SAMPLE

Matrix: blood
Sample preparation: 1-2 mL Plasma + 1 mL 1 M pH 10 bicarbonate buffer + 8 mL n-
hexane: ethyl acetate 70:30, shake for 10 min, centrifuge. Remove the organic layer and
evaporate it to dryness under a stream of nitrogen, reconstitute the residue in 200 \xL
mobile phase, inject an aliquot.
HPLC VARIABLES
Column: 250 X 4.6 Ultrasphere ODS
Mobile phase: Gradient. A was MeCN:buffer 30:70. B was MeCN:buffer 70:30. A:B from
85:15 to 40:60 over 10 min (Waters curve no. 5), to 0:100 over 5 min (Waters curve no.
1), return to initial conditions over 10 min (Waters curve no. 1). (Buffer was 10 mM pH
3.35 NaH
2
PO
4
.)
FIo
1
W rate: 1
Detector: UV 229
CHROMATOGRAM
Internal standard: lorazepam
OTHER SUBSTANCES
Extracted: desmethyldiazepam, diazepam
KEYWORDS
plasma; lorazepam is IS
REFERENCE
Caraco, Y.; Tateishi, T.; Wood, A.J.J. Interethnic difference in omeprazole's inhibition of diazepam me-
tabolism. Clin.Pharmacol.Ther., 1995, 58, 62-72
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 |xL mobile phase, centrifuge at 2800 g for 5 min,
inject a 50 JJLL 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 ^m NovaPack C18
Mobile phase: MeOH:THF:buffer 65:5:30 (Buffer was 0.68 g/L (10 mM (sic)) KH
2
PO
4
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
Lorazepam
Molecular formula: C
15
H
10
CI
2
N
2
O
2
Molecular weight: 321.2
CAS Registry No.: 846-49-1
Injection volume: 50
Detector: UV 230
CHROMATOGRAM
Retention time: 4.19
Limit of detection: <120 ng/mL
OTHER SUBSTANCES
Extracted: acebutolol, acenocoumarol, acepromazine, aceprometazine, acetaminophen,
aconitine, ajmaline, albuterol, alimemazine, alminoprofen, alpidem, alprenolol, amisul-
pride, amitriptyline, amodiaquine, amoxapine, aspirin, astemizole, atenolol, benazepril,
benperidol, benzocaine, benzoylecgonine, bepridil, betaxolol, bisoprolol, bromazepam,
brompheniramine, bumadizone, bupivacaine, buprenorphine, buspirone, caffeine, carba-
mazepine, carbinoxamine, carpipramine, carteolol, eeliprolol, cetirizine, chlorambucil,
chlordiazepoxide, chlormezanone, chlorophenacinone, chloroquine, chlorpheniramine,
chlorpromazine, chlorpropamide, cibenzoline, cicletanine, clemastine, clobazam, clomipra-
mine, clonazepam, clonidine, clorazepate, clozapine, cocaine, codeine, colchicine, cyame-
mazine, cyclizine, cyproheptadine, cytarabine, dacarbazine, daunorubicin, debrisoquine,
demexiptiline, desipramine, dextromethorphan, dextromoramide, dextropropoxyphene,
diazepam, diazoxide, diclofenac, dihydralazine, diltiazem, diphenhydramine, dipyrida-
mole, disopyramide, dosulepine, doxepin, doxylamine, droperidol, ephedrine, estazolam,
etodolac, fenfluramine, fenoprofen, fentiazac, flecainide, floctafenine, fiumazenil, flunitra-
zepam, fluoxetine, fiuphenazine, flurbiprofen, fluvoxamine, glibenclamide, glibornuride,
glipizide, glutethimide, haloperidol, histapyrrodine, hydroxychloroquine, hydroxyzine,
ibuprofen, imipramine, indomethacin, iproniazid, ketoprofen, labetalol, levomepromazine,
lidocaine, lidoflazine, lisinopril, loperamide, loprazolam, loratadine, loxapine, maprotiline,
medazepam, medifoxamine, mefenamic acid, mefenidramine, mefloquine, melphalan, me-
peridine, mephenesin, mephentermine, mepivacaine, metapramine, metformin, metha-
done, methocarbamol, methotrexate, metipranolol, metoclopramide, mexiletine, mianser-
ine, midazolam, minoxidil, moclobemide, moperone, morphine, nadolol, nalbuphine,
nalorphine, naloxone, naltrexone, naproxen, nialamide, nicardipine, niflumic acid, nimo-
dipine, nitrazepam, nitrendipine, nizatidine, nomifensine, nortriptyline, omeprazole, op-
