitivity is high, result is accurate, it is reproducible,
The advantages of false positive is few, but certain shortcoming is still suffered from, such as sample pretreatment process is complicated, and instrumentation degree is high and expensive,
Analyze speed is slow, is not suitable for carrying out extensive Site Detection etc..
The content of the invention
It is an object of the invention to provide a kind of streptomysin detection method, with asking for solving to propose in above-mentioned background technology
Topic.
To achieve the above object, the present invention provides following technical scheme：
A kind of streptomysin detection method, comprises the following steps：（1）Sample is extracted, successively with the various concentrations containing acid
Acetonitrile-aqueous solution carries out mechanical shaking extraction, centrifugation to sample in three times, merges No. three extract solutions, obtains sample extracting solution；（2）To sample
Product extract solution is saltoutd, grease removal, adds methyl alcohol and 0.2% ~ 0.5% ammoniacal liquor to be saltoutd in the sample extracting solution, is centrifuged,
N-hexane is added in gained supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic
Solvent obtained solution A, alkali soluble is in water obtained solution B；（3）Solution A and solution B are mixed with the standard liquid of various concentrations respectively
Close, vibrate, water layer is pipetted after stratification, measure fluorescence intensity, set up related bent between fluorescence intensity and concentration of standard solution
Line；（4）Solution A and solution B are mixed with testing sample solution, are vibrated, water layer is pipetted after stratification, measure fluorescence intensity,
With step（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.
As further scheme of the invention：The Chemical Luminescent Probes are 9,10- phenanthrenequione.
As further scheme of the invention：In the step（2）In, the organic solvent is immiscible with water and close
Degree is bigger than water.
As further scheme of the invention：The acetonitrile-aqueous solution concentration of the various concentrations containing acid is 5% ~ 15%.
As further scheme of the invention：The volume ratio of the solution A, solution B and standard liquid and the solution
A, solution B are identical with the volume ratio of the testing sample solution.
Compared with prior art, the beneficial effects of the invention are as follows：Streptomysin detection method of the present invention has easy to operate, fast
The advantages of mechanical sensing equipment that is fast, not needing complexity.
Specific embodiment
The technical scheme in the embodiment of the present invention is clearly and completely described below.
In the embodiment of the present invention, a kind of streptomysin detection method comprises the following steps：（1）Sample is extracted, successively
Carry out mechanical shaking extraction, centrifugation in three times to sample with the acetonitrile-aqueous solution of the various concentrations containing acid, merge No. three extract solutions, obtain
Sample extracting solution；（2）Sample extracting solution is saltoutd, grease removal, in the sample extracting solution add methyl alcohol and 0.2% ~ 0.5%
Ammoniacal liquor is saltoutd, centrifugation, and n-hexane is added in gained supernatant, and vortex vibration, grease removal obtains standard liquid；（2）To change
Learn fluorescence probe and be dissolved in organic solvent obtained solution A, alkali soluble is in water obtained solution B；（3）By solution A and solution B respectively with difference
The standard liquid mixing of concentration, vibration, pipette water layer after stratification, measure fluorescence intensity, set up fluorescence intensity molten with standard
Correlation curve between liquid concentration；（4）Solution A and solution B are mixed with testing sample solution, is vibrated, water is pipetted after stratification
Layer, measures fluorescence intensity, with step（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.Describedization
Fluorescence probe is 9,10- phenanthrenequione.In the step（2）In, the organic solvent is immiscible with water, and density ratio water is big.
The acetonitrile-aqueous solution concentration of the various concentrations containing acid is 5% ~ 15%.The volume ratio of the solution A, solution B and standard liquid
Volume ratio with the solution A, solution B and the testing sample solution is identical.
Embodiment 1
Streptomysin detection method of the present invention, comprises the following steps：（1）Sample is extracted, successively with the various concentrations containing acid
Acetonitrile-aqueous solution sample is carried out in three times mechanical shaking extraction, centrifugation, merge No. three extract solutions, obtain sample extracting solution；（2）It is right
Sample extracting solution is saltoutd, grease removal, adds methyl alcohol and 0.2%% ammoniacal liquor to be saltoutd in the sample extracting solution, is centrifuged,
N-hexane is added in gained supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic molten
Agent obtained solution A, alkali soluble is in water obtained solution B；（3）Solution A and solution B are mixed with the standard liquid of various concentrations respectively,
Vibration, pipettes water layer after stratification, measure fluorescence intensity, sets up the correlation curve between fluorescence intensity and concentration of standard solution；
（4）Solution A and solution B are mixed with testing sample solution, is vibrated, water layer is pipetted after stratification, fluorescence intensity is measured, with step
Suddenly（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.The Chemical Luminescent Probes are 9,10- luxuriant and rich with fragrance
Quinone.In the step（2）In, the organic solvent is immiscible with water, and density ratio water is big.The various concentrations containing acid
Acetonitrile-aqueous solution concentration is 5%.The volume ratio of the solution A, solution B and standard liquid and the solution A, solution B and described
The volume ratio of testing sample solution is identical.
