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A Review: Current Analytical Methods for Determination of Ketoconazole in
Pharmaceutical and Biological Samples

Olga Popovska
1
*, Vesna Rafajlovska
1
, Zoran Kavrakovski
2

1
Ss. Cyril and Methodius University in Skopje, Faculty of Technology and Metallurgy
Rudjer Boskovic 16, 1000 Skopje, REPUBLIC OF MACEDONIA
2
Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy
Vodnjanska 17, 1000 Skopje, REPUBLIC OF MACEDONIA

*Corresponding Author: Olga Popovska



























ABSTRACT:
Ketoconazole is an imidazole antifungal drug which is available in the different
pharmaceutical dosage forms through various routes of administration, such as oral and topical. The
drug is a broad-spectrum antifungal agent and it is used in the treatment of superficial or systemic
fungal infections. This article reviews the current analytical methods for identification and
quantitative determination of ketoconazole in samples. The most commonly used methods for the
determination of ketoconazole presented in this article are chromatographic methods: high-pressure
liquid chromatography (HPLC) and thin-layer chromatography (TLC); electroanalytical methods:
voltammetry and potentiometry; capillary electrophoresis (CE); spectrofluorimetric and
spectrophotometric methods. Recent preferences in the analysis of ketoconazole samples prove
primacy of HPLC and confirm general trends moving towards more sensitive methods, with higher
resolution potential, consuming small quantities of samples and reagents and require less time of the
analysis.
Keywords: Ketoconazole, Antifungal drug, UV-Vis spectroscopy, Chromatography
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Olga Popovska et al Page 2

1. INTRODUCTION
Ketoconazole (cis-1-acetyl-4-[4-[[2-(2,4-dichlorophenyl)-2-(1H-imidazol-1-ylmethyl)-1,3-dioxolan-4-
yl]methoxy]phenyl]piperazine) is a lipophilic imidazole derivative appears as white to off white crystalline powder
(Figure 1). The drug is practically insoluble in water, soluble in strong bases and sparingly soluble in strong acid.
Ketoconazole is a weak base with pKa values of 6.51 and 2.94, contains two nitrogen atoms in the five-membered
azole ring.
1,2


Figure

1: Ketoconazole enantiomer chemical structures.

Cl
O
O
N
N
*
H
O N N
*
Cl
*
H
O N N
*
Cl
O
O
N
N
CH
3
CH
3
O
O
Cl
(+)-(2R,4S)
(-)-(2S,4R)


Ketoconazole is a chiral drug administered as a racemic (1:1) mixture of enantiomers of the cis
configuration.
3,4
It is a broad-spectrum antifungal imidazole agent that has been shown to be efficient in the treatment
of human systemic fungal infections, against Candida species, Cryptococcus neoformans, histoplasmosis, tinea
corporis, tinea cruris, tinea manuum and tinea pedis, onychomycosis, seborrheic dermatitis, possessing anti-
inflammatory and some antibacterial activities with topical and systemic action.
5-15
Ketoconazole undergoes very easily
on chemical degradation. The stressed degradation of ketoconazole drug substance into imidazole and acetamide
radical species were performed under acid, base, thermal, photo and oxidative stress conditions.
16,17
The potency losing
and forming harmful degradation products are the most common consequences of the degradation of the drug.
2

Ketoconazole was formulated into several pharmaceutical forms through various routes of administration such
as: tablets, topical creams, ointments, gels and antidandruff shampoo.
2,13,18-23
The role of ketoconazole as tablet has
been limited to few indications due to the variable bioavailability, liver toxicity and inhibition of steroid
biosynthesis.
15,24-26
Nowadays, no specific evidence of development of resistance or cross-resistance of fungi to
ketoconazole used in cosmetic dandruff shampoo at concentrations up to 2 %.
27

The mechanism of attacks and changes in the structure and function of the cell membrane permeability is the
most important factor in the ketoconazole action of the fungi.
28
The primary mechanism of action of ketoconazole as an
azole derivative is the inhibition of sterol 14-α-demethylase, a microsomal cytochrome P450 dependent enzyme system
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which converts lanosterol into ergosterol required for fungal cell membrane synthesis.
29-34
Ketoconazole prevents the
converting of the lanosterol in ergosterol and disrupts the integrity of membrane-bound enzymes and fungal cell
membranes. This ketoconazole action increases membrane permeability and the leakage of small ions, amino acids and
proteins from the fungi,
32
leading to cell death.
35

