Method for quantitatively detecting chloramphenicol by utilizing photobacterium leiognathus
A quantitative detection and chloramphenicol technology, which is applied in the determination/inspection of microorganisms, chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problems of environmental pollutant detection without luminescent bacteria
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[0009] Example 1
[0010] A single colony of P.leiognathi YL was inoculated in 150mL liquid medium (medium composition: peptone 5g, yeast extract 1g, FePO 4 0.1g, NaCl 20g, glycerol 3ml, distilled water 1000ml), 26°C, 150r / m shaker for about 6h, until the luminescence value reaches 2.0×10 5 -4.0×10 5 When the concentration of chloramphenicol is 0, 0.1ng / mL, 0.2ng / mL, 0.4ng / mL, 0.6ng / mL, 0.8ng / mL, and 1.0ng / mL, respectively, the bacterial solution was transferred to the test tube. 5mL of chloramphenicol and 5mL of bacterial solution. After mixing evenly, incubate at 26°C, 150r / m shaker for 30min, and measure the luminescence value. Draw a standard curve for the detection of chloramphenicol ( figure 1 )
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[0011] Example 2
[0012] fish pretreatment method
[0013] (1) Take out the fish viscera, peel off the fish skin, and homogenize it. Divide 20.0g of fish paste per bag into small packaging bags and set aside.
[0014] (2) Thaw the frozen fish, weigh 5.0g into a 50mL centrifuge tube, and add 50uL of standard CAP solution (concentration of 10ng / mL) to the centrifuge tube, which is equivalent to the concentration of chloramphenicol in the fish after adding the standard to 0.1ng / g, do three parallel groups.
[0015] (3) Mix the samples in each centrifuge tube for 1 min.
[0016] (4) Add 20 mL of ethyl acetate to each centrifuge tube, mix for 1 min, sonicate for 10 min, and centrifuge for 5 min (4000 r / m).
[0017] (5) Pour the supernatant into a chicken heart flask, and rotate to dryness in a water bath at 40°C.
[0018] (6) Add 1 mL of methanol and vortex to dissolve the residue.
[0019] (7) Add 15 mL of n-hexane and mix evenly, then add 25 mL of 4% NaCl solution, shake ...
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[0024] Example 3
[0025] Thaw the frozen fish meat, weigh 5.0 g into a 50 mL centrifuge tube, and add 50 μL, 100 μL, 200 μL, 30 μL, 400 μL, and 500 μL of standard CAP solution (concentration: 10 ng / mL) to the centrifuge tube in turn. The concentrations of chloramphenicol were 0.1ng / g, 0.2ng / g, 0.4ng / g, 0.6ng / g, 0.8ng / g, 1.0ng / g, and six parallel groups were made for each concentration.
[0026] After adding chloramphenicol solution to the sample, pre-treatment was carried out according to Example 2.
[0027] A single colony of P. leiognathi YL was inoculated into 150 mL of liquid medium, and cultured in a shaker. The parameters of the shaker were set to 26°C, 150 r / m. The luminescence intensity of the bacterial solution in P.leiognathi YL was at 3.0×10 5 When left and right, add 5 mL of bacterial solution to each test tube containing 5 mL of chloramphenicol solution recovered by standard addition, and make a set of standard solution blank control for each treatment method, ...
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