Atropine

Molecular formula: C17H23NO3 Molecular weight: 289.4 CAS Registry No.: 51-55-8 (atropine), 52-88-0 (atropine methylnitrate), 101-31-5 (hyoscyamine), 306-03-6 (hyoscyamine hydrobromide), 6835-16-1 (hyoscyamine sulfate dihydrate), 620-61-1 (hyoscyamine sulfate)

SAMPLE Matrix: blood Sample preparation: 1 mL Plasma + 100 |xL 5 ng/mL scopolamine in MeOH, vortex briefly, add 50 |JLL 1 M ammonium hydroxide, mix, add 5 mL dichloromethane, shake horizontally for 5 min, centrifuge at 2500 rpm for 5 min. Remove the organic layer and evaporate it to dryness under a stream of nitrogen at 40, reconstitute the residue in 100 |JLL mobile phase, inject a 20 JULL aliquot. HPLCVARIABLES Guard column: 10 X 2 5 |xm BDS C18 (Keystone) Column: 50 X 3 3 |xm BDS C18 (Keystone) Mobile phase: MeCN: MeOH: 10 mM ammonium acetate 62.5:37.5:15 Flow rate: 0.5 Injection volume: 20 Detector: MS, Perkin Elmer Sciex API Ill-Plus triple quadrupole, APCI, nebulizer 400 and 80 psi, auxiliary nitrogen 1.2 L/min, curtain gas 1.2 L/min, interface 55, collision gas argon, electron multiplier 3000 V, declustering potential 35 V, collision energy 35 eV CHROMATOGRAM Retention time: 1.2 Internal standard: scopolamine (0.8) Limit of quantitation: 20 pg/mL KEYWORDS plasma; protect from light REFERENCE
Xu, A.; Havel, J.; Linderholm, K.; Hulse, J. Development and validation of an LC/MS/MS method for the determination of L-hyoscyamine in human plasma. J.Pharm.Biomed.AnaL, 1996, 14, 33-42

SAMPLE Matrix: blood Sample preparation: Condition a Sep-Pak C18 SPE cartridge with water, MeOH, and 100 mM ammonium acetate. Add 200 jxL plasma to the SPE cartridge, wash with 100 mM ammonium acetate, elute with MeOH: 100 mM ammonium acetate 3:1. Evaporate the eluate to dryness under reduced pressure, dissolve the residue in 200 |xL mobile phase, inject a 20 |xL aliquot. HPLCVARIABLES Column: 150 X 4.6 Hitachi gel 3056 octadecylsilica Mobile phase: MeOH: 100 mM ammonium acetate 60:40 Flow rate: 1 Injection volume: 20 Detector: MS, Hitachi M1000, APCI, nebulizer 260, vaporizer 399

CHROMATOGRAM Retention time: 4.3 Limit of detection: 0.5-2.5 ng/mL OTHER SUBSTANCES Simultaneous: atipamezole, butorphanol, flumazenil, ketamine, medetomidine, midazolam, xylazine KEYWORDS plasma; SPE; dog REFERENCE
Kanazawa, H.; Nagata, Y.; Matsushima, Y; Takai, N.; Uchiyama, H.; Nishimura, R.; Takeuchi, A. Liquid chromatography-mass spectrometry for the determination of medetomidine and other anaesthetics in plasma. J .Chromatogr., 1993, 631, 215-220

SAMPLE Matrix: blood Sample preparation: 2 mL Plasma + 100 \xL 1 M NaOH + 10 mL chloroform, shake for 30 s, remove a 9 mL aliquot of the organic phase and add it to 1 mL 100 mM HCl, extract. Remove a 900 |xL aliquot of the aqueous layer and add it to 100 |xL 5 M NaOH, heat at 38 for 3 h, acidify with 5 M HCl, add 8 mL dichloromethane, vortex for 2 min, centrifuge. Remove the organic layer and evaporate it to dryness under a stream of nitrogen at 4045, add 3 mg solid potassium bicarbonate, add 100 |xL 10 |xg/mL mandelic acid in MeCN, add 300 |xL 50 |xg/mL 4-bromomethyl-7-methoxycoumarin in MeCN, add 100 |JLL 15 |xg/mL 18-crown-6 in MeCN, vortex for 15 s, heat at 70 for 45 min, inject a 25 |JLL aliquot. HPLCVARIABLES Column: 150 X 4.6 5 |xm ODS (Dupont) Mobile phase: MeCN: buffer 33:67 (Buffer was 10 mM (NH4)H2PO4 adjusted to pH 5.0.) Column temperature: 40 Flow rate: 2 Injection volume: 25 Detector: F ex 328 em 389 (cutoff filter) CHROMATOGRAM Retention time: 10 Internal standard: mandelic acid (7) Limit of detection: 108 ng/mL Limit of quantitation: 125 ng/mL KEYWORDS plasma; derivatization; atropine determined after hydrolysis to tropic acid REFERENCE
Li, S.; Wahba Khalil, S.K. An HPLC method for determination of atropine in human plasma. J.Liq.Chromatogr., 1990, 13, 1339-1350

