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Amitriptyline
Molecular formula: C20H23N Molecular weight: 277.4 CAS Registry No.: 50-48-6 (amitriptyline), 549-18-8 (amitriptyline hydrochloride)

SAMPLE Matrix: blood Sample preparation: 1 mL Serum + 500 jxL 750 mM pH 10 sodium bicarbonate/carbonate buffer + 50 |xL IS in EtOH: water 50:50 + 8 mL heptane: isoamyl alcohol 98:2, shake at 250 cycles/min for 5 min, centrifuge at 1500 g for 10 min, freeze in dry ice/EtOH. Remove the organic layer and add it to 150 |xL 22 mM pH 2.5 KHaPCVphosphoric acid buffer, shake at 250 cycles/min for 5 min, centrifuge at 1500 g for 10 min, freeze in dry ice/EtOH. Discard the organic layer, inject a 65 |xL aliquot of the aqueous layer. HPLCVARIABLES Column: 250 X 4.6 Supelco C18 Mobile phase: MeCN: buffer 45:55 (Buffer was 44 mM KH2PO4 containing 1.5 mL/L triethylamine, adjusted to pH 2.5 with phosphoric acid.) Flow rate: 1.5 Injection volume: 65 Detector: UV 240 CHROMATOGRAM Retention time: 11.5 Internal standard: 1 - (3- (dimethylamino)propyl) -1 - (p-chlorophenyl) -1,3-dihydroisobenzofuran-5-carbonitrile (LU 10-202) (Lundbeck, Copenhagen) (8.33) OTHER SUBSTANCES Extracted: citalopram, nortriptyline Simultaneous: chlorprothixene, clomipramine, clozapine, flupenthixol, haloperidol, levomepromazine, perphenazine, zuclopenthixol Noninterfering: benzodiazepines Interfering: didesmethylclomipramine, levomepromazine KEYWORDS serum REFERENCE
Olesen, O.V.; Linnet, K. Simplified high-performance liquid chromatographic method for the determination of citalopram and desmethylcitalopram in serum without interference from commonly used psychotropic drugs and their metabolites. J.Chromatogr.B, 1996, 675, 83-88

SAMPLE Matrix: blood Sample preparation: 1 mL Plasma -I- 500 |xL 1 M sodium carbonate, vortex for 30 s, add 5 mL diethyl ether, vortex for 2 min, centrifuge at 4000 rpm for 10 min. Remove the organic layer and evaporate it to dryness under a stream of nitrogen at 40°, reconstitute the residue in 250 fxL mobile phase, vortex for 30 s, centrifuge at 12000 rpm for 5 min, inject an aliquot of the supernatant. HPLCVARIABLES Column: 300 X 3.9 10 |xm jxBondapak C18

Remove the lower organic layer and evaporate it to dryness under vacuum . Niazy.1 Internal standard: amitriptyline OTHER SUBSTANCES Extracted: pheniramine Simultaneous: chlorpheniramine.2 Detector: UV 262 CHROMATOGRAM Retention time: 6. S. 35. centrifuge at 2800 g for 10 min. flurbiprofen.Mobile phase: MeOH: water 62:38 adjusted to pH 3. Chaleby. A. J.PharmacoL. D. Raines.5:2.Liq.03% phosphoric acid. J. A..M. High-performance liquid chromatographic method for the quantitative determination of pheniramine in plasma.Chromatogr. 18. mebeverine. metoclopramide. itraconazole. Remove the organic phase and add it to 0.6) OTHER SUBSTANCES Extracted: fluoxetine. phenylbutazone KEYWORDS dog. 763—777 SAMPLE Matrix: blood Sample preparation: 1 mL Plasma + doxepin + NaOH -f hexane: isoamyl alcohol 98:2. extract.5 Detector: UV 220 CHROMATOGRAM Retention time: 17. ketoprofen... Gad.8 Internal standard: doxepin (11. Steady-state kinetics of fluoxetine and amitriptyline in patients treated with a combination of these drugs as compared with those treated with amitriptyline alone. Khidir. E. ibuprofen.. diltiazem. K.A. amitriptyline is IS REFERENCE El-Sayed. diazepam. 1995. extract. HPLCVARIABLES Guard column: C18 Column: 100 X 8 10 jxm Resolve C8 (Waters) Mobile phase: MeCN: MeOH: 56 mM ammonium acetate :1 M ammonium hydroxide 100: 10:4. YM..Clin. 17-21 SAMPLE Matrix: blood Sample preparation: 2 mL Whole blood or plasma + 2 mL buffer + 5 mL chloroform: isopropanol: n-heptane 60:14:26.H.5 with phosphoric acid Column temperature: 40 Flow rate: 1. nortriptyline KEYWORDS plasma REFERENCE el-Yazigi. plasma. norfluoxetine. 1995.6 Flow rate: 2.. inject an aliquot of the aqueous phase. shake gently horizontally for 10 min.

