I want to quantify OP pesticide contents in animal tissues using HPLC-UV technique.
Please suggest suitable protocol(s) of the sample preparation for the above purpose with references.
with thanks
prem
No. The levels required (sub ppb or ppt) necessitate the use of a GC. See the United States EPA and USDA methods.
Bruce Neagle is absolutely right, GC/NPD are the top of the top, but maybe you don't need such analytical power, which concentration range are you going to investigate?
Sorry Bruce and Andrea, y'all are both really, really wrong. HPLC-MS/MS is by far the most sensitive technique for o-P pesticides. However, HPLC-UV simply won't work. You'll get way too many interferences extracted from the animal tissue unless you go through extensive and exhaustive cleanup procedures. Move to HPLC-MS/MS and use conventional extraction and cleanup procedures. You'll need a relatively polar column (we use the Restek Ultra Aqueous) to get the front end pesticides separated, and it will help with the remainder of the o-P pests.
Hello Mark, I'm glad you got better sensitivity by GC/MS. I'm not sure what detector Bruce was referring to... He did not mentioned it, I think.
Hello Rajak,
I agree with my colleagues that you will probably need a more sensitive detector than UV (e.g. QqQ). Regarding sample preparation, the Quecher method could be a good choice for animal tissue. In the following links you can find some examples:
-https://link.springer.com/article/10.1007/s11356-014-3361-2
- https://www.researchgate.net/publication/264831846_Determination_of_Organophosphorus_Pesticides_in_Stomach_Contents_of_Postmortem_Animals_by_QuEChERS_and_Gas_Chromatography
-http://www.waters.com/webassets/cms/library/docs/720004456en.pdf
Article Determination of Organophosphorus Pesticides in Stomach Cont...
I would assume that you want to analyze OP with the instrument you have. I agree, the ideal instrument is either GC/MS or LC/MS, but if you don't have it then it is no use. You can ride a bike from India to China, but it will take a while. Depending on the level and I hope that it is high enough. You have to do a lot of sample cleanup to get rid of interference that have similar UV absorbance characteristics as your OP. I found a good SPE MIP that will trap only OP and release other stuff. You may use QuEChERS extraction and cleanup first for simplicity. However, it is too dirty for UV, so you need the SPE MIP to make the extract cleaner.
Good luck
I do agree with all Colleagues,
HPLC-UV wouldn't work in your case. It is recommended to move to either GC or GC/MS.
regards
mufeed
Article Multiresidue Analysis of Pesticides in Soil Profile from the...
Thanks everyone for their valuable comments.
I too understand that, it is difficult to estimate OP contents in animal tissues using HPLC-UV, as there would be several other components which may have similar absorbance at a particular wave length. Clearing is an issue. Therefore, it may produce erroneous output. After going through comments and several papers, I think GC-MS would be more sensitive and a better choice to estimate OP contents at ppt or sub-ppt levels than HPLC-UV.
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