SAMPLE Matrix: bulk, plants Sample preparation: Place 0.5 g powdered crude drug in 25 mL mobile phase, reflux 30 min, cool, centrifuge at 1600 g, decant wash residue twice with 10 mL portions of mobile phase, combine extracts and washings, make up to 50 mL with mobile phase, inject 10 |xL aliquot.

HPLCVARIABLES

Column: 150 X 4 5 ^m TSK gel 120A ODS Mobile phase: MeCN: 67 mM pH 2.5 phosphate buffer 35:65, containing 17.5 mM sodium dodecylsulfate Column temperature: 35 Flow rate: 1.5 Injection volume: 10 Detector: UV 210
CHROMATOGRAM Retention time: 15 OTHER SUBSTANCES Simultaneous: scopolamine REFERENCE
Oshima, T.; Sagara, K.; Tong, Y.Y.; Zhang, G.; Chen, YH. Application of ion-pair high performance liquid chromatography for analysis of hyoscyamine and scopolamine in solanaceous crude drugs. Chem.Pharm.BulL, 1989, 37, 2456-2458

SAMPLE

Matrix: formulations Sample preparation: Injections and ophthalmic solutions. Dilute with water to an atropine concentration of 80 |xg/mL, inject a 20 |xL aliquot. Ointment. Weigh out ointment equivalent to about 4 mg atropine sulfate, add 10 mL THF: water 80:20, sonicate and swirl until the ointment is completely dispersed, make up to 50 mL with water, filter (0.45 |xm), inject a 20 |xL aliquot
HPLCVARIABLES

Column: 250 X 4.6 5 |xm Spherisorb CN Mobile phase: MeCN: 50 mM NaH2PO4 10:90, pH adjusted to 4.0 with 10% phosphoric acid Flow rate: 1 Injection volume: 20 Detector: UV 220
CHROMATOGRAM Retention time: 8 OTHER SUBSTANCES

Simultaneous: phenol, tropic acid Noninterfering: benzalkonium chloride, benzyl alcohol, chlorobutanol, methylparaben
KEYWORDS

injections; ophthalmic solutions; ointments

REFERENCE
Lehr, G.J.; Yuen, S.M.; Lawrence, G.D. Liquid chromatographic determination of atropine in nerve gas antidotes and other dosage forms. J.AOAC Int., 1995, 78, 339-343

SAMPLE

Matrix: formulations Sample preparation: Inject a 20 |xL aliquot.

HPLCVARIABLES

Column: 100 X 4.6 Spheri-5 EPS

Mobile phase: MeCN: buffer 40:60 (Buffer was 10 mM KH2PO4 adjusted to pH 4.0 with 1 M KOH.) Flow rate: 1 Injection volume: 20 Detector: UV 254 CHROMATOGRAM Retention time: 7.8 Limit of detection: 8.6 jig/mL OTHER SUBSTANCES Simultaneous: ondansetron Noninterfering: degradation products KEYWORDS injections; saline REFERENCE
Venkateshwaran, T.G.; King, D.T.; Stewart, J.T. HPLC determination of ondansetron-atropine and ondansetron-glycopyrrolate mixtures in 0.9% sodium chloride injection. J.Liq.Chromatogr., 1995, 18, 2647-2659

SAMPLE Matrix: formulations Sample preparation: Dilute with water, inject a 20 |xL aliquot. HPLC VARIABLES Guard column: 12.5 X 4 5 urn Zorbax RX-C18 Column: 250 X 4.6 5 ^m Zorbax RX-C18 Mobile phase: MeCN: buffer 20:80 (Buffer was 50 mM NaH2PO4 + 1 mM tetramethylammonium chloride + 0.5 mM 1-octanesulfonic acid adjusted to pH 3.5 with concentrated orthophosphoric acid.) Column temperature: 25 Flow rate: 1 Injection volume: 20 Detector: UV 203 CHROMATOGRAM Retention time: 9 OTHER SUBSTANCES Simultaneous: HI-6, obidoxime, phenol, tropic acid Also analyzed: pralidoxime chloride KEYWORDS nerve agent antidote mixtures REFERENCE
Paddle, B.M.; Dowling, M.H. Simple high-performance liquid chromatographic method for assessing the deterioration of atropine-oxime mixtures employed as antidotes in the treatment of nerve agent poisoning. J.Chromatogr., 1993, 648, 373-380

SAMPLE Matrix: formulations Sample preparation: Dilute with water, inject an aliquot.