sotalol. clozapine. dextromoramide. methotrexate. fluvoxamine. zopiclone. carteolol. zorubicine Interfering: etodolac. thioridazine. sultopride. flecainide. fluphenazine. amisulpride. quinupramine. buprenorphine. re-equilibrate for 30 min. nicardipine. ritodrine. cyclizine. amodiaquine.68 g/L (10 mM (sic)) KH2PO4 adjusted to pH 2. terfenadine. cytarabine. toloxatone. nimodipine. fenoprofen. nalbuphine. lisinopril. pipamperone. methaqualone. tioclomarol KEYWORDS whole blood. oxazepam.8 Injection volume: 50 Detector: UV 240 CHROMATOGRAM Retention time: 9. imipramine. phenol. protriptyline. ajmaline. nitrendipine. phencyclidine. propranolol. pentazocine. tetracaine. hydroxyzine. flumazenil.at 45°. cocaine. medifoxamine. reconstitute the residue in 100 |xL mobile phase. histapyrrodine. loprazolam. bumadizone. glibornuride. temazepam. acenocoumarol. strychnine. fluoxetine. cyproheptadine. acetaminophen. chlorambucil. moclobemide.) (At the end of each session wash the column with water for 1 h and MeOH for 1 h.5 with 25% ammonia solution. aceprometazine. moperone. clonazepam. ketamine. brompheniramine. haloperidol. alprenolol. celiprolol. yohimbine. thiopental. desipramine. fentiazac. diazepam. niflumic acid. diphenhydramine. quinidine. nortriptyline. nadolol. glipizide. chlorpropamide. diazoxide. alpidem. glibenclamide. ibuprofen.) Column temperature: 30 Flow rate: 0. chlordiazepoxide. clomipramine. loperamide. penfluridol. trazodone. benzoylecgonine. mefenamic acid. triprolidine. buspirone. ramipril. loxapine. proguanil. bromazepam. procainamide. pyrimethamine. glutethimide. labetalol. ticlopidine. clobazam. atenolol. lorazepam. bisoprolol. cicletanine. plasma . verapamil. mefenidramine. codeine. metoclopramide. naproxen. acepromazine. trifluperidol. iproniazid. warfarin. carbinoxamine. floctafenine. chlormezanone. aconitine. suriclone. flurbiprofen.6 with concentrated orthophosphoric acid. alimemazine. nizatidine. reserpine. carpipramine. prilocaine. phenobarbital. fenfluramine. dipyridamole. loratadine. naltrexone. vindesine. meperidine. benzocaine. clonidine. lidoflazine. triazolam. carbamazepine. trifluoperazine. tenoxicam. minoxidil. vinblastine. prazepam. cycloguanil. astemizole. mexiletine. bepridil. betaxolol.23 Limit of detection: <120 ng/mL OTHER SUBSTANCES Extracted: acebutolol. oxprenolol. dextromethorphan. demexiptiline. omeprazole. clemastine. metapramine. cibenzoline. doxylamine. bupivacaine. alminoprofen. methadone. tiaprofenic acid. droperidol. tolbutamide. aspirin. nitrazepam.) HPLCVARIABLES Column: 300 X 3. alprazolam. albuterol. chlorophenacinone. metformin. trimipramine. zolpidem. thioproperazine. amoxapine. mefloquine. mianserine. opipramol. colchicine. secobarbital. benperidol. dihydralazine. pimozide. debrisoquine. ephedrine. nomifensine. melphalan. piroxicam. lidocaine. midazolam. tianeptine. metipranolol. estazolam. promethazine. ranitidine. prazosin. tetrazepam. benazepril. maprotiline. naloxone. dextropropoxyphene. chlorpheniramine. clorazepate. dosulepine. vincristine. tofisopam. phenylbutazone. tiapride. dacarbazine. centrifuge