ipramol, oxazepam, oxprenolol, penbutolol, penfluridol, pentazocine, phencyclidine, phen-
obarbital, phenol, phenylbutazone, pimozide, pindolol. pipamperone, prazepam, prazosin,
prilocaine, procainamide, procarbazine, proguanil, promethazine, propafenone, propran-
olol, protriptyline, pyrimethamine, quinidine, quinupramine, ramipril, ranitidine, re-
serpine, ritodrine, secobarbital, sotalol, strychnine, sulfinpyrazole, sulpride, sultopride,
suriclone, temazepam, tenoxicam, terbutaline, terfenadine, tetracaine, tetrazepam, thio-
pental, thioproperazine, thioridazine, tianeptine, tiapride, tiaprofenic acid, ticlopidine, ti-
molol, tioclomarol, tofisopam, tolbutamide, toloxatone, trazodone, triazolam, trifluopera-
zine, trifluperidol, trimipramine, triprolidine, tropatenine, verapamil, viloxazine,
vinblastine, vincristine, vindesine, warfarin, yohimbine, zolpidem, zopiclone, zorubicine
Interfering: alprazolam, cycloguanil, ketamine, methaqualone, metoprolol, nifedipine, pi-
roxicam, quinine, sulindac
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: Make 1 mL serum alkaline with borate buffer, extract with
cyclohexane: dichloromethane 60:40. Remove the organic layer and evaporate it to dry-
ness, reconstitute the residue in mobile phase, inject an aliquot
HPLC VARIABLES
Column: C18 DB (Supelco)
Mobile phase: MeCN :pH 2.5 phosphate buffer 37:63
Detector: UV 254
OTHER SUBSTANCES
Extracted: bromazepam, clobazam, diazepam, fluvoxamine, oxazepam
KEYWORDS
serum
REFERENCE
Vandenberghe, H.; MacDonald, J.C. Analysis of fluvoxamine, clobazam and other benzodiazepines on
the same HPLC system (Abstract 40). Ther.Drug Monit, 1995, 17, 393
SAMPLE
Matrix: blood
Sample preparation: Inject 100-200 JJLL plasma onto column A with mobile phase A and
elute to waste, after 5 min backflush the contents of column A onto column B with mobile
phase B, after 5 min remove column A from the circuit, elute column B with mobile phase
B, monitor the effluent from column B. Wash column A with MeCN: water 60:40 at 1
mL/min for 6 min then re-equilibrate with pH 7.5 buffer for 10 min.
HPLC VARIABLES
Column: A 45 X 4 12 ^m TSK-gel G 3 PW (Tosohass); B 75 X 4.6 Ultrasphere ODS C18 3
|jim
Mobile phase: A 50 mM pH 7.5 phosphate buffer; B Gradient. A was MeCN. B was 65 mM
KH
2
PO
4
+ 1% diethylamine adjusted to pH 5.4 with phosphoric acid. A:B 22:78 for 5
min, to 25:75 over 10 min, to 60:40 over 15 min.
Flow rate: 1
Injection volume: 100-200
Detector: UV 230
CHROMATOGRAM
Retention time: 22
OTHER SUBSTANCES
Extracted: alprazolam, bromazepam, chlordiazepoxide, clobazam, clorazepate, clotiaze-
pam, desmethylclobazam, desmethyldiazepam, diazepam, estazolam, flunitrazepam, lof-
lazepate, medazepam, nitrazepam, oxazepam, prazepam, temazepam, tetrazepam, tofi-
sopam, triazolam
Noninterfering: carbamazepine, ethosuximide, phenobarbital, phenytoin, primidone, val-
proic acid
Interfering: clonazepam
KEYWORDS
plasma; column-switching
REFERENCE
Lacroix, C; Wojciechowski, R; Danger, P. Monitoring of benzodiazepines (clobazam, diazepam and their
main active metabolites) in human plasma by column-switching high-performance liquid chromatog-
raphy. J.Chromatogr., 1993, 617, 285-290
SAMPLE
Matrix: blood
Sample preparation: 0.5 mL Plasma +10 |JLL 250 |JLg/mL 1-acetamidopyrene in MeOH +
200 |xL 1 M ammonium sulfate + 800 ^xL cold MeCN, vortex for 30 s, store at -20 for at
least 30 min, vortex, centrifuge at 1500 g for 30 min. Remove 400 jxL of the upper organic
layer and evaporate it under a stream of nitrogen. Reconstitute with 100 jxL mobile phase,
vortex for 30 s, inject a 75 |xL aliquot.