HPLCVARIABLES Column: 250 X 4.6 10 |xm Whatman PXS ODS-3 C18 Mobile phase: MeCN: Buffer 25:75 (Buffer was 1.08 g sodium octanesulfonate in 900 mL of water adjusted to pH 3.5 with glacial acetic acid and diluted to 1 L with water.) Flow rate: 2 Injection volume: 20 Detector: UV 229 CHROMATOGRAM Retention time: 5 OTHER SUBSTANCES Simultaneous: milrinone KEYWORDS injections; 10% calcium chloride; 7.5% sodium bicarbonate; stability-indicating REFERENCE
Wilson, T.D.; Forde, M.D. Stability of milrinone and epinephrine, atropine sulfate, lidocaine hydrochloride, or morphine sulfate injection. Am.J.Hosp.Pharm., 1990, 47, 2504-2507

SAMPLE Matrix: formulations Sample preparation: Tablets. Crush tablet, add 2 mL water, add 500 JULL 0.1 M NaOH, if not basic to litmus add 0.1 M NaOH dropwise until it is. Extract with 10 mL chloroform then with three 5 mL portions of chloroform. Pass all extracts through a sodium sulfate column and then wash column with 2 mL chloroform. Evaporate all organic extracts with heating under a stream of air. Take up residue in 1 mL MeOH, inject a 100 |xL aliquot. Injections. Evaporate 2 mL to dryness on a steam bath, take up residue in 5 mL water, add 500 |JLL 0.1 M NaOH, if not basic to litmus add 0.1 M NaOH dropwise until it is. Extract with 10 mL chloroform then with three 5 mL portions of chloroform. Pass all extracts through a sodium sulfate column and then wash column with 2 mL chloroform. Evaporate all organic extracts with heating under a stream of air. Take up residue in 2 mL MeOH, inject a 100 |xL aliquot. (Sodium sulfate column was a 300 X 20 glass chromatography column containing 1Og anhydrous sodium sulfate washed with 10 mL chloroform before use.) HPLC VARIABLES Column: 250 X 4.6 Alltech C18 Mobile phase: MeOH: 20 g/L sodium pentanesulfonate 95:5 Flow rate: 1 Injection volume: 100 Detector: F ex 255 em 285 CHROMATOGRAM Retention time: 3.5 OTHER SUBSTANCES Simultaneous: scopolamine Noninterfering: ergotamine, phenobarbital KEYWORDS tablets; injections REFERENCE
Cieri, U.R. Determination of small quantities of atropine in commercial preparations by liquid chromatography with fluorescence detection. J.Assoc.Off.Anal.Chem., 1985, 68, 1042-1045

SAMPLE Matrix: formulations Sample preparation: Dilute 10 mL eye drops to 100 mL with water, inject a 20 |xL aliquot. HPLC VARIABLES Column: 150 X 3.9 5 |xm Perkin-Elmer C18 Mobile phase: MeOH: water: heptanesulfonic acid solution 500:500:25 (Heptanesulfonic acid solution was 5 g heptanesulfonic acid + 20 mL glacial acetic acid diluted to 150 mL with water.) Flow rate: 1.8 Injection volume: 20 Detector: UV 225 CHROMATOGRAM Retention time: 4 Limit of detection: 5 |xg/mL OTHER SUBSTANCES Simultaneous: tropic acid KEYWORDS eye drops REFERENCE
Richard, A.; Andermann, G. Simultaneous determination of atropine sulphate and tropic acid by reversed phase high-pressure liquid chromatography. Pharmazie, 1984, 39, 866867

