Ondansetron

Molecular formula: C18H19N3O

Molecular weight: 293.4
CAS Registry No.: 99614-02-5 (ondansetron), 116002-70-1
(ondansetron), 99614-01-4 (ondansetron hydrochloride dihydrate),
103639-04-9 (ondansetron hydrochloride dihydrate)

SAMPLE
Matrix: beverages
Sample preparation: Juice, soft drinks. Inject a 10 |xL aliquot. Tea. Cool 60 mL tea in an
ice bath to about 0 in 1 min. Remove a 10 mL aliquot, add 1 drop concentrated HCl, add
10 mL MeCN, mix, inject a 10 |xL aliquot.
HPLCVARIABLES
Guard column: 30 X 4.6 5 |xm C18 (Phase Separations)
Column: 250 X 4.5 5 |xm Spherisorb cyanopropyl
Mobile phase: MeCN: 20 mM pH 5.4 phosphate buffer 60:40
Flow rate: 2
Injection volume: 10
Detector: UV 216
CHROMATOGRAM
Retention time: 6.5
KEYWORDS
tea; apple juice; soft drinks; stability indicating
REFERENCE
Yamreudeewong, W.; Danthi, S.N.; Hill, R.A.; Fox, J.L. Stability of ondansetron hydrochloride injection
in various beverages. Am.J.Health-Syst.Pharm., 1995, 52, 2011-2014

SAMPLE
Matrix: blood
Sample preparation: Condition a 1 mL 100 mg cyanopropyl SPE cartridge (J & W Scientific) with 2 volumes of MeOH and 2 volumes of water, do not allow to dry. 1 mL Serum
+ 30 JJIL 10 |jLg/mL prazosin in EtOH, vortex for 10 s, add to SPE cartridge, wash with 1
volume MeOH: water 10:90, dry under full vacuum for 20 min, elute with four 250 |JLL
aliquots of MeOH: triethylamine 999:1. Evaporate the eluate to dryness under a stream
of nitrogen at room temperature, dissolve the residue in 250 jxL mobile phase, inject a
100 |xL aliquot.
HPLCVARIABLES
Guard column: 500 X 4.6 10 |xm Chiralcel OD precolumn (sic)
Column: 250 X 4.6 10 ^m Chiralcel OD
Mobile phase: Hexane: 95% EtOH: isopropanol: MeCN 65:25:10:1
Flow rate: 1
Injection volume: 100
Detector: UV 216
CHROMATOGRAM
Retention time: 9.96 (R-(-)), 11.56 (S-(+))
Internal standard: prazosin (7.99)

Limit of detection: 2.5 ng/mL

Limit of quantitation: 10 ng/mL
KEYWORDS
serum; SPE; chiral
REFERENCE
Kelly, J.W.; He, L.; Stewart, J.T. High-performance liquid chromatographic separation of ondansetron
enantiomers in serum using a cellulose-derivatized stationary phase and solid-phase extraction.
J.Chromatogr., 1993, 622, 291-295

SAMPLE
Matrix: blood
Sample preparation: Condition a 100 mg Bond Elut CN SPE cartridge with two 1 mL
portions of MeOH, two 1 mL portions of isopropanol: ammonia 99:1, and two 1 mL portions of water, do not allow to dry. Add 50 |xL 0.5 M HCl then 1 mL plasma to the SPE
cartridge, dry the cartridge with a vacuum, wash with two 1 mL portions of water, allow
to dry, wash with two 1 mL portions of MeCN, allow to dry, elute with two 1 mL portions
of isopropanol: ammonia 99:1, evaporate the eluate at 45 under vacuum, reconstitute in
200 |xL mobile phase, sonicate for 5 min, vortex, inject a 100 JJLL aliquot.
HPLCVARIABLES
Column: 100 X 4.6 3 |jim Spherisorb S3W silica
Mobile phase: MeCN: buffer 40:60 (Buffer was 25 mM sodium acetate adjusted to pH 4.2
with glacial acetic acid.)
Column temperature: 35
Flow rate: 1
Injection volume: 100
Detector: UV 305
CHROMATOGRAM
Retention time: 4-5
Limit of quantitation: 1 ng/mL
OTHER SUBSTANCES
Simultaneous: metabolites, fluorouracil, melphalan
Noninterfering: cisplatin
KEYWORDS
SPE; plasma
REFERENCE
Colthup, P.V.; Felgate, C C ; Palmer, J.L.; Scully, N.L. Determination of ondansetron pharmacokinetics
in the young and elderly. J.Pharm.ScL, 1991, 80, 868-871