HPLC VARIABLES
Column: 300 X 3.9 10 |xm fxBondapak C18
Mobile phase: MeCN: buffer 47:53 (Buffer was 6.805 g potassium monophosphate in 1 L
water, adjust pH to 6.00 with 10 M NaOH.)
Flow rate: 1
Injection volume: 75
Detector: UV 214
CHROMATOGRAM
Retention time: 6.1
Internal standard: 1-acetamidopyrene (9.7)
Limit of detection: 0.781 ng/mL
OTHER SUBSTANCES
Simultaneous: antipyrine, indocyanine green
Noninterfering: adenosine, albuterol, alphenal, aspirin, caffeine, carbamazepine, cefazo-
lin, cephalexin, cephalothin, cimetidine, ciprofloxacin, claforan, desipramine, enoxacin,
fleroxacin, furosemide, hydralazine, hydrochlorothiazide, minoxidil, norfloxacin, pheny-
toin, propafenone, sulindac, teicoplanin, theophylline, vancomycin
Interfering: some indocyanine green impurities
KEYWORDS
plasma
REFERENCE
Awni, W.M.; Bakker, L.J. Antipyrine, indocyanine green, and lorazepam determined in plasma by high-
pressure liquid chromatography. Clin.Chem., 1989, 35, 2124-2126
SAMPLE
Matrix: blood
Sample preparation: 1 mL Plasma + 100 |xL 1 fxg/mL diazepam in MeOH + 2 mL phos-
phate buffer, vortex for 30 s, add 8 mL hexane: isoamyl alcohol 95:5, shake gently by
hand for 10 min, vortex for 1 min, centrifuge at 400 g at 4 for 10 min. Remove organic
layer and add it to 2 mL 6 M HCl, shake for 10 min, vortex for 1 min, centrifuge at 400
g for 10 min. Remove the aqueous layer and slowly add about 2 mL 6 M NaOH to it to
achieve a pH greater than 7.0, add 2 mL phosphate buffer, vortex for 10 s, add 8 mL
hexane: isoamyl alcohol 95:5, shake for 10 min, vortex for 1 min, centrifuge at 400 g for
10 min. Remove the organic layer and evaporate it to dryness at 40 with nitrogen, rinse
residue from sides with hexane: isoamyl alcohol 95:5, again dry with nitrogen, take up
residue in 40 \xL MeOH, inject a 20-40 |xL aliquot. (Phosphate buffer was 136.1 g KH
2
PO
4
in 1 L water, adjusted to pH 7 with 1 M K
2
HPO
4
.)
HPLCVARIABLES
Column: 300 X 3.9 10 |xm fxBondapak C18
Mobile phase: MeOH: MeCN: 10 mM sodium acetate 40:12.5:47.5, at pH 4.6
Flow rate: 2.2
Injection volume: 20-40
Detector: UV 254
CHROMATOGRAM
Retention time: 5.2
Internal standard: diazepam (9.2)
Limit of quantitation: 2.5 ng/mL
OTHER SUBSTANCES
Simultaneous: midazolam, flunitrazepam, clonazepam, flurazepam, temazepam, nitraze-
pam
Interfering: oxazepam
KEYWORDS
plasma
REFERENCE
Egan, J.M.; Abernethy, D.R. Lorazepam analysis using liquid chromatography: improved sensitivity for
single-dose pharmacokinetic studies. J.Chromatogr., 1986, 380, 196-201
SAMPLE
Matrix: blood
Sample preparation: 0.5 mL Plasma + 0.5 mL water + 0.5 mL 0.25 M NaOH, vortex,
allow to stand at room temperature for 20 min, add 20 JJLL of 100 |xg/mL phenacetin and
3 |xg/mL flunitrazepam, vortex, add 5 mL diethyl ether, vortex for 30 s, centrifuge at 900
g for 5 min, freeze in acetone/dry ice for 5 min. Remove the supernatant and dry it under
nitrogen. Reconstitute in 115 |JLL MeCN: 0.1% pH 3 sodium phosphate buffer 30:70, inject
an aliquot.