The use of the ketoconazole as a drug essential in pharmaceutical formulations highlights the requirement for
its determination and quantification with appropriate analytical methods. This paper gives an overview of the analytical
techniques that are available and nowadays have been used for determination of ketoconazole in pharmaceutical and
biological samples.
2. METHODS FOR THE DETERMINATION OF KETOCONAZOLE
The analytical methods that are currently used for determination of ketoconazole in pharmaceutical samples
(tablets, shampoos, creams and ointments) and biological samples (plasma, serum, urine, saliva, tissues of lung, liver,
muscles) are: spectrophotometric methods, as well as chromatographic methods (thin-layer chromatography, TLC and
high-pressure liquid chromatography, HPLC), capillary electrophoresis (CE), spectrofluorimetry and electroanalytical
methods (differential pulse polarography, adsorptive stripping differential pulse voltammetry, rotating disk electrode
voltammetry, coulometry and potentiometry). The analytical methods which are already published usually require
sample preparation, including extraction and clean-up, as well as the subsequent instrumental determination of
ketoconazole from the matrixes with the other azole derivatives. The developed method for determination of the
ketoconazole should be selective, sensitive and reproducible, with less consumption of solvent and time for analysis, as
well as they should be validated in order to prove that the developed procedure is suitable for intended analytical
purpose and give accurate results.

A. Sample Procedures:
A number of extraction techniques such as liquid-liquid extraction (LLE) and solid-phase extraction (SPE) are
available for isolation of the ketoconazole from the pharmaceutical and biological matrixes.
31,36-40
Newly extraction
technologies such as ultrasound-enhanced surfactant-assisted dispersive liquid-liquid microextraction (UESA-
DLLME)
41
and supercritical fluid extraction (SFE)
42
were introduced, also.
The application of the liquid-liquid extraction technique was tested in the ketoconazole isolation from tablets
or creams using chloroform
43-45
; dichloromethane for creams and tablets
46
; tetrachloromethane for creams
47
and toluene
for tablets.
48
The frequently used organic solvents in LLE for the analyses of ketoconazole in human and rat plasma
are: acetonitrile and 1-chlorobutane in mixture ratio of 1:4,
49
diethyl ether
38,41
; chloroform
38,41
and ethyl acetate.
39

In the extraction step of analyzing ketoconazole in samples, buffers were used, also. The citrate buffer (pH 2)
and phosphate buffer (pH 3) were used in the procedure for the ketoconazole determination in tablets and creams,
43,44

while for the plasma analyses phosphate buffer (pH 2) or dihydrogen phosphate-sodium hydroxide buffer solution (pH
6).
40,41
The solid-phase extraction of ketoconazole from pharmaceutical samples and endogenous substances was
performed using SDS-coated Al
2
O
3
for tablet, cream and shampoo samples,
31
Sep-Pak C18 cartridges for serum
36,50
and
Bond Elute Plexa cartridge for plasma samples,
40
followed by elution with methanol
36,40
or with ethanol.
50

B. Spectrophotometric Methods:
The quantitative determination of ketoconazole with UV-Vis spectrophotometric methods was based on
forming coloured complexes
43,46,51-53
or stabilizing diprotonated form of ketoconazole in HCl
54,55
followed by
measuring of the absorbance maximum.
The ketoconazole in pharmaceutical samples was quantified by the method based on the formation of ion-pair
complexes of the ketoconazole with reagents like bromocresol green, bromocresol purple, bromophenol blue and
bromophenol red in acidic medium.
56
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An analytical procedure for the determination of ketoconazole in bulk drugs and tablets was developed by the
forming of pink complex from the reaction between ketoconazole and iron(III) chloride in the presence of potassium
thiocyanate absorbed at 510 nm.
51
Two procedures for spectrophotometric determination of three piperazine derivatives: ketoconazole, piribedil
and prazosin hydrochloride as a result of charge-transfer and ion-pair complexation reactions was described in the
literature.
52
The reaction of the drugs with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone in acetonitrile was studied. The
maximum absorbance of formed orange-red complex was measured at 460 nm. A stable yellow ion-pair complex
which absorbs at 410 nm was constituted in the interaction of the drugs in chloroform with bromophenol blue in
acetonitrile. The two methods were applied for determination of the piperazine derivatives contents in tablets.
A highly colored product was produced in the reaction of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone and
ketoconazole. The spectrophotometric method was developed for determination of the six drugs with different n-
donating groups: clotrimazole, clozapine, ketoconazole, oxamniquine and pimozide in form of referent material and
pharmaceutical dosage forms.
53
The formation of a charge-transfer complex between antifungal drugs from the pharmaceutical dosage forms:
clotrimazole, econazole, ketoconazole and miconazole as n-donor and iodine as acceptor were described in the
literature.
57