SAMPLE
Matrix: blood
Sample preparation: Condition a 100 mg Bondelut CN SPE cartridge with 2 mL MeOH,
2 mL isopropanol: ammonia 99:1, and 2 mL water. Mix 1 mL plasma with 50 |xL 0.5 M
HCl and add to the SPE cartridge, wash with 2 mL water, wash with 2 mL MeCN, elute
with 2 mL isopropanol: ammonia 99:1, evaporate the eluate, dissolve the residue in 200
(xL mobile phase, inject a 100 JULL aliquot.
HPLCVARIABLES
Column: 100 X 4.6 3 ^m Spherisorb S3W silica
Mobile phase: MeCN: 25 mM pH 4.2 sodium acetate 40:60

Column temperature: 35
Injection volume: 100
Detector: UV 305
CHROMATOGRAM
Limit of quantitation: 1 ng/mL
KEYWORDS
plasma; SPE; pharmacokinetics
REFERENCE
Colthup, RV.; Palmer, J.L. The determination in plasma and pharmacokinetics of ondansetron.
Eur.J.Cancer Clin.OncoL, 1989, 25 Suppl 1, S71-S74

SAMPLE
Matrix: formulations
Sample preparation: Dilute with mobile phase, inject a 20 |xL aliquot.
HPLCVARIABLES
Guard column: ODS
Column: 250 X 4.6 5 jxm Spherisorb ODS-I
Mobile phase: MeCN: 100 mM pH 4.5 KH2PO4 40:60
Flow rate: 1.2
Injection volume: 20
Detector: UV 210
CHROMATOGRAM
Retention time: 11.6
Limit of detection: 70 ng/mL
OTHER SUBSTANCES
Simultaneous: diphenhydramine, methyl paraben, propyl paraben
KEYWORDS
injections; saline
REFERENCE
Ye, L.; Stewart, J.T. HPLC determination of an ondansetron and diphenhydramine mixture in 0.9%
sodium chloride injection. J.Liq.Chromatogr.& Rel.TechnoL, 1996, 19, 711-718

SAMPLE
Matrix: formulations
Sample preparation: Dilute with an equal volume of saline (if necessary), filter (0.2 jxm),
inject a 50 |xL aliquot of the nitrate.
HPLCVARIABLES
Column: 250 X 4.6 5 jim Accubond CN (J & W)
Mobile phase: MeCN:buffer 60:40 adjusted to pH 6.0 with 1 M NaOH (Buffer was 20 mM
KH2PO4 containing 5 mM octanesulfonic acid.)
Flow rate: 1.5
Injection volume: 50
Detector: UV 305
CHROMATOGRAM
Retention time: 8-9

KEYWORDS

saline; injections; stability-indicating

REFERENCE
Chung, K.C.; Moon, Y.S.K.; Chin, A.; Ulrich, R.W.; Gill, M.A. Compatibility of ondansetron hydrochloride
and piperacillin sodium tazobactam sodium during simulated Y-site administration. Am.J.HealthSyst.Pharm., 1995, 52, 1554-1556

SAMPLE

Matrix: formulations
Sample preparation: Filter (0.22 |xm), inject a 20 |xL aliquot.
HPLCVARIABLES

Column: 250 X 4.6 5 juim Ultrasphere reverse-phase

Mobile phase: MeCN: water 40:60
Flow rate: 1.5
Injection volume: 20
Detector: UV 200
CHROMATOGRAM

Retention time: 3.3

OTHER SUBSTANCES
Simultaneous: cyclophosphamide
KEYWORDS

injections; saline; 5% dextrose

REFERENCE
Fleming, R.A.; Olsen, D.J.; Savage, RD.; Fox, J.L. Stability of ondansetron hydrochloride and cyclophosphamide in injectable solutions. Am.J.Health-Syst.Pharm., 1995, 52, 514-516

