Retinoic Acid

Molecular formula: C20H28O2

Molecular weight: 300.4
CAS Registry No.: 302-79-4 (tretinoin (all-trans)), 4759-48-2 (isotretinoin (13-cis))

SAMPLE
Matrix: blood
Sample preparation: Condition a 100 mg methyl-Cl Accubond SPE cartridge (J&W) with
three 1 mL portions of MeOH and three 1 mL portions of 1% ammonium acetate. 500 |xL
Plasma + 20 p,L 1 \xg/mL acitretin in MeCN containing 10 mM BHT + 1 mL 10 mM
BHT in isopropanol, vortex, rotate for 15 min, centrifuge at 16000 g for 10 min. Remove
the supernatant and add it to 11 mL 1% ammonium acetate, add to the SPE cartridge,
wash with 1 mL 0.1% ammonium acetate, wash with 1 mL MeOH: 0.1% ammonium ac-
etate 50:50, dry under vacuum for 30 s, elute with 1.5 mL 10 mM BHT in MeCN. Add
10 |xL pentafluorobenzyl bromide and 10 |xL 10 mg/mL potassium carbonate in MeCN:
water 50:50 to the eluate, vortex, let stand at room temperature for 1 h, evaporate to
dryness under reduced pressure for 2 h, reconstitute with 20-100 \xh 10 mM BHT in
MeCN, inject a 20 |xL aliquot.

HPLCVARIABLES
Column: 150 X 3.9 Nova-Pak C18 + 75 X 3.9 Nova-Pak C18 (in series)
Mobile phase: Gradient. MeCN:buffer 80:20 for 10 min, to 90:10 (step gradient). (Buffer
was 100 mM ammonium acetate adjusted to pH 5.0 with acetic acid.)
Column temperature: 40
Injection volume: 20
Detector: UV 369; MS Hewlett-Packard model 5988A, particle beam interface nebulizer
60, helium 35 psi, m/z 299

CHROMATOGRAM
Retention time: 26 (isotretinoin), 28.2 (tretinoin)
Internal standard: acitretin (m/z 325) (16)
Limit of detection: 0.05 ng/mL

OTHER SUBSTANCES
Extracted: 9-cis-retinoic acid

KEYWORDS
plasma; protect from light; derivatization; SPE

REFERENCE
Lehman, P.A.; Franz, T.J. A sensitive high-pressure liquid chromatography/particle beam/mass spec-
trometry assay for the determination of all-trans-vetinoic acid and 13-ds-retinoic acid in human
plasma. J.Pharm.Sci., 1996, 85, 287-290

SAMPLE
Matrix: blood
Sample preparation: 10 fxL Serum + 30 JULL 200-500 ng/mL retinyl acetate in
isopropanol: dichloroethane 2:1 + 5 |xL glacial acetic acid, vortex for 30 s, centrifuge for
1 min, inject a 10-20 |xL aliquot.

HPLCVARIABLES
Guard column: C18 (Upchurch)
Column: 150 X 4.6 Ultracarb 5 ODS30 (Phenomenex)
Mobile phase: MeCN: dichloromethane: MeOH 85:12:3 containing 0.1% ammonium ace-
tate (dissolve ammonium acetate in MeOH first)
Flow rate: 1
Injection volume: 10-20
Detector: UV 335

CHROMATOGRAM
Retention time: 9 (tretinoin)
Internal standard: retinyl acetate (5.5)
Limit of detection: 10 ng/mL
Limit of quantitation: 50 ng/mL

OTHER SUBSTANCES
Extracted: vitamin A

KEYWORDS
protect from light; serum

REFERENCE
Barua, A.B.; Kostic, D.; Barua, M.; Olson, J.A. Determination of retinol and retinoic acid in capillary
blood by high-performance liquid chromatography. J.Liq.Chromatogr., 1995, 18, 1459-1471

SAMPLE
Matrix: blood
Sample preparation: Condition a Bakerbond SPE octadecyl SPE cartridge with 2 mL
MeOH and 2 mL 1 M acetic acid. 1 mL Serum + 3 mL 1 M acetic acid + 50 |xL 10 |xM
IS in DMSO, mix, add to the SPE cartridge, wash with 2 mL acetone: 1 M acetic acid
50:50, dry under vacuum for 15 min, elute with 500 JJLL MeCN. Evaporate the eluate and
take up the residue in 200 |ULL mobile phase, inject a 50 JULL aliquot.

