CN103076319A - Method for detecting residual pesticidal toxicity in fruits and vegetables - Google Patents

Method for detecting residual pesticidal toxicity in fruits and vegetables Download PDF

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Publication number
CN103076319A
CN103076319A CN2012105814276A CN201210581427A CN103076319A CN 103076319 A CN103076319 A CN 103076319A CN 2012105814276 A CN2012105814276 A CN 2012105814276A CN 201210581427 A CN201210581427 A CN 201210581427A CN 103076319 A CN103076319 A CN 103076319A
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toxicity
fruits
slant medium
vegetables
luminous intensity
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CN103076319B (en
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黄山梅
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SGS-CSTC STANDARDS TECHNICAL SERVICES Co Ltd
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Abstract

The invention relates to the field of food safety, in particular to a method for detecting residual pesticidal toxicity in fruits and vegetables. The method for detecting the residual pesticidal toxicity in the fruits and the vegetables is characterized by comprising the following steps: preparing luminescent bacteria, preparing a detection sample, performing luminescent detection on the luminescent bacteria, mixing a fruit and vegetable sample to be detected with strain liquid of a vibrio qinghaiensis Q67 strain, and judging the residual pesticidal toxicity in the detected fruit and vegetable sample. By the method, the detection time of the residual pesticidal toxicity in the detected fruit and vegetable sample is shortened and the stable luminescent time of the vibrio qinghaiensis Q67 strain is prolonged.