A procedure with picric acid for determination of ketoconazole in tablets and creams, followed by reading the
absorbance at 410 nm was introduced in the literature.
45
A spectrophotometric method for the determination of clotrimazole, econazole nitrate, ketoconazole,
miconazole and tolnaftate based on the reaction between antifungal agents and 2,3-dichloro-5,6-dicyano-1,4-
benzoquinone in methanol or with p-chloranilic acid in acetonitrile by subsequently measured the absorbance at 460
nm and 520 nm, respectively was introduced in the literature.
47
Blue-colored stable complexes were formed when ketoconazole as a ligand reacted quantitatively with
copper(II) and cobalt(II). The values of the absorbance of the Cu(II) or Co(II) complexes were measured at 720 nm and
612 nm, respectively.
46
The spectrophotometric method based on the coupled redox-complexation reactions which proceed in the
ketoconazole-iron(III) and 1,10-phenanthroline systems was described for determination of ketoconazole in samples of
tablets, creams and shampoos.
58

The reactions of a triiodide ion and alizarin red S with clotrimazole and ketoconazole were studied with aim to
analyze the studied drugs in pure forms and formulated in commercial preparations. Highly stable products were
formed with the reaction of protonated forms of ketoconazole and clotrimazole with triiodide ion. The absorption
maximum was achieved at 425 nm.
44

A spectrophotometric method for determination of three pharmaceutical piperazine derivatives: ketoconazole,
piribedil and trimetazidine hydrochloride was introduced in the literature.
59
The yellow orange complexes were
obtained in the reaction between iron(III) chloride and the pharmaceutical piperazine derivatives.
A kinetic-spectrophotometric method for determination of the ketoconazole in tablets, creams and shampoo
samples was presented.
31
The manganate ion formed by the alkaline oxidation of ketoconazole with potassium
permanganate in micellar SDS medium was measured at 610 nm.
Zero-crossing and the first derivative ultraviolet spectrophotometric method with methanol as background
solvent for quantitative determination of ketoconazole in commercial and simulated emulsion formulations were used
in the literature.
20,21
A method for quantitative determination of seven azole antifungal agents was introduced in the literature.
60

Analysis was performed directly by using zero-order (fluconazole), first derivative (bifonazole, clotrimazole,
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econazole, itraconazole, miconazole) and second derivative (ketoconazole) UV spectrometry. The antifungals were
dissolved in methanol. The UV absorption spectra in range 200-400 nm were recorded for solutions of the standards
and the analysed samples.
The absorbance of the red complex formed in the reaction between ketoconazole from the pharmaceutical
preparations and iron(III) ions in an acid medium was measured at 495 nm.
61

The procedure for determination of ketoconazole, clotrimazole and fluconazole in their pure forms and
pharmaceutical preparations in form of ion-pair yellow complex obtained with bromothymol blue and picric acid was
reported.
43
The absorbances were recorded at 410 nm and 415 nm for ketoconazole, 410 nm and 413 nm for
clotrimazole and at 373 nm and 415 nm for fluconazole using picric acid and bromothymol blue, respectively.
A method for determination of the ketoconazole in tablets based on amplification reactions was described in
the literature.
48
In the first stage, ketoconazole was oxidized with periodate in KC
2+
and iodate ions. The iodate was
treated with iodide to release iodine, after masking the excess periodate with molybdate. The absorbance was measured
at 535 nm.