SAMPLE Matrix: formulations Sample preparation: Tablets, capsules. Powder tablets or remove contents of capsules, weigh out amount equivalent to about 600 |xg hyoscyamine sulfate-atropine sulfate, add 25 mL 25 mM sulfuric acid, shake for 15 min, centrifuge at 3000 rpm for 5 min. Remove 5 mL of the supernatant and extract it twice with 30 mL portions of dichloromethane, discard the organic phase, add 2 mL buffer to the aqueous phase, extract with four 30 mL portions of dichloromethane, filter extracts through dichloromethane-rinsed glass wool, add 3 mL 2.25 |xg/mL theophylline in dichloromethane, distil off the dichloromethane through a Snyder column by using a steam bath, when the volume reaches 10 mL rinse the column with 1-2 mL dichloromethane, continue distillation to 0.5-1 mL, remove the column and rinse the concentrator tube-column junction with 1 mL dichloromethane, evaporate to 1 mL with a stream of air at 40, add 100 jxL 1% concentrated HCl in MeOH, mix, evaporate to dryness with a stream of air at 40, rinse the sides of the concentrator tube with 500 |xL MeOH, evaporate to dryness with a stream of air at 40, dissolve the residue in 300 |xL water, inject a 20 |xL aliquot. Elixirs. Add an amount equivalent to about 600 |xg hyoscyamine sulfate-atropine sulfate to a 150 mL beaker, warm at 40 with a current of air for 30 min to remove alcohol, cool, make up to 25 mL with water, remove 5 mL of this solution, add 2 mL 100 mM sulfuric acid, extract twice with 30 mL portions of dichloromethane, discard the organic phase, add 2 mL buffer to the aqueous phase, extract with four 30 mL portions of dichloromethane, filter extracts through dichloromethane-rinsed glass wool, add 3 mL 2.25 [xg/mL theophylline in dichloromethane, distil off the dichloromethane through a Snyder column by using a steam bath, when the volume reaches 10 mL rinse the column with 1-2 mL dichloromethane, continue distillation to 0.5-1 mL, remove the column and rinse the concentrator tube-column junction with 1 mL dichloromethane, evaporate to 1 mL with a stream of air at 40, add 100 JJLL 1% concentrated HCl in MeOH, mix, evaporate to dryness with a stream of air at 40, rinse the sides of the concentrator tube with 500 |xL MeOH, evaporate to dryness with a stream of air at 40, dissolve the residue in 300 \L water, inject a 20 |xL aliquot. (Buffer was 5.3 g

anhydrous sodium carbonate and 4.2 g sodium bicarbonate in 100 mL water, pH 9.4. Pass dichloromethane through 75 g basic aluminum oxide, Brockmann Activity Grade 1, store over 25 g alumina/4 L.)
HPLCVARIABLES

Column: 250 X 4 5 jxm Spherisorb ODS Mobile phase: MeOH: buffer 250:525 (The 50 mM tetramethylammonium phosphate buffer was prepared from 500 mL water + 23 mL 20% tetramethylammonium hydroxide in MeOH + 10 mL concentrated phosphoric acid, adjust to pH 2.0 with concentrated phosphoric acid, make up to 1 L with water. Flow rate: 0.8 Injection volume: 20 Detector: UV 220
CHROMATOGRAM

Retention time: 7.5 Internal standard: theophylline (6.5)

OTHER SUBSTANCES

Simultaneous: hyoscyamine, phenobarbital, scopolamine

KEYWORDS

tablets; capsules; elixirs

REFERENCE
Pennington, L.J.; Schmidt, W.F. Belladonna alkaloids and phenobarbital combination Pharmaceuticals analysis I: High-performance liquid chromatographic determinations of hyoscyamine-atropine and scopolamine. J.Pharm.Sci., 1982, 71, 951-953

SAMPLE

Matrix: plants Sample preparation: 100 mg Freeze-dried powdered plant leaves + 10 mL mobile phase, heat at 40 for 15 min, filter, inject a 20 |xL aliquot.
HPLCVARIABLES

Column: 150 X 4 4 |xm Novapack C18 Mobile phase: MeCN-.water 12.5:87.5 with 0.3% phosphoric acid adjusted to pH 2.2 with triethylamine Flow rate: 0.8 Injection volume: 20 Detector: UV 204
CHROMATOGRAM

Retention time: 11.7 Limit of detection: 50 jxg/g

OTHER SUBSTANCES

Simultaneous: tropic acid, scopolamine

KEYWORDS

freeze-dried plant leaves

REFERENCE
Fliniaux, M.-A.; Manceau, F.; Jacquin-Dubreuil, A. Simultaneous analysis of 1-hyoscyamine, 1-scopolamine and dl-tropic acid in plant material by reversed phase high-performance liquid chromatography. J.Chromatogr., 1993, 644, 193-197

SAMPLE Matrix: plants Sample preparation: Extract 0.1 g dry plant material with 10 mL MeOH for 10 min under reflux, filter, inject aliquot. HPLCVARIABLES Guard column: 40 X 4 10 |xm Hypersil ODS Column: 250 X 4 10 jim Hypersil ODS Mobile phase: MeOH-.water 45:55 containing 0.1% phosphoric acid adjusted to pH 7 with triethylamine Flow rate: 1 Detector: UV 229 CHROMATOGRAM Retention time: 20.7 OTHER SUBSTANCES Simultaneous: scopolamine REFERENCE
Hagemann, K.; Piek, K.; Stockigt, J.; Weiler, E.W. Monoclonal antibody-based enzyme immunoassay for the quantitative determination of the tropane alkaloid, scopolamine. Planta Med., 1992, 58, 68-72

