Fluoxetine

Molecular formula: C17H18F3NO Molecular weight: 309.3 CAS Registry No.: 54910-89-3, 59333-67-4 (HCI)

SAMPLE Matrix: blood Sample preparation: 1 mL Plasma + doxepin + NaOH + hexane: isoamyl alcohol 98:2, extract. Remove the organic phase and add it to 0.03% phosphoric acid, extract, inject an aliquot of the aqueous phase. HPLC VARIABLES Guard column: C18 Column: 100 X 8 10 fjtm Resolve C8 (Waters) Mobile phase: MeCN: MeOH: 56 mM ammonium acetate :1 M ammonium hydroxide 100: 10:4.5:2.6 Flow rate: 2.5 Detector: UV 220 CHROMATOGRAM Retention time: 16 Internal standard: doxepin (11.6) OTHER SUBSTANCES Extracted: amitriptyline, norfluoxetine, nortriptyline KEYWORDS plasma REFERENCE
el-Yazigi, A.; Chaleby, K.; Gad, A.; Raines, D.A. Steady-state kinetics of fluoxetine and amitriptyline in patients treated with a combination of these drugs as compared with those treated with amitriptyline alone. J.CUn.Pharmacol, 1995, 35, 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, shake gently horizontally for 10 min, centrifuge at 2800 g for 10 min. Remove the lower organic layer and evaporate it to dry ness under vacuum at 45, reconstitute the residue in 100 |xL mobile phase, centrifuge at 2800 g for 5 min, inject a 50 |JLL aliquot of the supernatant. (Buffer was saturated ammonium chloride solution 25% diluted with water, adjusted to pH 9.5 with 25% ammonia solution.) HPLCVARIABLES Column: 300 x 3.9 4 |xm NovaPack C18 Mobile phase: MeOH:THF:buffer 65:5:30 (Buffer was 0.68 g/L (10 mM (sic)) KH2PO4 adjusted to pH 2.6 with concentrated orthophosphoric acid.) (At the end of each session wash the column with water for 1 h and MeOH for 1 h, re-equilibrate for 30 min.) Column temperature: 30 Flow rate: 0.8 Injection volume: 50 Detector: UV 226