C. Chromatographic Methods:
The high-pressure liquid chromatography (HPLC) and the thin-layer chromatography (TLC) are widely used
chromatographic methods in the analysis of imidazole derivatives including ketoconazole.
I . Thin-layer chromatography
The quantification of azole derivatives in pharmaceutical creams and ointments using thin-layer
chromatography (TLC) densitometric methods was reported.
62
The method was used for separation, qualitative and
quantitative analysis of pure ketoconazole or in mixture with the clotrimazole and miconazole. A precoated silica gel
and solvent system consist of n-hexane-chloroform-methanol-diethylamine (50:40:10:1 v/v) were applied in the
analysis. A high-pressure thin-layer chromatographic method for determination of ketoconazole in shampoos and
creams was reported in the literature.
63
The samples were separated on a silica gel plate and developed in mobile phase
consist of ethanol-acetone-1 M H
2
SO
4
in ratio of 80:10:10 v/v.
I I . High-pressure liquid chromatography
HPLC was commonly used technique in developing and validating assay methods for determination of the
ketoconazole in biological samples and pharmaceutical formulations where it was introduced as a pure drug or together
with others azole derivatives. The detection modes extensively applied in HPLC analysis of ketoconazole were: UV
(ultra-violet) and DAD (diode array detector); spectrofluorimetric and detector with electrochemical detection.
64,65
On
the other hand, mass spectrometry (MS) coupled with HPLC or with liquid chromatography (LC) as tandem methods
were more often used in the direct analysis of ketoconazole and azole substances in the procedure for their
identification.
66
The new trends in mass spectrometry involved multiple steps in tandem MS-MS with intention to
increase the selectivity and sensitivity of the determination resulting in far better limits of detection than those achieved
by using LC-MS.
38,66
The reversed-phase column and isocratic elution with mobile phase consist of two solvents or a
mixture of solvents with buffers were reported in the literature as the most appropriate conditions for quantification of
ketoconazole in pharmaceutical and biological samples (Table 1). The internal standards used in HPLC methods for
quantification of the ketoconazole were phenothiazine,
67
clotrimazole,
37,49
terconazole,
20,68
benzafibrate,
69
R51012,
38
9-
acetylanthracene,
70
paclitaxel
39
and linezolid.
40
In the literature simultaneous determination of ketoconazole and other
azole derivatives in pharmaceutical formulations, as well as in biological samples was reported, also.
40,41,66,71-74
The
separation methods were based on adjusting parameters such as mobile phase, pH, polarity and flow speed (Table 2).


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Table 1: Conditions for HPLC analysis of ketoconazole in samples

Detector
type
Sample matrix
Chromatographic
column
Mobile phase Reference
UV (225 nm) Commercial and
simulated emulsion
formulations
LiChrospher® 100 RP-18
(125 mm x 4 mm, 5 µm
particle size)

Triethylamine in methanol
(1:500 v/v) and
ammonium acetate
solution in water (1:200
w/v), 75:25 v/v
20
DAD-UV
(238 nm)


Bulk drug Promosil C-18
(250 mm x 4.6 mm, 5 µm
particle size)



water-acetonitrile-buffer
(51:45:4 v/v; pH 6.8)




33



UV (225 nm)




Shampoo




LiChrospher® 100 RP-8
(150 mm x 4.6 mm, 5 µm
particle size)


Monoisopropylamine-
methanol (2:500 v/v) and
ammonium acetate-water
(1:200 w/v), 7:3 v/v
with pH adjusted to 5.5
with pH adjusted to 5.5



34





UV (231 nm)


Serum (human)


MicroPak MCH-10
(30 cm x 4 mm, 10 µm
particle size)


Methanol-phosphate
buffer
(75:25 v/v; pH 7.5)


36



No data


Lung, liver, plasma and
adrenal gland (rat)


Novapak C-18


Methanol-acetonitrile-
phosphate buffer
(35:30:35 v/v; pH 6.8)




37


LC-MS-MS



Plasma (human)



BDS
Hypersil C-18
(50 mm x 3 mm, 5 µm
particle size)
Acetonitrile-water-formic
acid (75:25:1 v/v)


38



LC-MS-MS




Plasma
(rat)
(ketoconazole,
docetaxel)


Cosmosil C-18
(150 mm x 2 mm)



Formic acid and methanol
(gradient elution)



39



DAD-UV
(224 nm)



Creams, tablets and
shampoos
(ketoconazole,
clotrimazole)



C-18
(150 mm x 4.6 mm)



Methanol-water-
diethylamine-glacial acetic
acid (80:20:0.3:0.2 v/v; pH
7)



