Paclitaxel

Molecular formula: C47H51NO14

Molecular weight: 853.9
CAS Registry No.: 33069-62-4

SAMPLE
Matrix: blood
Sample preparation: 1 mL Plasma + 300 ng N-nitrosodiphenylamine (Caution! N-Nitrosodiphenylamine is a carcinogen!) + 400 fxL 500 mM Na2HPO4, vortex for 5 s, add 5 mL
ethyl acetate, mix for 1 min, centrifuge at 3000 rpm for 10 min. Remove the organic layer
and evaporate it to dryness under a stream of nitrogen at room temperature, reconstitute
the residue in 200 jxL mobile phase, vortex for 30 s, sonicate for 30 s, inject a 100 jxL
aliquot.
HPLC VARIABLES
Guard column: Guard-PAK C18 (Waters)
Column: 100 X 8 4 jim Nova Pak C18 radial compression
Mobile phase: MeCN:buffer 45.5:55.5 (Buffer was 1 mM Na2HPO4 adjusted to pH 5 with
phosphoric acid.)
Flow rate: 4.5
Injection volume: 100
Detector: UV 227
CHROMATOGRAM
Retention time: 5.26
Internal standard: N-nitrosodiphenylamine (6.45)
Limit of quantitation: 10 ng/mL
OTHER SUBSTANCES
Simultaneous: carmustine, 5-fluorouracil, hydrocortisone, teniposide, thiotepa
Noninterfering: acetaminophen, aspirin, bleomycin, busulfan, carboplatin, cyclophosphamide, cytarabine, dacarbazine, hydroxyurea, ifosfamide, methotrexate, mitomycin, mitoxantrone, procarbazine
KEYWORDS
plasma; pharmacokinetics
REFERENCE
el-Yazigi, A.; Yusuf, A. Expedient liquid chromatographic assay for paclitaxel in plasma after its administration to cancer patients. Ther.Drug Monit., 1995, 17, 511-515

SAMPLE
Matrix: blood
Sample preparation: Condition a 1 mL cyano Bond Elut SPE cartridge with 2 mL MeOH
and 2 mL 10 mM pH 5.0 ammonium acetate buffer. 1 mL Plasma + 1 mL 200 mM
ammonium acetate, mix, add a 1 mL aliquot to the SPE cartridge, wash with 2 mL 10
mM pH 5.0 ammonium acetate, wash with 1 mL MeOH: 10 mM pH 5.0 ammonium acetate 20:80, wash with 1 mL hexane, dry under vacuum for 1 min, elute with 2 mL
MeCN: triethylamine 100:0.1, evaporate the eluate to dryness under a stream of nitrogen
at 30, reconstitute with 200 JULL mobile phase, vortex for 30 s, inject a 50 |xL aliquot.

HPLCVARIABLES
Guard column: 4 X 4 5 |xm LiChrospher RP-8
Column: 150 X 4.6 5 |xm APEX-octyl (Jones Chromatography)
Mobile phase: MeCN: MeOH: 20 mM pH 5.0 ammonium acetate buffer 40:10:50
Injection volume: 50
Detector: UV 227
CHROMATOGRAM

Retention time: 10.1

Limit of quantitation: 10 ng/mL
OTHERSUBSTANCES

Extracted: metabolites
KEYWORDS
plasma; SPE
REFERENCE
Huizing, M.T.; Sparreboom, A.; Rosing, H.; van Tellingen, 0.; Pinedo, H.M.; Beijnen, J.H. Quantification
of paclitaxel metabolites in human plasma by high-performance liquid chromatography.
J.Chromatogr.B, 1995, 674, 261-268

SAMPLE
Matrix: blood
Sample preparation: Plasma. 500 |xL Plasma + 20 |xL 10 |xg/mL diethylstilbestrol in
MeOH, vortex for 20 s, add 5 mL MTBE, vortex for 20 s, centrifuge at 170 g for 5 min,
freeze in dry ice/acetone. Remove the organic layer and put it into a clean tube (twice),
evaporate it to dryness under a stream of nitrogen at room temperature, reconstitute the
residue in 200 |xL MeCN: water 45:55, inject a 50 |xL aliquot. Microsomal incubations.
250 |xL Microsomal incubation + 5 mL MTBE, vortex for 20 s, centrifuge at 170 g for 5
min, freeze in dry ice/acetone. Remove the organic layer and put it into a clean tube
(twice), evaporate it to dryness under a stream of nitrogen at room temperature, reconstitute the residue in 200 |xL MeCN: water 45:55, inject a 50 fxL aliquot.
HPLCVARIABLES
Guard column: 30 X 3.2 5 jjim Hypersil C8
Column: 150 X 3.2 5 jxm Hypersil C8
Mobile phase: MeOH: 30 mM pH 6 potassium phosphate buffer 58:42 (After 25 min wash
column with MeOH: water 95:5 for 7 min, re-equilibrate for 13 min. Wash is not necessary
for microsomal incubations.)
Flow rate: 0.4
Injection volume: 50
Detector: UV 227
CHROMATOGRAM

