Sandfly, Phlebotomus papatasi Scopoli was considered one of the dangerous medical insects that transmits a lot of diseases such as Leishmaniasis disease around the world. In Egypt, synthetic pyrethroids and organophosphorous insecticides were the main chemicals control methods for P. papatasi. So, the samples of adults and 4th larvae were collected from two different localities (10th of Ramadan city and Al- Quareen city), in addition, lab. strain. The common insecticides used synthetic pyrethroids (deltamethrin and lambda-cyhalothrin) and organophosphorous (chlorpyrifos and malathion) and diagnosed this tolerance biochemically. Respecting the biochemical diagnoses, electrophoresis protein separation utilizing sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE), some target and detoxifying enzymes acetylcholinesterase (AChE), general esterases (GES), cytochrome-P450 (C-450), glutathione-S-transferase (GST)} were characterized. Also, (amylase, invertase, alkaline phosphatase, acid phosphatase and total lipids) were determined. The obtained results showed that the 4th larvae of P. papatasi collected from the 10th of Ramadan locality was the most tolerant to cholorpyrifos (2.79- fold), deltamethrin (2.21 - fold), malathion (1.76- fold) and lambda-cyhalothrin (1.16- fold) compared with the laboratory strain, while in adults the most tolerant was lambda-cyhalothrin (2.81-fold), cholorpyrifos (2.16-fold), deltamethrin (1.65-fold) and malathion (1.43). It was observed that there was a significant difference between the insect populations, the presence of different proteins bands in 10th of Ramadan population tolerant to these insecticides, followed by Al-Quareen city population compared with laboratory strain. Regarding the target and metabolic enzymes, data found that there was a highly significant increase in these metabolic enzymes activities (GES, P-450 and GST) in 10th of Ramadan locality, followed by Al- Quareen locality compared with laboratory strain in adults and larvae, while, other biochemical constituents, Total lipids, trehalase, alkaline phosphatase, acid phosphatase, amylase, invertase activities and their roles in energies for larvae and adults of P. papatasi showed a reduction. Generally, the results showed that there was a highly significant reduction in most of these energies enzymes. So, the current study recommended avoiding intensive insecticides used and randomly used, which led to increasing insecticides tolerance/ insecticides development in these the dangerous medical pests transmitting the epidemic diseases and human disaster.