CN106404732A - Detection method for biotoxin in Islamic beef and mutton - Google Patents
Detection method for biotoxin in Islamic beef and mutton Download PDFInfo
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- CN106404732A CN106404732A CN201610782625.7A CN201610782625A CN106404732A CN 106404732 A CN106404732 A CN 106404732A CN 201610782625 A CN201610782625 A CN 201610782625A CN 106404732 A CN106404732 A CN 106404732A
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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"
Abstract
The invention discloses a detection method for biotoxin in Islamic beef and mutton. The detection method comprises the following steps: mincing c beef and mutton to obtain meat paste liquid, adding 5 ml of purified water, carrying out mixing under stirring, and then carrying out centrifugation at a rotation speed of 1000 rpm for 10 min; adding supernatant into an aseptic centrifuge tube, carrying out centrifugation at a rotation speed of 1000 rpm for 10 min and discarding obtained supernatant; subjecting a precipitate obtained in the previous step suspension with 540 [mu]L of aseptic peptone water so as to prepare a to-be-detected solution; mixing rhodamine B with hydrazine hydrate according to a mol ratio of 1: 5 to 1: 20 until a red color disappears, evaporating a solvent so as to obtain N-amino-rhodamine amide, then dissolving benzaldehyde equimolar to rhodamine B in ethanol, carrying out a reaction and then carrying out cooling to room temperature so as to obtain a detecting solution; adding the to-be-detected solution into the detecting solution, adding double distilled water to allow the volume of the mixed solution to reach 3 mL, carrying out color development for 15 min, then carrying out determination, and subjecting a reaction system to fluorescent scanning to determine biotoxin. According to the detection method provided by the invention, the detecting solution reacts with biotoxin in beef and mutton in a short period of time, so detection accuracy is improved, detection time is greatly shortened, and the demand for rapid detection of beef and mutton is met.
Description
Technical field
The invention belongs to technical field of food detection, it is related to a kind of food detection method, specifically a kind of Islamic beef and mutton
Biotoxin detection method.
Background technology
Biotoxin be by animal, plant, microorganism etc. produce under certain condition toxic to other biological species
Not reproducible chemical substance, is the important natural pollutant of a class., also known as natural toxin, species is various for biotoxin, point
Cloth is extensive. and the biotoxin of known chemical constitution has thousands of kinds, and basis source is divided into bacteriotoxin, fungi poison biotoxin
Element, Algae toxins, zootoxin, phytotoxin etc..Biotoxin can cause the acute poisoning of humans and animals, carcinogenic, teratogenesis or
Mutagenesis.
China is beef and mutton big producing country, and with the change of the consumption of meat structure of urban and rural residents, beef and mutton consumption figure is not
Disconnected raising, beef and mutton is consumed in shared proportion in consumption of meat structure and is totally also improving, the cattle and sheep sarcobiont of listing supply
Toxin controlling unit is weak, and the biotoxin pollution of beef and mutton is commonplace.
Fundamentally to solve this kind of food-safety problem, be necessary for the biotoxin of beef and mutton is detected, and mesh
Front traditional biotoxin detection method mainly includes high performance liquid chromatography, mass spectrum and immunological detection etc., mostly need according to
Bad antibody, testing equipment is expensive, complex operation and very time-consuming it is very difficult to meet quick to modern food safety detection technology, clever
Quick, easily require, at present, be badly in need of establishing a kind of quick, accurate, sensitive, easily novel detection method be used for biological poison
Element detection.
Content of the invention
It is an object of the invention to provide a kind of Islamic beef and mutton biotoxin detection method.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of Islamic beef and mutton biotoxin detection method, this detection method comprises the steps：
Prepare solution to be measured：Take 20g beef and mutton to rub and form meat gruel liquid, add purified water 5ml mix and blend, 1000rpm
Centrifugation 10min；Take supernatant in sterile centrifugation tube, 1000rpm is centrifuged 10min, abandons supernatant；The precipitation aseptic egg of 540 μ L
White peptone water suspends, and to be measured solution is obtained；
Preparation detection solution：Mol ratio is 1:5-1:20 rhodamine B is mixed with hydrazine hydrate, continues the condition at 80 DEG C
Under be heated to reflux 8-10 hour, until red disappear, evaporation solvent, obtain N- amino-rhodamine acid amides, then will be with rhodamine
The equimolar benzaldehyde of B is dissolved in ethanol, and under conditions of 100 DEG C, heating response 10 minutes, are cooled to room temperature after reaction, are examined
Survey solution；
Add solution to be measured in detection solution, supply 3mL with two double distilled waters, colour developing measured, to anti-after 15 minutes
Answer system to carry out fluorescent scanning and measure biotoxin.