HPLCVARIABLES
Guard column: 23 X 3.9 37-50 fxm juuBondapak phenyl
Column: 300 X 3.9 10 |jim jjiBondapak phenyl
Mobile phase: Gradient. A was MeCN: 0.1% pH 3 sodium phosphate buffer 5:95. B was
MeCN:0.1% pH 3 sodium phosphate buffer 70:30. A:B 80:20 for 2.5 min, then to 45:55
over 20 min, then to 25:75 over 3 min, then to 80:20 over 3 min, equilibrate at 80:20
for 7 min.
Column temperature: 40
Flow rate: 2
Injection volume: 200
Detector: UV 229
CHROMATOGRAM
Retention time: 15.11
Internal standard: flunitrazepam (17.90)
Limit of quantitation: 10.5 ng/mL
OTHER SUBSTANCES
Simultaneous: antipyrine (at 254 nm)
Noninterfering: acetaminophen, allopurinol, indocyanine green, sulfamethoxazole, tri-
methoprim
KEYWORDS
plasma
REFERENCE
Riley, CA.; Evans, W.E. Simultaneous analysis of antipyrine and lorazepam by high-performance liquid
chromatography J.Chromatogr., 1986, 382, 199-205
SAMPLE
Matrix: blood, CSF, gastric fluid, urine
Sample preparation: 200 |xL Serum, urine, CSF, or gastric fluid + 300 \xL reagent. Flush
column A to waste with 500 |xL 500 mM ammonium sulfate, inject sample onto column
A, flush column A to waste with 500 JJLL 500 mM ammonium sulfate, backflush the con-
tents of column A onto column B with mobile phase, monitor the effluent from column B.
(Reagent was 8.05 M guanidine hydrochloride and 1.02 M ammonium sulfate in water.)
HPLC VARIABLES
Column: A 40 |xm preparative grade C18 (Analytichem); B 75 X 2.1 pellicular C18 (What-
man) + 250 X 4.6 5 |xm C8 end-capped (Whatman)
Mobile phase: Gradient. A was 50 mM pH 4.5 KH
2
PO
4
. B was MeCN: isopropanol 80:20.
A:B 90:10 for 1 min, to 30:70 over 20 min.
Column temperature: 50
Flow rate: 1.5
Detector: UV 220
CHROMATOGRAM
Retention time: 12.57
Internal standard: heptanophenone (19)
OTHER SUBSTANCES
Extracted: acetaminophen, allobarbital, azinphos, barbital, brallobarbitone, bromazepam,
butethal, caffeine, carbamazepine, carbaryl, cephaloridine, chloramphenicol, chlordiaze-
poxide, chlorothiazide, chlorvinphos, clothiapine, cocaine, coomassie blue, desipramine,
diazepam, diphenhydramine, dipipanone, ethylbromphos, flufenamic acid, formothion,
griseofulvin, indomethacin, lidocaine, malathion, medazepam, midazolam, oxazepam, par-
aoxon, penicillin G, pentobarbital, prazepam, propoxyphene, prothiophos, quinine, sali-
cylic acid, secobarbital, strychnine, sulfamethoxazole, theophylline, thiopental, thiorida-
zine, trimethoprim
KEYWORDS
serum; column-switching
REFERENCE
Kruger, RB.; Albrecht, C.F.De V.; Jaarsveld, P.P. Use of guanidine hydrochloride and ammonium sulfate
in comprehensive in-line sorption enrichment of xenobiotics in biological fluids by high-performance
liquid chromatography. J.Chromatogr., 1993, 612, 191-198
SAMPLE
Matrix: formulations
Sample preparation: Dilute with mobile phase, inject an aliquot.
HPLC VARIABLES
Column: 250 X 4.6 5 |xm cyano
Mobile phase: MeCN: 100 mM NaH
2
PO
4
20:80 adjusted to pH 4.2 with phosphoric acid
Flow rate: 1.5
Injection volume: 20
Detector: UV 220
CHROMATOGRAM
Retention time: 8.72
OTHER SUBSTANCES
Simultaneous: granisetron (UV 300)
KEYWORDS
stability-indicating; injections; saline
REFERENCE
Mayron, D.; Gennaro, A.R. Stability and compatibility of granisetron hydrochloride in i.v. solutions and
oral liquids and during simulated Y-site injection with selected drugs. Am.J.Health-Syst.Pharm.,
1996, 53, 294-304
SAMPLE
Matrix: formulations
Sample preparation: Inject a 20 |xL aliquot.