42



UV (206 nm) Plasma (human) Inertsil ODS-80A
(150 mm x 4.6 mm, 5 µm
particle size)
Water-acetonitrile-
tetrahydrofuran-
ammonium hydroxide-
triethylamine
(45:50.2:2.5:0.1:0.1 v/v)
49

UV (225 nm)



Tablets and creams



Merck LiChrospher® 100
RP-18 (5 µm particle size)




Diisopropylamine-
methanol (1:500) and
ammonium acetate (1:200)
(8:2)

54




Electrochemical Plasma and saliva µBondapak Formic acid and
65
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detection (human) octadecylsilane
(3.9 mm x 300 mm)
dibutylamine in methanol
(pH 3)
UV (231 nm)



Serum, plasma,
cerebrospinal and
synovial fluid (human)



C-18



No data




67




No data Plasma and tablets µBondapak Methanol-water-glacial
acetic acid (67.5:32:0.5
v/v)
69
UV
(240 nm)
Plasma
(dog)
Hypersil BDS RP-C-18
(250 mm x 4.6 mm, 5 µm
particle size)
Methanol-water-
diethylamine (74:26:0.1
v/v)
70
Fluorimetric
Detection
(260 nm and
(375 nm)
Plasma
(human)
Metaphase
KR100-5-C-18
(250 mm x 4.6mm, 5 µm
particle size)
Disodium hydrogen
orthophosphate-
acetonitrile
(50:50 v/v; pH 6)
75
UV Shampoo
(ketoconazole at 250
nm, formaldehyde at
345 nm after
derivatisation with 2,4-
dinitrophenylhydrazine)
Interchrom Nucleosil C-8
(250 mm x 4.6 mm, 5 µm
particle size)
Acetonitrile-phosphate
buffer
(45:55 v/v; pH 4)
76

UV (225 nm) Tablets and creams Lichrosorb® RP-18
(250 mm x 4 mm, 5 µm
particle size)
Methanol-ammonium
acetate
(80:20 v/v)
77

Table 2: Conditions for HPLC analysis for simultaneous determination of azole antifungal agents in samples

Detector
type
Sample matrix
Chromatographic
column
Mobile phase Reference
DAD-UV
(260 nm)
Plasma
(fluconazole,
posaconazole,
voriconazole,
itraconazole and
its metabolite
hydroxyl-
itraconazole,
ketoconazole)
Gemini C-6-Phenyl
(150 mm x 4.6 mm, 5 µm
particle size)

Phosphate buffer pH 7 and
acetonitrile
(gradient elution)
40
DAD-UV
(220 nm)
Blood (human)
(ketoconazole,
econazole nitrate)
Agilent HC-C-18
(250 mm x 4.6 mm, 5 µm
particle size)
Acetonitrile-water (75:25 v/v)
41
UV (212
nm)
Residues
(cow milk)
(ketoconazole,
clotrimazole)
Zorbax XDB C-18
(250 mm x 4.6mm, 5 µm
particle size)
Acetonitrile-sodium acetate buffer
(85:15 v/v; pH 4.6)
50
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LC-MS-
MS
Liver and
muscles (residue,
chickens)
(voriconazole,
griseofulvin,
clotrimazole,
bifonazole,
econazole,
ketoconazole,
itraconazole,
miconazole,
terconazole,
fluconazole)
LC BEH C18
(2.1 mm x 100 mm, 1.7
µm particle size)

Water–methanol,
water–acetonitrile, water containing
0.1% formic
acid–methanol, and water containing
0.1% formic
acid–acetonitrile
(gradient elution)
66
UV (260
nm)
Pharmaceutical
formulations
(ketoconazole,
clotrimazole,
fluconazole)
Bondapak C-18
(25 cm x 4.6 mm, 10 µm
particle size)
Acetonitrile-
tris(hydroxymethyl)aminomethane
(55:45 v/v) in phosphate buffer
(pH 7)
72
UV (254
nm)
Pharmaceutical
formulations
(ketoconazole,
isoconazole,
miconazole)
C-18-BDS
(100 mm x 4.6 mm)
Acetonitrile-ammonium acetate
buffer (70:30 v/v), with pH
adjusted to 6 with phosphoric acid
74

