Oxycodone

Molecular formula: C18H21NO4

Molecular weight: 315.4
CAS Registry No.: 76-42-6, 124-90-3 (HCI), 64336-55-6 (terephthalate)

SAMPLE
Matrix: bile, blood
Sample preparation: 0.5 mL Blood or bile + 10 (blood) or 15 (bile) |JLL 100 |xg/mL nalorphine in MeOH + 300 jxL 1.1 M pH 5.0 sodium acetate buffer + 3000-3500 U of Patella
vulgata glucuronidase, incubate at 55 overnight, add 0.5 mL borate buffer to achieve a
pH of 8.3-8.5. Add 8 mL chloroform: isopropanol 90:10, gently rotate for 30 min, centrifuge at 3500 rpm for 10 min, remove aqueous layer. Wash organic layer (twice for blood,
three times for bile) with 3 mL 100 mM pH 9.9 sodium phosphate buffer with gentle
rotation for 10 min and centrifugation each time. Add organic layer to 200 (blood) or 400
(bile) (JLL 0.2% phosphoric acid, gently rotate for 30 min, discard organic layer, inject 50
IxL of the acid layer. (Borate buffer was 50 mM boric acid and 43 mM sodium tetraborate,
adjusted to pH 9.8.)
HPLCVARIABLES
Guard column: Nova-Pak phenyl guard column
Column: 150 X 3.9 5 |xm Nova-Pak phenyl
Mobile phase: MeCN: 10 mM pH 6.6 NaH2PO4 10:90
Flow rate: 1.2
Injection volume: 50
Detector: UV 210; F ex 220 em 370 (cut-off)
CHROMATOGRAM
Retention time: 21.4
Internal standard: nalorphine (23.5)
OTHER SUBSTANCES
Simultaneous: codeine, dihydrocodeine, hydrocodone, 6-monoacetylmorphine, morphine
Noninterfering: acetylcodeine, amitriptyline, amphetamine, diamorphine, diazepam, dothiepin, doxepin, ephedrine, ephedrine, hydromorphone, mesoridazine, methadone, methamphetamine, 3-monoacetylmorphine, nordiazepam, norpropoxyphene, nortriptyline,
oxazepam, propoxyphene, pseudoephedrine, quinidine, quinine, sulfamethoxazole, sulforidazine, thioridazine
KEYWORDS
UV and F detection used together
REFERENCE
Crump, K.L.; Mclntyre, LM.; Drummer, O.H. Simultaneous determination of morphine and codeine in
blood and bile using dual ultraviolet and fluorescence high-performance liquid chromatography.
JAnal.ToxicoL, 1994, 18, 208-212

SAMPLE
Matrix: blood
Sample preparation: 1 mL Plasma + 60 |xL 1 |xg/mL codeine in water + 100 |xL 1 M
phosphoric acid + 5 mL butyl chloride, mix for 1.5 min, centrifuge at 1500 g for 3 min,
discard upper organic layer. To the aqueous layer add 500 |xL pH 10 1 M carbonate buffer

and 5 mL butyl chloride, mix for 1.5 min, centrifuge at 1500 g for 3 min, remove organic
layer and repeat extraction. Combine butyl chloride layers and evaporate them to dryness
under a stream of air at 40. Reconstitute the residue in 200 |xL mobile phase, inject a
150 |JLL aliquot.
HPLCVARIABLES
Guard column: Novapak C18 guard column
Column: 4 |xm Novapak C18 in a Waters RCM 8 X 10 radial compression unit
Mobile phase: MeOH: MeCN: 10 mM pH 7 phosphate buffer 230:20:1000, containing 40
mg/L cetyltrimethylammonium bromide (cetavlon)

Flow rate: 2
Injection volume: 150
Detector: E, Waters Model 460, working electrode 1.10 V
CHROMATOGRAM
Retention time: 8.5
Internal standard: codeine (13.5)
Limit of detection: 5 ng/mL
OTHER SUBSTANCES
Noninterfering: acetaminophen, amitriptyline, aspirin, atenolol, camazepam, carbamazepine, chlorimipramine, chlorthalidone, clonazepam, cortisone, desipramine, diazepam,
halazepam, hydrochlorothiazide, hydrocortisone, imipramine, lorazepam, maprotiline,
meperidine, methylphenobarbital, methylprednisolone, metoclopramide, midazolam, morphine, nalorphine, naloxone, nitrazepam, nortriptyline, oxazepam, oxprenolol, phenobarbital, phenytoin, pindolol, prazepam, prednisolone, prednisone, primidone, prochlorperazine, propranolol, salicylic acid, temazepam
KEYWORDS
plasma
REFERENCE
Smith, M.T.; Watt, J.A.; Mapp, G.P.; Cramond, T. Quantitation of oxycodone in human plasma using
high-performance liquid chromatography with electrochemical detection. Ther.Drug Monit., 1991,13,
126-130