CHROMATOGRAM Retention time: 9.14 Limit of detection: <120 ng/mL OTHER SUBSTANCES Extracted: acebutolol, acenocoumarol, acepromazine, aceprometazine, acetaminophen, aconitine, ajmaline, albuterol, alimemazine, alminoprofen, alpidem, alprazolam, alprenolol, amisulpride, amodiaquine, amoxapine, aspirin, astemizole, atenolol, benazepril, benperidol, benzocaine, benzoylecgonine, bepridil, betaxolol, bisoprolol, bromazepam, brompheniramine, bumadizone, bupivacaine, buprenorphine, buspirone, caffeine, carbamazepine, carbinoxamine, carpipramine, carteolol, celiprolol, cetirizine, chlorambucil, chlordiazepoxide, chlormezanone, chlorophenacinone, chloroquine, chlorpheniramine, chlorpromazine, chlorpropamide, cibenzoline, cicletanine, clemastine, clobazam, clomipramine, clonazepam, clonidine, clorazepate, clozapine, cocaine, codeine, colchieine, cyamemazine, cyclizine, cycloguanil, cyproheptadine, cytarabine, dacarbazine, debrisoquine, demexiptiline, desipramine, dextromethorphan, dextromoramide, dextropfopoxyphene, diazepam, diazoxide, diclofenac, dihydralazine, diltiazem, diphenhydramine, dipyridamole, disopyramide, dosulepine, doxepin, doxylamine, droperidol, ephedrine, estazolam, fenfluramine, fenoprofen, fentiazac, flecainide, floctafenine, flumazenil, flunitrazepam, fluphenazine, flurbiprofen, fluvoxamine, glibenclamide, glibornuride, glipizide, glutethimide, haloperidol, histapyrrodine, hydroxychloroquine, hydroxyzine, ibuprofen, imipramine, iproniazid, ketamine, ketoprofen, labetalol, levomepromazine, lidocaine, Iidoflazine, lisinopril, loperamide, loprazolam, loratadine, lorazepam, loxapine, medazepam, medifoxamine, mefenamic acid, mefenidramine, mefloquine, melphalan, meperidine, mephenesin, mephentermine, mepivacaine, metapramine, metformin, methadone, methaqualone, methocarbamol, methotrexate, metipranolol, metoclopramide, metoprolol, mexiletine, mianserine, midazolam, minoxidil, moclobemide, moperone, morphine, nadolol, nalbuphine, nalorphine, naloxone, naltrexone, naproxen, nialamide, nicardipine, nifedipine, niflumic acid, nimodipine, nitrazepam, nitrendipine, nizatidine, nomifensine, omeprazole, opipramol, oxazepam, oxprenolol, penbutolol, penfluridol, pentazocine, phencyclidine, phenobarbital, phenol, phenylbutazone, pimozide, pindolol, pipamperone, piroxicam, prazepam, prazosin, prilocaine, procainamide, procarbazine, proguanil, promethazine, propafenone, propranolol, protriptyline, pyrimethamine, quinidine, quinine, quinupramine, ramipril, ranitidine, reserpine, ritodrine, secobarbital, sotalol, strychnine, sulfinpyrazole, sulindac, sulpride, sultopride, suriclone, temazepam, tenoxicam, terbutaline, terfenadine, tetracaine, tetrazepam, thiopental, thioproperazine, thioridazine, tianeptine, tiapride, tiaprofenic acid, ticlopidine, timolol, tofisopam, tolbutamide, toloxatone, trazodone, triazolam, trifluoperazine, trifluperidol, trimipramine, triprolidine, verapamil, viloxazine, vinblastine, vincristine, vindesine, warfarin, yohimbine, zolpidem, zopiclone, zorubicine Interfering: amitriptyline, daunorubicin, etodolac, indomethacin, maprotiline, nortriptyline, tioclomarol, tropatenine KEY WORDS whole blood; plasma REFERENCE
Tracqui, A.; Kintz, P.; Mangin, P. Systematic toxicological analysis using HPLC/DAD. J.Forensic Sci., 1995, 40, 254-262

SAMPLE Matrix: blood Sample preparation: Automated SPE by ASPEC system. Condition a C18 Clean-Up SPE cartridge (CEC 18111, Worldwide Monitoring) with 2 mL MeOH then 2 mL water. 1 mL Plasma + 1 mL 400 ng/mL protriptyline in water, vortex, add to the SPE cartridge, wash with 3 mL water, wash with 3 mL 750 mL/L methanol. Elute with three aliquots of 300 |xL 0.1 M ammonium acetate in MeOH. Add 0.5 mL 0.5 M NaOH and 4 mL 50 mL/L

isopropanol in heptane to eluate, mix thoroughly. Allow 5 min for phase separation. Remove upper heptane phase and add it to 300 |xL 0.1 M phosphoric acid (pH 2.5), mix, separate, inject a 100 |xL aliquot of the aqueous phase. HPLC VARIABLES Guard column: LC-8-DB (Supelco) Column: 150 X 4.6 LC-8-DB (Supelco) Mobile phase: MeCN: buffer 35:65 (Buffer was 10 mL/L triethylamine in water adjusted to pH 5.5 with glacial acetic acid.) Flow rate: 2 Injection volume: 100 Detector: UV 228 CHROMATOGRAM Retention time: 6.5 Internal standard: protriptyline (4) OTHER SUBSTANCES Extracted: acetazolamide, amitriptyline, chlordiazepoxide, chlorimipramine, chlorpromazine, desipramine, dextromethorphan, diazepam, diphenhydramine, doxepin, encainide, fentanyl, flecainide, haloperidol, ibuprofen, imipramine, lidocaine, maprotiline, methadone, methaqualone, mexiletine, midazolam, nordiazepam, nordoxepin, norfluoxetine, nortriptyline, norverapamil, pentazocine, promazine, propafenone, propoxyphene, propranolol, protriptyline, quinidine, temazepam, trazodone, trimipramine, verapamil Noninterfering: acetaminophen, acetylmorphine, amiodarone, amobarbital, amphetamine, bendroflumethiazide, benzocaine, benzoylecgonine, benzthiazide, butalbital, carbamazepine, chlorothiazide, clonazepam, cocaine, codeine, cotinine, cyclosporine, cyclothiazide, desalkylflurazepam, diamorphine, dicumerol, ephedrine, ethacrynic acid, ethanol, ethchlorvynol, ethosuximide, furosemide, glutethimide, hydrochlorothiazide, hydrocodone, hydroflumethiazide, hydromorphone, lorazepam, mephentermine, meprobamate, methamphetamine, metharbital, methoxsalen, methoxyphenteramine, methsuximide, methylcyclothiazide, metoprolol, MHPG, monoacetylmorphine, morphine, normethsuximide, oxazepam, oxycodone, oxymorphone, pentobarbital, phencyclidine, phenteramine, phenylephrine, phenytoin, polythiazide, primidone, prochlorperazine, salicylic acid, sulfanilamide, THC-COOH, theophylline, thiazolam, thiopental, thioridazine, tocainide, trichloromethiazide, trifluoperazine, valproic acid, warfarin Interfering: flurazepam, hydroxyethylflurazepam, norchlorimipramine KEYWORDS plasma; SPE REFERENCE
Nichols, J.H.; Charlson, J.R.; Lawson, G.M. Automated HPLC assay of fluoxetine and norfluoxetine in serum. Clin.Chem., 1994, 40, 1312-1316

