Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

1.0 Purpose

1.1 This procedure describes collection of data required to verify the purity and potency of reference materials and the concentrations of standard and control solutions prior to use in the Harris County Institute of Forensic Sciences’ Toxicology Laboratory

2.0 Scope

2.1 This procedure is applicable to the verification of externally obtained or internally prepared reference materials, standard solutions, and control solutions used in conjunction with analytical procedures performed by the Toxicology Laboratory.

2.2 The procedures may also be applied to revalidation ofCopy expired Reference Materials and solutions.

2.3 Reference materials and solutions are acceptable for use if a positive match is obtained to published or library MS or UV spectra and greater than 90 percent purity can be demonstrated by UV or chromatographic methods

3.0 Definitions and Abbreviations

3.1 No method-specific or non-standard terms are used in this procedure.

4.0 Materials

4.1 Equipment

4.1.1 Agilent 8453 UV- Visible Spectrophotometer

4.2 Reagents

4.2.1 Refer to individual analytical procedures

Uncontrolled4.3 Stock Standards

4.3.1 Refer to individual analytical procedures.

4.4 Working Standards and Controls

4.4.1 Refer to individual analytical procedures

5.0 Procedure

Page 1 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

5.1 Verification of concentration can be performed on newly prepared and existing solutions.

5.2 Most reference materials are supplied as acid or base salts while solution concentrations are stated as mg/L or mg/L of the free drug. If a solution has been prepared previously, the appropriate conversion factor can be obtained from the Toxicology Drug Inventory Log. To confirm the calculation or to calculate the amount required for new drugs apply Formula 1. This weighing factor (F) will never be less than 1.0.

5.2.1 Formula 1:

Molecular weight of drug as supplied = Weighing factor (F) Molecular weight of free base (acid) drugCopy

5.2.2 Weight required (generally 2.5 or 10.0 mg) = 2.5 (or 10.0) mg X F

5.3 Preparation of solutions:

5.3.1 Prepare solutions of sodium or other alkaline metal salts in deionized water. 5.3.2 Prepare solutions of all other drugs in methanol or acetonitrile unless otherwise instructed. 5.3.3 To prepare working solutions: A. From 10.0 mg equivalents of drug. a. Dissolve the weighed amount of drug into 10.0 mL of solvent to obtain a 1.0 mg/mL stock solution. b. To prepare 0.1 and 0.01 mg/mL working solutions perform serial 1:10 dilutions into the same solvent (dilution 1 = 0.1 mg/mL, dilution 2 = 0.01 mg/mL). c. Alternatively, prepare the 0.01 mg/mL working solution by diluting 0.10 mL of the 1.0 mg/mL stock solution to 10.0 mL. B. From 2.5 mg equivalents of drug. Uncontrolleda. 0.1 mg/mL: Dissolve the weighed amount of drug into 25.0 mL of the preferred solvent. b. 0.01 mg/mL: Transfer 1.0 mL of the 0.1 mg/mL solution to a 10.0 mL flask and dilute to 10.0 mL with the preferred solvent.

5.4 Perform necessary maintenance

5.4.1 Refer to TOX08.5029 procedure

5.5 Purity and strength verification:

Page 2 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

5.5.1 Ultraviolet Spectrometry A. Determine the ultraviolet spectra using the 0.01 mg/mL solution. If the expected absorbance is less than 100 use the 0.1 mg/mL solution. B. Record the absorbance at the wavelength specified in the published or supplier reference. C. Calculate the equivalent absorbance for a 1 g % w/v solution (1 g/dL).

To convert from To convert to Multiply absorbance by 0.10 mg/mL 1.0 g / dL (1g%) 100 0.01 mg/mL 1.0 g / dL (1g%) 1000

D. Compare the corrected absorbance to literature reference values or to values listed in Table 1. Copy E. Solutions are accepted if the determined absorbance is within the range from 90 to 110 % of target. F. When the absorbance is outside the acceptable range: a. If the 0.01 mg/mL dilution was tested, repeat the test on a fresh dilution. b. If the absorbance remains outside the acceptable range reweigh the test material and prepare new solutions. c. If the 0.10 mg/mL solution was initially tested, reweigh the sample. G. If absorbance is within specifications accept the solutions for analytical purposes. H. Substances that do not have significant ultraviolet absorbance may be checked by GC/MS, GC/MS/MS, LC/MS/MS, or HPLC.

6.0 Data Analysis / Interpretation/ Documentation

6.1 Place the UV spectra into the Reference Material Validation Log.

7.0 Acceptance Criteria

Uncontrolled7.1 Described in Section 5.0.

8.0 References

8.1 The Merck Index, Thirteenth Edition, M.J. O’Neil, Senior Editor. 2001.

8.2 Clarke’s Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Post-Mortem Materials. 2nd Edition, A.C. Moffat Editor. 1986.