Description

A kind of detection method of fruits and vegetables Pesticide Residues toxicity
Technical field
The present invention relates to the food security field, relate in particular to a kind of detection method of fruits and vegetables Pesticide Residues toxicity.
Background technology
The Detecting Pesticide of vegetables and fruit generally adopts vapor-phase chromatography, liquid phase chromatography and By Gas Chromatography-mass Spectrometry etc. at present, uses the chromatograph in laboratory, and analytical work needs the personnel of specialty to finish.The detection of pesticide residue toxicity, generally adopt at present spectrophotometric method, this method more complicated that operates, obtain analysis result and often need a few hours from the processing of sampling, sample, even time a couple of days, and the spectrophotometer complex structure, can't be applicable to fast qualitative detection of scene.Photobacteria is a class can be launched visible fluorescence under normal physiological condition bacterium, photobacteria is with after external tested material contacts, because poisonous substance has the light action of pressing down, the luminous intensity of photobacteria namely changes to some extent, the degree that changes and the toxicity size of tested material are correlationship, and the variation of luminous intensity can be measured with luminometer.But the stabilized illumination limited time of Qinghai Vibrion Q67 bacterial strain affects the detection of fruits and vegetables Pesticide Residues toxicity to a certain extent.
Summary of the invention
For above-mentioned technical matters, the present invention has designed and developed a kind of detection method of fruits and vegetables Pesticide Residues toxicity, and purpose is to shorten the detection time of fruits and vegetables Pesticide Residues toxicity, prolongs the stabilized illumination time of Qinghai Vibrion Q67 bacterial strain, improves accuracy in detection.
Technical scheme provided by the invention is:
A kind of detection method of fruits and vegetables Pesticide Residues toxicity may further comprise the steps:
Step 1, NaCl 2.48~4.06g, NaH 2PO 40.07~0.25g, MgSO 47H 2O 0.13~0.32g, MgCl 26H 2O 0.29~0.40g, CaCl 20.03~0.16g, NaHCO 32.27~4.19g, MgCO 30.34~0.62g, MgBr 20.01~0.23g, KCl 0.01~0.05g, yeast extract 3~6g, tryptone 3~6g, agar 18~25g, glycerine 2~7g is dissolved in the 1000mL distilled water, adjusts pH 4~5, make first generation slant medium, 121 ℃ of sterilization 20min, second generation slant medium adds Na again except the identical component of first generation slant medium 2HPO 40.07~0.25g adjusts 8~9,121 ℃ of sterilizations of pH 20min;
Step 2, the bacterial classification of Qinghai Vibrion Q67 bacterial strain moved receive on the above-mentioned first generation slant medium, 15~25 ℃ of lower 12~18h that cultivate, with physiological saline lawn is brushed down from first generation slant medium upper punch, be transferred on the second generation slant medium, the same terms is cultivated, and obtains test bacterium liquid;
Step 3, smash into fruits and vegetables to be measured to pieces juice, add distilled water and also stir, get wherein 10mL as testing sample, contrast select the national quality system certification without agricultural chemicals All Pure Nature fruits and vegetables, same method is processed, and gets wherein in contrast sample of 10mL;
Step 4, get testing sample and control sample 1mL respectively and place two cuvettes, add simultaneously 1mL concentration and be 1.2%~1.8% NaNO 3Solution adds the bacterium liquid on probation for the treatment of that makes in the 0.1mL step 2 again, places 30min, measures every glass luminous intensity with luminous detector, is calculated as follows relative luminous intensity
Relative luminous intensity=testing sample luminous intensity/control sample luminous intensity
If relative luminous intensity is between 90%~110%, then the pesticide residue toxicity of testing sample is safety; If relative luminous intensity is more than 110% or between 80%~90%, then the pesticide residue toxicity of testing sample is suspicious; If relative luminous intensity is below 80%, then the pesticide residue toxicity of testing sample is dangerous.
Preferably, in the detection method of described fruits and vegetables Pesticide Residues toxicity, NaH in the described step 1 first generation slant medium 2PO 4The pH that solution is adjusted first generation slant medium is 4.5, NaH in the second generation slant medium 2PO 4Solution and Na 2HPO 4The pH that the buffer solution of solution composition is adjusted second generation slant medium is 8.5.
Preferably, in the detection method of described fruits and vegetables Pesticide Residues toxicity, in the described step 2 bacterial classification of Qinghai Vibrion Q67 bacterial strain on two generation slant mediums all in 20 ℃ of lower 16h that cultivate.
Preferably, in the detection method of described fruits and vegetables Pesticide Residues toxicity, NaNO in the described step 4 3The concentration of solution is 1.5%.
Preferably, in the detection method of described fruits and vegetables Pesticide Residues toxicity, the physiological saline in the described step 2 is that concentration is 0.85% NaCl solution.
In the detection method of fruits and vegetables Pesticide Residues toxicity of the present invention, use NaH 2PO 4The pH that solution is adjusted first generation slant medium uses NaH 2PO 4Solution and Na 2HPO 4The buffer solution of solution composition is adjusted the pH of second generation slant medium, and Qinghai Vibrion Q67 bacterial strain is cultivated on two generation slant mediums in succession, and to add simultaneously concentration be 1.2%~1.8% NaNO detecting 3Solution has prolonged stabilized illumination time of Qinghai Vibrion Q67 bacterial strain, be conducive to fruits and vegetables Pesticide Residues toxicity fast, accurately detect.The Qinghai Vibrion Q67 bacterial strain that this method adopts all has sensitive reaction to all kinds of poisonous substances; even the new chemosynthesis poisonous substance that never occurred; as long as poisonous to biosome; be bound to photobacteria is exerted an influence; and reflect from the luminous power of photobacteria delicately, effectively avoid undetected.
Description of drawings
Fig. 1 is the schematic flow sheet of the detection method of fruits and vegetables Pesticide Residues toxicity of the present invention.
Embodiment
The present invention is described in further detail below in conjunction with accompanying drawing, can implement according to this with reference to the instructions literal to make those skilled in the art.
The invention provides a kind of detection method of fruits and vegetables Pesticide Residues toxicity, may further comprise the steps:
Step 1, NaCl 2.48~4.06g, NaH 2PO 40.07~0.25g, MgSO 47H 2O 0.13~0.32g, MgCl 26H 2O 0.29~0.40g, CaCl 20.03~0.16g, NaHCO 32.27~4.19g, MgCO 30.34~0.62g, MgBr 20.01~0.23g, KCl 0.01~0.05g, yeast extract 3~6g, tryptone 3~6g, agar 18~25g, glycerine 2~7g is dissolved in the 1000mL distilled water, adjusts pH 4~5, make first generation slant medium, 121 ℃ of sterilization 20min, second generation slant medium adds Na again except the identical component of first generation slant medium 2HPO 40.07~0.25g adjusts 8~9,121 ℃ of sterilizations of pH 20min;
Step 2, the bacterial classification of Qinghai Vibrion Q67 bacterial strain moved receive on the above-mentioned first generation slant medium, 15~25 ℃ of lower 12~18h that cultivate, with physiological saline lawn is brushed down from first generation slant medium upper punch, be transferred on the second generation slant medium, the same terms is cultivated, and obtains test bacterium liquid;
Step 3, smash into fruits and vegetables to be measured to pieces juice, add distilled water and also stir, get wherein 10mL as testing sample, contrast select the national quality system certification without agricultural chemicals All Pure Nature fruits and vegetables, same method is processed, and gets wherein in contrast sample of 10mL;
Step 4, get testing sample and control sample 1mL respectively and place two cuvettes, add simultaneously 1mL concentration and be 1.2%~1.8% NaNO 3Solution adds the bacterium liquid on probation for the treatment of that makes in the 0.1mL step 2 again, places 30min, measures every glass luminous intensity with luminous detector, is calculated as follows relative luminous intensity
If relative luminous intensity=testing sample luminous intensity/control sample luminous intensity relative luminous intensity is between 90%~110%, then the pesticide residue toxicity of testing sample is safety; If relative luminous intensity is more than 110% or between 80%~90%, then the pesticide residue toxicity of testing sample is suspicious; If relative luminous intensity is below 80%, then the pesticide residue toxicity of testing sample is dangerous.
In the detection method of described fruits and vegetables Pesticide Residues toxicity, NaH in the described step 1 first generation slant medium 2PO 4The pH that solution is adjusted first generation slant medium is 4.5, NaH in the second generation slant medium 2PO 4Solution and Na 2HPO 4The pH that the buffer solution of solution composition is adjusted second generation slant medium is 8.5.
In the detection method of described fruits and vegetables Pesticide Residues toxicity, in the described step 2 bacterial classification of Qinghai Vibrion Q67 bacterial strain on two generation slant mediums all in 20 ℃ of lower 16h that cultivate.
In the detection method of described fruits and vegetables Pesticide Residues toxicity, NaNO in the described step 4 3The concentration of solution is 1.5%.
In the detection method of described fruits and vegetables Pesticide Residues toxicity, the physiological saline in the described step 2 is that concentration is 0.85% NaCl solution.
Embodiment one
NaCl 3.67g, NaH 2PO 40.18g, MgSO 47H 2O 0.18g, MgCl 26H 2O 0.39g, CaCl 20.07g, NaHCO 33.41g, MgCO 30.53g, MgBr 20.07g, KCl 0.03g, yeast extract 5g, tryptone 5g, agar 20g, glycerine 3g is dissolved in the 1000mL distilled water, and adjusting pH is 4.5, make first generation slant medium, 121 ℃ of sterilization 20min, second generation slant medium adds Na again except the identical component of first generation slant medium 2HPO 40.18g adjusting pH is 8.5,121 ℃ of sterilization 20min; The bacterial classification of Qinghai Vibrion Q67 bacterial strain moved receive on the above-mentioned first generation slant medium; 20 ℃ of lower 16h that cultivate, the physiological saline with 0.85% brushes down lawn from the slant medium upper punch, then be transferred on the second generation slant medium; same method is cultivated, and obtains testing the bacterium liquid of usefulness.The common cucumber 10g that will buy from the market of farm produce smashs into juice to pieces, and adds 40mL distilled water, and centrifugal 5min gets supernatant for subsequent use under the rotating speed of 10000rpm; The contrast select the national quality system certification without agricultural chemicals All Pure Nature cucumber, process equally with said method, get supernatant for subsequent use.Get respectively cucumber sample supernatant to be measured and contrast cucumber sample supernatant 1mL places two cuvettes, add simultaneously 1mL concentration and be 1.5% NaNO 3Solution adds the bacterium liquid on probation for the treatment of that 0.1mL makes again, places 30min, and with the luminous intensity of every glass of luminous detector mensuration, the relative luminous intensity of cucumber sample to be measured is 115% in the present embodiment, and pesticide residue toxicity is dangerous.
Embodiment two
NaCl 3.15g, NaH 2PO 40.24g, MgSO 47H 2O 0.29g, MgCl 26H 2O 0.31g, CaCl 20.14g, NaHCO 32.72g, MgCO 30.47g, MgBr 20.19g, KCl 0.02g, yeast extract 4g, tryptone 4g, agar 23g, glycerine 5g is dissolved in the 1000mL distilled water, and adjusting pH is 4, make first generation slant medium, 121 ℃ of sterilization 20min, second generation slant medium adds Na again except the identical component of first generation slant medium 2HPO 40.24g adjusting pH is 8,121 ℃ of sterilization 20min; The bacterial classification of Qinghai Vibrion Q67 bacterial strain moved receive on the above-mentioned first generation slant medium; 23 ℃ of lower 15h that cultivate, the physiological saline with 0.85% brushes down lawn from the slant medium upper punch, then be transferred on the second generation slant medium; same method is cultivated, and obtains testing the bacterium liquid of usefulness.The common apple 10g that will buy from the market of farm produce smashs into juice to pieces, and adds 40mL distilled water, and centrifugal 5min gets supernatant for subsequent use under the rotating speed of 10000rpm; Processing equally with said method without agricultural chemicals All Pure Nature apple of national quality system certification selected in contrast, gets supernatant for subsequent use.Get respectively apple sample supernatant to be measured and contrast apple sample supernatant 1mL places two cuvettes, add simultaneously 1mL concentration and be 1.3% NaNO 3Solution adds the bacterium liquid on probation for the treatment of that 0.1mL makes again, places 30min, and with the luminous intensity of every glass of luminous detector mensuration, the relative luminous intensity of apple sample to be measured is 98% in the present embodiment, and pesticide residue toxicity is safety.
Although embodiment of the present invention are open as above, but it is not restricted to listed utilization in instructions and the embodiment, it can be applied to various suitable the field of the invention fully, for those skilled in the art, can easily realize other modification, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend of describing.