SAMPLE Matrix: plants Sample preparation: Dissolve plant extract in 1 mL MeOH, add 40 ng homatropine, inject aliquot. HPLCVARIABLES Column: 150 X 4.1 5 [xm Hamilton PRP-I Mobile phase: MeCN: 100 mM pH 10.4 ammonium acetate 30:70 Flow rate: 1 Injection volume: 20 Detector: MS thermospray, VG Trio-2, ion source 150, vaporizer tip 170, repeller electrode 150 V, m/z 290 CHROMATOGRAM Internal standard: homatropine (m/z 276) Limit of detection: 2.5 ng/mL OTHER SUBSTANCES Simultaneous: scopolamine KEYWORDS total run time 6 min REFERENCE
Auriola, S.; Martinsen, A.; Oksman-Caldentey, K.M.; Naaranlahti, T. Analysis of tropane alkaloids with thermospray high-performance liquid chromatography-mass spectrometry. J.Chromatogr., 1991,562, 737-744

SAMPLE Matrix: solutions

HPLCVARIABLES Column: 250 X 4.1 6 |xm PolyEncap ODS (n-octadecylacrylate copolymerized with vinyl silica in heptane, carrier Ultrasep ES 100; preparation described in paper) Mobile phase: MeCN :pH 2.2 phosphate buffer 20:80 Flow rate: 1 Detector: UV 220 CHROMATOGRAM Retention time: 2.7 OTHER SUBSTANCES Simultaneous: barbituric acid, codeine, diphenhydramine, noscapine, papaverine REFERENCE
Engelhardt, H.; Cufiat-Walter, M.A. Polymer encapsulated stationary phases with improved efficiency. Chromatographia, 1995, 40, 657-661

SAMPLE Matrix: solutions HPLCVARIABLES Column: 150 X 2 PRP-I (Keystone) Mobile phase: MeCN :2-butanone: 100 mM pH 7.5 phosphate buffer 40:20:40 Flow rate: 0.15 Injection volume: 1 Detector: Chemiluminescence following post-column reaction. A 1 mM solution of Ru(2,2'bipyridine)32+ in 50 mM sodium sulfate (continuously sparged with helium) was oxidized to Ru(2,2'-bipyridine)33+ using a Princeton Applied Research Model 174A polarographic analyzer with a platinum gauze working electrode, a platinum wire auxiliary electrode, and a silver wire reference electrode. The Ru solution pumped at 0.3 mL/min mixed with the column effluent in the flow cell of the detector, a fluorescence detector with the light source removed. CHROMATOGRAM Retention time: 3 Limit of detection: 0.1-1 |xg/mL OTHER SUBSTANCES Simultaneous: cyclobenzaprine, cyclopentolate, dicyclomine, procyclidine KEYWORDS post-column reaction REFERENCE
Holeman, J.A.; Danielson, N.D. Microbore liquid chromatography of tertiary amine anticholinergic pharmaceuticals with tris(2,2'-bipyridine)ruthenium(III) chemiluminescence detection. J.Chromatogr. ScL, 1995, 33, 297-302

SAMPLE Matrix: solutions HPLCVARIABLES Column: 250 X 4.6 5 jim Supelcosil LC-DP (A) or 250 X 4 5 |xm LiChrospher 100 RP-8 (B) Mobile phase: MeCN:0.025% phosphoric acid:buffer 25:10:5 (A) or 60:25:15 (B) (Buffer was 9 mL concentrated phosphoric acid and 10 mL triethylamine in 900 mL water, adjust pH to 3.4 with dilute phosphoric acid, make up to 1 L.)