SAMPLE Matrix: blood Sample preparation: 1 mL Serum or plasma + 30 |JLL trimipramine in MeOH + 200 |JLL 0.33 M NaOH, shake 5 s, add 7 mL n-hexane: iso-amyl alcohol 985:15, shake 20 min, centrifuge at 2100 g for 5 min. Remove organic phase and add 200 |xL 0.1 M HCl to it, shake for 1 min, discard organic phase, inject 30 JAL of aqueous phase. HPLCVARIABLES Guard column: 10 mm 10 |j,m Bischoff C18 Column: 125 X 4 5 |xm Ecotube Nucleosil C8 Mobile phase: MeCN: water: diethylamine: PicB5 370:630:0.4:25 (PicB5 is water-MeOH1-pentanesulfonic acid.)

Column temperature: 55 Flow rate: 1.7 Injection volume: 30 Detector: UV 230

CHROMATOGRAM

Retention time: 5.6 Internal standard: trimipramine (8.4) Limit of detection: 10 ng/mL
OTHER SUBSTANCES

Simultaneous: norfluoxetine Noninterfering: alprazolam, amitriptyline, bromazepam, clomipramine, clorazepate, desipramine, diazepam, flunitrazepam, fluvoxamine, imipramine, lorazepam, nortriptyline, oxazepam, triazolam
KEYWORDS

serum; plasma
REFERENCE
el Maanni, A.; Combourieu, L; Bonini, M.; Creppy, E.E. Fluoxetine, an antidepressant, and norfluoxetine, its metabolite, determined by HPLC with a C8 column and ultraviolet detection. Clin.Chem., 1993, 39, 1749-1750

SAMPLE

Matrix: blood Sample preparation: Condition a 1 mL BondElut C18 SPE cartridge with 1 mL 1 M HCl5 1 mL MeOH, 1 mL water, and 1 mL 1% potassium carbonate. 700 JJLL Serum + 50 |xL 5 jjLg/mL protriptyline in 5% potassium bicarbonate + 700 J U L L MeCN, vortex, centrifuge at 1500 g for 5 min, add supernatant to SPE cartridge (at ca. 1 mL/min). Wash with 2 mL water and 1 mL MeCN, elute with 250 jxL MeOH: 35% perchloric acid 20:1 by gravity (10 min) then centrifuge for 20 s to remove rest of eluate, inject a 50 |xL aliquot of the eluate.
HPLC VARIABLES

Guard column: 15 mm 7 (xm Brownlee RP-8 Column: 150 X 4.6 5 |xm Ultrasphere Octyl Mobile phase: MeCN.water 37.5:62.5 containing 0.5 g/L tetramethylammonium perchlorate and 0.5 mL/L 7% perchloric acid Flow rate: 1.5 Injection volume: 50 Detector: UV 226
CHROMATOGRAM

Retention time: 10.9 Internal standard: protriptyline (6.6) Limit of quantitation: 5 ng/mL
OTHER SUBSTANCES