HPLC VARIABLES
Column: 250 X 4 5 |xm Lichrospher Si-60
Mobile phase: Hexane: dichloromethane :l,4-dioxane 78:18:4 containing 1% acetic acid
Flow rate: 0.8
Injection volume: 50
Detector: UV 360

CHROMATOGRAM
Retention time: 6 (isotretinoin), 7 (tretinoin)
Internal standard: (all-E)-3-methyl-7-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-
2,4,6-octanoic acid (Ro 13-6307) (7.5)
Limit of detection: 1.5 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
protect from light; serum; SPE; normal phase; pharmacokinetics
REFERENCE
Lefebvre, P.; Agadir, A.; Cornic, C; Gourmel, B.; Hue, B.; Dreux, C; Degos, L.; Chomienne, C. Simul-
taneous determination of a\\-trans and 13-cis retinoic acids and their 4-oxo metabolites by adsorption
liquid chromatography after solid-phase extraction. J.Chromatogr.B, 1995, 666, 55-61

SAMPLE
Matrix: blood
Sample preparation: 500 |xL Plasma + 1 mL IS in EtOH, vortex for 30 s, add 5 mL water,
add 7.5 mL n-hexane, add 300 uX, 2 M HCl, rotate for 10 min, centrifuge at 1250 g for 8
min. Remove the organic layer and evaporate it at room temperature under a stream of
nitrogen. Dissolve the residue in 100 |xL mobile phase, inject a 50 |xL aliquot.

HPLCVARIABLES
Column: 150 X 4.6 5 |xm Spherisorb S5W
Mobile phase: n-Hexane:isopropanol:acetic acid 200:0.7:0.135
Flow rate: 0.9
Injection volume: 50
Detector: UV 350

CHROMATOGRAM
Retention time: 8 (isotretinoin), 11 (tretinoin)
Internal standard: Ro 15-1570 (22)
Limit of detection: 0.5 ng/mL

OTHER SUBSTANCES
Extracted: retinol

KEYWORDS
plasma; normal phase

REFERENCE
Meyer, E.; Lambert, W.E.; De Leenheer, A.P. Simultaneous determination of endogenous retinoic acid
isomers and retinol in human plasma by isocratic normal-phase HPLC with ultraviolet detection.
Clin.Chem., 1994, 40, 48-51

SAMPLE
Matrix: blood
Sample preparation: 200 jxL Serum + 2 mL diluting agent (amber tube), mix for 10 s,
add 5 mL n-hexane, vortex for 30 s, centrifuge at 1600 g at 4 for 5 min. Remove the
organic phase and concentrate to < 1 mL under vacuum below 30, evaporate the rest of
the solvent under nitrogen, dissolve the residue in 25 |xL MeCN: MeOH 2:1, mix for 30
s, centrifuge at 8000 g for 1 min, inject a 20 |xL aliquot. (Diluting agent was MeCN: 100
mM ammonium acetate 25:75, pH adjusted to 5.5 with acetic acid.)