HPLCVARIABLES
Column: 150 X 4.6 5 |xm Spherisorb ODS-I
Mobile phase: MeOH: 50 mM (NH
4
)H
2
PO
4
50:50, adjusted to pH 7.22
Flow rate: 1.3 for 12 min then 2.1 for 14 min
Injection volume: 20
Detector: UV 240
CHROMATOGRAM
Retention time: 9
OTHER SUBSTANCES
Simultaneous: degradation products
KEYWORDS
stability-indicating; 5% dextrose; saline; injections
REFERENCE
Mancano, M.A.; Boullata, J.I.; Gelone, S.P.; Zitterman, R.E.; Borenstein, M.R. Availability of lorazepam
after simulated administration from glass and polyvinyl chloride containers. Am.J.Health-
Syst.Pharm., 1995, 52, 2213-2216
SAMPLE
Matrix: formulations
Sample preparation: Dilute with water.
HPLC VARIABLES
Column: Waters C18 column (PN 86344)
Mobile phase: MeOH: 50 mM (NH
4
)H
2
PO
4
50:50, adjusted to pH 6.5 with ammonium
hydroxide
Flow rate: 2
Injection volume: 20
Detector: UV 240
CHROMATOGRAM
Retention time: 21.1
KEYWORDS
saline; injections; stability-indicating
REFERENCE
Stiles, M.L.; Allen, L.V., Jr.; Prince, S.J.; Holland, J.S. Stability of dexamethasone sodium phosphate,
diphenhydramine hydrochloride, lorazepam, and metoclopramide hydrochloride in portable infusion-
pump reservoirs. Am.J.Hosp.Pharm., 1994, 51, 514-517
SAMPLE
Matrix: formulations
Sample preparation: Inject an aliquot directly.
HPLC VARIABLES
Column: 250 X 4.6 5 |xm Vydac 208TP54 C8
Mobile phase: MeOH: 50 mM (NH
4
)H
2
PO
4
57:43, pH adjusted to 6.5 with ammonium
hydroxide
Flow rate: 1
Detector: UV 254
CHROMATOGRAM
Retention time: 6.1
KEYWORDS
injections; saline; 5% dextrose; lactated Ringer's; stability-indicating
REFERENCE
Trissel, L.A.; Pearson, S.D. Storage of lorazepam in three injectable solutions in polyvinyl chloride and
polyolefin bags. Am.J.Hosp.Pharm., 1994, 51, 368-372
SAMPLE
Matrix: formulations
Sample preparation: Dilute with saline, inject a 10 |JLL aliquot.
HPLC VARIABLES
Column: 250 X 4.6 Lichrosorb 10 RP 8
Mobile phase: MeOH: THF: water 50:5:50
Flow rate: 3
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 8
OTHER SUBSTANCES
Simultaneous: diazepam, thiopental
KEYWORDS
injections; saline
REFERENCE
Martens, H.J.; de Goede, RN.; van Loenen, A.C. Sorption of various drugs in polyvinyl chloride, glass,
and polyethylene-lined infusion containers. Am.J.Hosp.Pharm., 1990, 47, 369-373
SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 150 X 3.9 4 |xm Nova pack C18
Mobile phase: MeOH: water 52:48
Column temperature: 48
Flow rate: 0.8
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 6
OTHER SUBSTANCES
Simultaneous: bromazepam, chlordiazepoxide, clobazam, clorazepate, diazepam, flunitra-
zepam, nitrazepam, oxazepam, tofisopam
REFERENCE
Guillaume, Y.; Guinchard, C. Thermodynamic behavior of mixed benzodiazepines by a new liquid chro-
matographic method. Chromatographia, 1995, 40, 193-196
SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 150 X 3.9 4 |xm Nova pak C18
Mobile phase: MeCN:water 57:43
Column temperature: 44
Flow rate: 1.1
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 6.3
OTHER SUBSTANCES
Simultaneous: bromazepam, chlordiazepoxide, clobazam, clorazepate, diazepam, flunitra-
zepam, nitrazepam, oxazepam, tofisopam
REFERENCE
Guillaume, Y; Guinchard, C. Marked difference between acetonitrile/water and methanol/water mobile
phase systems on the thermodynamic behavior of benzodiazepines in reversed phase liquid chro-
matography. Chromatographia, 1995, 41, 8487
SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 250 X 4.6 5 ^m Supelcosil LC-DP (A) or 250 X 4 5 jim 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: 6.14 (A), 5.78 (B)
