Buspirone

Molecular formula: C2IH31N5O2 Molecular weight: 385.5 CAS Registry No.: 36505-84-7 (buspirone), 33386-08-2 (buspirone hydrochloride)

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 dryness under vacuum at 45, reconstitute the residue in 100 jxL. mobile phase, centrifuge at 2800 g for 5 min, inject a 50 |xL 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 236 CHROMATOGRAM Retention time: 5.39 Limit of detection: <120 ng/mL OTHER SUBSTANCES Also analyzed: acebutolol, acenocoumarol, acepromazine, aceprometazine, acetaminophen, aconitine, ajmaline, albuterol, alimemazine, alminoprofen, alpidem, alprazolam, alprenolol, amisulpride, amitriptyline, 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, colchicine, cyamemazine, cyclizine, cycloguanil, cyproheptadine, cytarabine, dacarbazine, daunorubicin, debrisoquine, demexiptiline, desipramine, dextromethorphan, dextromoramide, dextropropoxyphene, diazepam, diazoxide, diclofenac, dihydralazine, diltiazem, diphenhydramine, dipyridamole, disopyramide, dosulepine, doxepin, doxylamine, droperidol, ephedrine, estazolam, etodolac, fenfluramine, fenoprofen, fentiazac, flecainide, floctafenine, flumazenil, flunitrazepam, fluoxetine, fluphenazine, flurbiprofen, fluvoxamine, glibenclamide, glibornuride, glipizide, glutethimide, haloperidol, histapyrrodine, hydroxychloroquine, hydroxyzine, ibuprofen, imipramine, indomethacin, iproniazid, ketamine, ketoprofen, labetalol, levomepromazine, lidocaine, lidoflazine, lisinopril, loperamide, loprazolam, loratadine, lorazepam, loxapine, maprotiline, 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, ni-

flumic acid, nimodipine, nitrazepam, nitrendipine, nizatidine, nomifensine, nortriptyline, 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, tioclomarol, tofisopam, tolbutamide, toloxatone, trazodone, triazolam, trifluoperazine, trifluperidol, trimipramine, triprolidine, tropatenine, verapamil, viloxazine, vinblastine, vincristine, vindesine, warfarin, yohimbine, zolpidem, zopiclone, zorubicine KEYWORDS 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: Condition a 3 mL Baker carboxylic acid SPE cartridge with 5 mL 2 M HCl and 10 mL water. Mix 500 |JLL plasma + 5 mL water and add to column. Wash with 5 mL water and elute with 2 mL 1 M formic acid. Evaporate eluate to dryness under vacuum and dissolve residue in 200 |xL mobile phase, inject a 20 |JLL aliquot. HPLCVARIABLES Guard column: 5 |xm Cyanonitrile Column: 33 X 4.6 Supelcosil LC-CN Mobile phase: MeCN: 20 mM pH 7 potassium phosphate buffer 43:57 adjusted to pH 7.34 with 0.5 M KOH Flow rate: 0.4 Injection volume: 20 Detector: E, ESA Coulochem model 5100 A, two porous graphite working electrodes and associated palladium reference electrodes, +0.55 V for first electrode, +0.70 V for second (monitoring) electrode CHROMATOGRAM Retention time: 10 Limit of detection: 1 ng/mL OTHER SUBSTANCES Extracted: metabolites, l-(2-pyrimidinyl)piperazine KEYWORDS plasma; mouse; SPE REFERENCE
Betto, P.; Meneguz, A.; Ricciarello, G.; Pichini, S. Simultaneous high-performance liquid chromatographic analysis of buspirone and its metabolite l-(2-pyrimidinyl)-piperazine in plasma using electrochemical detection. J.Chromatogr., 1992, 575, 117-121

SAMPLE Matrix: blood

Sample preparation: Condition a 1 mL C18 Supelco SPE cartridge with two 1 mL portions of MeOH and two 1 mL portions of 50 mM KH2PO4 adjusted to pH 7.2 with 2 M NaOH, add 2 mL serum and flush through 2-5 mL air. Wash with two 1 mL portions of 50 mM KH2PO4 adjusted to pH 7.2 with 2 M NaOH, wash with two 1 mL and one 0.5 mL portions of MeOH: water 1:1 and flush through with 2-5 mL of air. Elute with 1 mL MeCN: triethylamine 99:1. Evaporate solvent at 37 under a stream of air and dissolve residue in 100 IJLL of MeCN: 5 mM KH2PO4 + 0.1% triethylamine adjusted to pH 2.5 with orthophosphoric acid 45:55, centrifuge at 5000 g for 10 min, inject a 70 |xL aliquot onto column A with mobile phase A and elute to waste, switch a 0.7 min fraction onto column B (retention time is ca. 3.6 min) and chromatograph on column B with mobile phase B, monitor the effluent from column B. HPLCVARIABLES Column: A 150 X 4.6 5 jxm Spherisorb ODS2; B 150 X 4.6 5 fxm Spherisorb ODS2 Mobile phase: A MeCN: 5 mM KH2PO4 + 0.1% triethylamine adjusted to pH 2.5 with orthophosphoric acid 45:55; B MeCN: 5 mM KH2PO4 + 0.2% triethylamine adjusted to pH 2.5 with orthophosphoric acid 55:45, containing 5 mM sodium lauryl sulfate Flow rate: 1.2 Injection volume: 70 Detector: UV 235 CHROMATOGRAM Retention time: 7 (on column B) Limit of detection: 0.2 ng/mL KEYWORDS serum; column-switching; heart-cut; SPE; pharmacokinetics REFERENCE
Kristjansson, F. Sensitive determination of buspirone in serum by solid-phase extraction and two-dimensional high-performance liquid chromatography. J.Chromatogr., 1991, 566, 250-256