SAMPLE
Matrix: blood
Sample preparation: 2 mL Plasma + 50 |xL 2 |xg/mL methadone in MeOH + 500 |xL pH
9.6 carbonate buffer + 6 mL butyl chloride, shake on a mechanical shaker at 100 rpm
for 15 min, centrifuge at 2000 g for 5 min. Remove the organic layer and add it to 3 mL
200 mM HCl. Shake for 15 min, centrifuge, remove aqueous layer. Add aqueous layer to
3 drops 60% NaOH and 6 mL butyl chloride, shake for 15 min, centrifuge. Remove organic
layer and evaporate it to dryness at 50 under a stream of nitrogen, reconstitute residue
in 60 |xL mobile phase, inject a 40 u,L aliquot.
HPLCVARIABLES
Column: 10 u,m Brownlee RP-8
Mobile phase: MeCN: MeOH: 10 mM KH2PO4 50:30:20

Flow rate: 1
Injection volume: 40

Detector: E, Bioanalytical Systems, glassy carbon working electrode 1.20 V, Ag/AgCl reference electrode
CHROIUIATOGRAM
Retention time: 6.4

Internal standard: methadone (16)

Limit of quantitation: 2 ng/mL
OTHER SUBSTANCES
Simultaneous: fentanyl, meperidine, phenoperidine
KEYWORDS
plasma
REFERENCE
Schneider, J.J.; Triggs, E.J.; Bourne, D.W.; Stephens, LD.; Haviland, A.M. Determination of oxycodone
in human plasma by high-performance liquid chromatography with electrochemical detection.
J.Chromatogr., 1984, 308, 359-362

SAMPLE
Matrix: solutions
Sample preparation: Inject a 10 JULL aliquot of a 50 |xg/mL solution in water.
HPLCVARIABLES
Column: 150 X 4.6 5 jim Supelcosil LC-ABZ
Mobile phase: MeCN: 25 mM pH 7.5 potassium phosphate buffer 35:65
Flow rate: 1.5
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 3.6
OTHER SUBSTANCES
Simultaneous: codeine, meperidine
REFERENCE
Supelco Chromatography Products, Supelco, Inc., Beliefonte PA, 1996, p. A23

SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 250 X 4.6 5 |xm 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: 6.51 (A), 5.82 (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, 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, levorphanol, lidocaine, loratadine, lorazepam, lovastatin, loxapine, mazindol,
mefenamic acid, meperidine, mephenytoin, mepivacaine, mesoridazine, metaproterenol,
metformin, 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,
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
Flow 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, 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, 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-fiuorouracil, 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, 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, triprolidine, 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: 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: 2.670

OTHER SUBSTANCES
Also analyzed: 6-acetylmorphine, amiloride, amphetamine, benzocaine, benzoylecgonine,
caffeine, cocaine, codeine, doxylamine, fluoxetine, glutethimide, hexobarbital, hypoxanthine, levorphanol, LSD, meperidine, mephobarbital, methadone, methylphenidate, methyprylon, N-norcodeine, oxazepam, 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: solutions
HPLCVARIABLES
Column: 150 X 3.9 ixBondapak C18
Mobile phase: MeCN: phosphate buffer 36:64, pH adjusted to 4.0
Flow rate: 1.5
Injection volume: 100
Detector: UV 254
CHROMATOGRAM
Retention time: 2.5
Limit of detection: 50 ng/mL
REFERENCE
Tien, J.H. Transdermal-controlled administration of oxycodone. J.Pharm.ScL, 1991, 80, 741-743

SAMPLE
Matrix: solutions
Sample preparation: Dissolve in MeOH at a concentration of 1 mg/mL, inject a 20 |JLL
aliquot.
HPLCVARIABLES
Column: 250 X 5 Spherisorb S5W
Mobile phase: MeOH.buffer 90:10 (Buffer was 94 mL 35% ammonia and 21.5 mL 70%
nitric acid in 884 mL water, adjust the pH to 10.1 with ammonia.)
Flow rate: 2
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 2.44 (tails)
OTHER SUBSTANCES
Simultaneous: benzphetamine, benzylmorphine, bromo-STP, buprenorphine, caffeine, codeine, codeine-N-oxide, dextromoramide, dextropropoxyphene, diamorphine, diethylpropion, dihydrocodeine, dihydromorphine, dimethylamphetamine, dipipanone, ephedrine,
epinephrine, ethoheptazine, ethylmorphine, etorphine, fenethyline, fentanyl, hydrocodone, 4-hydroxyamphetamine, hydroxypethidine, levallorphan, levorphanol, mazindol,
meperidine, mephentermine, mescaline, methadone, methamphetamine, methylephedrine, methylphenidate, morphine, morphine-3-glucuronide, morphine-N-oxide, nalorphine, naloxone, norcodeine, norlevorphanol, normetanephrine, normethadone, normorphine, norpethidine, norpipanone, noscapine, papaverine, pemoline, pentazocine,
phenazocine, phendimetrazine, phenelzine, 2-phenethylamine, phenoperidine, phenylephrine, pholcodeine, pipradol, piritramide, prolintane, pseudoephedrine, STP, tranylcypromine, trimethoxyamphetamine, tyramine

Noninterfering: dopamine, levodopa, methyldopa, methyldopate, norepinephrine

Interfering: acetylcodeine, amphetamine, chlorphentermine, fencamfamin, fenfluramine,
methylenedioxyamphetamine, monoacetylmorphine, norpseudoephedrine, phentermine,
phenylpropanolamine, thebacon, thebaine
REFERENCE
Law, B.; Gill, R.; Moffat, A.C. High-performance liquid chromatography retention data for 84 basic drugs
of forensic interest on a silica column using an aqueous methanol eluent. J.Chromatogr., 1984, 301,
165-172