at 2800 g for 5 min. flunitrazepam. viloxazine. sulindac. methocarbamol. pindolol. disopyramide. terbutaline. ketoprofen. cyamemazine. penbutolol. (Buffer was saturated ammonium chloride solution 25% diluted with water. diclofenac. adjusted to pH 9. cetirizine. nalorphine. medazepam. sulfinpyrazole. mephentermine. inject a 50 |xL aliquot of the supernatant. morphine. chlorpromazine. diltiazem. nialamide. chloroquine. daunorubicin.9 4 |xm NovaPack C18 Mobile phase: MeOH:THF:buffer 65:5:30 (Buffer was 0. propafenone. mephenesin. metoprolol. mepivacaine. levomepromazine. caffeine. nifedipine. hydroxychloroquine. tropatenine. timolol. sulpride. indomethacin. doxepin. quinine. procarbazine.

1 M ammonium acetate in MeOH.5 with glacial acetic acid. separate. 1995.2 potassium phosphate buffer 60:15:25'(Prepare buffer by mixing 194 mL 1. Systematic toxicological analysis using HPLC/DAD. Potential interference of cyclobenzaprine and norcyclobenzaprine with HPLC measurement of amitriptyline and nortriptyline: resolution by GC-MS analysis. J. inject a 100 |xL aliquot of the aqueous phase. 19. Remove the upper organic layer and evaporate it to dryness under a stream of air at 40°. Mangin.C. reconstitute the residue in 200 |JLL MeOH. J. inject a 50 |xL aliquot. Add 0. mix. HPLCVARIABLES Guard column: LC-8-DB (Supelco) Column: 150 X 4.1) OTHER SUBSTANCES Extracted: norcycyclobenzaprine. 254-262 SAMPLE Matrix: blood Sample preparation: 1 mL Serum + 500 |JLL 250 |xg/mL protriptyline hydrochloride + 1 mL 500 mM NaOH + 4 mL toluene: n-hexane: isoamyl alcohol 77:22:3. Worldwide Monitoring) with 2 mL MeOH then 2 mL water.) Flow rate: 2 Injection volume: 100 Detector: UV 228 .Forensic ScL. add to SPE cartridge. Koenig.C. Turk. A. mix for 10 min.74 g/L K2HPO4. J. J. E.8 Internal standard: protriptyline (8. 1995. centrifuge at 3000 rpm for 5 min.5 M NaOH and 4 mL 50 mL/L isopropanol in heptane to eluate. P. Condition a C18 Clean-Up SPE cartridge (CEC 18111.1 M phosphoric acid (pH 2.. 1 mL Plasma + 1 mL 400 ng/mL protriptyline in water. Allow 5 min for phase separation.36 g/L KH2PO4 with 274 mL 1.. P.MeOH.REFERENCE Tracqui. wash with 3 mL water. nortriptyline Interfering: cyclobenzaprine KEYWORDS serum REFERENCE Wong.) Flow rate: 2 Injection volume: 50 Detector: UV 254 CHROMATOGRAM Retention time: 3.5).. 218-224 SAMPLE Matrix: blood Sample preparation: Automated SPE by ASPEC system. vortex. wash with 3 mL 750 mL/L methanol.10 mM pH 7.5 mL 0. Remove upper heptane phase and add it to 300 jxL 0.ToxicoL. Elute with three aliquots of 300 IxL 0. HPLCVARIABLES Column: 150 X 4. Kintz.Anal. mix thoroughly.7 5 jxm Supelcosil LC-PCN cyanopropyl Mobile phase: MeCN.6 LC-8-DB (Supelco) Mobile phase: MeCN: buffer 35:65 (Buffer was 10 mL/L triethylamine in water adjusted to pH 5. 40.