SAMPLE Matrix: blood Sample preparation: 2 mL Plasma + 50 |xL 400 ng/mL gepirone in EtOH + 0.5 mL 1 M NaOH + 4 mL ethyl acetate, shake slowly mechanically for 10 min, centrifuge at 400 g for 10 min. Freeze lower plasma layer in dry ice/acetone and decant ethyl acetate into a centrifuge tube containing 2 mL 50 mM HGl, shake 10 min, centrifuge 400 g 10 min, remove, discard organic phase. Add 500 |xL 4 M ammonia solution and 400 |xL butyl acetate to the aqueous phase, mix for 15-20 sec, centrifuge at 400 g for 10 min. Remove organic layer and evaporate it to dryness at 30 under vacuum. Reconstitute in 90 \xL mobile phase, inject an aliquot. HPLCVARIABLES Guard column: 5 |xm Cyanonitrile (Anachem) Column: 10 X 4.6 5 |xm cyanonitrile (Brownlee) Mobile phase: MeCN: 40 mM potassium phosphate buffer adjusted to pH 6.6 with 2 M KOH 34:66 (Columns were initially conditioned with 50 mL water; then 5 mM pH 4.8 sodium acetate buffer; then MeCN: 5 mM pH 4.8 sodium acetate buffer 40:60; then 5 mM pH 4.8 potassium acetate; then mobile phase.) Flow rate: 1.5 Injection volume: 50 Detector: E, ESA Coulochem model 5100 A, Model 5020 guard cell, guard cell 0.3 V, detector 0.55 V for first electrode and 0.70 V for second electrode CHROMATOGRAM Retention time: 7.3

Internal standard: gepirone (5.4) Limit of detection: 0.5 ng/mL OTHER SUBSTANCES Simultaneous: l-(2-pyrimidinyl)piperazine, metabolites, amitriptyline, chlorpromazine, clomipramine, fluphenazine, imipramine Noninterfering: caffeine, diazepam, desipramine, mianserin, zimeldine Interfering: haloperidol KEYWORDS plasma REFERENCE
Franklin, M. Determination of plasma buspirone by high-performance liquid chromatography with coulometric detection. J.Chromatogr., 1990, 526, 590-596

SAMPLE Matrix: blood Sample preparation: 2 mL Plasma + 100 |xL 50 |xg/mL 1-phenylpiperazine + 1 mL pH 10 borate buffer + 5 mL chloroform: MeCN 8:2, agitate, centrifuge, repeat extraction. Evaporate organic layers under reduced pressure below 40, dissolve residue in 100 |xL mobile phase, inject a 20 |JLL aliquot. HPLCVARIABLES Guard column: 23 X 3.6 37-50 |xm CN/Corasil Column: 250 X 4.6 5 |xm Spherisorb CN Mobile phase: MeOH: 5 mM KH2PO4 pH 7.4 35:65 Flow rate: 1.7 Injection volume: 20 Detector: UV 254 CHROMATOGRAM Retention time: 10 Internal standard: 1-phenylpiperazine (13) Limit of detection: 5 ng/mL OTHER SUBSTANCES Extracted: l-(2-pyrimidinyl)piperazine, metabolites KEYWORDS plasma; rat REFERENCE
Diaz-Marot, A.; Puigdellivol, E.; Salvatella, C; Cornelias, L.; Gassiot, M. Determination of buspirone and l-(2-pyrimidinyl)piperazine in plasma samples by high-performance liquid chromatography. J.Chromatogr., 1989, 490, 470-473

SAMPLE Matrix: solutions HPLCVARIABLES Column: 250 X 4.6 5 |xm Supelcosil LC-DP (A) or 250 X 4 5 jjim 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: 9.07 (A)5 4.98 (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, 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.Chromatogr.A, 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 FIo1W rate: 2 Detector: UV 210 OTHER SUBSTANCES Also analyzed: acepromazine, acetaminophen, acetophenazine, albuterol, aminophylline, amitriptyline, amobarbital, amoxapine, amphetamine, amylocaine, antipyrine, aprobarbital, aspirin, atenolol, atropine, avermectin, barbital, benzocaine, benzoic acid, benzotropine, benzphetamine, berberine, bibucaine, bromazepan, brompheniramine, buprenorphine, 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, diethylprorjion, diethylstilbestrol, diflunisal, digitoxin, digoxin, diltiazem, diphenhydramine, diphenoxylate, 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


ANNOTATED BIBLIOGRAPHY

Gil, M.S.; Ochoa, C; Vega, S. High performance liquid chromatography of new potential anxiolytic drugs and related benzodiazepines: A comparative study of hydrophobicity. J.Liq.Chromatogr., 1991, 14, 2141-2156 [also chlordiazepoxide, diazepam] Sarati, S.; Guiso, G.; Spinelli, R.; Caccia, S. Determination of piribedil and its basic metabolites in plasma by high-performance liquid chromatography. J.Chromatogr., 1991, 563, 323-332 [plasma; rat; extracted piribedil; buspirone is IS; gradient]