8.3 Bexar County Forensic Toxicology Laboratory, UV Absorbance Data, In-house Page 3 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

modifications.

8.4 Normative reference: ABFT E 13

9.0 Revision History

Revision Description of Change Reviewed By Date 0 Original Document T. Danielson 11/02 1 Reformatted, deleted GC/MS and HPLC verification I. Bermudez 0907 2 2008 annual review C. Young 0808 3 Name changed to Institute of Forensic Sciences T. Cao 0410 4 Added maintenance reference CopyA. Beard 1212 5 Updated final approver and added raised change A. Beard 1213

10.0 Attachment(s) or Appendices

10.1 Table 1. Specific absorbances for UV-Vis comparison

Clarke's Values Bexar County Medical Examiner's Office Acid Acid Methanol* Analyte Sa nm sa nm sa Nm N-Acetylprocainamide HCI Xx xx 633 267 779 270 Albuterol Sulfate Xx xx 69 276 81 278 Alprazolam Xx 260 473 264 1346 222 Amantadine HCI Nus nus nus nus nus Nus Amitriptyline HCI 504a 239 512 240 502 240 Amoxapine 362a 252 374 252 394 252 Amphetamine HCI 14a 257 12 258 11 258 Antipyrine xx xx 575 230 476 244 AtenololUncontrolled 48b 274 45 274 51 276 Atropine 6.3c 258 6 258 6 258 Benztropine Mesylate 14.5a 259 14 260 15 260 Brompheniramine Maleate 272a 265 265 265 244 262 Bupropion HCI xx xx 59 298 56 296 Bupropion Mtb HCI xx xx 12 268 11 268 Caffeine Alkaloid 504a 273 469 273 480 272 Carbamazepine 490a 285 491 285 489 285 Carbinoxamine Maleate 323a 263 346 263 190 262 Chlordiazepoxide HCI 1112a 246 1110 246 1089 264

Page 4 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

Chloroquine Diphosphate 600b 329 534 256 500 257 Chlorpheniramine Maleate 302a 265 302 266 202 262 Chlorpromazine HCI 1025a 255 1040 255 1105 258 Cinnarizine 584a 254 20 292 39 284 Citalopram HBr xx xx 55 272 55 272 Demethycitalopram HCI xx xx 59 272 58 272 Clomipramine HCI 253a 251 242 251 256 252 Clonazepam 645b 273 645b 273 462 245 Clozapine xx xx 729 242 345 296 Cocaine HCI 430a 233 433 233 466 230 Codeine Alkaloid 55a 285 52 285 54 284 Conessine xx xx nus nus nus nus Cyclobenzaprine HCI 407b 290 385 Copy290 396 290 Desipramine HCI 308a 250 313 252 367 252 Desmethyldoxepin HCI xx xx 138 . 292 171 296 N-Desmethylsertraline Maleate xx xx insol. nus 41 266 Dextromethorphan HBr 70a 278 70 278 78 280 Diazepam 1020a 242 995 242 1177 230 Dicyclomine HCI xx xx nus nus nus nus Diethylpropion HCI 673a 253 617 253 598 248 Diltiazem HCI xx xx 550 238 609 242 Diphenhydramine HCI 17a 257 18 258 16 260 Dipyrone Sodium 266a 258 266 258 236 266 Disopyramide Phosphate 199a 269 185 269 112 262 Doxapram HCI 12c 259 11 260 11 262 Doxepin HCI 131a 292 135 294 156 296 Doxylamine Succinate 335a 261 319 262 158 262 (-)Ephedrine HCl 10.3 257 10 257 10 258 Fenfluramine HCI 22a 264 24 264 25 264 Fentanyl Citrate 13a 257 13 257 12 258 Flecainide Acetate xx xx 52 296 74 298 Fluoxetine HCI xx xx 425 228 421 228 Flurazepam 2HCI 503 236 503 236 734 232 FluvoxamineUncontrolled Maleate xx xx 352 247 372 256 Hydrocodone Bitartrate 41a 280 43 280 46 284 Hemihydrate Hydromorphone HCI 50a 280 49 280 49 284 Hydroxychloroquine Sulfate 560a 235 612 235 511 256 Hydroxyzine HCI 416a 232 442 232 385 230 Imipramine HCI 298a 251 276 251 290 252 Ketamine HCI 25a 269 27 269 23 270 Laudanosine xx xx 159 280 172 282 Lidocaine HCI 19a 263 19 263 16 264 Page 5 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