Extracted: amitriptyline, clomipramine, desipramine, doxepin, fluvoxamine, imipramine, maprotiline, nortriptyline, trimipramine Interfering: desmethylclomipramine
KEYWORDS

serum; SPE

REFERENCE
Gupta, R.N. An improved solid phase extraction procedure for the determination of antidepressants in serum by column liquid chromatography. J.Liq.Chromatogr., 1993, 16, 2751-2765

SAMPLE Matrix: blood Sample preparation: Add 10 |xL 20 jxg/mL oxaprotiline in MeOH to 990 JJLL plasma or serum. Inject 100 |xL plasma or serum onto column A with mobile phase A and elute to waste, after 15 min elute column A onto column B with mobile phase B for 2 min. Remove column A from circuit and re-equilibrate it with mobile phase A for 5 min. Chromatograph on column B with mobile phase B. HPLCVARIABLES Column: A 20 X 4.6 10 jim Hypersil MOS C8; B 20 X 4.6 5 |jim Hypersil CPS CN + 250 X 4.6 5 |xm Nucleosil 100 CN Mobile phase: A MeOH: water 5:95; B MeOH: MeCN: 10 mM pH 6.8 potassium phosphate buffer 188:578:235 Flow rate: 1.5 Injection volume: 100 Detector: UV 214 CHROMATOGRAM Retention time: 8.7 Internal standard: oxaprotiline (9.5) Limit of detection: 20 ng/mL OTHER SUBSTANCES Simultaneous: clozapine, desipramine, doxepin, fluvoxamine, imipramine, maprotiline, metoclopramide, norfluoxetine, nortriptyline Noninterfering: carbamazepine, chlordiazepoxide, clobazam, diazepam, flurazepam, fluspirilene, haloperidol, lorazepam, nitrazepam, nordiazepam, oxazepam, perazine, pimozide, spiroperidol, trifluperidol Interfering: amitriptyline, clomipramine KEYWORDS plasma; serum; column-switching REFERENCE
Hartter, S.; Wetzel, H.; Hiemke, C. Automated determination of fluvoxamine in plasma by columnswitching high-performance liquid chromatography. Clin.Chem., 1992, 38, 2082-2086

SAMPLE Matrix: blood Sample preparation: 1 mL Plasma + 1 mL 0.6 M pH 9.8 carbonate buffer + 40 /ULL 5 |xg/mL maprotiline in 10 mM HCl + 5 mL 200 g/L ethyl acetate in n-heptane, mix by rocking for 10 min, centrifuge at 1500 g for 10 min. Remove organic layer and add it to 150 |xL 100 mM HCl, mix 10 min, centrifuge at 1500 g for 10 min. Discard organic layer and evaporate aqueous layer at 45 in a vacuum centrifuge for 1 h. Take up residue in 50 |xL 1 M pH 10.3 carbonate buffer and 25 |xL 10 mg/mL dansyl chloride in MeCN, vortex, allow to react at room temperature for 45 min, evaporate at 45 in a vacuum centrifuge for 20 min, reconstitute in 125 |xL MeCN: water 75:25, vortex, centrifuge for 3-5 min, inject a 25-40 jiL aliquot. HPLCVARIABLES Column: 250 X 4.6 5 |xm Supelcosil LC-18

Mobile phase: MeCN: 25 mM KH2PO4 75:25 containing 500 |xL/L orthophosphoric acid and 600 |xL/L n-butylamine Flow rate: 2 Injection volume: 25-40 Detector: F ex 235 em 470 (cut-off)
CHROMATOGRAM

Retention time: 10.4 Internal standard: maprotiline (12.8) Limit of quantitation: 3 ng/mL
OTHER SUBSTANCES

Simultaneous: amoxapine, clovoxamine, desipramine, fenfluramine, fluvoxamine, norfluoxetine, nortriptyline, propranolol, protriptyline, sertraline Noninterfering: amitriptyline, atenolol, bupropion, carbamazepine, chlordiazepoxide, citalopram, clomipramine, clozapine, cyclobenzaprine, doxepin, imipramine, loxapine, metoprolol, mianserin, moclobemide, nomifensine, pindolol, thioridazine, tranylcypromine, trazodone, trimipramine
KEYWORDS

plasma
REFERENCE
Suckow, R.F.; Zhang, M.R; Cooper, T.B. Sensitive and selective liquid-chromatographic assay of fluoxetine and norfluoxetine in plasma with fluorescence detection after precolumn derivatization. Clin.Chem., 1992, 38, 1756-1761

