Terbinafine

Molecular formula: C21H25N

Molecular weight: 291.4
CAS Registry No.: 91161-71-6

SAMPLE
Matrix: blood
Sample preparation: 500 |xL Plasma + 100 |xL 5 |xg/mL IS in water + 1 mL 200 mM pH
9 borate buffer + 8 mL n-hexane, shake horizontally at 200 rpm for 25 min, centrifuge
at 2000 g for 10 min. Remove 7 mL of the supernatant and add it to 1 mL 500 mM
sulfuric acid: isopropanol 85:15, shake for 15 min, centrifuge at 2000 g for 5 min, inject
a 250 |xL aliquot of the aqueous phase.

HPLCVARIABLES
Column: 100 X 4.6 3 |xm Pecospher C18
Mobile phase: MeCN-.buffer 50:50 (Buffer was 3.35 mL triethylamine and 2.65 mL ortho-
phosphoric acid in 2 L water.)
Flow rate: 1
Injection volume: 250
Detector: UV 224

CHROMATOGRAM
Retention time: 9.9
Internal standard: 2-(l-naphthyl)-l-(3-phenylprop-2-enyl)piperidine hydrochloride (IW 85-
190) (12.4)
Limit of quantitation: 2 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
plasma; pharmacokinetics

REFERENCE
Denouel, J.; Keller, H.P.; Schaub, P.; Delaborde, C; Humbert, H. Determination of terbinafine and its
desmethyl metabolite in human plasma by high-performance liquid chromatography. J.Chroma-
togr.B, 1995, 663, 353-359

SAMPLE
Matrix: blood
Sample preparation: Add IS to plasma, adjust pH to 9, extract with n-hexane. Add the
organic layer to 500 mM sulfuric acid: isopropanol 85:15, extract, inject an aliquot of the
aqueous layer.

HPLCVARIABLES
Column: 100 X 4.6 Spherisorb RP-18/5
Mobile phase: MeCN: 10 mM K2HPO4: triethylamine 60:40:0.01
Column temperature: 50
Flow rate: 1
Detector: UV 224
CHROMATOGRAM
Internal standard: 2-(l-naphthyl)-l-(3-phenylprop-2-enyl)piperidine hydrochloride (SDZ
85-190)
Limit of quantitation: 2 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
plasma; pharmacokinetics

REFERENCE
Kovarik, J.M.; Kirkesseli, S.; Humbert, H.; Grass, P.; Kutz, K. Dose-proportional pharmacokinetics of
terbinafine and its N-demethylated metabolite in healthy volunteers. Br.J.DermatoL, 1992, 126
Suppl39, 8-13

SAMPLE
Matrix: blood, milk
Sample preparation: Plasma. 100 jxL Plasma + 300 |xL 4 g/L ammonium acetate in
MeOH, centrifuge, inject a 200 |xL aliquot of the supernatant. Milk. 100 |xL Milk + 100
\LL 100 mM NaCl (?) + 800 |xL MeCN, centrifuge, inject a 100 |xL aliquot of the
supernatant.

HPLCVARIABLES
Column: 250 X 4.6 10 |xm RP-8 (Merck)
Mobile phase: MeOH: 10 mM pH 7 phosphate buffer 77:23 containing 4 g/L ammonium
acetate
Column temperature: 35
Flow rate: 1.5
Injection volume: 100-200
Detector: E, Metrohm 656, glassy carbon electrode + 1.1 V

CHROMATOGRAM
Retention time: 10.50 (plasma), 12.03 (milk)
Limit of detection: 50 ng/mL (plasma); 150 ng/mL (milk)

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
plasma; human; rat; dog

REFERENCE
Schatz, R; Haberl, H. Analytical methods for the determination of terbinafine and its metabolites in
human plasma, milk and urine. Arzneimittelforschung, 1989, 39, 527-532

