IOSR Journal of Applied Chemistry (IOSR-JAC) e-ISSN: 2278-5736.Volume 14, Issue 3 Ser. I (March. 2021), PP 31-38 www.iosrjournals.org DOI: 10.9790/5736-1403013138 www.iosrjournals.org 31 |Page GC-FID without derivatization method for the quantitative determination of Drug-Facilitated Crimes (Diazepam) in Bulk, Pharmaceuticals, urine, blood, biscuits and beverages. Moftah A. Moustafa 1,† , Saadeldin E. Taher 1 , Awatef A. Massoud 1 and Ali A. Khaled 2 1 Chemistry Department, Faculty of Science, Tobruk University, Tobruk, Libya 2 Chemistry Department, Faculty of Education, Tobruk University, Tobruk, Libya † Corresponding author: Moftah A. Moustafa Abstract Diazepam (DZ) drug-facilitated crime is frequently encountered in clinical and forensic toxicology cases due to misuse or abuse. This paper reports simple, sensitive, selective, reliable, rapid and expeditious GC–FID method for the quantitative analysis of diazepam (DZ) drug (1,4-benzodiazepines) in pure form, pharmaceutical preparations, urine, blood, biscuits and beverages, 1μg/ml clonazepam (CZ) as internal standard (I.S.) has been developed for different forensic toxicology laboratories. The chromatographic system of GC–FID was carried out without derivatization using a GC- 17AGas Chromatograph SHIMDZU in conjunction with a column (DP – 5, length: 30 meter and ID: 0.25 ml) was validated. The proposed method has been validated as per US-FDA bioanalytical guidelines in terms of linearity, accuracy, precision, matrix effects, stability, selectivity, and recovery. The method was linear over the concentration range of 0.8‒1000 μg ml −1 with limits of detection ad quantification of 0.0308 and 0.128 μg ml −1 , respectively. The intraday and interday precisions and accuracy expressed by the relative standard deviation and the relative standard error were both less than 6.12 % and 1.8 %, respectively. The proposed method was successfully applied in pure form, pharmaceutical preparations, urine, blood, biscuits and beverages, 100 μg ml -1 clonazepam (CZ) as internal standard (I.S.) has been developed for different forensic toxicology laboratories. The stability of DZ in the presence of all endogenous applications components was studied and the new GC–FID method was successfully employed. Keywords: Diazepam; Clonazepam; GC–FID; Extraction; Stability; Urine, Blood, Biscuits and Beverages. --------------------------------------------------------------------------------------------------------------------------------------- Date of Submission: 04-03-2021 Date of Acceptance: 18-03-2021 --------------------------------------------------------------------------------------------------------------------------------------- I. Introduction Benzodiazepines identification and confirmation their presences in human body are important in emergency rooms and toxicology laboratory; various reliable and specific techniques are required (1). Benzodiazepines remain the primary targets in clinical and forensic urine drug testing as it's related to suicide attempts, road traffic offenses, drug facilitated sexual assault, sudden deaths, car accidents, rapes, burglaries and robbery (2–5). Diazepam (1,4-benzodiazepines) is one of the most frequently observed compound in cases of drug-facilitated crime (DFC) and drug-facilitated sexual assaults (DFSA) (6–10). This is due to Diazepam (D Z) has become the most commonly used drugs for their anti-convulsant, anaesthetic, anti-depressive, hypnotic, tranquilizer and sedative properties. They are also used as pre-medication and for induction or general anaesthesia and are widely prescribed throughout the world (11–14). Apart from their therapeutic applications, benzodiazepines are often abused by drug addicts; as a consequence, these drugs are frequently involved in both clinical and forensic cases (15). Diazepam (DZ) and clonazepam (CZ) as internal standard (I.S.) are considered the most important 1,4-benzodiazepines (Figure 1) where diazepam (7-chloro-1,3-dihydro-1-methyl-5-phenyl- 2H- 1,4-benzodiazepin-2-one) drug enhances the activity of gamma-aminobutyric acid, the most common inhibitory neurotransmitter in the central nervous system which an abused drug in which sudden withdrawal, particularly from high dosage, carries the risk of epileptic seizures that used in the treatment of severe anxiety disorders, as a hypnotic in the short-term management of insomnia, as a sedative and premedicant, as an anticonvulsant, in the management of alcohol withdrawal syndrome (16–19). Hepatic N-demethylation results in the formation of an active metabolite, desmethyldiazepam (nordiazepam). This metabolite is hydroxylated to form oxazepam, which is also active and is metabolised to form oxazepam glucuronide. Aminor active metabolite is temazepam, which can be in turn demethylated to oxazepam or glucuronidated (20). Various analytical methods have been described for the determination of diazepam (DZ) in pharmaceutical preparations,