Determination of the abused intravenously self-administered madness drops (Tropicamide) by liquid chromatography in rat plasma; an application to pharmacokinetic study and greenness profile assessment

•Two novel chromatographic methods developed for determination of the TPC abuse in rat plasma.•This is the first article related to TPC abuse for detecting traces of the studied drug in rat plasma.•Pharmacokinetic behaviors of tropicamide have been studied for the first time.•Greenness assessment of the proposed methods was investigated and evaluated.•These relied on the simulation of the behavior of abusers for further use in toxicological field. Tropicamide (TPC) is an antimuscarinic ophthalmic solution. Recently, it is reported to be used intravenously for recreational purposes. Misuse of the drug may leads to several effects such as slurred speech, unconsciousness, hallucinations, dysphoria, suicidal feelings, convulsions, and tachycardia. Two rapid and sensitive green chromatographic methods were newly established for detection and quantification of TPC abuse in rat plasma. The proposed TLC-densitometric method relied on the separation of TPC and the internal standard paracetamol (PCM) using a developed system of ethyl acetate: ethanol (9:1, v/v) on TLC silica gel 60 F254 plates and detection was performed at 210.0 nm. Linearity was achieved over a concentration range of 0.20–5.00 µg/band for TPC using 0.50 µg/band of PCM. On the other hand, the developed RP-HPLC method was based on the separation of TPC and sofosbuvir (SOF) as an internal standard using a mobile phase consisting of ethanol and water in the ratio of (45: 55, v/v) on ZORBAX Eclipse Plus® C18 column (250 × 4.6 mm, 5 µm) at a flow rate of 1.8 mL/ min and UV detection at 210.0 nm. Linearity was obtained over the concentration range of 0.10–30.00 µg/mL for TPC using 10.00 µg/mL of sofosbuvir (SOF) as an internal standard. Different evaluation tools assessed and measured the greenness profile of the established methods. On the validation of developed methods FDA recommendation was followed and all the results obtained agreed with acceptance criteria. The suggested methods were successfully used to study the pharmacokinetic parameters of TPC in rat plasma. This is the first article related to TPC abuse for determining the traces of the studied drug in rat plasma. The developed methods can be used for further monitoring of TPC abuse in addicted people plasma.