Retention time: 22
Internal standard: diethylstilbestrol (17)
Limit of detection: 2OnM
OTHER SUBSTANCES

Extracted: metabolites
KEYWORDS
microsomal incubations; plasma; human; liver; pharmacokinetics

REFERENCE
Sonnichsen, D.S.; Liu, Q.; Schuetz, E.G.; Schuetz, J.D.; Pappo, A.; Relling, M.V. Variability of human
cytochrome P450 paclitaxel metabolism. J.Pharm.Exp.Ther., 1995, 275, 566-575

SAMPLE
Matrix: blood
Sample preparation: Dilute 50-500 |xL mouse plasma to 500 |xL with water, add 25 jxL
40 |xg/mL N-octylbenzamide in MeOH, mix, add 4 mL MTBE, vortex for 30 s, centrifuge
at 500 g at 4 for 10 min. Remove 3 mL of the organic layer and evaporate it to dryness
under a stream of nitrogen, reconstitute the residue in 100 |xL MeOH, inject a 70 |xL
aliquot. (Full details for the synthesis of N-octylbenzamide are given in the paper.)
HPLCVARIABLES
Guard column: 6.6 X 3 10 (xm ixBondapak C18
Column: 300 X 3.9 10 |xm ^Bondapak C18
Mobile phase: MeOH: water 70:30
Flow rate: 2
Injection volume: 70
Detector: UV 227
CHROMATOGRAM
Retention time: 7.35
Internal standard: N-octylbenzamide (13.00)
Limit of quantitation: 0.15 nM
KEYWORDS
plasma; mouse
REFERENCE
Sharma, A.; Conway, W.D.; Straubinger, R.M. Reversed-phase high-performance liquid chromatographic
determination of taxol in mouse plasma. J.Chromatogr.B, 1994, 655, 315-319

SAMPLE
Matrix: blood
Sample preparation: Condition a 1 mL Bond Elut cyano SPE cartridge with 2 mL MeOH
and 2 mL 10 mM pH 5.0 ammonium acetate, do not allow to dry. 500 (xL Plasma + 500
IJLL 200 mM pH 5.0 ammonium acetate, vortex for 20 s, add to the SPE cartridge, wash
with 2 mL 10 mM pH 5.0 ammonium acetate, wash with 2 mL MeOH: 10 mM pH 5.0
ammonium acetate 20:80, wash with 1 mL hexane, dry under vacuum for 1 min. Elute
with two 1 mL volumes of 0.1% triethylamine in MeCN, evaporate the eluate to dryness
at 30 under nitrogen, reconstitute the residue in 200 |xL MeCN: MeOH: water 40:10:50
containing 10 mM ammonium acetate adjusted to pH 5.0 with glacial acetic acid, vortex
for 30 s, inject a 100 |xL aliquot.
HPLCVARIABLES
Column: 250 X 4.6 5 |xm octyl (Jones Chromatography)
Mobile phase: MeCN: MeOH: water :1 M ammonium acetate adjusted to pH 5.0 with glacial acetic acid 40:10:49:1
FIo5W rate: 1
Injection volume: 100
Detector: UV 227
CHROMATOGRAM
Retention time: 10.6
Limit of quantitation: 10 ng/mL

OTHER SUBSTANCES
Simultaneous: amsacrine, baccatin III, cephalomannine, daunorubicin, epipaclitaxel, paclitaxel C
KEYWORDS
plasma; SPE; pharmacokinetics; methyl paclitaxel can be used as IS
REFERENCE
Willey, T.A.; Bekos, E.J.; Gaver, R.C.; Duncan, G.F.; Tay, L.K.; Beijnen, J.H.; Farmen, R.H. High-performance liquid chromatographic procedure for the quantitative determination of paclitaxel (Taxol) in
human plasma. J.Chromatogr., 1993, 621, 231-238