SAMPLE

Matrix: blood Sample preparation: 2 mL Plasma + 160 |xL 10 |xg/mL clomipramine in MeOH, vortex, add 2 mL 1 M NaOH, vortex, add 5 mL n-hexane: isoamyl alcohol 99:1, rotate for 5 min, centrifuge for 10 min. Remove organic layer and add it to 200 |xL 0.05% phosphoric acid, rotate for 5 min, centrifuge for 10 min, remove lower aqueous layer and inject a 25-50 IxL aliquot of it.
HPLCVARIABLES

Guard column: Bondapak/Corasil C18 Column: 300 X 4.6 jxBondapak C18 Mobile phase: MeCN: 50 mM KH2PO4 adjusted to pH 4.7 with KOH 40:60 Column temperature: 50 Flow rate: 2 Injection volume: 25-50 Detector: UV 214
CHROMATOGRAM Retention time: 7

Internal standard: clomipramine (9) Limit of detection: 6 ng/mL

OTHER SUBSTANCES

Simultaneous: amoxapine, chlordiazepoxide, chlorpromazine, cimetidine, desipramine, diazepam, doxepin, flurazepam, imipramine, lorazepam, norfluoxetine, nortriptyline, oxazepam, pentobarbital, perphenazine, phenobarbital, phenytoin, prochlorperazine, secobarbital, thioridazine, trifluoperazine Noninterfering: acetaminophen, codeine, meperidine Interfering: amitriptyline, propoxyphene

KEYWORDS plasma REFERENCE

Wong, S.H.; Dellafera, S.S.; Fernandes, R.; Kranzler, H. Determination of fluoxetine and norfluoxetine by high-performance liquid chromatography. J.Chromatogr., 1990, 499, 601-608

SAMPLE Matrix: blood Sample preparation: 1 mL Serum H - 50 |xL 1 mg/mL reduced haloperidol in water H -2 mL 0.5 M NaH2PO4 adjusted to pH 10 with 10 M NaOH, vortex 3-5 s, add 5 mL hexane: isoamyl alcohol 97:3, shake 20 min, centrifuge at 1000 g for 10 min. Remove organic layer and add 1 mL 0.1 M HCl to it, shake for 20 min, centrifuge for 10 min, discard organic layer. Add 1 mL 0.2 M NaOH to aqueous layer, add 5 mL hexane: isoamyl alcohol 97:3, shake for 20 min, centrifuge for 10 min. Remove organic layer, add 1 drop of 0.3 M HCl in MeOH, evaporate under nitrogen at 40, reconstitute with 250 jxL mobile phase, vortex, inject 100 |JLL aliquot. HPLCVARIABLES Guard column: C18 Guard-Pak (Waters no. 88070) Column: Nova-Pak phenyl (Waters no. 10656) Mobile phase: MeCN: buffer 40:60 (Buffer was 600 mL water H - 1 mL triethylamine, adjusted to pH 5.5 with acetic acid.) Flow rate: 1.7 Injection volume: 100 Detector: UV 226 CHROMATOGRAM Retention time: 7.3 Internal standard: reduced haloperidol (3.9) Limit of detection: 15 ng/mL OTHER SUBSTANCES Simultaneous: alprazolam, amitriptyline, amoxapine, chlordiazepoxide, chlorimipramine, clonazepam, demoxepam, diazepam, doxepin, halazepam, haloperidol, lorazepam, maprotiline, norfluoxetine, nortriptyline, oxazepam, temazepam, trazodone, trimipramine Interfering: desipramine, imipramine, loxapine, protriptyline KEYWORDS serum REFERENCE
Orsulak, RJ.; Kenney, J.T.; Debus, J.R.; Crowley, G.; Wittman, RD. Determination of the antidepressant fluoxetine and its metabolite norfluoxetine in serum by reversed-phase HPLC with ultraviolet detection. Clin.Chem., 1988, 34, 1875-1878