OTHER SUBSTANCES
Also analyzed: acebutolol, acepromazine, acetaminophen, acetazolamide, acetophenazine,
albuterol, alprazolam, amitriptyline, amobarbital, amoxapine, antipyrine, atenolol, atro-
pine, azatadine, baclofen, benzocaine, bromocriptine, brompheniramine, brotizolam,
bupivacaine, buspirone, butabarbital, butalbital, caffeine, carbamazepine, cetirizine,
chlorcyclizine, chlordiazepoxide, ehlormezanone, chloroquine, chlorpheniramine,
chlorpromazine, chlorpropamide, chlorprothixene, chlorthalidone, chlorzoxazone, cimeti-
dine, cisapride, clomipramine, clonazepam, clonidine, clozapine, cocaine, codeine, colchi-
cine, cyclizine, cyclobenzaprine, dantrolene, desipramine, diazepam, diclofenac, diflunisal,
diltiazem, diphenhydramine, diphenidol, diphenoxylate, dipyridamole, disopyramide, do-
butamine, doxapram, doxepin, droperidol, encainide, ethidium bromide, ethopropazine,
fenoprofen, fentanyl, flavoxate, fluoxetine, fluphenazine, flurazepam, flurbiprofen, fluvox-
amine, furosemide, glutethimide, glyburide, guaifenesin, haloperidol, homatropine, hy-
dralazine, hydrochlorothiazide, hydrocodone, hydromorphone, hydroxychloroquine, hydro-
xyzine, ibuprofen, imipramine, indomethacin, ketoconazole, ketoprofen, ketorolac,
labetalol, levorphanol, lidocaine, loratadine, lovastatin, loxapine, mazindol, mefenamic
acid, meperidine, mephenytoin, mepivacaine, mesoridazine, metaproterenol, methadone,
methdilazine, methocarbamol, methotrexate, methotrimeprazine, methoxamine, methyl-
dopa, methylphenidate, metoclopramide, metolazone, metoprolol, metronidazole, mida-
zolam, moclobemide, morphine, nadolol, nalbuphine, naloxone, naphazoline, naproxen,
nifedipine, nizatidine, norepinephrine, nortriptyline, oxazepam, oxycodone, oxymetazo-
line, paroxetine, pemoline, pentazocine, pentobarbital, pentoxifylline, perphenazine,
pheniramine, phenobarbital, phenol, phenolphthalein, phentolamine, phenylbutazone,
phenyltoloxamine^ phenytoin, pimozide, pindolol, piroxicam, pramoxine, prazepam, pra-
zosin, probenecid, procainamide, procaine, prochlorperazine, procyclidine, promazine, pro-
methazine, propafenone, propantheline, propiomazine, propofol, propranolol, protripty-
line, quazepam, quinidine, quinine, racemethorphan, ranitidine, remoxipride, risperidone,
salicylic acid, scopolamine, secobarbital, sertraline, sotalol, spironolactone, sulfinpyra-
zone, sulindac, temazepam, terbutaline, terfenadine, tetracaine, theophylline, thiethyl-
perazine, thiopental, thioridazine, thiothixene, timolol, tocainide, tolbutamide, tolmetin,
trazodone, triamterene, triazolam, trifluoperazine, triflupromazine, trimeprazine, tri-
methoprim, 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
Sample preparation: Prepare a 100 |JLM solution in buffer, inject a 20 JULL aliquot.
HPLCVARIABLES
Column: 100 X 4.6 column containing riboflavin binding proteins (Prepare as follows. Add
riboflavin to saturate protein of egg yolk, homogenize with 3 volumes buffer, centrifuge,
add the supernatant to a 500 X 30 column of DEAE-cellulose (Whatman) equilibrated
with buffer, wash extensively with buffer to remove bound protein, elute riboflavin bind-
ing proteins (RFBP) with buffer containing 200 mM NaCl (RFBP has intense yellow color,
absorption at 455 nm). Purify RFBP on a Sephadex G-100 column with 50 mM pH 7.5
Tris-HCl buffer as eluent, remove the bound riboflavin by extensive dialysis at pH 3.0.
