[ APPLICATION NOTE ] ANALYSIS OF DRUGS OF ABUSE USING XBRIDGE SHIELD RP18 TEST CONDITIONS: COMPOUNDS: Column: XBridge™ Shield RP18 , 4.6 X 100 mm, 3.5 μm Part Number 186003044 0 0 Mobile Phase A: H2O Mobile Phase B: ACN 0 / 0 )/ Mobile Phase C: 100 mM NH4HCO3, pH 9.6 ) Flow Rate: 0.6 mL/min Gradient: Time Profile 1. MDMA 2. MDA (min) %A %B %C 0.0 63 32 5 0 3.0 63 32 5 0 )/ )/ 7.0 45 50 5 0) 9.0 45 50 5 / / 10.0 63 32 5 #S $M $M 12.0 63 32 5 / Injection Volume: 10 μL Sample Concentration 4. Lorazepam and Diluent: 10 μg/mL in H2O 3. Bromazepam Temperature: 40 ˚C Detection: UV @ 210 Sampling Rate: 5 points/second 0 0 )/ / Time Constant: 1.0 Needle Wash: 5/95 MeOH/H2O / / Instrument: Alliance® 2695 with 2996 PDA 0/ $M 0/ ' System Suitability Parameters 5. Clonazepam 6. Flunitrazepam Retention USP Tailing Width at USP Time (min) Factor 4.4% Resolution Benzodiazepines, such as clonazepam, bromazepam, lorazepam and MDA 3.32 0.98 0.181 flunitrazepam are a class of drugs with sedative, hypnotic, anxiolytic, MDMA 3.66 1.21 0.190 2.14 anticonvulsant, amnestic and muscle relaxant properties. They are considered minor tranquilizers and have a high potential for abuse. MDA Bromazepam 5.16 1.05 0.223 8.28 is the parent drug of MDMA, also known as Ecstasy. They are commonly Lorazepam 8.13 0.97 0.209 16.68 abused, amphetamine-derived recreational drugs. Clonazepam 8.60 0.97 0.209 2.81 Flunitrazepam 9.14 0.98 0.208 3.28 6 0.15 45 0.10 3 1 2 AU 0.05 Compounds: 1. MDA 2. MDMA 0.00 3. Bromazepam 0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 4. Lorazepam 5. Clonazepam Minutes 6. Flunitrazepam [ APPLICATION NOTE ] Sales Offices: India and India Subcontinent Austria and European Export 91 80 2837 1900 (Central South Eastern Europe, CIS Ireland 353 1 448 1500 and Middle East) 43 1 877 18 07 Italy 39 02 27 421 1 Australia 61 2 9933 1777 Japan 81 3 3471 7191 Belgium 32 2 726 1000 Korea 82 2 820 2700 Brazil 55 11 5094-3788 Mexico 52 55 5200 1860 Canada 1 800 252 4752 x2205 The Netherlands 31 76 508 7200 China 86 10 8586 8899 Norway 47 6 384 60 50 CIS/Russia +7 495 3367000 Poland 48 22 833 4400 Czech Republic 420 2 617 1 1384 Puerto Rico 1 787 747 8445 Denmark 45 46 59 8080 Singapore 65 6273 1221 Finland 09 5659 6288 Spain 34 93 600 9300 France 33 1 30 48 72 00 Sweden 46 8 555 11 500 Germany 49 6196 400600 Switzerland 41 56 676 70 00 Hong Kong 852 29 64 1800 Taiwan 886 2 2543 1898 Hungary 36 1 350 5086 United Kingdom 44 208 238 6100 Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. Waters, XBridge, Alliance and The Science of What’s Possible are trade- T: 1 508 478 2000 marks of Waters Corporation. All other trademarks are acknowledged. ©2007 Waters Corporation. Printed in the U.S.A. F: 1 508 872 1990 June 2007 WA60202 MC-PDF www.waters.com.