Flow rate: 0.6 Injection volume: 25 Detector: UV 229 CHROMATOGRAM Retention time: 6.05 (A), 3.75 (B) OTHER SUBSTANCES Also analyzed: acebutolol, acepromazine, acetaminophen, acetazolamide, acetophenazine, albuterol, alprazolam, amitriptyline, amobarbital, amoxapine, antipyrine, atenolol, azatadine, baclofen, benzocaine, bromocriptine, brompheniramine, brotizolam, bupivacaine, buspirone, butabarbital, butalbital, caffeine, carbamazepine, cetirizine, chlorcyclizine, chlordiazepoxide, chlormezanone, chloroquine, chlorpheniramine, chlorpromazine, chlorpropamide, chlorprothixene, chlorthalidone, chlorzoxazone, cimetidine, cisapride, clomipramine, clonazepam, clonidine, clozapine, cocaine, codeine, colchicine, cyclizine, cyclobenzaprine, dantrolene, desipramine, diazepam, diclofenac, diflunisal, diltiazem, diphenhydramine, diphenidol, diphenoxylate, dipyridamole, disopyramide, dobutamine, doxapram, doxepin, droperidol, encainide, ethidium bromide, ethopropazine, fenoprofen, fentanyl, flavoxate, fluoxetine, fluphenazine, flurazepam, flurbiprofen, fluvoxamine, furosemide, glutethimide, glyburide, guaifenesin, haloperidol, homatropine, hydralazine, hydrochlorothiazide, hydrocodone, hydromorphone, hydroxychloroquine, hydroxyzine, ibuprofen, imipramine, indomethacin, ketoconazole, ketoprofen, ketorolac, labetalol, Ievorphanol, lidocaine, loratadine, lorazepam, lovastatin, loxapine, mazindol, mefenamic acid, meperidine, mephenytoin, mepivacaine, mesoridazine, metaproterenol, methadone, methdilazine, methocarbamol, methotrexate, methotrimeprazine, methoxamine, methyldopa, methylphenidate, metoclopramide, metolazone, metoprolol, metronidazole, midazolam, moclobemide, morphine, nadolol, nalbuphine, naloxone, naphazoline, naproxen, nifedipine, nizatidine, norepinephrine, nortriptyline, oxazepam, oxycodone, oxymetazoline, paroxetine, pemoline, pentazocine, pentobarbital, pentoxifylline, perphenazine, pheniramine, phenobarbital, phenol, phenolphthalein, phentolamine, phenylbutazone, phenyltoloxamine, phenytoin, pimozide, pindolol, piroxicam, pramoxine, prazepam, prazosin, probenecid, procainamide, procaine, prochlorperazine, procyclidine, promazine, promethazine, propafenone, propantheline, propiomazine, propofol, propranolol, protriptyline, quazepam, quinidine, quinine, racemethorphan, ranitidine, remoxipride, risperidone, salicylic acid, scopolamine, secobarbital, sertraline, sotalol, spironolactone, sulfinpyrazone, sulindac, temazepam, terbutaline, terfenadine, tetracaine, theophylline, thiethylperazine, thiopental, thioridazine, thiothixene, timolol, tocainide, tolbutamide, tolmetin, trazodone, triamterene, triazolam, trifluoperazine, triflupromazine, trimeprazine, trimethoprim, trimipramine, verapamil, warfarin, xylometazoline, yohimbine, zopiclone KEYWORDS some details of plasma extraction REFERENCE
Koves, E.M. Use of high-performance liquid chromatography-diode array detection in forensic toxicology.

J.ChromatogrA, 1995, 692, 103-119 SAMPLE Matrix: solutions HPLCVARIABLES Column: 250 x 4.6 Zorbax RX Mobile phase: Gradient. A was 10 mL concentrated orthophosphoric acid and 7 mL triethylamine in 1 L water. B was 10 mL concentrated orthophosphoric acid and 7 mL triethylamine in 200 mL water, make up to 1 L with MeCN. A:B from 100:0 to 0:100 over 30 min, maintain at 0:100 for 5 min. Column temperature: 30