HPLCVARIABLES
Column: 150 X 4.6 5 |xm Chemcosorb 5-ODS-H
Mobile phase: MeCN: MeOH: 100 mM ammonium acetate 46.7:23.3:30, pH adjusted to
7.0
Column temperature: 50
Flow rate: 1
Injection volume: 20
Detector: UV 340
CHROMATOGRAM
Retention time: 14.9 (isotretinoin), 17.0 (tretinoin)
Limit of quantitation: 0.5 ng/mL

OTHER SUBSTANCES
Extracted: vitamin A (retinol)

KEYWORDS
serum

REFERENCE
Takeda, N.; Yamamoto, A. Simultaneous determination of 13-cis- and all-trans-retinoic acids and retinol
in human serum by high-performance liquid chromatography. J.Chromatogr.B, 1994, 657, 53-59

SAMPLE
Matrix: blood
Sample preparation: 500 JJLL Plasma or serum + 100 /JLL 6 jxg/mL 9-methylanthracene in
MeOH + 1.5 mL MeCN + 100 \xL 100 mM perchloric acid, flush headspace of vial with
argon, vortex, centrifuge, inject a 50 |xL aliquot of the supernatant. Sonicate serum, sol-
vents, and mobile phase under vacuum before use. Use low-actinic glassware and yellow
light.

HPLCVARIABLES
Column: 250 X 4.6 5 |xm Zorbax C18
Mobile phase: MeCN: 0.5% acetic acid 85:15 containing 0.05% sodium hexanesulfonate
Flow rate: 2
Injection volume: 50
Detector: UV 365

CHROMATOGRAM
Retention time: 6 (isotretinoin), 8 (tretinoin)
Internal standard: 9-methylanthracene (5)
Limit of detection: 12 ng/mL

OTHER SUBSTANCES
Simultaneous: 4-oxo-13-cis-retinoic acid, retinol

KEYWORDS
plasma; serum

REFERENCE
Gadde, R.R.; Burton, RW. Simple reversed-phase high-performance liquid chromatographic method for
13-cis-retinoic acid in serum. J.Chromatogr., 1992, 593, 41-46

SAMPLE
Matrix: blood
Sample preparation: 0.5-2 mL Plasma + 100 |xL pH 7 phosphate buffer + 2 mL diethyl
ether:ethyl acetate 50:50, vortex gently for 5 min, centrifuge at 2000 g for 10 min. Re-
move the organic layer and evaporate it to dryness under a stream of nitrogen, reconsti-
tute the residue in 30-100 |xL MeOH, inject a 25 |xL aliquot.

HPLCVARIABLES
Column: 250 X 4.6 5 jjim Nucleosil C18
Mobile phase: MeOH: 1% aqueous acetic acid 85:15
Flow rate: 1.5
Injection volume: 25
Detector: UV 350

CHROMATOGRAM
Retention time: 12 (isotretinoin), 15 (tretinoin)
Limit of detection: 2 ng/mL

OTHER SUBSTANCES
Extracted: acitretin, 13-cis-acitretin, etretinate, 4-oxo-13-cis-retinoic acid
Noninterfering: antidepressants, benzodiazepines, psoralen

KEYWORDS
plasma; handle under yellow light

REFERENCE
Bun, H.; al-Mallah, N.R.; Aubert, C; Cano, J.R High-performance liquid chromatography of aromatic
retinoids and isotretinoin in biological fluids. Methods EnzymoL, 1990, 189, 167-172

SAMPLE
Matrix: blood
Sample preparation: 500 |xL Plasma + 750 JJLL 100 ng/mL acitretin in MeCN: 9 mM NaOH
20:80, centrifuge at 1500 g for 3 min, inject a 500 jxL aliquot onto column A with mobile
phase A and elute for 7 min, elute column A in backflush mode with mobile phase A for
3 min, backflush contents of column A onto column B with mobile phase B and start the
gradient for mobile phase B. At the end of the process flush the lines with component B
of mobile phase B, re-equilibrate columns for 4 min. (Keep sample at 10 in the
autosampler.)

