Method for preparing fast detecting test paper of illegal cooking oil
A technology for detecting test strips and waste oil. It is used in measurement devices, instruments, scientific instruments, etc. It can solve the problems of cumbersome, requiring very professional personnel to operate, not suitable for rapid on-site screening, and prone to false positives. Simple, reduced detection cost, and high sensitivity
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Embodiment 1
[0016] Example 1 Preparation of immunochromatic latex microspheres.
[0017] 1. Pretreatment of colored latex microspheres.
[0018] Take red, yellow, blue, and green four-color latex microsphere solutions with a diameter of about 100nm, uniform particle size, and a coefficient of variation <10%. After 5 minutes of ultrasonic treatment at 80Hz, adjust the concentration to 1% with ultrapure water, 10000g ×10min centrifugation, the precipitate was washed twice with 25mM, pH6.0 MES buffer for later use. Use pre-cooled 25mM, pH6.0 MES buffer solution to prepare EDC and NHS into 50mg / ml solution for later use, add 165μl DEC solution and 165μl NHS solution per ml latex, add EDC and NHS to the washed latex . Mix slowly at room temperature for 30 min; centrifuge at 10,000 g × 10 min after incubation, and wash the precipitate twice with an equal volume of 25 mM, pH 6.0 MES buffer.
[0019] 2. Antibody labeling.
[0020] Dilute antibody A (mouse anti-aflatoxin monoclonal ant...
Embodiment 2
[0021] Example 2 Performance evaluation of color latex microsphere immunochromatographic detection test strip.
[0022] 1. Determination of the minimum detection limit of the test strip.
[0023] At room temperature, prepare aflatoxin B1 standard, benzopyrene standard, anisidine standard and DBS standard into solutions with a series of concentrations (1-243ng / ml), and add 100 μl to the microwells of the microtiter plate , use waste oil color latex microsphere immunochromatography test strips to analyze in turn, repeat 3 times; through visual observation, the minimum standard concentration when the color of the test line of the test strip is obviously lighter than that of the negative control strip, is The detection limit of the test strip is determined by naked eyes; the detection limit of aflatoxin is 7ppb, the detection limit of benzopyrene is 9ppb, the detection limit of anisidine is 9ppb, and the detection limit of DBS is 5ppb.
[0024] The results are shown in Table 1:...
Embodiment 3
[0030] Example 3 Detection of samples and judgment of results.
[0031] 1. Processing of samples to be tested.
[0032] use figure 2 The shown device 1 draws the edible oil sample, squeezes it gently, squeezes out 2 drops of the oil sample into the No. 1 tube of the device 3, and uses the device 2a to absorb the redistilled petroleum ether in the No. 2 tube of the device 3 and squeezes out 4 drops Put it into tube No. 1, use device 1 to blow and mix the liquid in tube 1, then use device 2a to suck up the 80% methanol water (v / v) solution of tube 3 in device 3 and squeeze out 4 drops to tube 1 In the same way as above, mix evenly, let it stand until the liquid is clearly separated, use the device 2a to carefully absorb the upper liquid and discard it, use the device 2b to absorb the pure water from the No. 4 tube of the device 3, and squeeze out 16 drops to the No. 1 tube. Mixed to be tested.
[0033] 2. Sample testing.
[0034] Insert the end of a test strip with the ma...
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