SAMPLE
Matrix: blood, culture medium, tissue
Sample preparation: Plasma, culture medium. 0.2-1 mL plasma or culture medium + 100
U.L 8.7 |xg/mL cephalomannine in MeOH + three volumes ethyl acetate, extract, repeat
extraction. Combine extracts and centrifuge them at 2000 g for 5 min. Remove the supernatant and evaporate it to dryness under a stream of nitrogen, reconstitute the residue
in 100-150 IJLL MeCN: water 37.5:62.5, inject a 10-60 |xL aliquot onto column A with
mobile phase A and elute to waste. After 8 min direct the effluent from column A onto
column B. After another 7 min elute column B with mobile phase B, monitor the effluent
from column B, re-equilibrate column A with mobile phase A. Tissue. 40 mg Dog bladder
tissue + 100 |xL 8.7 |xg/mL cephalomannine in MeOH + 3-4 mL ethyl acetate or MeCN,
homogenize (Tekmar) for 1 min, wash homogenizer probe with 3-4 mL same solvent for
15-20 s. Combine organic layers and centrifuge them at 2000 g for 5 min. Remove the
supernatant and evaporate it to dryness under a stream of nitrogen, reconstitute the
residue in 100-150 |xL MeCN: water 37.5:62.5, inject a 10-60 jxL aliquot onto column A
with mobile phase A and elute to waste. After 8 min direct the effluent from column A
onto column B. After another 7 min elute column B with mobile phase B, monitor the
effluent from column B, re-equilibrate column A with mobile phase A.
HPLCVARIABLES
Column: A Nova-Pak C8 guard column + 75 X 3.9 3 juim Nova-Pak C8; B 250 X 4.6 5 jmm
Bakerbond C18
Mobile phase: A MeCN: water 37.5:62.5; B MeCN: water 49:51
Flow rate: A 1; B 1.2
Injection volume: 10-60
Detector: UV 229
CHROMATOGRAM
Retention time: 21.35
Internal standard: cephalomannine (20.11)
Limit of detection: 5 ng/mL (plasma); 5 ng/inj. (tissue)
KEYWORDS
plasma; column-switching; dog; bladder; heart-cut
REFERENCE
Song, D.; Au, J.L.-S. Isocratic high-performance liquid chromatographic assay of taxol in biological fluids
and tissues using automated column switching. J.Chromatogr.B, 1995, 663, 337-344

SAMPLE
Matrix: blood, feces, tissue, urine
Sample preparation: Condition a 1 mL 100 mg Bond Elut cyano SPE cartridge with 2 mL
MeOH and 2 mL 10 mM pH 5.0 ammonium acetate buffer. Dilute urine with five volumes
of human plasma. Homogenize (Biospec) tissue or feces in 5-10 volumes of 4% Boseral in

water at 4. 200 (Feces, plasma), 200-1000 (tissue), or 1000 (urine) ^LL + 25 |xL 20 |jig/mL
2'-methylpaclitaxel in MeOH H- 4 mL diethyl ether, mix vigorously for 5 min, centrifuge
at 460 g for 5 min, freeze in EtOH/dry ice, repeat the extraction. Combine the organic
layers and evaporate them to dryness under vacuum at 43, reconstitute in 250 jxL human
plasma, add 300 |xL 200 mM pH 5.0 ammonium acetate buffer, mix, add 500 |xL to the
SPE cartridge, wash with 2 mL 10 mM pH 5.0 ammonium acetate buffer, wash with 1
mL MeOH: 10 mM pH 5.0 ammonium acetate buffer 20:80, elute with 500 |xL MeCN:
triethylamine 100:0.1. Evaporate the eluate to dryness under vacuum, reconstitute 200
jxL in MeCN: MeOH: water 40:10:50, vortex for 30 s, inject a 50 |xL aliquot.
HPLCVARIABLES
Column: 150 X 4.6 5 jim APEX-octyl (Jones Chromatography)
Mobile phase: MeCN: MeOH: 200 mM pH 5.0 ammonium acetate buffer 40:10:50
Flow rate: 1
Injection volume: 50
Detector: UV 227
CHROMATOGRAM
Retention time: 10.6
Internal standard: 2'-methylpaclitaxel (15.3)
Limit of detection: 15 ng/mL (plasma)
Limit of quantitation: 25-125 ng/g
OTHER SUBSTANCES
Extracted: metabolites
KEYWORDS
plasma; mouse; SPE; brain; fat; muscle; breast; colon; appendix; intestine; stomach; liver;
gall bladder; kidney; lung; spleen; heart; uterus; ovary; thymus; lymph nodes
REFERENCE
Sparreboom, A.; van Tellingen, O.; Nooijen, W.J.; Beijnen, J.H. Determination of paclitaxel and metabolites in mouse plasma, tissues, urine and faeces by semi-automated reversed-phase high-performance liquid chromatography. J.Chromatogr.B, 1995, 664, 383-391