HPLCVARIABLES
Column: A 14 X 4.6 37-50 u,m Bondapak C18 Corasil (column fitted with 3 |xm sieves not
glass fiber filters); B 30 X 4 5 |xm Spherisorb ODS 1 + 125 X 4 5 ^m Spherisorb ODS 1
+ 125 X 4 5 (Jim Spherisorb ODS 1
Mobile phase: AMeCN:l% ammonium acetate 10:90; B Gradient. A was MeCN-.water:
10% ammonium acetate:acetic acid 600:400:4:30. B was MeCN:water: 10% ammonium
acetate .acetic acid 850:146:4:10. A: B 100:0 to 70:30 over 6 min, then to 0:100 over 5
min, stay at 0:100 for 11 min.
Flow rate: A 1.5; B 1
Injection volume: 500
Detector: UV 360

CHROMATOGRAM
Retention time: 25 (isotretinoin), 27 (tretinoin)
Internal standard: acitretin (23)
Limit of detection: 0.5-1 ng/mL
Limit of quantitation: 2 ng/mL

OTHER SUBSTANCES
Extracted: metabolites, isotretinoin, 4-oxoisotretinoin, 4-oxotretinoin

KEYWORDS
plasma; column-switching

REFERENCE
Wyss, R. Determination of retinoids in plasma by high-performance liquid chromatography and auto-
mated column switching. Methods EnzymoL, 1990, 189, 146-155
SAMPLE
Matrix: blood
Sample preparation: 500 |xL Plasma + 50 fxL 5% perchloric acid, vortex for 30 s, add 500
|JLL ethyl acetate, whirl for 1 min, centrifuge at 13000 g for 1 min, inject a 50 |xL aliquot
of the organic layer.

HPLCVARIABLES
Guard column: present but not specified
Column: 250 X 4.6 5 jim Ultrasphere ODS + 300 X 4 10 jxm jjiBondapak in series
Mobile phase: MeCN: 1% ammonium acetate 95:5
Flow rate: 2.5
Injection volume: 50
Detector: UV 340; UV 365
CHROMATOGRAM
Retention time: 6.5 (isotretinoin), 8.2 (tretinoin)
OTHER SUBSTANCES
Extracted: vitamin A
KEYWORDS
protect from light; plasma
REFERENCE
Peng, Y-M.; Xu, M.-J.; Alberts, D.S. Analysis and stability of retinol in plasma. J.Nail.Cancer Inst.,
1987, 78, 95-99

SAMPLE
Matrix: blood
Sample preparation: 500 |xL Plasma + 100 fxL 5% perchloric acid, mix rapidly, add 500
jiL ethyl acetate, mix for 60-90 s, centrifuge at 13000 g for 5 min, inject an aliquot of the
supernatant.
HPLCVARIABLES
Column: Reversed-phase C18 (Waters or Bio-Rad)
Mobile phase: MeCN: 1% ammonium acetate 75:25
Flow rate: 2.5
Detector: UV 340
CHROMATOGRAM
Retention time: 3 (isotretinoin), 4 (tretinoin)
Limit of detection: 20 ng/mL
OTHER SUBSTANCES
Extracted: vitamin A

KEYWORDS
plasma; pharmacokinetics; protect from light
REFERENCE
Davis, T.R; Peng, Y-M.; Goodman, G.E.; Alberts, D.S. HPLC, MS, and pharmacokinetics of melphalan,
bisantrene and 13-cis retinoic acid. J.Chromatogr.ScL, 1982, 20, 511-516

SAMPLE
Matrix: blood, microsomal incubations
Sample preparation: 500 JJLL Plasma or 250 |xL microsomal incubation + 25 |xL 20 |xg/mL
IS in MeCN + 350 p,L l-butanol:MeCN 50:50, mix thoroughly, add 300 jxL saturated
K2HPO4, mix, centrifuge at 3000 g for 10 min, inject an aliquot of the organic layer.