Claims (5)

1. the detection method of a fruits and vegetables Pesticide Residues toxicity is characterized in that, may further comprise the steps:
Step 1, NaCl 2.48~4.06g, NaH 2PO 40.07~0.25g, MgSO 47H 2O 0.13~0.32g, MgCl 26H 2O 0.29~0.40g, CaCl 20.03~0.16g, NaHCO 32.27~4.19g, MgCO 30.34~0.62g, MgBr 20.01~0.23g, KCl 0.01~0.05g, yeast extract 3~6g, tryptone 3~6g, agar 18~25g, glycerine 2~7g is dissolved in the 1000mL distilled water, adjusts pH 4~5, make first generation slant medium, 121 ℃ of sterilization 20min, second generation slant medium adds Na again except the identical component of first generation slant medium 2HPO 40.07~0.25g adjusts 8~9,121 ℃ of sterilizations of pH 20min;
Step 2, the bacterial classification of Qinghai Vibrion Q67 bacterial strain moved receive on the above-mentioned first generation slant medium, 15~25 ℃ of lower 12~18h that cultivate, with physiological saline lawn is brushed down from first generation slant medium upper punch, be transferred on the second generation slant medium, the same terms is cultivated, and obtains test bacterium liquid;
Step 3, smash into fruits and vegetables to be measured to pieces juice, add distilled water and also stir, get wherein 10mL as testing sample, contrast select the national quality system certification without agricultural chemicals All Pure Nature fruits and vegetables, same method is processed, and gets wherein in contrast sample of 10mL;
Step 4, get testing sample and control sample 1mL respectively and place two cuvettes, add simultaneously 1mL concentration and be 1.2%~1.8% NaNO 3Solution adds the bacterium liquid on probation for the treatment of that makes in the 0.1mL step 2 again, places 30min, measures every glass luminous intensity with luminous detector, is calculated as follows relative luminous intensity
Relative luminous intensity=testing sample luminous intensity/control sample luminous intensity
If relative luminous intensity is between 90%~110%, then the pesticide residue toxicity of testing sample is safety; If relative luminous intensity is more than 110% or between 80%~90%, then the pesticide residue toxicity of testing sample is suspicious; If relative luminous intensity is below 80%, then the pesticide residue toxicity of testing sample is dangerous.
2. the detection method of fruits and vegetables Pesticide Residues toxicity as claimed in claim 1 is characterized in that, NaH in the described step 1 first generation slant medium 2PO 4The pH that solution is adjusted first generation slant medium is 4.5, NaH in the second generation slant medium 2PO 4Solution and Na 2HPO 4The pH that the buffer solution of solution composition is adjusted second generation slant medium is 8.5.
3. the detection method of fruits and vegetables Pesticide Residues toxicity as claimed in claim 1 is characterized in that, in the described step 2 bacterial classification of Qinghai Vibrion Q67 bacterial strain on two generation slant mediums all in 20 ℃ of lower 16h that cultivate.
4. the detection method of fruits and vegetables Pesticide Residues toxicity as claimed in claim 1 is characterized in that, NaNO in the described step 4 3The concentration of solution is 1.5%.
5. the detection method of fruits and vegetables Pesticide Residues toxicity as claimed in claim 4 is characterized in that, the physiological saline in the described step 2 is that concentration is 0.85% NaCl solution.
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CN107045026A (en) * 2017-03-28 2017-08-15 中国科学院华南植物园 A kind of method that utilization Qinghai Vibrion q67 detects mycotoxin
CN109548536A (en) * 2018-10-19 2019-04-02 权冉(银川)科技有限公司 A kind of de- green information fruit bag for managing and pesticide capable of being monitored of multifunctional self-regulating set

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CN103389301A (en) * 2013-08-01 2013-11-13 辛焕发 Method for fast detecting pesticide residue
CN107045026A (en) * 2017-03-28 2017-08-15 中国科学院华南植物园 A kind of method that utilization Qinghai Vibrion q67 detects mycotoxin
CN109548536A (en) * 2018-10-19 2019-04-02 权冉(银川)科技有限公司 A kind of de- green information fruit bag for managing and pesticide capable of being monitored of multifunctional self-regulating set

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