Beneficial effects of the present invention：The present invention passes through to detect that solution is anti-in the short period of time with biotoxin in beef and mutton
Should, thus reaching the purpose of quick detection, improve the accuracy of detection, detection time is significantly shortened, meet to beef and mutton
Quick detection.
Specific embodiment
Below in conjunction with embodiment, technical scheme is clearly and completely described with the reality it is clear that described
Applying example is only a part of embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, this area is general
All other embodiment that logical technical staff is obtained under the premise of not making creative work, broadly falls into present invention protection
Scope.
Embodiment 1
A kind of Islamic beef and mutton biotoxin detection method, this detection method comprises the steps：
First, prepare solution to be measured：Take 20g beef and mutton to rub and form meat gruel liquid, add purified water 5ml mix and blend,
1000rpm is centrifuged 10min；Take supernatant in sterile centrifugation tube, 1000rpm is centrifuged 10min, abandons supernatant；Precipitate with 540 μ L
Sterile peptone aqueous suspension, is obtained solution to be measured；
Secondly, preparation detection solution：Mol ratio is 1:5 rhodamine B is mixed with hydrazine hydrate, continues the condition at 80 DEG C
Under be heated to reflux 10 hours, until red disappear, evaporation solvent, obtain N- amino-rhodamine acid amides, then will be with rhodamine B
Equimolar benzaldehyde is dissolved in ethanol, and under conditions of 100 DEG C, heating response 10 minutes, are cooled to room temperature after reaction, are examined
Survey solution；
Finally, detect, to 2ml (0.5 μm of ol), the solution to be measured adding 20mL, 40mL in solution, supplied with two double distilled waters
To 3mL, measure after developing the color 15 minutes, fluorescent scanning mensure carried out to serial reaction system, draws in excitation wavelength 365nm,
Launch wavelength is accordingly strong at 590nm.
Embodiment 2
A kind of Islamic beef and mutton biotoxin detection method, this detection method comprises the steps：
First, prepare solution to be measured：Take 15g beef and mutton to rub and form meat gruel liquid, add purified water 6ml mix and blend,
1500rpm is centrifuged 10min；Take supernatant in sterile centrifugation tube, 1000rpm is centrifuged 10min, abandons supernatant；Precipitate with 540 μ L
Sterile peptone aqueous suspension, is obtained solution to be measured；
Secondly, preparation detection solution：Mol ratio is 1:20 rhodamine B is mixed with hydrazine hydrate, continues the bar at 80 DEG C
It is heated to reflux under part 8 hours, until red disappear, evaporation solvent, obtain N- amino-rhodamine acid amides, then will be with rhodamine B
Equimolar benzaldehyde is dissolved in ethanol, and under conditions of 100 DEG C, heating response 10 minutes, are cooled to room temperature after reaction, are examined
Survey solution；
Finally, detect, to 2ml (0.5 μm of ol), the solution to be measured adding 80mL, 160mL in solution, mended with two double distilled waters
Foot arrives 3mL, and colour developing measures after 15 minutes, carries out fluorescent scanning mensure to serial reaction system, draw in excitation wavelength
365nm, launch wavelength is accordingly strong at 590nm.