SAMPLE
Matrix: blood, urine
Sample preparation: 800 |JLL Plasma or urine H- 200 JXL 7.5 |xM N-cyclohexylbenzamide
in MeCN, vortex, add 3 mL ethyl acetate, shake vigorously for 2 min, centrifuge at 150
g for 2 min. Remove the organic layer and evaporate it under nitrogen at room temperature, dissolve the residue in 100 fiL MeCN, inject an aliquot.
HPLCVARIABLES
Guard column: C-18 Guard-Pak
Column: 100 X 8 10 ^m C18 Radial-Pak
Mobile phase: MeCN: water from 35:65 to 100:0 exponentially over 20 min (Waters exponential gradient curve no. 9), keep at 100.0 for 7 min, re-equilibrate at 35:65 for 5 min.
Flow rate: 2.5
Detector: UV 227
CHROMATOGRAM
Retention time: 19.5
Internal standard: N-cyclohexylbenz amide (12.5)
Limit of quantitation: 30 nM
KEYWORDS
plasma

REFERENCE
Longnecker, S.M.; Donehower, R.C.; Cates, A.E.; Chen, T.-L.; Brundrett, R.B.; Grochow, L.B.; Ettinger,
D.S.; Colvin, M. High-performance liquid chromatographic assay for taxol in human plasma and
urine and pharmacokinetics in a phase I trial. Cancer Treat.Rep., 1987, 71, 53-59

SAMPLE
Matrix: cell cultures
Sample preparation: Filter (Miracloth, Calbiochem). Evaporate 1 mL of filtrate to dryness
under vacuum, reconstitute in 200 |xL MeOH: acetic acid 99.99:0.01, sonicate for 1 min,
centrifuge at 16000 g for 15 min, filter (0.2 \xm PVDF), inject an aliquot of the filtrate.
HPLCVARIABLES
Guard column: Upchurch ODS
Column: 250 X 4.6 4 |xm Curosil G (Phenomenex)
Mobile phase: MeCN-.water 52.5:47.5
Flow rate: 1
Detector: UV 228
CHROMATOGRAM
Retention time: 18
OTHER SUBSTANCES
Simultaneous: impurities, related compounds
REFERENCE
Ketchum, R.E.B.; Gibson, D.M. A novel method of isolating taxanes from cell suspension cultures of yew
(Taxus spp.). J.Liq.Chromatogr., 1995, 18, 1093-1111

SAMPLE
Matrix: cell cultures
Sample preparation: Homogenize (Omni-mix) 50 mg bark or foliage in 1.5 mL MeOH for
2 min, sonicate for 5 min, centrifuge, filter (0.2 |xm) the supernatant, inject an aliquot of
the nitrate.
HPLCVARIABLES
Guard column: phenyl (Rainin)
Column: 250 X 4.6 8 |xm Dynamax 60 A phenyl (Rainin)
Mobile phase: MeOH: MeCN: 50 mM pH 4.4 acetate buffer 20:39:41
Flow rate: 1
Detector: UV 227
CHROMATOGRAM
Retention time: 19
OTHER SUBSTANCES
Extracted: cephalomannine
REFERENCE
Wickremesinhe, E.R.M.; Arteca, R.N. Methodology for the identification and purification of taxol and
cephalomannine from Taxus callus cultures. J.Liq.Chromatogr., 1993, 16, 3263-3274

SAMPLE
Matrix: formulations
Sample preparation: Dilute with mobile phase, inject an aliquot.