HPLCVARIABLES
Column: Adsorbosphere C18
Mobile phase: Gradient. MeCN: 10 mM ammonium acetate from 50:50 to 95:5 over 10
min
Flow rate: 1.5
Detector: UV 365

CHROMATOGRAM
Internal standard: Ro 23-4736
Limit of detection: 10 ng/mL

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
plasma; protect from light; human; liver; for tretinoin; pharmacokinetics

REFERENCE
Schwartz, E.L.; Hallam, S.; Gallagher, R.E.; Wiernik, RH. Inhibition of all-rcms-retinoic acid metabo-
lism by fluconazole in vitro and in patients with acute promyelocytic leukemia. Biochem.Pharmacol.,
1995, 50, 923-928

SAMPLE
Matrix: blood, tissue
Sample preparation: Plasma. Add 5 volumes of MeOH to plasma, cool to -20, centrifuge
at 16500 g for 5 min. Remove the supernatant and evaporate it to dryness under reduced
pressure, reconstitute the residue in mobile phase, centrifuge at 16500 g for 5 min, inject
a 10-250 |JLL aliquot. Liver. Homogenize (Kinematica-Polytron with an Aggregat PTA 10TS
generator) rat liver with 5 volumes of MeOH at 20000 rpm for 20 s, let stand at -20
overnight, centrifuge at 0-4 at 3200 g for 10 min. Remove the supernatant and evaporate
it to dryness under reduced pressure, reconstitute the residue in mobile phase, centrifuge
at 16500 g for 5 min, inject a 10-250 JULL aliquot.

HPLCVARIABLES
Guard column: 20 X 4 5 ^m C18 (Hewlett-Packard or Alltech)
Column: 250 X 4.6 5 |xm Microsorb-MV
Mobile phase: MeCNrIO mM ammonium acetate: glacial acetic acid 80:20:1
Column temperature: 40
Flow rate: 1
Injection volume: 10-250
Detector: UV 348

CHROMATOGRAM
Retention time: 13.6 (isotretinoin), 16.3 (tretinoin)

OTHER SUBSTANCES
Extracted: metabolites

KEYWORDS
rat; plasma; liver
REFERENCE
Shirley, M.A.; Bennani, Y.L.; Boehm, M.F.; Breau, A.R; Pathirana, C; UIm, E.H. Oxidative and reductive
metabolism of 9-ds-retinoic acid in the rat. Identification of the 13,14-dihydro-9-czs-retinoic acid and
its taurine conjugate. Drug Metab.Dispos., 1996, 24, 293-302

SAMPLE
Matrix: culture media
Sample preparation: 100 |xL Culture media + 200 |xL ice-cold EtOH, mix thoroughly, let
stand for 15 min, centrifuge at 12000 g for 15 min, inject an aliquot of the supernatant.
HPLCVARIABLES
Guard column: Whatman CO: PELL ODS guard column
Column: 100 X 8 5 |xm Nova-Pak C18 (radial-packed)
Mobile phase: MeOH: 100 mM pH 7.0 ammonium acetate 90:10
Flow rate: 1
Detector: UV 340
CHROMATOGRAM
Retention time: 9.44 (tretinoin)
OTHER SUBSTANCES
Extracted: acitretin, etretinate, isotretin, motretinid, retinal, vitamin A
REFERENCE
Kochhar, D.M.; Penner, J.D.; Minutella, L.M. Biotransformation of etretinate and developmental toxicity
of etretin and other aromatic retinoids in teratogenesis bioassays. Drug Metab.Dispos., 1989, 17,
618-624

SAMPLE
Matrix: solutions
Sample preparation: Inject an aliquot of a solution in the mobile phase.
HPLC VARIABLES
Column: 250 X 4.6 5 |xm Zorbax Rx-SiI
Mobile phase: Heptane: THF: acetic acid 96.5:3.5:0.015
Flow rate: 1.4
Detector: MS, Finnigan 4023, particle beam interface (Vestec universal interface model
700), electron impact mode 77 eV, scan m/z 200-350 (positive ion), multiplier voltage 1200
V, source 300; UV 365
CHROMATOGRAM
Retention time: 13 (isotretinoin), 16 (tretinoin)
OTHER SUBSTANCES
Simultaneous: degradation products
KEYWORDS
normal phase
REFERENCE
Bempong, D.K.; Honigberg, LL.; Meltzer, N.M. Normal phase LC-MS determination of retinoic acid
degradation products. J.Pharm.Biomed.Anal, 1995, 13, 285-291

SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 250 X 4.6 8 \xm Unisphere-PBD (polybutadiene on alumina) (Biotage, Charlottes-
viUe, VA)
Mobile phase: MeOH: water 92:8
Flow rate: 1
Detector: UV 330

CHROMATOGRAM
Retention time: 3.5 (tretinoin)

OTHER SUBSTANCES
Simultaneous: retinol acetate, vitamin A, vitamin E
REFERENCE
Jedrejewski, RT.; Taylor, L.T. Comparison of silica-, alumina-, and polymer-based stationary phases for
reversed-phase liquid chromatography. J.Chromatogr.ScL, 1995, 33, 438-445

SAMPLE
Matrix: solutions

HPLCVARIABLES
Column: 250 X 4.6 5 |xm Suplex-pKb-100 (Supelco)
Mobile phase: MeOH: acetic acid 99.95:0.05
Flow rate: 1
Detector: UV 350

CHROMATOGRAM
Retention time: 13.5 (isotretinoin), 22 (tretinoin)
OTHER SUBSTANCES
Simultaneous: other isomers

KEYWORDS
photoisomerization solutions
REFERENCE
Sundquist, A.R.; Stahl, W.; Steigel, A.; Sies, H. Separation of retinoic acid all-trans, mono-eis and poly-
cis isomers by reversed-phase high-performance liquid chromatography. J.Chromatogr., 1993, 637,
201-205

SAMPLE
Matrix: tissue
Sample preparation: 100-120 mg Frog embryos + 100 JJLL isopropanol, sonicate on ice,
vortex for 1 min, centrifuge at 4 at 4000 g for 20 min, inject an aliquot of the supernatant.

HPLCVARIABLES
Guard column: 20 X 4 10 |xm LiChrosorb RB 18
Column: 125 X 4.6 3 |xm Spherisorb ODS II
Mobile phase: Gradient. MeOH: 40 mM pH 7.3 ammonium acetate from 55:45 to 100:0
over 18 min.
Flow rate: 1.6
Detector: UV 354

CHROMATOGRAM
Retention time: 12.5 (isotretinoin), 12.7 (9-cis-retinoic acid), 13.5 (tretinoin)
OTHER SUBSTANCES
Extracted: metabolites, vitamin A

KEYWORDS
frog; embryo

REFERENCE
Creech Kraft, J.; Juchau, M.R. Xenopus laevis: A model system for the study of embryonic retinoid
metabolism. III. Isomerization and metabolism of all-rcms-retinoic acid and 9-ds-retinoic acid and
their dysmorphogenic effects in embryos during neurulation. Drug Metab.Dispos., 1995, 23, 1058
1071

SAMPLE
Matrix: tissue
Sample preparation: 100-120 mg Tadpole embryos + 100 mL isopropanol, sonicate on ice,
vortex for 1 min, centrifuge at 4000 g at 4 for 20 min, inject an aliquot of the supernatant.

HPLCVARIABLES
Guard column: 20 X 4 10 jxm LiChrosorb RB 18
Column: 125 X 4.6 3 |jim Spherisorb ODS II
Mobile phase: Gradient. MeOH: 40 mM pH 7.3 ammonium acetate from 55:45 to 100:0
over 20 min
Flow rate: 1.6
Detector: UV 354

CHROMATOGRAM
Retention time: 11.8 (isotretinoin), 12.2 (tretinoin)

OTHER SUBSTANCES
Extracted: metabolites, tretinoin, vitamin A

KEYWORDS
handle under yellow light; tadpoles; embryos

REFERENCE
Creech Kraft, J.; Kimelman, D.; Juchau, M.R. Xenopus Laevis: A model system for the study of embry-
onic retinoid metabolism. I. Embryonic metabolism of 9-cis- and all-rarcs-retinals and retinols and
their corresponding acid forms. Drug Metab.Dispos., 1995, 23, 72-82