R. polythiazide.6 Internal standard: protriptyline (4) OTHER SUBSTANCES Extracted: chlordiazepoxide. nordoxepin. hydrochlorothiazide. trichloromethiazide. 1994. benzthiazide. phenylephrine. diphenhydramine.5 mL/L 7% perchloric acid Flow rate: 1. butalbital. centrifuge at 1500 g for 5 min. salicylic acid. phenytoin. add supernatant to SPE cartridge (at ca. nortriptyline. chlorothiazide.H.5 containing 0. flecainide. quinidine. ethanol. haloperidol. methadone. 40.Chem. sulfanilamide. oxazepam. methsuximide.. codeine. warfarin Interfering: acetazolamide. thiazolam. nordiazepam. chlorpromazine. amobarbital.3 Internal standard: trimipramine (9. benzoylecgonine. amphetamine. mexiletine. ephedrine. methaqualone. pentazocine. glutethimide.5 g/L tetramethylammonium perchlorate and 0. cyclosporine. promazine. lidocaine. encainide. plasma REFERENCE Nichols. vortex.. lorazepam. theophylline. prochlorperazine. inject a 50 jxL aliquot of the elu HPLCVARIABLES Guard column: 15 mm 7 jxm Brownlee RP-8 Column: 150 X 4.6) Limit of quantitation: 5 ng/mL . Charlson. methoxyphenteramine. desipramine. doxepin. amiodarone. benzocaine. trazodone. phenteramine. phencyclidine. dextromethorphan. protriptyline. 1 mL water. propranolol. maprotiline. THC-COOH. norfluoxetine. monoacetylmorphine.M.6 5 \xm Ultrasphere Octyl Mobile phase: MeCN: water 37. propoxyphene. imipramine. valproic acid. 1 mL/min). metoprolol. MHPG. oxycodone. hydroxyethylfiurazepam. G. meprobamate. metharbital. norchlorimipramine. propafenone. primidone. Wash with 2 mL water and 1 mL MeCN. hydromorphone. clonazepam. hydrocodone. furosemide.5:62. trimipramine KEYWORDS SPE. pentobarbital. cocaine. 1 mL MeOH. methylcyclothiazide. norverapamil. oxymorphone.CHROMATOGRAM Retention time: 5. and 1 mL 1% potassium carbonate. J. carbamazepine. Clin. dicumerol. cyclothiazide. methoxsalen. ethchlorvynol. acetylmorphine.. ibuprofen. fluoxetine. verapamil Noninterfering: acetaminophen. temazepam.5 Injection volume: 50 Detector: UV 215 CHROMATOGRAM Retention time: 8. diazepam. Automated HPLC assay of fluoxetine and norfluoxetine in serum. diamorphine. chlorimipramine. mephentermine. midazolam. flurazepam. thiopental. normethsuximide. fentanyl. hydroflumethiazide. Lawson. 700 JJLL Serum + 50 |xL 5 |xg/mL trimipramine in 5% potassium bicarbonate + 700 |xL MeCN. desalkylflurazepam. elute with 250 |JLL MeOH: 35% perchloric acid 20:1 by gravity (10 min) then centrifuge for 20 s to remove rest of eluant. bendroflumethiazide. tocainide. cotinine. J. 1312-1316 SAMPLE Matrix: blood Sample preparation: Condition a 1 mL BondElut C18 SPE cartridge with 1 mL 1 M HCl. ethosuximide. ethacrynic acid. morphine. methamphetamine. trifluoperazine. thioridazine.