SAMPLE

Matrix: formulations
Sample preparation: Filter (0.22 jxm), inject a 20 |xL aliquot.
HPLCVARIABLES

Column: 300 X 4.6 5 |jim Spherisorb nitrile

Mobile phase: MeCN: 20 mM pH 5.4 phosphate buffer 60:40
Flow rate: 1.2
Injection volume: 20
Detector: UV 216
CHROMATOGRAM

Retention time: 9.3

OTHER SUBSTANCES
Noninterfering: cyclophosphamide
KEYWORDS

injections; saline; 5% dextrose; stability-indicating

REFERENCE
Fleming, R.A.; Olsen, D.J.; Savage, RD.; Fox, J.L. Stability of ondansetron hydrochloride and cyclophosphamide in injectable solutions. Am.J.Health-Syst.Pharm., 1995, 52, 514-516

SAMPLE

Matrix: formulations
Sample preparation: Dilute with 2 volumes water (if necessary), filter (0.45 jxm), inject
an aliquot.
HPLCVARIABLES

Column: 220 X 4.6 10 fxm Spheri-10

Mobile phase: MeCN:DMF:pH 6.5 sodium acetate 15:4:85
Flow rate: 1
Detector: UV 305
KEYWORDS

total parental nutrient; injections; stability-indicating

REFERENCE
Kirkham, J.C.; Rutherford, E.T.; Cunningham, G.N.; Daneshmand, K.A.; Falls, A.L. Stability of ondansetron hydrochloride in a total parenteral nutrient admixture. Am.J.Health-Syst.Pharm., 1995, 52,
1557-1558

SAMPLE

Matrix: formulations
Sample preparation: Inject a 20 jxL aliquot.
HPLC VARIABLES

Column: 100 X 4.6 Spheri-5 RP-8

Mobile phase: MeCN: buffer 45:55 (Buffer was 10 mM KH2PO4 adjusted to pH 4.0 with 1
M KOH.)
Flow rate: 1
Injection volume: 20
Detector: UV 254
CHROMATOGRAM

Retention time: 5.9

Limit of detection: 90 ng/mL
OTHER SUBSTANCES
Simultaneous: glycopyrrolate
Noninterfering: degradation products
KEYWORDS

injections; saline
REFERENCE
Venkateshwaran, T.G.; King, D.T.; Stewart, J.T. HPLC determination of ondansetron-atropine and ondansetron-glycopyrrolate mixtures in 0.9% sodium chloride injection. J.Liq.Chromatogr., 1995, 18,
2647-2659

SAMPLE

Matrix: formulations
HPLCVARIABLES

Column: 250 X 4.6 5 |xm Zorbax Rx-C8 base deactivated octylsilane (Mac-Mod Analytical)
Mobile phase: MeCN: 10 mM KH2PO4 adjusted to pH 4.0 with 1 M KOH 23:77
Flow rate: 1

Injection volume: 20
Detector: UV 273
CHROMATOGRAM

Retention time: 14.8

Limit of detection: 20 ng/mL
OTHER SUBSTANCES

Simultaneous: metoclopramide
KEYWORDS

saline; injections
REFERENCE
Venkateshwaran, T.G.; King, D.T.; Stewart, J.T. HPLC determination of a metoclopramide and ondansetron mixture in 0.9% sodium chloride injection. J.Liq.Chromatogr., 1995, 18, 117-126

SAMPLE

Matrix: formulations
Sample preparation: 1 mL Sample + 50 \xL 150 ng/mL cimetidine, inject a 10 |xL aliquot.
HPLCVARIABLES

Column: 250 X 4.8 Spherisorb S5CN

Mobile phase: MeCN-.buffer 50:50 (Buffer was 20 mM KH2PO4 adjusted to pH 5.4 with 1
M NaOH.)
Flow rate: 1.5
Injection volume: 10
Detector: UV 216
CHROMATOGRAM

Retention time: 9.5

Internal standard: cimetidine (4.5)
KEYWORDS

injections; stability-indicating; 5% dextrose

REFERENCE
Bosso, J.A.; Prince, R.A.; Fox, J.L. Compatibility of ondansetron hydrochloride with fluconazole, ceftazidime, aztreonam, and cefazolin sodium under simulated Y-site conditions. Am.J.Hosp.Pharm.,
1994, 51, 389-391