Add 4.5 g N, N-disuccinylimidyl carbonate to 3 g Nucleosil 5NH2 slurried in MeCN, filter,
wash with MeCN, wash with 50 mM pH 7.5 phosphate buffer. Suspend 300 mg RFBP in
50 mM phosphate buffer, add the activated silica, mix gently for 2 h using a rotary evap-
orator, filter, wash with sterile water, wash with isopropanol: water 1:2, pack in a 100 X
4.6 column.) (Buffer was 100 mM pH 5.3 sodium acetate.)
Mobile phase: EtOH: 50 mM pH 5.5 KH
2
PO
4
5:95
Flow rate: 0.8
Injection volume: 20
Detector: UV
CHROMATOGRAM
Retention time: k' 4.02
OTHER SUBSTANCES
Simultaneous: bepridil, manidipine, nicardipine
Interfering: oxazepam
KEYWORDS
chiral; a = 1.63
REFERENCE
Massolini, G.; De Lorenzi, E.; Ponci, M.C.; Gandini, C; Caccialanza, G.; Monaco, H.L. Egg yolk riboflavin
binding protein as a new chiral stationary phase in high-performance liquid chromatography.
J.Chromatogr.A, 1995, 704, 55-65
SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 150 X 4.6 cellulose 3,5-dimethylphenylcarbamate/10-undecenoate bonded to
allylsilica
Mobile phase: Heptane: isopropanol 90:10
Flow rate: 1
Injection volume: 1000
Detector: UV 254
CHROMATOGRAM
Retention time: k' 9.84
KEYWORDS
chiral; a 1.64
REFERENCE
Oliveros, L.; Lopez, P.; Minguillon, C; Franco, P. Chiral chromatographic discrimination ability of a
cellulose 3,5-dimethylphenylcarbamate/10-undecenoate mixed derivative fixed on several chromato-
graphic matrices. J.Liq.Chromatogr., 1995, 18, 1521-1532
SAMPLE
Matrix: solutions
Sample preparation: Dissolve the compound, S-trolox methyl ether (Fluka), dicyclohex-
ylcarbodiimide, and 4-dimethylaminopyridine in dichloromethane, stir at room tempera-
ture for 1 h, filter (0.45 |xm), inject an aliquot.
HPLCVARIABLES
Column: 300 X 0.32 5 jim LiChrosorb Diol
Mobile phase: Carbon dioxide:MeOH 91.5:8.5
Column temperature: 80
Injection volume: 0.2
Detector: UV 254
CHROMATOGRAM
Retention time: 50.2 (second peak)
KEYWORDS
derivatization; subcritical fluid chromatography; chiral; density of mobile phase 0.62 g/mL;
resolution (R
8
) 1.2
REFERENCE
Almquist, S.R.; Petersson, P.; Walther, W.; Markides, K.E. Direct and indirect approaches to enantio-
meric separation of benzodiazepines using micro column techniques. J.Chromatogr.A, 1994, 679,
139-146
SAMPLE
Matrix: solutions
Sample preparation: Dilute in MeOH to a concentration of 10-80 mg/mL, inject an aliquot
HPLCVARIABLES
Column: 150 X 3.9 5 fim Nova pak RP 18
Mobile phase: MeOH: water 50:50
Column temperature: 50
Flow rate: 0.82
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 8.5
OTHER SUBSTANCES
Simultaneous: bromazepam, chlorazepate, chlordiazepoxide, clobazam, diazepam, flunitra-
zepam, nitrazepam, oxazepam, tofisopam
KEYWORDS
conditions are optimized
REFERENCE
Guillaume, Y; Guinchard, C. Study and optimization of column efficiency in HPLC: Comparison of two
methods for separating ten benzodiazepines. J.Liq.Chromatogr., 1994, 17, 1443-1459
SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 250 X 4.6 Zorbax RX
Mobile phase: Gradient. A was 10 mL concentrated orthophosphoric acid and 7 mL trie-
thylamine in 1 L water. B was 10 mL concentrated orthophosphoric acid and 7 mL trie-
thylamine 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, aprobar-
bital, aspirin, atenolol, atropine, avermectin, barbital, benzocaine, benzoic acid, benzotro-
pine, benzphetamine, berberine, bibucaine, bromazepan, brompheniramine, buprenor-
phine, buspirone, butabarbital, butacaine, butethal, caffeine, carbamazepine, carbromal,