SAMPLE Matrix: blood, tissue Sample preparation: Blood or serum. 1 mL Blood or serum + 1 |xg cianopramine + 1 mL water, vortex, add 1 mL 200 mM sodium carbonate, vortex, add 6 mL hexane: 1-butanol 95:5, gently agitate for 30 min, centrifuge at 2500 g for 5 min. Remove the organic layer and add it to 100 |xL 0.2% phosphoric acid, agitate gently for 30 min, centrifuge for 5 min. Remove the organic layer and inject a 30 u,L aliquot of the aqueous layer. Liver homogenate. 0.5 mL Liver homogenate + 10 juig cianopramine + 500 JULL 2% sodium tetraborate + 8 mL hexane: 1-butanol 95:5, gently agitate for 30 min, centrifuge at 2500 g for 5 min. Remove the organic layer and add it to 400 |xL 0.2% phosphoric acid, agitate

gently for 30 min, centrifuge for 5 min. Remove the organic layer and inject a 30 |xL aliquot of the aqueous layer. HPLCVARIABLES Guard column: 15 X 3.2 7 jjim RP-18 Newguard (Applied Biosystems) Column: 100 X 4.6 5 |xm Brownlee Spheri-5 RP-18 Mobile phase: MeCN: 100 mM NaH2PO4: diethylamine 40:57.5:2.5 Flow rate: 2 Injection volume: 30 Detector: UV 220 CHROMATOGRAM Retention time: 6.70 Internal standard: cianopramine (8.93) Limit of detection: 100 ng/mL OTHER SUBSTANCES Extracted: amitriptyline, benztropine, brompheniramine, chlorpheniramine, chlorpromazine, clomipramine, cyproheptadine, desipramine, diphenhydramine, dothiepin, doxepin, haloperidol, imipramine, loxapine, maprotiline, meperidine, mesoridazine, methadone, metoclopramide, mianserin, moclobemide, nomifensine, nordoxepin, norfluoxetine, norpropoxyphene, northiaden, nortriptyline, pentobarbital, pheniramine, promethazine, propoxyphene, propranolol, protriptyline, quinidine, quinine, sulforidazine, thioridazine, thiothixene, tranylcypromine, trazodone, trihexyphenidyl, trimipramine, triprolidine Noninterfering: dextromethorphan, norphetidine, phenoxybenzamine, prochlorperazine, trifluoperazine Interfering: amoxapine KEYWORDS serum; whole blood; liver REFERENCE
Mclntyre, LM.; King, C.V.; Skafidis, S.; Drummer, O.H. Dual ultraviolet wavelength high-performance liquid chromatographic method for the forensic or clinical analysis of seventeen antidepressants and some selected metabolites. J.Chromatogr., 1993, 621, 215-223

SAMPLE Matrix: blood, tissue Sample preparation: Homogenize (Brinkman Polytron) tissue with 5-10 volumes of water. 500 |xL Plasma or tissue homogenate + 500 \xL water + 50 JULL 2 [xg/wiL IS in water + 100 |JLL 1 M NaOH, vortex gently, add 5 mL hexane: butanol 99.7:0.3, shake mechanically at 125-150 cycles/min for 30 min, centrifuge at 2000 g for 15 min. Remove the organic layer and mix it with 100 |mL 200 JULM R-(-)-l-(l-naphthyl)ethyl isocyanate in hexane, evaporate to dryness at 50-55 over 20-30 min, dry more vigorously when all the hexane is gone, reconstitute with 200 jxL mobile phase, inject a 75 |xL aliquot. HPLCVARIABLES Column: 250 X 4.6 5 \m Apex silica (Jones Chromatography) Mobile phase: Isooctane: THF 70:30 Column temperature: 35 Flow rate: 1 Injection volume: 75 Detector: F ex 218 em 333 CHROMATOGRAM Retention time: 8.3 (S), 9.3 (R)

Internal standard: S-nornisoxetine (15) Limit of detection: 5 ng/mL (plasma); 25 ng/g (tissue) OTHER SUBSTANCES Extracted: metabolites, norfluoxetine KEYWORDS chiral; derivatization; normal phase; plasma; silylate all glassware REFERENCE
Potts, B.D.; Parli, CJ. Analysis of the enantiomers of fluoxetine and norfluoxetine in plasma and tissue using chiral derivatization and normal-phase liquid chromatography. J.Liq.Chromatogr., 1992, 15, 665-681