HPLCVARIABLES
Column: 250 X 4.6 5 |xm C18
Mobile phase: MeCN: water 40:60
Flow rate: 2.25
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 4.95
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: Dilute 1:4, inject a 20 |ULL aliquot.
HPLCVARIABLES
Column: 250 X 4.6 5 |xm Adsorbosphere C18
Mobile phase: MeCN: 12.5 mM ammonium phosphate 60:40, pH adjusted to 4.5 with 1 M
HCl
Flow rate: 1
Injection volume: 20
Detector: UV 227
CHROMATOGRAM
Retention time: 10.0
OTHER SUBSTANCES
Noninterfering: ondansetron, ranitidine
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
Sample preparation: Filter (5 jxm), dilute 1 mg/mL solutions tenfold with the same solvent, inject an aliquot.
HPLC VARIABLES
Column: 250 X 4.6 5 |xm Vydac C18
Mobile phase: MeCN: water 53:47
Flow rate: 1.5
Detector: UV 254

CHROMATOGRAM

Retention time: 6.09

KEYWORDS

injections; 5% dextrose; saline; stability-indicating

REFERENCE
Xu, Q.; Trissel, L.A.; Martinez, J.F. Stability of paclitaxel in 5% dextrose injection or 0.9% sodium
chloride injection at 4, 22, or 32C. Am.J.Hosp.Pharm., 1994, 51, 3058-3060

SAMPLE

Matrix: microsomal incubations

Sample preparation: Extract microsomal incubation with 5 volumes ethyl acetate, evaporate the organic layer to dryness under a stream of nitrogen, reconstitute the residue in
500 fxL mobile phase, inject an aliquot.
HPLCVARIABLES

Column: 250 X 3.2 6 |xm Curosil G (Phenomenex)

Mobile phase: MeCN: water 35:65
Flow rate: 0.6
Detector: UV 229
CHROMATOGRAM

Retention time: 58
OTHER SUBSTANCES

Extracted: metabolites
KEYWORDS

rat; liver
REFERENCE
Anderson, CD.; Wang, J.; Kumar, G.N.; McMillan, J.M.; Walle, U.K.; Walle, T. Dexamethasone induction
of taxol metabolism in the rat. Drug Metab.Dispos., 1995, 23, 1286-1290

SAMPLE

Matrix: plants
Sample preparation: Extract needles with MeOH, concentrate the extract under reduced
pressure at <30, partition concentrate with 0.8 volumes of water and 0.8 volumes of
chloroform, repeat the extraction with 0.6 volumes chloroform then 0.4 volumes chloroform, concentrate under reduced pressure, dry under vacuum at 35-40, dissolve in
MeCN/water, inject an aliquot.
HPLCVARIABLES

Column: 250 X 4.6 5 jim Partisil C8

Mobile phase: MeCN: MeOH: water 50:10:40
Flow rate: 0.5
Detector: UV 254
KEYWORDS

needles; details of preparative HPLC also in paper

REFERENCE
Rao, K.V.; Bhakuni, R.S.; Juchum, J.; Davies, R.M. A large scale process for paclitaxel and other taxanes
from the needles of Taxus x media Hicksii and Taxus floridana using reverse phase column chromatography. J.Liq.Chrom.Rel.TechnoL, 1996, 19, 427-447

SAMPLE
Matrix: plants
Sample preparation: Shake 2 g ground plant material and 100 mL MeOH on a flat-bed
orbital shaker for 12-16 h, allow to settle. Remove a 10 mL aliquot and add it to 10 mL
5% NaCl solution, wash twice with 10 mL portions of hexane, discard the hexane washes,
extract four times with 10 mL portions of dichloromethane. Filter the dichloromethane
extracts through 3 g anhydrous sodium sulfate, evaporate the filtrate to dryness, reconstitute with 2 mL dichloromethane, add to a 250 X 10 column dry packed with 1 g 149250 |xm Davsil silica (Alltech), wash with two 2 mL portions of dichloromethane, wash
with two 5 mL portions of acetone: dichloromethane 4:96, elute with two 5 mL portions
of acetone: dichloromethane 50:50. Combine the eluates and evaporate them to dryness
under a stream of nitrogen at 35, reconstitute the residue in 1 mL MeOH, inject a 10
IxL aliquot.
HPLCVARIABLES
Column: 150 X 4.6 5 |xm Zorbax SB-C8
Mobile phase: Gradient. A was MeCN. B was MeOH: water 20:80. A:B 20:80 for 5 min,
to 25:75 over 0.1 min, maintain at 25:75 for 8.9 min, to 35:65 over 10 min, maintain at
35:65 for 17 min, to 65:35 over 3 min, maintain at 65:35 for 11 min, return to initial
conditions over 5 min, re-equilibrate for 20 min.
Column temperature: 35
Flow rate: 1
Injection volume: 10
Detector: UV 227
CHROMATOGRAM
Retention time: 42.5
Limit of detection: 660 ng/g
OTHER SUBSTANCES
Extracted: baccatin III, cephalomannine
KEYWORDS
SPE; leaves; twigs
REFERENCE
Lauren, D.R.; Jensen, D.J.; Douglas, J.A. Analysis of taxol, 10-deacetylbaccatin III and related compounds in Taxus baccata. J.Chromatogr.A, 1995, 712, 303-309

