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Tramadol

Molecular formula: C16H25NO2


Molecular weight: 263.4
CAS Registry No.: 27203-92-5 (tramadol), 22204-88-2 (tramadol
hydrochloride)

SAMPLE
Matrix: solutions
Sample preparation: Dissolve in mobile phase.
HPLCVARIABLES
Guard column: 15 X 3.2 7 |xm Applied Biosystems pre-column
Column: 100 X 2 10 (xm ixPorasil
Mobile phase: MeCN: 5 mM pH 3.75 sodium acetate 80:20
Flow rate: 1
Injection volume: 200
Detector: UV 214
CHROMATOGRAM
Retention time: 11.0
Limit of detection: 6.3 ng/mL
OTHER SUBSTANCES
Simultaneous: buprenorphine, butorphanol, codeine, ethylmorphine, fentanyl, morphine,
nalbuphine
Noninterfering: atropine, diazepam, neostigmine, pancuronium, succinylcholine, thiopen-
tal
Interfering: meperidine
REFERENCE
Ho, S.-T.; Wang, J.-J.; Ho, W.; Hu, O.Y.-P. Determination of buprenorphine by high-performance liquid
chromatography with fluorescence detection: application to human and rabbit pharmacokinetic stud-
ies. J.Chromatogr., 1991, 570, 339-350

SAMPLE
Matrix: urine
Sample preparation: Adjust pH of 1 mL urine to >10 with 25% aqueous ammonia, add 4
mL n-hexane, shake for 15 min, centrifuge at 3000 g for 15 min. Remove the organic layer
and evaporate it to dryness under a stream of nitrogen, reconstitute the residue in 100
|JLL mobile phase, inject a 50 jxL aliquot.
HPLC VARIABLES
Guard column: 25 X 4.6 10 |xm Chiralpak AD amylose tris-3,5-dimethylphenyl carbamate
Column: 250 X 4.6 10 jmm Chiralpak AD amylose tris-3,5-dimethylphenyl carbamate
Mobile phase: n-Hexane:isopropanol:diethylamine 97.5:2.5:0.01
Flow rate: 1
Injection volume: 50
Detector: UV 270
CHROMATOGRAM
Retention time: 10 (+), 15 (-)

OTHER SUBSTANCES
Extracted: metabolites
KEYWORDS
chiral; pharmacokinetics
REFERENCE
Elsing, B.; Blaschke, G. Achiral and chiral high-performance liquid chromatographic determination of
tramadol and its major metabolites in urine after oral administration of racemic tramadol.
J.Chromatogr., 1993, 612, 223-230

SAMPLE
Matrix: urine
Sample preparation: Adjust pH of 1 mL urine to 9 with 50-100 |xL 50 mM pH 10.5 Tris
buffer, add 50 |xL 50 |xg/mL IS in water, add 4 mL n-hexane: ethyl acetate 80:20, shake
for 15 min, centrifuge at 3000 g for 15 min, repeat extraction. Combine the organic layers
and evaporate them to dryness under a stream of nitrogen, reconstitute the residue in
100 |xL mobile phase, inject a 50 |xL aliquot.

HPLCVARIABLES
Guard column: 25 X 4 5 |xm RP Select B (Merck)
Column: 250 X 4 5 jxm RP Select B (Merck)
Mobile phase: MeOH: 50 mM pH 3.0 phosphate buffer 30:70
Flow rate: 1
Injection volume: 50
Detector: UV 270

CHROMATOGRAM
Retention time: 12.8
Internal standard: l-(m-ethoxyphenyl)-2-(dimethylaminomethyl)cyclohexan-l-ol (24.9)
Limit of detection: 80 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
pharmacokinetics
REFERENCE
Elsing, B.; Blaschke, G. Achiral and chiral high-performance liquid chromatographic determination of
tramadol and its major metabolites in urine after oral administration of racemic tramadol.
J.Chromatogr., 1993, 612, 223-230

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