There are many literature for extracting pesticides from tissues and then analyzing on HPLC, but each of them describe different ways for separation of the pesticides from animal tissues. Kindly give some most suitable and easily applicable method, starting from extraction to analysis of pesticide.
The methods that I know for pyrethroids and pesticides in animal tissues are going in GC-ECD with sample preparations based on MSPD o QueChERS, and can obtain LoD < 5ug/Kg, but if you don´t have GC-ECD and you don´t need a very low LoD, try to development a method based on NP-HPLC, using a -NO2 column with hexane + 0,25% 2-propanol as mobile phase and for the sample preparation:
5g of homogenized sample
15mL acetonitrile
10min of Shaken
10min of sonicated
add MgSO4/CH3COONa (4:1) both anhydrous salts, mixed
Repeat the shaken stage
Centrifugate 10 min and recuperate the solvent
Evaporate to dryness
and reconstituted with 1mL of the mobile phase
Try and tell me if you worked
Best Regards
well my first answer is . don't use HPLC for that purpose, it does not make sense. If you reall really have to Lucianos approach might work, but I am nearly 100% certain, that you will neither be able to controll environmental nor food safety issues.
If you want to work with HPLC, you might also want to remove lipids, size exclusion chroamtography is usually a good idea.
Thank you Bester. I appreciate your suggestion. Actually I wanted to measure the pesticide levels in fish which I have collected from local ponds situated in agricultural fields. These fishes are regularly consumed by farmers and sold in nearby rural and urban markets. I welcome if you could provide a different and more appropriate methods for measuring the tissue pesticide levels. Kindly give a link or attachment for the protocol. Thanks.
well there is a can do nearly all approach.
1) vigorous extraction either by soxhlet, aceton/DCM/water partitioning of a slurry or ASE
2) Size exclusion chromatography to remove lipids and co-extracted proteins
3) Silica microcolumn clean up
4) GC-MS or GC-ECD or GC-AFID (N/P-selective detector
the approach is used all over Europe and the US for controlling pesticde residues in food.
I have not invented this approach, but we are using it for multiple purposes.
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