Embodiment 2
Streptomysin detection method of the present invention, comprises the following steps：（1）Sample is extracted, successively with the various concentrations containing acid
Acetonitrile-aqueous solution sample is carried out in three times mechanical shaking extraction, centrifugation, merge No. three extract solutions, obtain sample extracting solution；（2）It is right
Sample extracting solution is saltoutd, grease removal, adds methyl alcohol and 0.3% ammoniacal liquor to be saltoutd in the sample extracting solution, is centrifuged,
N-hexane is added in gained supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic molten
Agent obtained solution A, alkali soluble is in water obtained solution B；（3）Solution A and solution B are mixed with the standard liquid of various concentrations respectively,
Vibration, pipettes water layer after stratification, measure fluorescence intensity, sets up the correlation curve between fluorescence intensity and concentration of standard solution；
（4）Solution A and solution B are mixed with testing sample solution, is vibrated, water layer is pipetted after stratification, fluorescence intensity is measured, with step
Suddenly（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.The Chemical Luminescent Probes are 9,10- luxuriant and rich with fragrance
Quinone.In the step（2）In, the organic solvent is immiscible with water, and density ratio water is big.The various concentrations containing acid
Acetonitrile-aqueous solution concentration is 10%.The volume ratio of the solution A, solution B and standard liquid and the solution A, solution B and described
The volume ratio of testing sample solution is identical.
Embodiment 3
Streptomysin detection method of the present invention, comprises the following steps：（1）Sample is extracted, successively with the various concentrations containing acid
Acetonitrile-aqueous solution sample is carried out in three times mechanical shaking extraction, centrifugation, merge No. three extract solutions, obtain sample extracting solution；（2）It is right
Sample extracting solution is saltoutd, grease removal, adds methyl alcohol and 5% ammoniacal liquor to be saltoutd in the sample extracting solution, is centrifuged, in institute
N-hexane is added in supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic solvent
Obtained solution A, alkali soluble is in water obtained solution B；（3）Solution A and solution B are mixed with the standard liquid of various concentrations respectively, is shaken
Swing, water layer is pipetted after stratification, measure fluorescence intensity, set up the correlation curve between fluorescence intensity and concentration of standard solution；（4）
Solution A and solution B are mixed with testing sample solution, is vibrated, water layer is pipetted after stratification, fluorescence intensity is measured, with step
（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.The Chemical Luminescent Probes are 9,10- luxuriant and rich with fragrance
Quinone.In the step（2）In, the organic solvent is immiscible with water, and density ratio water is big.The various concentrations containing acid
Acetonitrile-aqueous solution concentration is 15%.The volume ratio of the solution A, solution B and standard liquid and the solution A, solution B and described
The volume ratio of testing sample solution is identical.
Embodiment 4
Streptomysin detection method of the present invention, comprises the following steps：（1）Sample is extracted, successively with the various concentrations containing acid
Acetonitrile-aqueous solution sample is carried out in three times mechanical shaking extraction, centrifugation, merge No. three extract solutions, obtain sample extracting solution；（2）It is right
Sample extracting solution is saltoutd, grease removal, adds methyl alcohol and 4% ammoniacal liquor to be saltoutd in the sample extracting solution, is centrifuged, in institute
N-hexane is added in supernatant, vortex vibration, grease removal obtains standard liquid；（2）Chemical Luminescent Probes are dissolved in organic solvent
Obtained solution A, alkali soluble is in water obtained solution B；（3）Solution A and solution B are mixed with the standard liquid of various concentrations respectively, is shaken
Swing, water layer is pipetted after stratification, measure fluorescence intensity, set up the correlation curve between fluorescence intensity and concentration of standard solution；（4）
Solution A and solution B are mixed with testing sample solution, is vibrated, water layer is pipetted after stratification, fluorescence intensity is measured, with step
（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.The Chemical Luminescent Probes are 9,10- luxuriant and rich with fragrance
Quinone.In the step（2）In, the organic solvent is immiscible with water, and density ratio water is big.The various concentrations containing acid
Acetonitrile-aqueous solution concentration is 12%.The volume ratio of the solution A, solution B and standard liquid and the solution A, solution B and described
The volume ratio of testing sample solution is identical.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be in other specific forms realized.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit requires to be limited rather than described above, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.Moreover, it will be appreciated that although the present specification is described in terms of embodiments, not each
Implementation method only includes an independent technical scheme, and this narrating mode of specification is only this area for clarity
Specification an as entirety, the technical scheme in each embodiment should can also be formed this by technical staff through appropriately combined
Art personnel may be appreciated other embodiment.