chloramphenicol, chlordiazepoxide, chloroquine, chlorothiazide, chloroxylenol, chlorphe-
nesin, 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, dex-
tromethorphan, dextropropoxyphene, diamorphine, diazepam, diclofenac, diethylpropion,
diethylstilbestrol, diflunisal, digitoxin, digoxin, diltiazem, diphenhydramine, diphenoxy-
late, diprenorphine, dipyrone, disulfiram, dopamine, doxapram, doxepin, dronabinol,
ephedrine, epinephrine, epinine, estradiol, estriol, estrone, ethacrynic acid, ethosuximide,
etonitazene, etorphine, eugenol, famotidine, fenbendazole, fencamfamine, fenoprofen, fen-
proporex, fentanyl, flubendazole, flufenamic acid, flunitrazepam, 5-fluorouracil, fluo-
xymesterone, fluphenazine, furosemide, gentisic acid, gitoxigenin, glipizide, glunixin,
glutethimide, glybenclamide, guaiacol, halazepam, haloperidol, hydrochlorothiazide,
hydrocodone, hydrocortisone, hydromorphone, hydroxyquinoline, ibogaine, ibuprofen, im-
inostilbene, imipramine, indomethacin, isocarbostyril, isocarboxazid, isoniazid, isoproter-
enol, isoxsuprine, ivermectin, ketamine, ketoprofen, kynurenic acid, levorphanol, lido-
caine, lormetazepam, loxapine, mazindol, mebendazole, meclizine, meclofenamic acid,
medazepam, mefenamic acid, megestrol, mepacrine, meperidine, mephentermine, me-
phenytoin, mephesin, mephobarbital, mepivacaine, mescaline, mesoridazine, methadone,
methamphetamine, methapyrilene, methaqualone, methazolamide, methocarbamol,
methoxamine, methsuximide, methyl salicylate, methyldopa, methyldopamine, methyl-
phenidate, methylprednisolone, methyltestosterone, methyprylon, metoprolol, mibole-
rone, morphine, nadolol, nalorphine, naloxone, naltrexone, naphazoline, naproxen, nefo-
pam, niacinamide, nicotine, nicotinic acid, nifedipine, niflumic acid, nitrazepam,
norepinephrine, nortriptyline, noscapine, nylidrin, oxazepam, oxycodone, oxymorphone,
oxyphenbutazone, oxytetracycline, papaverine, pargyline, pemoline, pentazocine, pento-
barbital, persantine, phenacetin, phenazocine, phenazopyridine, phencyclidine, phendi-
metrazine, phenelzine, pheniramine, phenobarbital, phenothiazine, phensuximide, phen-
termine, phenylbutazone, phenylephrine, phenylpropanolamine, piperocaine, prazepam,
prednisolone, primidone, probenecid, progesterone, propiomazine, propranolol, propylpar-
aben, pseudoephedrine, puromycin, pyrilamine, pyrithyldione, quazepam, quinaldic acid,
quinidine, quinine, ranitidine, recinnamine, reserpine, resorcinol, saccharin, albuterol,
salicylamide, salicylic acid, scopolamine, scopoletin, secobarbital, strychnine, sulfacetam-
ide, sufadiazine, sulfadimethoxine, sulfaethidole, sulfamerazine, sulfamethazine, sulfa-
methoxizole, 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, tri-
amcinolone, tribenzylamine, trichloromethiazide, trifluoperazine, trihexyphenidyl, tri-
methoprim, 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: 250 x 4.6 10 jim Chiralpak OD (Daicel)
Mobile phase: Carbon dioxide: MeCN: EtOH: diethylamine 69.5:15:15:0.5
Column temperature: 30
Flow rate: 2
Detector: UV 220
CHROMATOGRAM
Retention time: 5.2, 7 (enantiomers)
KEYWORDS
SFC; chiral; pressure 200 bar
REFERENCE
Kot, A.; Sandra, P.; Venema, A. Sub- and supercritical fluid chromatography on packed columns: A
versatile tool for the enantioselective separation of basic and acidic drugs. J.Chromatogr.Sci., 1994,
32, 439-448

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flurazepam, glutethimide, hydroxyamoxapine, imipramine, maprotiline, meperidine, metabolites,
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