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generated using the peak area ratio counts of methamphetamine and internal standard versus the theoretical calibrator concentrations, and r2 > 0.9981 for all curves. The limit of quantitation was administratively set to 100 ng/mL. Six sets of controls were included with each analytical batch: Limit of Detection QC (100 ng/mL), Low QC (150 ng/mL), Mid QC (600 ng/mL), High QC (1500 ng/mL), a blank, and a double blank. Intraday precision and bias were determined by the largest percent coefficient of variation (% CV) and by the largest percent difference of the five runs (N=5). Carryover was assessed by injecting the lowest quality control (150 ng/mL) following the injection of the high-quality control (1500 ng/mL). (Figure 5). A method for the quantitation of methadone was validated using an Agilent 6890N Gas Chromatograph coupled to a 5973 Mass Selective Detector per SWGTOX guidelines. Chromatographic separation was performed on a HP-5MS column 30 m x 0.25 mm id x 0.25 mm (Agilent, Santa Clara, CA). The GC/MS was operated in a split mode at a ratio of 20:1, with 1 µL injection volume. The carrier gas was helium at a linear velocity of 20 mL/min. The GC had a front inlet temperature of 275 °C. The temperature program began with the oven temperature of 225 °C, and then programmed at a rate of 15 °C/min to 285 °C. The MSD was operated in SIM mode with 72, 223, 294, and 309 m/z as the selected ions for methadone and 78, 226, 303, and 318 m/z for methadone-d9. A seven-point calibration curve ranging from 100-5000 ng/mL of methadone was prepared along with a blank, a double blank control, and controls containing methadone and its internal standard were analyzed. The controls were run in triplicate with Limit of Detection/Quantitation control at 100 ng/mL; a low-control at 150 ng/mL; a mid-control at 1000 ng/mL; and a high-control at 4500 ng/mL. The limit of detection was arbitrarily set to match the limit of quantitation at 100 ng/mL. An internal standard (500 ng/mL methadone-d9) was 8 This resource was prepared by the author(s) using Federal funds provided by the U.S. Department of Justice. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. added to each calibrator, blank, controls, and sample. The coefficient of determination (r2 ) was 0.9985 for all calibration curves. The bias for all the controls (n=3) for methadone ranged from 5- 15% with coefficient of variance (% CV) of 20%. Carryover was assessed by injecting the lowest quality control following the injection of the high-quality control. Experiment 4. Develop a dynamic headspace GCMS (DHS GCMS) method for the direct analysis of e-cigarette components. Challenge: The DHS GC-MS had significant equipment failures over many months. A method for headspace analysis used in the analysis of fire debris evidence was concept tested to analyze atomizers on e-cigarettes. It was demonstrated that this method could be adopted for nicotine analysis. However, this method did not work for the analysis of methamphetamine. Static headspace with the charcoal strip used in nicotine analysis was evaluated a number of times with a number of extraction time periods and temperature parameters for methamphetamine analysis. Methamphetamine was demonstrated to aerosolize; however, it was not extracted from the aerosol onto the charcoal test strip or it was not possible to extract it from the charcoal test strip. The conclusion is that the charcoal test strip was not suitable for some drugs. Aim 2 Characterize representative electronic cigarettes available for consumers to purchase, to include efficacy of drug delivery. Experiment 1. Characterize major classes of e-cigarette devices available for consumers to purchase and describe the function of each component. Goal: The three major types of e-cigarettes were characterized (Figure 6-9). In order to understand the relationship between voltage, resistance, heat, and concentration of drugs in the aerosol, coil temperatures were assessed at various voltages and various gauges. Method: Two popular types of wire used within the e-cigarette community are Kanthal A-1 and Nichrome 80:20. Coils can be either contact or non-contact. Contact coils are tightly wrapped and the wire touches each wrap. Non-contact coils do not touch and typically have the wraps evenly spaced apart from one another. The temperature of the coils for the dry burns and wet burns when the coil is wrapped to the same resistance and burned at the same voltages were assessed. Both metal types will be evaluated at the same resistance (1.8 Ω), wet and dry, contact and noncontact, with wire gauge ranging from 26 to 36 and at 3.5, 3.7, 4.2, 4.5, 5.0, 5.5, and 6.0 volts, with replicates of at