Flow rate: 2 Detector: UV 210 OTHER SUBSTANCES Also analyzed: acepromazine, acetaminophen, acetophenazine, albuterol, aminophylline, amitriptyline, amobarbital, amoxapine, amphetamine, amylocaine, antipyrine, aprobarbital, aspirin, atenolol, avermectin, barbital, benzocaine, benzoic acid, benzotropine, benzphetamine, berberine, bibucaine, bromazepan, brompheniramine, buprenorphine, buspirone, butabarbital, butacaine, butethal, caffeine, carbamazepine, carbromal, chloramphenicol, chlordiazepoxide, chloroquine, chlorothiazide, chloroxylenol, chlorphenesin, chlorpheniramine, chlorpromazine, chlorpropamide, chlortetracycline, cimetidine, cinchonidine, cinchonine, clenbuterol, clonazepam, clonixin, clorazepate, cocaine, codeine, colchicine, cortisone, coumarin, cyclazocine, cyclobenzaprine, cyclothiazide, cyheptamide, cymarin, danazol, danthron, dapsone, debrisoquine, desipramine, dexamethasone, dextromethorphan, dextropropoxyphene, diamorphine, diazepam, diclofenac, diethylpropion, diethylstilbestrol, diflunisal, digitoxin, digoxin, diltiazem, diphenhydramine, diphtenoxylate, diprenorphine, dipyrone, disulfiram, dopamine, doxapram, doxepin, dronabinol, ephedrine, epinephrine, epinine, estradiol, estriol, estrone, ethacrynic acid, ethosuximide, etonitazene, etorphine, eugenol, famotidine, fenbendazole, fencamfamine, fenoprofen, fenproporex, fentanyl, flubendazole, flufenamic acid, flunitrazepam, 5-fluorouracil, fluoxymesterone, fluphenazine, furosemide, gentisic acid, gitoxigenin, glipizide, glunixin, glutethimide, glybenclamide, guaiacol, halazepam, haloperidol, hydrochlorothiazide, hydrocodone, hydrocortisone, hydromorphone, hydroxyquinoline, ibogaine, ibuprofen, iminostilbene, imipramine, indomethacin, isocarbostyril, isocarboxazid, isoniazid, isoproterenol, isoxsuprine, ivermectin, ketamine, ketoprofen, kynurenic acid, levorphanol, Iidocaine, lorazepam, lormetazepam, loxapine, mazindol, mebendazole, meclizine, meclofenamic acid, medazepam, mefenamic acid, megestrol, mepacrine, meperidine, mephentermine, mephenytoin, mephesin, mephobarbital, mepivacaine, mescaline, mesoridazine, methadone, methamphetamine, methapyrilene, methaqualone, methazolamide, methocarbamol, methoxamine, methsuximide, methyl salicylate, methyldopa, methyldopamine, methylphenidate, methylprednisolone, methyltestosterone, methyprylon, metoprolol, mibolerone, morphine, nadolol, nalorphine, naloxone, naltrexone, naphazoline, naproxen, nefopam, niacinamide, nicotine, nicotinic acid, nifedipine, niflumic acid, nitrazepam, norepinephrine, nortriptyline, noscapine, nylidrin, oxazepam, oxycodone, oxymorphone, oxyphenbutazone, oxytetracycline, papaverine, pargyline, pemoline, pentazocine, pentobarbital, persantine, phenacetin, phenazocine, phenazopyridine, phencyclidine, phendimetrazine, phenelzine, pheniramine, phenobarbital, phenothiazine, phensuximide, phentermine, phenylbutazone, phenylephrine, phenylpropanolamine, piperocaine, prazepam, prednisolone, primidone, probenecid, progesterone, propiomazine, propranolol, propylparaben, pseudoephedrine, puromycin, pyrilamine, pyrithyldione, quazepam, quinaldic acid, quinidine, quinine, ranitidine, recinnamine, reserpine, resorcinol, saccharin, albuterol, salicylamide, salicylic acid, scopolamine, scopoletin, secobarbital, strychnine, sulfacetamide, sufadiazine, sulfadimethoxine, sulfaethidole, sulfamerazine, sulfamethazine, sulfamethoxizole, sulfanilamide, sulfapyridine, sulfasoxizole, sulindac, tamoxifen, temazepam, testosterone, tetracaine, tetracycline, tetramisole, thebaine, theobromine, theophylline, thiabendazole, thiamine, thiamylal, thiobarbituric acid, thioridazine, thiosalicylic acid, thiothixene, thymol, tolazamide, tolazoline, tobutamide, tolmetin, tranylcypromine, triamcinolone, tribenzylamine, trichloromethiazide, trifluoperazine, trihexyphenidyl, trimethoprim, tripelennamine, triproilidine, tropacocaine, tyramine, verapamil, vincamine, warfarin, yohimbine, zoxazolamine REFERENCE
Hill, D. W.; Kind, A. J. Reversed-phase solvent gradient HPLC retention indexes of drugs. J.Anal.ToxicoL, 1994, 18, 233-242

SAMPLE Matrix: solutions

HPLCVARIABLES Column: 250 X 4.6 cellulose tris(3,5-dimethylphenylcarbamate) Mobile phase: Hexane: isopropanol: diethylamine 80:20:0.1 Flow rate: 0.5 Detector: UV CHROMATOGRAM Retention time: k' 0.72 (of first (+) enantiomer) KEYWORDS chiral; a 1.62 REFERENCE
Okamoto, Y; Aburatani, R.; Hatano, K.; Hatada, K. Optical resolution of racemic drugs by chiral HPLC on cellulose and amylose tris(phenylcarbamate) derivatives. J.Liq.Chromatogr., 1988, 11, 2147-2163