SAMPLE
Matrix: plants
Sample preparation: Homogenize (Omni-mix) 50 mg bark or foliage in 1.5 mL MeOH for
2 min, sonicate for 5 min, centrifuge, filter (0.2 |xm) the supernatant, inject an aliquot of
the filtrate.
HPLCVARIABLES
Column: 1000 X 4.7 7 |xm porous graphitized carbon (Shandon)
Mobile phase: Dioxane: water 46:54 (Caution! Dioxane is a carcinogen!)
Flow rate: 0.8
Injection volume: 20
Detector: UV 228
CHROMATOGRAM
Retention time: 18
Limit of detection: 30 ng/mL

KEYWORDS

bark; foliage
REFERENCE
Forgacs, E. Use of porous graphitized carbon column for determining taxol in Taxus baccata. Chromatographia, 1994, 39, 740-742

SAMPLE

Matrix: plants
Sample preparation: Grind yew needles to <3 mm in a blender. Weigh out 3-4 g, add 100
mL MeOH, shake for 16 h on a wrist-action shaker, filter (Whatman 1 or 2 paper), wash
the solid with 25 mL MeOH. Evaporate the nitrate to dryness under reduced pressure at
40-43, reconstitute the residue in 10 mL MeOH and 1 mL water. Condition a 47 mm
Empore SPE extraction disk (3M Corp.) with 15 mL ethyl acetate, 15 mL MeOH and 15
mL water. Use a 47 mm polypropylene separator with 10 |xm pore size (Gelman 61757)
in front of the extraction disk. Pass 10 mL water and 7 mL crude extract through the
disk, wash with 15 mL water, wash with 15 mL MeOH: water 20:80, 15 mL MeOH: water
45:55, elute with 20 mL MeOH-.water 80:20, filter (2 jim) the eluate, inject a 10 \xh
aliquot.
HPLCVARIABLES

Guard column: Taxsil guard cartridge (0335-CS) (MetaChem)

Column: 250 X 4.6 5 |xm Taxsil (0335-250 X 046) (MetaChem)
Mobile phase: Gradient. A was MeCN. B was MeOH: water 30:70. A:B 41:59 for 15 min,
to 65:35 over 5 min, maintain at 65:35 for 10 min, to 41:59 over 5 min, maintain at
41:59 for 5 min.
Column temperature: 32
Flow rate: 1
Injection volume: 10
Detector: UV 230
CHROMATOGRAM

Retention time: 10
KEYWORDS

SPE
REFERENCE
Mattina, M.J.I.; MacEachern, G.J. Extraction, purification by solid-phase extraction and high-performance liquid chromatographic analysis of taxanes from ornamental Taxus needles. J .Chromatogr.A,
1994, 679, 269-275

SAMPLE
Matrix: solutions
HPLCVARIABLES

Column: 250 X 4.6 5 jjim LiChrospher diol

Mobile phase: Gradient. MeOH: carbon dioxide 8:92 for 3 min, to 28:72 over 25 min, to
35:65 over 5.7 min, maintain at 35:65 for 4 min.
Column temperature: 30
Flow rate: 2
Detector: UV 227
CHROMATOGRAM

Retention time: 15.77

OTHER SUBSTANCES
Simultaneous: impurities, degradation products
KEYWORDS
SFC; pressure 150 bar
REFERENCE
Jagota, N.K.; Nair, J.B.; Frazer, R.; Klee, M.; Wang, M.Z. Supercritical fluid chromatography of paclitaxel. J.ChromatogrA, 1996, 721, 315-322

