Objective: This study aims to develop and validate the analytical method to determine 3,4-Methylenedioxy-N-ethylamphetamine (MDEA) in DBS using GC-MS. Methods: This research used liquid-liquid micro-extraction for sample preparation and analysis was performed by GC-MS. In the method development, the optimized parameters were flow rate, column temperature, the spot of blood volume, % haematocrit, extraction and reconstitution of solvent volume, and sonication duration. Validation of the chosen method was performed based on EMEA bioanalytical guideline in 2011. Results: The optimum chromatographic conditions were obtained using HP-5 MS capillary columns (30 m x 0.25 mm i.d; 0.25 μm ); helium with 99.9% purity as a mobile phase; flow rate of 1.0 ml/min; column temperature was 250 °C; MS detection using 4 fragments at m/z values ​​of 72.00 and 44.00 for MDEA and 58.00 and 77.00 for ephedrine HCl as an internal standard. The DBS paper with the volume of blood spot 40 μl was then extracted using liquid-liquid micro-extraction with methanol 700 μl, sonication for 5 min, evaporated with nitrogen gas then reconstituted with 50 μl ethyl acetate. The validation results fulfilled the requirements based on the EMEA bioanalytical guideline in 2011. Conclusion: It can be concluded that the optimum condition of the analytical method by using GC-MS was obtained and fulfilled validation criteria with a range concentration of 15-250 ng/ml.

Objective: This research was carried out to obtain the MDMA bioanalysis method in DBS using liquid-liquid micro-extraction method using gas chromatography mass spectrometry (GC-MS) with the standard in Ephedrine. Methods: The blood was spotted on DBS paper and dried prior to analysis. The instrument used is Shimadzu GCMS-QP2010 Ultra Mass Chromatography Gas, with an HP-5 MS capillary column length of 30 m, 0.25 mm diameter and a film thickness of 0.25 μm. The mobile phase used is 99.99% Helium gas and split less injection mode. Results: The optimum chromatography condition for MDMA analysis in DBS is the HP-5 MS capillary column with a length of 30 m; inner diameter 0.25 mm; Helium mobile phase 99.999%; flow rate of 0.8 mL / minute; MS detection at m / z 58.00 and 135.00 and ephedrine HCl as internal standard. Sample preparation used a liquid-liquid micro-extraction method with methanol as solvent then it was sonicated or 15 minutes then dried with Nitrogen gas and reconstituted with 40 μL of ethyl acetate then centrifuged for 5 minutes with 3000 rpm. Conclusion: The developed method for determination of MDMA in DBS fulfilled validation based on EMEA bioanalytical guidelines in 2011 for linearity, LLOQ limits, accuracy and precision, selectivity, carry over, matrix effect and stability. The method obtained was linear in the concentration range 25.0-500.0 ng / mL with r> 0.999.

Methamphetamine or known as sabu is one of the amphetamine groups that give a very strong central nervous system stimulant effect. Nowadays, the method used to test the specimens to prove a person who had methamphetamine, for example is urine and blood. This research was aimed to develop methamphetamine analysis method in Dried Blood Spot (DBS). This research used non-invasive bio sampling method namely DBS using Gas Chromatography-Mass Spectrometry (GC-MS) because it is suitable for compounds that are stable to high temperatures and present in small levels in the body. This research started from optimization of gas chromatography condition-optimization mass spectrometry, optimization of sample preparation method from DBS, until validation of bio analysis method. The optimum chromatographic conditions were MS-5 capillary DB columns with a length of 30 m; 0.25 mm inner diameter; Helium gas phase 99.999%; 1.0 mL / min flow rate; detection of MS at m / z value 58.00; 91.00; and 77,00 with ephedrine HCl as an internal standard. Sample preparation using liquid-liquid micro-extraction method (LLM) using methanol solvent then residue dried under the flow of N2 gas and reconstituted with 50 μL ethyl acetate. The method obtained was linear in the concentration range 1.750-35.000 ng / mL with r > 0.9800. (y =-0.0001 + 0.0004x withr = 0.9987). The validation results of the methamphetamine analysis method performed met the validation requirements based on the EMEA Bio analytical Guideline of 2011. This method can be applied in the detection and analysis of amphetamine in drug abuse.