Claims (5)
1. a kind of streptomysin detection method, it is characterised in that comprise the following steps：（1）Sample is extracted, successively with containing acid
Various concentrations acetonitrile-aqueous solution sample is carried out in three times mechanical shaking extraction, centrifugation, merge No. three extract solutions, obtain sample and carry
Take liquid；（2）Sample extracting solution is saltoutd, grease removal, add methyl alcohol and 0.2% ~ 0.5% ammoniacal liquor to enter in the sample extracting solution
Row is saltoutd, and is centrifuged, and n-hexane is added in gained supernatant, and vortex vibration, grease removal obtains standard liquid；（2）By chemiluminescence
Probe is dissolved in organic solvent obtained solution A, and alkali soluble is in water obtained solution B；（3）By solution A and solution B respectively with various concentrations
Standard liquid mixing, vibration, pipette water layer after stratification, measure fluorescence intensity, set up fluorescence intensity and concentration of standard solution
Between correlation curve；（4）Solution A and solution B are mixed with testing sample solution, is vibrated, water layer is pipetted after stratification, measured
Fluorescence intensity, with step（3）The curve set up is compared, and calculates the streptomycin sulphate content of sample.
2. streptomysin detection method according to claim 1, it is characterised in that the Chemical Luminescent Probes are 9,10-
Phenanthrenequione.
3. streptomysin detection method according to claim 1, it is characterised in that in the step（2）In, it is described organic molten
Agent is immiscible with water, and density ratio water is big.
4. streptomysin detection method according to claim 1, it is characterised in that the acetonitrile of the various concentrations containing acid-
Concentration of aqueous solution is 5% ~ 15%.
5. streptomysin detection method according to claim 1, it is characterised in that the solution A, solution B and standard liquid
Volume ratio it is identical with the volume ratio of the solution A, solution B and the testing sample solution.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023615A2 (en) * | 1998-10-15 | 2000-04-27 | Bioimage A/S | Method for extracting quantitative information relating to an influence on a cellular response |
| CN104535664A (en) * | 2014-03-31 | 2015-04-22 | 中华人民共和国北京出入境检验检疫局 | Method for simultaneously detecting a plurality of mycotoxins in sesame paste |
| CN104568925A (en) * | 2014-12-30 | 2015-04-29 | 中国农业科学院农产品加工研究所 | Streptomycin sulfate detection method |
| US20160282357A1 (en) * | 2012-07-11 | 2016-09-29 | Nubad, LLC | Methods and compositions related to nucleic acid binding assays |
-
2016
- 2016-12-05 CN CN201611103945.1A patent/CN106770102A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000023615A2 (en) * | 1998-10-15 | 2000-04-27 | Bioimage A/S | Method for extracting quantitative information relating to an influence on a cellular response |
| US20160282357A1 (en) * | 2012-07-11 | 2016-09-29 | Nubad, LLC | Methods and compositions related to nucleic acid binding assays |
| CN104535664A (en) * | 2014-03-31 | 2015-04-22 | 中华人民共和国北京出入境检验检疫局 | Method for simultaneously detecting a plurality of mycotoxins in sesame paste |
| CN104568925A (en) * | 2014-12-30 | 2015-04-29 | 中国农业科学院农产品加工研究所 | Streptomycin sulfate detection method |
Non-Patent Citations (3)
| Title |
|---|
| 朱葆佺: "《新编药物合成反应路线图设计与制备工艺新技术》", 31 March 2005, 天津电子出版社 * |
| 林荆,张金虎等: "高效液相色谱法测定水产品中氟苯尼考和甲苯咪唑的残留量", 《农药》 * |
| 黄雅丽: "固相萃取-高效液相色谱-串联质谱法测定禽畜和水产品中沃尼妙林和泰妙菌素", 《分析实验室》 * |
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