SAMPLE
Matrix: solutions
Sample preparation: Inject a 20 |xL aliquot.
HPLCVARIABLES
Guard column: C-18 Guard Pak
Column: 100 X 8 10 (xm Resolve C-18 radial compression (Waters)
Mobile phase: Gradient. MeCN.water from 45:55 to 100:0 over 20 min (exponential gradient), maintain at 100:0 for 5 min, re-equilibrate at initial conditions for 5 min.
Column temperature: 21
Flow rate: 2.5
Injection volume: 20
Detector: UV 227
CHROMATOGRAM
Retention time: 9.33
REFERENCE
Wenk, M.R.; Fahr, A.; Reszka, R.; Seelig, J. Paclitaxel partitioning into lipid bilayers. J.Pharm.ScL,
1996, 85, 228-231

SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 250 X 4.6 5 |xm Partisil C8
Mobile phase: MeCN: MeOH: water 50:10:40
Flow rate: 0.5
Detector: UV 254
CHROMATOGRAM
Retention time: 17
OTHER SUBSTANCES
Simultaneous: impurities
KEYWORDS
details of preparative chromatography
REFERENCE
Rao, K.V.; Hanuman, J.B.; Alvarez, C; Stoy, M.; Juchum, J.; Davies, R.M.; Baxley, R. Anew large-scale
process for taxol and related taxanes from Taxus brevifolia. Pharm.Res., 1995, 12, 1003-1010

SAMPLE
Matrix: urine

Sample preparation: 19 mL Urine + 1 mL Cremophor EL: EtOH 50:50 (Sigma), mix. 500
|xL Sample + 500 jxL 200 mM pH 5.0 ammonium acetate buffer + 5 mL n-butyl chloride,
rotate slowly for 10 min, centrifuge at 2500 g for 10 min, freeze in dry ice/acetone. Remove
the organic layer and evaporate it to dryness under a stream of nitrogen at 40, reconstitute the residue in 200 |xL MeCN: MeOH: water 40:50:10 (40:10:50 ?), vortex for 30
s, inject a 50 |xL aliquot.
HPLCVARIABLES
Guard column: 4 x 4 5 |xm LiChrospher RP-8
Column: 150 X 4.6 5 |xm APEX-octyl (Jones Chromatography)
Mobile phase: MeCN: MeOH: 20 mM pH 5.0 ammonium acetate buffer 40:10:50
Flow rate: 1
Injection volume: 50
Detector: UV 227
CHROMATOGRAM
Retention time: 10
Limit of quantitation: 250 ng/mL
REFERENCE
Huizing, M.T.; Rosing, H.; Koopman, F.; Keung, A.C.R; Pinedo, H.M.; Beijnen, J.H. High-performance
liquid chromatographic procedures for the quantitative determination of paclitaxel (Taxol) in human
urine. J.Chromatogr.B, 1995, 664, 373-382

SAMPLE
Matrix: urine
Sample preparation: Condition a 1 mL 100 mg Bond Elut cyano SPE cartridge with 2 mL
MeOH and 2 mL 10 mM pH 5.0 ammonium acetate buffer. 19 mL Urine + 1 mL Cremophor ELiEtOH 50:50 (Sigma), mix. 500 jxL Sample + 500 fxL 200 mM pH 5.0 ammonium acetate buffer, vortex for 20 s, add to the SPE cartridge, wash with 2 mL 10 mM
pH 5.0 ammonium acetate buffer, wash with 1 mL MeOH: 10 mM pH 5.0 ammonium
acetate buffer 20:80, wash with 1 mL hexane, dry under vacuum for 1 min, elute with 2
mL MeCN: triethylamine 100:0.1. Evaporate the eluate to dryness under a stream of
nitrogen at 30, reconstitute in 200 |xL MeCN: MeOH: water 40:50:10 (40:10:50 ?), vortex
for 30 s, inject a 50 (xL aliquot.
HPLC VARIABLES
Guard column: 4 X 4 5 |xm LiChrospher RP-8
Column: 150 X 4.6 5 |xm APEX-octyl (Jones Chromatography)
Mobile phase: MeCN: MeOH: 20 mM pH 5.0 ammonium acetate buffer 40:10:50
Flow rate: 1
Injection volume: 50
Detector: UV 227
CHROMATOGRAM
Retention time: 10
Limit of detection: 8 ng/mL
Limit of quantitation: 10 ng/mL
KEYWORDS
SPE
REFERENCE
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