SAMPLE
Matrix: blood, urine
Sample preparation: Plasma. 750 ^xL Plasma + 50 |xL 320 jxg/mL IS + 25 |xL 85% phos-
phoric acid + 750 |xL EtOH: isopropanol 75:25, vortex for 10 s, let stand on crushed ice
for 30 min, centrifuge at 1000 g for 15 min. Remove a 400 |xL aliquot of the supernatant
and add it to 400 |JLL 10 mM pH 5 phosphate buffer, vortex for 10 s, inject a 250 |xL
aliquot onto column A with mobile phase A and elute to waste. After 4 min backflush
column A with mobile phase A to waste, after 1 min backflush the contents of column A
onto column B with mobile phase B, after 10 min remove column A from the circuit, elute
 column B with mobile phase B and monitor the effluent. (Recondition column A with
mobile phase A for 30 min at 0.1 mL/min). Urine. 500 fxL Urine + 50 |JLL 220 jxg/mL IS
+ 200 |xL 10000 IU/mL p-glucuronidase (bovine liver, Sigma) in 10 mL 10 mM pH 5
phosphate buffer, heat at 37 for 18 h, add 300 |xL 10 mM pH 5 phosphate buffer, vortex
for 10 s, centrifuge at 2500 g for 18 min, inject a 100-250 |xL aliquot of the supernatant
onto column A with mobile phase A and elute to waste. After 4 min backflush column A
with mobile phase A to waste, after 1 min backflush the contents of column A onto column
B with mobile phase B, after 10 min remove column A from the circuit, elute column B
with mobile phase B and monitor the effluent. (Recondition column A with mobile phase
A for 30 min at 0.1 mL/min).

HPLC VARIABLES
Column: A 15 X 3.2 30-40 \xm Perisorb RP-2 (Merck); B 220 X 4.6 5 |xm Spheri-5 phenyl
Mobile phase: A 20 mM KH2PO4 containing 0.25% triethylamine, pH adjusted to 3.8 with
1 M phosphoric acid; B MeCN: buffer 55:45 (Buffer was 20 mM KH2PO4 containing
0.125% triethylamine, pH adjusted to 3.8 with 1 M phosphoric acid.)
Column temperature: 30-35
Flow rate: 1
Injection volume: 100-250
Detector: UV 224

CHROMATOGRAM
Retention time: 34-36
Internal standard: 2-(l-naphthyl)- l-(3-phenylprop-2-enyl)piperidine hydrochloride (IW 85-
190)
Limit of quantitation: 20 ng/mL (plasma); 100 ng/mL (urine)

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
plasma; column-switching; pharmacokinetics

REFERENCE
Zehender, H.; Denouel, J.; Roy, M.; Le Saux, L.; Schaub, P. Simultaneous determination of terbinafine
(Lamisil) and five metabolites in human plasma and urine by high-performance liquid chromatog-
raphy using on-line solid-phase extraction. J.Chromatogr.B, 1995, 664, 347-355

SAMPLE
Matrix: nails
Sample preparation: 10-20 mg Nail clippings + 1 mL 1 M NaOH, let stand at room
temperature for 48 h, adjust pH to 9 with 1 M HCl, adjust molarity to 0.2 with water,
add 100 |xL 10 mg/mL proteinase K (Type I fungal proteinase, P0390, Sigma), heat at 37
for 12-16 h, add 50 ng IS, vortex, add 3 mL hexane, vortex for 3 min, add 100 JULL MeOH,
centrifuge at 500 g for 10 min. Remove a 2.7 mL aliquot of the organic layer and add it
to 200 JJLL 500 mM sulfuric acid: propanol 85:15, vortex for 3 min, centrifuge, inject a 100
JULL aliquot of the aqueous layer.

HPLC VARIABLES
Guard column: 20 X 2 5-20 |xm Prepsil RPC8 (Apex)
Column: 250 mm long 5 |xm Spherisorb C18
Mobile phase: MeCN: 10 mM phosphate buffer: triethylamine 80:20:0.1, pH adjusted to
4.0 with 85% phosphoric acid
Flow rate: 2
Injection volume: 100
Detector: UV 224
CHROMATOGRAM
Retention time: 7.5
Internal standard: 2-(l-naphthyl)-l-(3-phenylprop-2-enyl)piperidine hydrochloride (San-
doz 85-190) (8.4)

OTHER SUBSTANCES
Extracted: metabolites

REFERENCE
Dykes, RJ.; Thomas, R.; Finlay, A.Y. Determination of terbinafine in nail samples during systemic treat-
ment for onychomycoses. Br.J.DermatoL, 1990, 123, 481-486