SAMPLE Matrix: solutions Sample preparation: Dissolve in MeOH or water to 0.1%, inject an aliquot HPLCVARIABLES Column: two 250 mm p-cyclodextrin bonded phase columns in series (Advanced Separation Technologies) Mobile phase: MeCN : 1 % pH 4.1 aqueous triethylammonium acetate 2:98 Flow rate: 0.5 Injection volume: 1 Detector: UV CHROMATOGRAM Retention time: k' 6.83 (d-isomer) KEYWORDS chiral; optical isomers are separated REFERENCE
Armstrong, D.W.; Han, S.M.; Han, Y.I. Separation of optical isomers of scopolamine, cocaine, homatropine, and atropine. Anal.Biochem., 1987, 167, 261-264

SAMPLE Matrix: solutions HPLCVARIABLES Column: 300 X 3.9 10 ^m LiChrosorb Si-60 Mobile phase: MeOH: water 60:40 containing 4 mM disodium citrate and 4 mM tetrabutylammonium bromide, pH 5.9 Flow rate: 1 Injection volume: 10 Detector: UV 254 CHROMATOGRAM Retention time: 13 OTHER SUBSTANCES Simultaneous: codeine, dansylamide, dansylcadaverine, doxorubicin, methylatropine, naphazoline, noscapine, xylometazoline

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REFERENCE
Lingeman, H.; van Munster, H.A.; Beynen, J.H.; Underberg, W.J.; Hulshoff, A. High-performance liquid chromatographic analysis of basic compounds on non-modified silica gel and aluminium oxide with aqueous solvent mixtures. J.Chromatogr., 1986, 352, 261-274

SAMPLE Matrix: solutions

Sample preparation: Dissolve in MeOH: water 1:1 at a concentration of 50 |xg/mL, inject a 10 jxL aliquot.
HPLCVARIABLES

Column: 300 X 3.9 10 |xm ixBondapak C18 Mobile phase: MeOH: acetic acid: triethylamine: water 15:1.5:0.5:83 Flow rate: 1.5 Injection volume: 10 Detector: UV 230
CHROMATOGRAM Retention time: 17 OTHER SUBSTANCES

Simultaneous: homatropine, methscopolamine, tropic acid, atropine methyl, scopolamine

REFERENCE
Roos, R.W.; Lau-Cam, CA. General reversed-phase high-performance liquid chromatographic method for the separation of drugs using triethylamine as a competing base. J.Chromatogr., 1986, 370, 403 -418

ANNOTATED BIBLIOGRAPHY

Theodoridis, G.; Papadoyannis, L; Vasilikiotis, G.; Tsoukali-Papadopoulou, H. Reversed-phase high-performance liquid chromatography-photodiode-array analysis of alkaloid drugs of forensic interest. J.Chromatogr.B, 1995, 668, 253-263 [also amphetamine, bamifylline, caffeine, cocaine, codeine, diamorphine, ethylmorphine, flufenamic acid, methadone, morphine, nalorphine, norcodeine, papaverine, quinine, scopolamine, strychnine, theobromine, theophylline, tolfenamic acid] Buch, U.; Isenberg, E.; Buch, H.P. HPLC assay for atropine in serum and protein solutions after in vitro addition of the tropane alkaloid. Methods Find.Exp.Clin.PharmacoL, 1994, 16, 361-365 Pohjola, J.; Harpf, M. Determination of atropine and obidoxime in automatic injection devices used as antidotes against nerve agent intoxication. J.Chromatogr., 1994, 686, 350-354 [simultaneous obidoxime; phenol (IS)] Schill, G.; Wainer, I.W.; Barkan, S.A. Chiral separation of cationic drugs on an al-acid glycoprotein bonded stationary phase. J.Liq.Chromatogr., 1986, 9, 641-666 [chiral; also bromdiphenhydramine, brompheniramine, bupivacaine, butorphanol, carbinoxamine, chlorpheniramine, clidinium, cocaine, cyclopentolate, dimethindene, diperidone, disopyramide, doxylamine, ephedrine, homatropine, labetalol B, labetalol, labetalol A, mepensolate, mepivacaine, methadone, methorphan, methylatropine, methylhomatropine, methylphenidate, metoprolol, nadolol, nadolol A, nadolol B, oxprenolol, oxyphencyclimine, phenmetrazine, phenoxybenzamine, promethazine, pronethalol, propoxyphene, propranolol, pseudoephedrine, terbutaline, tocainide, tridihexethyl] Achari, R.G.; Jacob, J.T. A study of the retention behavior of some basic drug substances by ion-pair HPLC. J.Liq.Chromatogr., 1980, 3, 81-92 [also N-acetylprocainamide, antazoline, caffeine, chlorpheniramine, codeine, ephedrine, epinephrine, naphazoline, papaverine, pheniramine, phenylephrine, phenylpropanolamine, procainamide, quinidine, scopolamine, xylocaine]