SAMPLE Matrix: bulk Sample preparation: Prepare a 2 mg/mL solution of fluoxetine hydrochloride in mobile phase, inject a 10 |xL solution. HPLCVARIABLES Column: 150 X 4.6 3 jim Exsil 100A/ODS-B octadecylsilane (Keystone) Mobile phase: MeCN: THF: buffer 15:10:75 (Buffer was 50 mM ammonium acetate adjusted to pH 5.75 with 50 mM acetic acid.) Flow rate: 1 Injection volume: 10 Detector: UV 214 CHROMATOGRAM Retention time: 31.9 OTHER SUBSTANCES Simultaneous: impurities, meta isomer REFERENCE
Lacroix, P.M.; Yat, RN.; Lovering, E.G. Liquid chromatographic methods for fluoxetine hydrochloride, its meta isomer, and related compounds in raw materials. J.AOAC Int., 1995, 78, 334-339

SAMPLE Matrix: bulk Sample preparation: Reflux 1.23 g fluoxetine with 788 mg (R)-(-)-l-(l-naphthyl)ethylisocyanate in 25 mL toluene for 2 h, evaporate to dryness under reduced pressure, reconstitute, inject an aliquot. HPLCVARIABLES Column: 250 X 4.6 silica (IBM) Mobile phase: Dichloromethane:MeOH 99.75:0.25 Flow rate: 2 Detector: UV 254 CHROMATOGRAM Retention time: 6.69 (S), 7.52 (R) KEYWORDS derivatization; chiral; normal phase

REFERENCE
Robertson, D.W.; Krushinski, J.H.; Fuller, R.W.; Leander, J.D. Absolute configurations and pharmacological activities of the optical isomers of fluoxetine, a selective serotonin-uptake inhibitor. J.Med.Chem., 1988, 31, 1412-1417

SAMPLE

Matrix: formulations Sample preparation: Weigh out 4 mL of 1 mg/mL solution and make up to 100 mL with mobile phase, inject 20 |xL aliquot.
HPLC VARIABLES

Column: 250 X 4.6 Zorbax cyano special Mobile phase: MeCN.buffer 50:50 (Buffer was 1% triethylamine, pH adjusted to 6 with concentrated phosphoric acid.) Flow rate: 1 Injection volume: 20 Detector: UV 215
CHROMATOGRAM

Retention time: 11
OTHER SUBSTANCES

Simultaneous: benzoic acid

KEYWORDS

syrup; elixir; stability-indicating

REFERENCE
Peterson, J.A.; Risley, D.S.; Anderson, RN.; Hostettler, K.F. Stability of fluoxetine hydrochloride in fluoxetine solution diluted with common pharmaceutical diluents. Am.J.Hosp.Pharm., 1994, 51, 13421345

SAMPLE

Matrix: solutions
HPLC VARIABLES

Column: 250 X 4.6 5 jjum Supelcosil LC-DP (A) or 250 X 4 5 ^m 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: 12.20 (A), 7.07 (B)

OTHER SUBSTANCES

Also analyzed: acebutolol, acepromazine, acetaminophen, acetazolamide, acetophenazine, albuterol, alprazolam, amitriptyline, amobarbital, amoxapine, antipyrine, atenolol, atropine, 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, colchi-

cine, cyclizine, cyclobenzaprine, dantrolene, desipramine, diazepam, diclofenac, diflunisal, diltiazem, diphenhydramine, diphenidol, diphenoxylate, dipyridamole, disopyramide, dobutamine, doxapram, doxepin, droperidol, encainide, ethidium bromide, ethopropazine, fenoprofen, fentanyl, flavoxate, 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.Chromatogr.A, 1995, 692, 103-119

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 150 X 4.6 Supelcosil LC-ABZ Mobile phase: MeCN: 25 mM pH 6.9 potassium phosphate buffer 35:65 Flow rate: 1.5 Injection volume: 25 Detector: UV 254
CHROMATOGRAM