D. Alternative Methods
I . Capillary zone electrophoresis
A method of capillary zone electrophoresis for simultaneous determinations of a mixture of antifungal drugs:
ketoconazole, clotrimazole and econazole in pharmaceutical forms were adapted.
78
The separation was achieved using
a fused-silica capillary column with acetic acid-Tris buffer at pH 5 and UV detection at 196 nm. A capillary zone
electrophoresis for determination of ketoconazole in tablets and creams with phosphate buffer adjusted with phosphoric
acid at pH 2.3 and UV detection at 225 nm was introduced in the literature.
77
I I . Electroanalytical methods
A non-aqueous potenciometric titration where the equivalence point of ketoconazole was potentiographically
located using D1 differential potentiograph was introduced.
79
The oxidation of ketoconazole by rotating disk electrode
voltammetry, cyclic voltammetry, and controlled potential coulometry in chloroform or acetonitrile at platinum, gold
and glassy carbon electrodes using tetrabutylammonium perchlorate as supporting electrolyte was introduced in the
literature.
80,81
The method was used for quantification of ketoconazole in tablets and cream. The oxidation of
ketoconazole on a bare carbon electrode voltammetrically was investigated.
82
The results obtained by determination of
ketoconazole in human urine and formulations indicated that the process was irreversible and controlled by an
adsorption-extraction process that allowed accumulation of the drug at the electrode surface. A simple potenciometric
method using liquid-plasticized PVC membrane sensor for determination of the ketoconazole in pharmaceutical
preparations and biological matrixes was described in the literature.
83
A water-insoluble ketoconazole-
tetraphenylborate ion pair took a role of an ion-exchanger. The electroanalytical behavior of ketoconazole in Briton-
Robinson buffer in a gel formulation and spiked urine samples was studied.
84
I I I . Proton nuclear magnetic resonance
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1H-NMR was used for determination of ketoconazole in pure form and in tablets involving integration of the
methyl protons-signal of ketoconazole (at 2.07 δ) relative to the benzocaine (at 1.30 δ) used as an internal standard.
79
I V. Spectrofluorimetric methods
Spectrofluorimetric methods used as an analytical technique are based on the measuring of the native
fluorescence of ketoconazole. A synchronous spectrofluorimetric determination of famotidine, fluconazole and
ketoconazole in bulk powder and pharmaceutical formulations was reported in the literature.
85
The fluorescence
intensity was recorded at 364 nm with excitation at 239 nm for ketoconazole, 384 nm with excitation at 284 nm for
famotidine and 285 nm with excitation at 255 nm for fluconazole. The quantitative determination of antifungal drugs:
clotrimazole, econazole nitrate, ketoconazole, miconazole and tolnaftate in pharmaceutical formulations using
spectrofluorimetric method was introduced in the literature.
47
The spectrofluorimetric method measured native
fluorescence of ketoconazole at 375 nm with excitation at 288 nm or the induced fluorescence after alkaline hydrolysis
of tolnaftate with NaOH solution at 420 nm with excitation at 344 nm.
V. Thermogravimetric method
A thermogravimetric method for quantification of the ketoconazole in raw materials and tablets was studied in
the literature.
86
The method included changes in physical and chemical properties of analytes as a function of
increasing temperature. The samples were analyzed by dynamic thermogravimetry in nitrogen and nitrogen-synthetic
air mixture at heating rates of 10, 20, 40, 60 and 80°C min
−1
. The concentrations of ketoconazole were obtained from
the vapour pressure curves.
3. CONCLUSION
Presented systematic review covers the current analytical methods for the determination of ketoconazole in
pharmaceutical and biological samples. The limitation of the reported methods requires developing new optimized
method which would be suitable for intended analytical purpose for analyzing the content of ketoconazole in
pharmaceutical, as well as in biological samples. The new trends and advances for quantification of ketoconazole are
based on using high-pressure liquid chromatography which is widely available and flexible method with the ability of
coupling with mass spectrometry. The HPLC method could be automated; there are different column fillings; different
solvents with different polarity as mobile phases and different detection modes. The faster time, high sensitivity,
specificity and better separation efficiency enable HPLC to be used frequently for the simultaneous qualitative and
quantitative determination of azole derivatives in the comparison with the other methods. The development of a new
established method should reduce existing analytical problems including many steps for the preparation of the sample,
as well as improving the resolution which can give accurate results for the concentration of all ketoconazole samples
and could be used for routine analysis.
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