Lorazepam xx xx 732 232 1205 230 Loxapine Succinate 346b 251 346 251 393 252 3,4-MDA HCI 216b 233 213 233 214 238 3,4-MDMA HCI xx xx 179 236 201 238 Meclizine HCI 2H20 xx xx 33 266 27 266 Meperidine HCI 8.5a 257 9 256 8 258 Mesoridazine Mesylate 773b 262 723 262 1080 262 Methadone HCI 18a 292 17 294 15 294 Methadone Mtb HI, EDDP xx xx nd nd nus nus (+) Methamphetamine HCI 12.1a 257 11 257 11 258 Methylphenidate HCI 9a 257 10 258 7 258 Metoclopramide HCI 467b 273 365 274 532 276 Metoprolol Tartrate 52a 274 52 Copy274 54 276 Mexiletine HCI 14a 260 14 262 14 262 Midazolam xx xx 363 260 879 224 Mirtazapine xx xx 362 316 220 294 Nalorphine HCI 49a 285 49 285 58 288 Naloxone xx 281 38 282 38 284 Nicotine Hydrogen Tartrate 338a 259 298 259 154 262 Norchlordiazepoxide xx xx nd 950 262 Nordiazepam 1140 238 1140 238 86 318 Norfluoxetine HCI xx xx nd nd 422 228 Normeperidine HCI xx xx nd nd 8 258 Norpropoxyphene Maleate xx xx 23 258 31 258 Nortriptyline HCI 517a 239 504 239 527 240 Norverapamil HCI xx xx nd nd 131 280 Olanzapine xx xx 675 259 607 272 Orphenadrine Citrate 24a 264 nd nd 20 264 Oxazepam xx 234:280 insol. nus 1318 230 Oxycodone HCI 44a 280 39 282 41 284 Papaverine HCI 1830a 250 1785 250 2158 240 Paroxetine HCI Hemihydrate xx xx 11 294 149 296 Pentazocine HCI 69a 278 65 278 77 282 PhencyclidineUncontrolled HCI 13.5a 263 13 263 13 264 Pheniramine Maleate 305a 261 nd nd 204 262 Phentermine HCI 13.9b 257 11 257 10 260 Phenylephrine HCI 110a 273 112 273 128 276 Phenylpropranolamine HCI 11.7a 257 12 257 11 258 Phenyltoloxamine Citrate 73b 270 72 270 71 272 Pramoxine HCI 80c 286 77 286 91 290 Prazepam (Toluene) 1760a 240 nd nd 59 316 Procainamide HCI 693a 308 nd nd 766 282 Prochlorperazine Edisylate 875a 254 784 254 91 258 Page 6 of 7 Harris County Institute of Forensic Sciences Section: Toxicology Approved by: Toxicology Manager Document Type: QA/QC Procedure No.: TOX07.1025 Title: Verification of reference materials and standard/control solutions Rev.: 5

Promethazine HCI 1032a 249 957 250 1149 255 Propoxyphene HCl (Acetonitrile) 12a 257 nd nd 12 258 Propranolol HCI 222a 288 217 288 239 290 Protriptyline HCI 530a 292 nd nd 525 292 (+) Pseudoephedrine HCI 11.9a 257 10 257 10 258 Pyrilamine Maleate xx xx 599 239 645 248 Quinidine Sulfate 2H20 959a 250 900 250 933 232 Quinine Sulfate 2H20 959a 250 914 250 1043 232 Sertraline HCI xx xx 19 274 29 274 Sibutramine HCI 2H20 xx xx nd nd 16 268 Strychnine 373a 254 363 254 362 256 Sufentanil Citrate xx xx 286 230 289 230 Temazepam 980b 237 nd Copynd 79 314 Thiethylperazine Malate 857b 255 101 309 99 316 Thioridazine HCI 987a 262 880 264 1031 264 Ticlopidine HCI xx xx 281 233 25 270 Tocainide HCI 22a 262 23 262 19 264 Tramadol HCI 70a 272 67 272 72 274 Tramadol Mtb. HCI xx xx nd nd 84 276 Trazodone HCI 314a 246 313 246 416 256 Triazolam xx xx 254 256 1206 222 Trifluoperazine 2HCI 743a 256 709 256 807 260 Trihexyphenidyl HCI xx xx 7 258 6 258 Trimethobenzamide HCI 312b 258 nd nd 322 262 Trimipramine Maleate 300a 250 268 250 321 250 Triprolidine HCI 347a 290 335 290 246 282 Venlafaxine HCI xx xx 44 274 49 276 O-Desmethylvenlafaxine xx xx nd nd 53 278 Verapamil HCI 127a 278 121 278 129 280 Zolpidem Hemitartrate xx xx 408 294 272 318

NUS = No Usable Spectra nd = Not Determined insol Uncontrolled= Analyte insoluble in matrix XX = Not Reported * ( ) = Parenthesis next to analyte describes organic solvent other than methanol

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