Instructions for organophosphorus pesticide residue kit
Organophosphorus Pesticide Residue Assay kit
(300 T)
This kit allows for the in vitro semi-quantitative determination of organophosphorus pesticide residue in fruit and vegetable.
Expiration date six months from the date of manufacture
FOR RESEARCH USE ONLY.
The kit determine organophosphorus pesticide residues in fruit and vegetable samples according to the change of acetylcholinesterase activity (organophosphorus pesticide will inhibit the activity of acetylcholinesterase). The result is calculated as inhibition rate.
REAGENT PREPARATION
1. Extract Buffer: Dilute the two small bags of Extract Powder with 1000mL distilled water, shake and dissolved. (The two small bags of Extract Powder are packed together in a bigger plastic bag) Store it at room temperature.
2. Enzyme Diluent: Add 3.1ml Extract Buffer to the bottle labeled "Enzyme Solution" and mix well. It can be stored at 4 ºC for 7 days. (Do not freeze it)
3. Substrate Diluent: Add 3.1ml distilled water to the bottle labeled "Substrate Solution" and mix well. It can be stored at 4 ºC for 3 days or at -20ºC for 15 days.
4. Chromogenic Diluent: Add 10.5ml Extract Buffer to the labeled "Chromogenic Solution" and mix well. Store it at 4 ºC for 10 days
ASSAY PROCEDURE
1. Sample extraction: Take 2g fruit and vegetable samples cutting into about 1 cm 2. Add 10 ml Extract Buffer, oscillating for 2min for extraction. Sta-nd for 3-5 minutes, then take 2.5ml supernatant as Sample to test.
2. Control test: Mix 100 μl Enzyme Diluent and 2.5 ml Extract Buffer well. Then add 100 μl Chromogenic Diluent and 100 μl Substrate Diluent respectively. Test immediately after mix well. (Spectrophotometer, set at 412nm) Record the change of the OD in 3min --- Δ K0
3. Sample test: Mix 100 μl Enzyme Diluent and 2.5 ml Sample well. Sta-nd for 10min. Then add 100 μl Chromogenic Diluent an-d100 μl Substrate Diluent respectively. Test immediately after mix well. (Spectrophotometer, set at 412nm) Record the change of the OD in 3min --- Δ Kt
twenty three
CALCULATION
Δ K0 -Δ Kt Δ K0 Y ---- Inhibition rate IF Y> 50% ---- Positive IF 40%
MATERIALS PROVIDED
Reagent
Quantity
Extract Powder
3
Enzyme Solution
10
Substrate Solution
10
Chromogenic Solution
3
Capable for testing 300 samples
DETECTION LIMIT
Name
Concentration (mg / kg)
Dichlorvos
0.1
Parathion
1.0
Phoxim
0.3
Methamidophos
2.0
Malathion
4.0
Dimethoate
3.0
Omethoate
0.8
Isofenphos methyl
5.0
Methomyl
0.1
Carbosulfan
0.05
Trichlorfon
0.2
Carbofura
0.05
TECHNICAL HINTS
1. The assay is recommended to operate under room temperature (20ºC-40ºC).
2. Store the Enzyme Diluent and Substrate Diluent at 4ºC right after use. Dont put them at room temperature for long time. There may be a little white precipitate in Chromogenic Diluent. It wont influence the assay.
3. Proper mixing and heating will help to dissolve Extract Powder.
4. Leek, ginger, onion, garlic, peppers, carrots and other vegetables contain the substance which can destroy enzyme activity. Therefore, for such samples, do not cut them into too small pieces, or extracting for too long time.
5. Once the reagent is removed out of the bottle, strictly forbidden to put the reagent back into the reagent bottle, even if it does not run out.
4
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