SAMPLE

Matrix: formulations
Sample preparation: Dilute 1:2, inject a 50 |xL aliquot.
HPLCVARIABLES

Column: 250 X 4.6 5 |xm Accubond CN (J & W)

Mobile phase: MeCN: 20 mM KH2PO4 and 5 mM octanesulfonic acid 50:50, pH adjusted
to 6.0 with 1 M NaOH
Flow rate: 1.5
Injection volume: 50
Detector: UV 305
CHROMATOGRAM

Retention time: 8.5

OTHER SUBSTANCES
Noninterfering: ranitidine, paclitaxel
KEYWORDS
injections; 5% dextrose; stability-indicating
REFERENCE
Burm, J.-R; Jhee, S.S.; Chin, A.; Moon, Y.S.K.; Jeong, E.; Nii, L.; Fox, J.L.; Gill, M.A. Stability of
paclitaxel with ondansetron hydrochloride or ranitidine hydrochloride during simulated Y-site administration. Am.J.Hosp.Pharm., 1994, 51, 1201-1204

SAMPLE
Matrix: formulations
HPLCVARIABLES
Column: 300 X 3.9 10 |xm ixBondapak phenyl
Mobile phase: MeCN:buffer 50:50 (Buffer was 20 mM KH2PO4 adjusted to pH 5.4 with 1
M NaOH.)
Flow rate: 1
Injection volume: 20
Detector: UV 233
CHROMATOGRAM
Retention time: 10.5
Limit of detection: 200 ng/mL
OTHER SUBSTANCES
Simultaneous: degradation products, doxorubicin, methyl paraben, vincristine
KEYWORDS
injections; saline
REFERENCE
King, D.T.; Venkateshwaran, T.G.; Stewart, J.T. HPLC determination of a vincristine, doxorubicin, and
ondansetron mixture in 0.9% sodium chloride injection. J.Liq.Chromatogr., 1994, 17, 1399 1411

SAMPLE
Matrix: formulations
Sample preparation: Dilute 1:10, inject a 10 (JLL aliquot.
HPLCVARIABLES
Column: 300 X 4.6 5 |xm Spherisorb CN
Mobile phase: MeCN: 20 mM KH2PO4 50:50, pH adjusted to 5.40 with 1 M NaOH
Flow rate: 1.5
Injection volume: 10
Detector: UV 216
CHROMATOGRAM
Retention time: 8.8-10.0
OTHER SUBSTANCES
Simultaneous: hydromorphone, morphine
KEYWORDS
injections; saline; stability-indicating

REFERENCE
Trissel, L.A.; Xu, Q.; Martinez, J.F.; Fox, J.L. Compatibility and stability of ondansetron hydrochloride
with morphine sulfate and with hydromorphone hydrochloride in 0.9% sodium chloride injection at
4, 22, and 32C. Am.J.Hosp.Pharm., 1994, 51, 2138-2142

SAMPLE
Matrix: formulations
HPLCVARIABLES
Column: 220 X 4.6 5 ynn silica (Brownlee)
Mobile phase: MeCN: 6.25 mM NaH2PO4 adjusted to pH 3.0 with concentrated phosphoric
acid 40:60
Flow rate: 1
Injection volume: 50
Detector: UV 216
CHROMATOGRAM
Retention time: 8.6
Limit of detection: 8 ng/mL
OTHER SUBSTANCES
Simultaneous: dacarbazine, doxorubicin
Noninterfering: degradation products
KEY WORDS
injections; 5% dextrose
REFERENCE
King, D.T.; Stewart, J.T. HPLC determination of dacarbazine, doxorubicin, and ondansetron mixture in
5% dextrose injection on underivatized silica with an aqueous-organic mobile phase.
J.Liq.Chromatogr., 1993, 16, 2309-2323