55. 50 mM ammonium phosphate. elute with mobile phase A for 6 min. R. On-line solid-phase extraction and high-performance liquid chromatographic determination of nortriptyline and amitriptyline in serum. centrifuge at 2500 g for 8 min. 1993. inject a 250 |JLL aliquot of the supernatant onto column A with mobile phase A.6 5 jxm Hypersil CPS CN + 250 X 4. pH 2. When not in use flush column A with mobile phase A. serum REFERENCE Dolezalova. Inject serum or plasma directly onto column A with mobile phase A.Liq.. doxepin. monitor the effluent from column B. After 15 min elute column A onto column B (foreflush) with mobile phase B. Re-equilibrate column A with mobile phase A.6 5 \xm Nucleosil 100 CN .Chromatogr. imipramine.. Every eight injections backflush column A with MeCN: water 70:30. An improved solid phase extraction procedure for the determination of antidepressants in serum by column liquid chromatography. serum REFERENCE Gupta. 2751-2765 SAMPLE Matrix: blood Sample preparation: 500 |xL Serum + 250 |xL 100 mM lauryl sulfate. B 150 X 3 7 jim Separon SGX CN (Tessek) Mobile phase: A MeCN: water 3:97.) Flow rate: 1 Injection volume: 250 Detector: UV 210 CHROMATOGRAM Retention time: 12 Limit of detection: 20-25 ng/mL OTHER SUBSTANCES Extracted: nortriptyline KEYWORDS column-switching. M. B MeCN: buffer 26:74 (Buffer was 50 mM phosphoric acid.6 10 |xm Hypersil MOS C8. elute with mobile phase A to waste. HPLCVARIABLES Column: A 20 X 4. After 2 min remove column A from the circuit.OTHER SUBSTANCES Extracted: clomipramine.Chromatogr. fluvoxamine. 16. desipramine. 1992. nortriptyline. maprotiline KEYWORDS SPE.N. J. B 20 X 4. 579. Elute column B with mobile phase B for 6 min and conduct analysis. backflush contents of column A onto column B with mobile phase B for 4 min. HPLCVARIABLES Column: A Guard-Pak 10 |xm Resolve CN (Waters). 291-297 SAMPLE Matrix: blood Sample preparation: For each 1 mL plasma or serum add 10 jxL 14 |xg/mL trimipramine in MeOH. J. elute column B with mobile phase B. protriptyline Interfering: desmethyltrimipramine. and 28 mM diethylamine. fluoxetine.