Embodiment 3
A kind of Islamic beef and mutton biotoxin detection method, this detection method comprises the steps：
First, prepare solution to be measured：Take 15g beef and mutton to rub and form meat gruel liquid, add purified water 5ml mix and blend,
1000rpm is centrifuged 10min；Take supernatant in sterile centrifugation tube, 1000rpm is centrifuged 10min, abandons supernatant；Precipitate with 520 μ L
Sterile peptone aqueous suspension, is obtained solution to be measured；
Secondly, preparation detection solution：Mol ratio is 1:10 rhodamine B is mixed with hydrazine hydrate, continues the bar at 80 DEG C
It is heated to reflux under part 9 hours, until red disappear, evaporation solvent, obtain N- amino-rhodamine acid amides, then will be with rhodamine B
Equimolar benzaldehyde is dissolved in ethanol, and under conditions of 100 DEG C, heating response 10 minutes, are cooled to room temperature after reaction, are examined
Survey solution；
Finally, detect, to 2ml, the solution to be measured adding 320mL, 640mL in solution, supply 3mL with two double distilled waters,
Colour developing measured after 19 minutes, carries out fluorescent scanning mensure to serial reaction system, draws in excitation wavelength 365nm, transmitted wave
Long accordingly strong at 590nm.
Above content is only citing made for the present invention and illustrates, affiliated those skilled in the art are to being retouched
The specific embodiment stated is made various modifications or supplements or substituted using similar mode, without departing from invention or super
More scope defined in the claims, all should belong to protection scope of the present invention.
Claims (1)
1. a kind of Islamic beef and mutton biotoxin detection method is it is characterised in that this detection method comprises the steps：
Prepare solution to be measured：Take 20g beef and mutton to rub and form meat gruel liquid, add purified water 5ml mix and blend, 1000rpm is centrifuged
10min；Take supernatant in sterile centrifugation tube, 1000rpm is centrifuged 10min, abandons supernatant；Precipitate with 540 μ L sterile peptone
Aqueous suspension, is obtained solution to be measured；
Preparation detection solution：Mol ratio is 1:5-1:20 rhodamine B is mixed with hydrazine hydrate, continues to add under conditions of 80 DEG C
Heat backflow 8-10 hour, until red disappear, evaporation solvent, obtain N- amino-rhodamine acid amides, then will be with rhodamine B etc.
Mole benzaldehyde be dissolved in ethanol, under conditions of 100 DEG C, heating response 10 minutes, are cooled to room temperature after reaction, are detected
Solution；
Add solution to be measured in detection solution, supply 3mL with two double distilled waters, colour developing measured, to reactant after 15 minutes
System carries out fluorescent scanning and measures biotoxin.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610782625.7A CN106404732A (en) | 2016-08-30 | 2016-08-30 | Detection method for biotoxin in Islamic beef and mutton |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610782625.7A CN106404732A (en) | 2016-08-30 | 2016-08-30 | Detection method for biotoxin in Islamic beef and mutton |
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| CN106404732A true CN106404732A (en) | 2017-02-15 |
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| CN201610782625.7A Pending CN106404732A (en) | 2016-08-30 | 2016-08-30 | Detection method for biotoxin in Islamic beef and mutton |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101413872A (en) * | 2008-10-27 | 2009-04-22 | 吉林大学 | Method for rapidly detecting microorganism viable bacteria number |
| CN101943660A (en) * | 2010-08-13 | 2011-01-12 | 浙江大学 | Method for detecting psychrophilic bacteria in crude milk by utilizing aminopeptidase method |
| CN105385439A (en) * | 2015-12-01 | 2016-03-09 | 东华大学 | Reaction type Rhodamine-class fluorescence probe for mercury ion detection and preparation as well as application thereof |
-
2016
- 2016-08-30 CN CN201610782625.7A patent/CN106404732A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101413872A (en) * | 2008-10-27 | 2009-04-22 | 吉林大学 | Method for rapidly detecting microorganism viable bacteria number |
| CN101943660A (en) * | 2010-08-13 | 2011-01-12 | 浙江大学 | Method for detecting psychrophilic bacteria in crude milk by utilizing aminopeptidase method |
| CN105385439A (en) * | 2015-12-01 | 2016-03-09 | 东华大学 | Reaction type Rhodamine-class fluorescence probe for mercury ion detection and preparation as well as application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 杨博 等: "具有荧光增强性能的罗丹明B衍生物的合成及其对汞离子的检测", 《影响科学与光化学》 * |
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