SAMPLE
Matrix: microsomal incubations
Sample preparation: 750 jxL Microsomal incubation + 1 . 5 mL MeCN, concentrate, inject
an aliquot.
HPLCVARIABLES
Column: 150 X 4.6 5 um Hypersil C8 BDS
Mobile phase: MeCN: 40 mM pH 4.5 ammonium acetate 20:80
Flow rate: 0.8
Detector: Radioactivity
CHROMATOGRAM
Retention time: 26
OTHER SUBSTANCES
Extracted: metabolites
KEYWORDS
human; liver; 14C labeled
REFERENCE
Dixon, CM.; Colthup, RV; Serabjit-Singh, C.J.; Kerr, B.M.; Boehlert, C C ; Park, G.R.; Tarbit, M.H.
Multiple forms of cytochrome P450 are involved in the metabolism of ondansetron in humans. Drug
Metab.Dispos., 1995,23, 1225-1230

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 250 X 4.6 10 |mm Partisil ODSl

Mobile phase: MeOH: 50 mM pH 3.0 phosphoric acid 40:60
Column temperature: 30
Flow rate: 1.5
Detector: Radioactivity
KEYWORDS
14
C labeled
REFERENCE
Collett, A.; Sims, E.; Walker, D.; He, Y-L.; Ayrton, J.; Rowland, M.; Warhurst, G. Comparison of HT2918-Ci and Caco-2 cell lines as models for studying intestinal paracellular drug absorption.
Pharm.Res., 1996, 13, 216-221

SAMPLE

Matrix: solutions
Sample preparation: Inject an aliquot of an aqueous solution.
HPLC VARIABLES

Column: 250 X 4 Nucleosil C18

Mobile phase: MeOH:THF:buffer 30:5:65 (Buffer was 100 mM triethylamine adjusted to
pH 3.0 with nitric acid.)
Flow rate: 0.8
Injection volume: 20
Detector: UV 254
CHROMATOGRAM

Retention time: 7.40

OTHER SUBSTANCES

Simultaneous: granisetron, tropisetron

REFERENCE
Barbato, R; Immacolata La Rotonda, M.; Quaglia, F. Retention behaviour of anti-emetic serotonin antagonists in reversed phase high performance liquid chromatography. Farmaco, 1995, 50, 875-880

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 250 X 4.6 5 ^m YMC-Pack ODS-A

Mobile phase: MeCN: MeOH: isopropanol: 20 mM NaH2PO4 50:15:5:30
Flow rate: 1
Detector: UV 216
CHROMATOGRAM

Retention time: 3.7

OTHER SUBSTANCES

Simultaneous: impurities

REFERENCE
Ong, CR; Chow, K.K.; Ng, CL.; Ong, RM.; Lee, H.K.; Li, S.RY. Use of overlapping resolution mapping
scheme for optimization of the high-performance liquid chromatographic separation of pharmaceuticals. J.Chromatogr.A, 1995, 692, 207-212

ANNOTATED BIBLIOGRAPHY
Rischer, V.; Vickers, A.E.M.; Heitz, R; Mahadevan, S.; Baldeck, J.-R; Minery, P.; Tynes, R. The polymorphic cytochrome P-4502D6 is involved in the metabolism of both 5-hydroxytryptamine antagonists, tropisetron and ondansetron. Drug Metab.Dispos., 1994,22, 269274 [microsomal incubations;
human; liver; extracted metabolites; gradient; column temp 40; radioactivity detection]
Pompilio, RM.; Fox, J.L.; Inagaki, K.; Burm, J.-R; Jhee, S.; Gill, M.A. Stability of ranitidine hydrochloride with ondansetron hydrochloride or fiuconazole during simulated Y-site administration.
Am.J.Hosp.Pharm., 1994, 51, 391394 [stability-indicating; injections; 5% dextrose]
Williams, CL.; Sanders, RL.; Laizure, S.C; Stevens, R.C; Fox, J.L.; Hak, L.J. Stability of ondansetron
hydrochloride in syrups compounded from tablets. Am.J.Hosp.Pharm., 1994, 51, 806-809 [stabilityindicating; syrup]
Graham, CL.; Dukes, G.E.; Kao, C-R; Bertsch, J.M.; Hak, L.J. Stability of ondansetron in large-volume
parenteral solutions. Ann.Pharmacother., 1992, 26, 768-771 [injections; saline; 5% dextrose; lactated
Ringer's solution; simultaneous degradation products]