2 mL/L butylamine. trifluperidol Interfering: clomipramine. reconstitute in 20 jxL mobile phase. evaporate to dryness under a stream of helium at 30°.. imipramine. 578.8 with 85% phosphoric acid. fluspirilene.) Flow rate: 1. clonazepam. desipramine.54 Internal standard: trimipramine (6. nortriptyline Noninterfering: chlordiazepoxide.5) Limit of detection: 5-10 ng/mL (1 ng/mL with 3 injections) OTHER SUBSTANCES Extracted: metabolites.. haloperidol. nitrazepam. Hiemke. nitrazepam. fluvoxamine. Column switching and high-performance liquid chromatography in the analysis of amitriptyline. spiroperidol. column-switching REFERENCE Hartter. C. triazolam KEYWORDS serum . diazepam. maprotiline. serum. centrifuge at 3000 g for 10 min. desipramine. B MeCN: MeOH: buffer 578:188:235 (Buffer was 10 mM K2HPO4 adjusted to pH 6.Mobile phase: A MeOH: water 5:95. flunitrazepam. lorazepam. clobazam. S. diazepam.) Flow rate: 1 Injection volume: 10 Detector: UV 254 CHROMATOGRAM Retention time: 9 Internal standard: clobazam (5) Limit of detection: 10 ng/mL OTHER SUBSTANCES Extracted: clomipramine. 273-282 SAMPLE Matrix: blood Sample preparation: 1 mL Serum + 3 (xL 20 ng/mL clobazam in methanol -I.1 mL saturated sodium borate (pH adjusted to 11 with 6 M NaOH) + 5 mL n-hexane. inject a 10 |xL aliquot. perazine. pH adjusted to 3 with phosphoric acid.5 Injection volume: 100 Detector: UV 214 CHROMATOGRAM Retention time: 8. clozapine. Separate organic phase. oxazepam. mix for 2 min. haloperidol. HPLCVARIABLES Guard column: 20 mm 40 jxm Pelliguard LC-8 (Supelco) Column: 150 X 4. doxepin KEYWORDS plasma. nortriptyline and hydroxylated metabolites in human plasma or serum. nortriptyline Simultaneous: alprazolam. pimozide. 1992. imipramine.6 Supelco 5 |xm C8 Mobile phase: MeCN: buffer 50:50 (Buffer was 10 mM NaH2PO4 + 1. maprotiline. J.Chromatogr. flurazepam.

B 150 X 4 TSKgel ODS-80TM (Tosoh) Mobile phase: A 50 mM pH 7. fluvoxamine.) Flow rate: 1 Injection volume: 20 Detector: UV 215 CHROMATOGRAM Retention time: 5.5 Internal standard: clovoxamine (3. Elute column B with mobile phase B and monitor the effluent.Chromatogr. R.P. 1991. Rapid and simple high-performance liquid chromatographic determination of tricyclic antidepressants for routine and emergency serum analysis.M. Remove the organic layer and add it to 100 |xL 100 mM HCl. C.. L.5:67. 1989. Lucarelli. 536. HPLCVARIABLES Column: 120 X 4. inject a 20 |xL aliquot of the aqueous layer. 295-302 SAMPLE Matrix: blood Sample preparation: Inject 200 |xL serum onto column A and elute with mobile phase A for 10 min then back-flush column A onto column B with mobile phase B for 4 min.5 potassium phosphate.REFERENCE Segatti. Determination of fluvoxamine in human plasma by high-performance liquid chromatography with ultraviolet detection. J..7 potassium phosphate 32. containing 0. trimipramine KEYWORDS plasma REFERENCE Foglia. Birder.5 with concentrated phosphoric acid. imipramine. G. shake for 30 min.. HPLCVARIABLES Column: A 40 X 4 TSKprecolumn PW (Tosoh). desipramine. B MeCN: 100 mM pH 2. Mangiarotti. M.3) OTHER SUBSTANCES Extracted: chlorimipramine. Grossi.. J. doxepin. centrifuge at 3000 g for 10 min.A. nortriptyline.2 g/L sodium 1-heptanesulfonate Flow rate: 1 Injection volume: 200 Detector: UV 210 CHROMATOGRAM Retention time: 17 Limit of detection: 10 ng/mL . J.Chromatogr. 495. M. Perel. 319-325 SAMPLE Matrix: blood Sample preparation: 1 mL Plasma + 20 |xL 10 |Jig/mL clovoxamine + 120 |xL 2 M NaOH + 4 mL heptane: isopropanol 98:2. Nisi. shake for 20 min. centrifuge at 3000 g for 10 min... Remove column A from circuit and wash with MeCN: water 60:40 for 6 min then with mobile phase A for 10 min. J..5.P.6 5 |xm Nucleosil C8 Mobile phase: MeCN:buffer 36:64 (Buffer was 16 mM KH2PO4 adjusted to pH 2.