ANNOTATED BIBLIOGRAPHY
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pharmacokinetic properties of orally administered all-trans-retinoic acid and 9-cis-retinoic acid in
the plasma of nude mice. Drug Metab.Dispos., 1994, 22, 451-458 [gradient; LOD 0.5 ng/mL; ex-
tracted metabolites, isotretinoin, tretinoin, vitamin A]
Eckhoff, C; Chari, S.; Kromka, M.; Staudner, H.; Juhasz, L.; Rudiger, H.; Agnish, N. Teratogenicity and
transplacental pharmacokinetics of 13-cis-retinoic acid in rabbits. Toxicol.Appl.Pharmacol., 1994,
125, 34-41 [plasma; acitretin (IS); SPE; column temp 35; tissue; extracted metabolites, isotretinoin,
tretinoin]
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trans-retinoic acid in human plasma by high-performance liquid chromatography. J.Chromatogr.B,
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Jiang, X.G.; Xi, N.Z. [A reversed-phase HPLC method for determining tretinoin]. Chung Kuo Yao Li
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0.25 ng; column temp 60; gradient; simultaneous metabolites, isotretinoin, tretinoin]
Ranalder, U.B.; Lausecker, B.B.; Huselton, C. Micro liquid chromatography-mass spectrometry with
direct liquid introduction used for separation and quantitation of all-rans- and 13-cis-retinoic acids
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ng/mL; extracted metabolites, isotretinoin, tretinoin]
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retinoic acid using microcalorimetry and HPLC analysis. Pharm.Res., 1992, 9, 1203-1208 [solutions;
simultaneous degradation products, isotretinoin, tretinoin]
Bryan, PD.; Honigberg, LL.; Meltzer, N.M. Electrochemical detection of retinoids using normal phase
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vitamin A palmitate]
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all-trans-retinoic acid, their 4-oxo- metabolites and retinol in plasma, amniotic fluid and embryo by
reversed-phase high-performance liquid chromatography with a precolumn switching technique.
J.Liq.Chromatogr., 1988, 11, 2051-2069 [gradient; LOD 2 ng/mL; mouse; extracted metabolites, is-
otretinoin, tretinoin, vitamin A]
Wyss, R.; Bucheli, F. Quantitative analysis of retinoids in biological fluids by high-performance liquid
chromatography using column switching. I. Determination of isotretinoin and tretinoin and their 4-
oxo metabolites in plasma. J.Chromatogr., 1988, 424, 303-314 [plasma; LOQ 2 ng/mL; extracted
metabolites, isotretinoin, tretinoin; etretin (IS)]
Furr, H.C; Amedee-Manesme, 0.; Olson, J.A. Gradient reversed-phase high-performance liquid chro-
matographic separation of naturally occurring retinoids. J.Chromatogr., 1984, 309, 299-307 [gradi-
ent; rat; human; pig; liver; kidney; extracted retinyl esters, tretinoin, vitamin A]
Shelley, R.; Price, J.C.; Jun, H.W.; Cadwallader, D.E.; Capomacchia, A.C. Improved and rapid high-
performance liquid chromatographic assay for 13-cis-retinoic acid or all-trans-retinoic acid.
J.Pharm.Sci., 1982, 71, 262264 [rat; serum; extracted isotretinoin, retinol acetate, tretinoin, vita-
min A; pharmacokinetics; LOQ 100 ng/mL]
Vane, F.M.; Stoltenborg, J.K.; Bugge, CJ. Determination of 13-cis-retinoic acid and its major metabolite,
4-oxo-13-cis-retinoic acid, in human blood by reversed-phase high-performance liquid chromatogra-
phy. J.Chromatogr., 1982, 227, 471-484 [gradient; whole blood; extracted metabolites, isotretinoin,
tretinoin, vitamin A; LOQ 10 ng/mL; pharmacokinetics]