Retention time: 11.460

OTHER SUBSTANCES

Also analyzed: 6-acetylmorphine, amiloride, amphetamine, benzocaine, benzoylecgonine, caffeine, cocaine, codeine, doxylamine, glutethimide, hexobarbital, hypoxanthine, levorphanol, LSD, meperidine, mephobarbital, methadone, methylphenidate, methyprylon, Nnorcodeine, oxazepam, oxycodone, phenylpropanolamine, prilocaine, procaine, terfenadine
REFERENCE
Ascah, T.L. Improved separations of alkaloid drugs and other substances of abuse using Supelcosil LCABZ column. Supelco Reporter, 1993, 12(3), 18-21

SAMPLE

Matrix: tissue

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Sample preparation: Homogenize 1 mL liver in 5 mL water. Centrifuge 1 mL homogenate, add the supernatant to 1.2 (xg clomipramine, add 75 |JLL MeOH, add 75 |JLL MeCN, add 100 \LL 1 M HCl, vortex, centrifuge, inject an aliquot of the supernatant.
HPLC VARIABLES

Column: 300 X 3.9 ^Bondapak C18 Mobile phase: MeCN: 50 mM potassium phosphate buffer 35:65 Flow rate: 1.3 Detector: UV 226
CHROMATOGRAM

Internal standard: clomipramine

OTHER SUBSTANCES

Extracted: metabolites, norfluoxetine Also analyzed: sertraline

KEYWORDS

mouse; liver
REFERENCE
von Moltke, L.L.; Greenblatt, D.J.; Cotreau-Bibbo, M.M.; Duan, S.X.; Harmatz, J.S.; Shader, R.I. Inhibition of desipramine hydroxylation in vitro by serotonin-reuptake-inhibitor antidepressants and by quinidine and ketoconazole: A model system to predict drug interactions in vivo. J.Pharmacol.Exp.Ther., 1994,268, 1278-1283

ANNOTATED BIBLIOGRAPHY

Hubmann, M.R.; Waschgler, R.; Moll, W.; Conca, A.; Konig, P. Simultaneous drug monitoring of citalopram, clozapine, fluoxetine, maprotiline, and trazodone by HPLC analysis (Abstract 41). Ther.Drug Monit., 1995, 17, 393 [simultaneous citalopram, clozapine, maprotiline, trazodone; LOQ 50 ng/mL] Joron, S.; Robert, H. Simultaneous determination of antidepressant drugs and metabolites by HPLC. Design and validation of a simple and reliable analytical procedure. Biomed.Chromatogr., 1994, 8, 158-164 [simultaneous amineptine, amitriptyline, amoxapine, clomipramine, demexiptiline, desipramine, dosulepine, doxepin, doxepine, fluvoxamine, imipramine, maprotiline, medifloxamine, mianserine, opipramol, quinupramine, tianeptine, toloxatone, trazodone, trimipramine, viloxazine; LOQ 3-17 ng/mL; plasma] Thomare, P.; Wang, K.; Van Der Meersch-Mougeot, V.; Diquet, B. Sensitive micromethod for column liquid chromatographic determination of fluoxetine and norfluoxetine in human plasma. J.Chromatogr., 1992, 583, 217221 [plasma; extracted metabolites; LOD 2 ng/mL; simultaneous amineptine, amitriptyline, chlordiazepoxide, chlorpromazine, clomipramine, clonazepam, clorazepate, desipramine, diazepam, doxepin, flunitrazepam, fluvoxamine, imipramine, levomepromazine, lorazepam, loxapine, maprotiline, mefloquine, nortriptyline, oxazepam, thioridazine] Peyton, A.L.; Carpenter, R.; Rutkowski, K. The stereospecific determination of fluoxetine and norfluoxetine enantiomers in human plasma by high-pressure liquid chromatography (HPLC) with fluorescence detection. Pharm.Res., 1991, 8, 1528-1532 Gupta, R.N.; Steiner, M. Determination of fluoxetine and norfluoxetine in serum by liquid chromatography with fluorescence detection. J.Liq.Chromatogr., 1990, 13, 3785-3798 [extracted norfluoxetine; serum; fluorescence detection; SPE; protriptyline (IS); simultaneous amitriptyline, nortriptyline]