ArticlePDF Available

Figures

Content may be subject to copyright.
~ 103 ~
The Pharma Innovation Journal 2019; 8(5): 103-108
ISSN (E): 2277- 7695
ISSN (P): 2349-8242
NAAS Rating: 5.03
TPI 2019; 8(5): 103-108
© 2019 TPI
www.thepharmajournal.com
Received: 21-03-2019
Accepted: 25-04-2019
Tomal Majumder
Primeasia University, Dhaka,
Bangladesh
Md. Razibul Hasan
State University of Bangladesh,
Dhaka, Bangladesh
Pritam Roy
State University of Bangladesh,
Dhaka, Bangladesh
Ratan Pramanik
State University of Bangladesh,
Dhaka, Bangladesh
Md. Nazmul Hasan
State University of Bangladesh,
Dhaka, Bangladesh
Correspondence
Tomal Majumder
Primeasia University, Dhaka,
Bangladesh
Method development and validation of RP-HPLC
method for estimation of luliconazole in marketed
formulation (Cream)
Tomal Majumder, Md. Razibul Hasan, Pritam Roy, Ratan Pramanik and
Md. Nazmul Hasan
Abstract
A simple, specific, accurate, precise, rapid, robust and selective stability indicating reverse phase high
performance liquid chromatography (RP-HPLC) method has been developed for assay and validated for
quantification of antifungal drug with its excipients in its topical dosage form. The mobile phase has been
used for separation consisting of Water: Acetonitrile (60:40). Column used was C18 (4.6 X150 mm, 5µm)
with flow rate of 2.0 ml/min. Detection wavelength for Luliconazole was 294 nm. The method has been
linear at 60-140% range with r2 0.999. Luliconazole has showed 97-103% recovery. The LOD and LOQ
were found to be 0.38µg/ml and 1.06µg/ml respectively. Methanol was used as solvent. The method have
been robust under various variation with flow rate, detection wavelength and column oven temperature.
Developed method can be used routinely for estimation of drug Luliconazole with its excipients in
dosage form and stability sample. The validation of method was carried out as per ICH Guidelines.
Keywords: luliconazole, assay, RP-HPLC, stability, antifungal, validation
Introduction
Luliconazole belongs to imidazole class of drug that possesses a wide spectrum of antifungal
activity and is very potent against dermatophytes. Luliconazole is chemically, (2E)-2- [(4R)-4-
(2, 4-dichlorophenyl)-1, 3-dithiolan-2-ylidene]-2-imidazol-1-yl-acetonitrile. Its structural
formula is:
Luliconazole
The molecular formula is C14H9Cl2N3S2 with a molecular weight of 354.28. Luliconazole is the
R enantiomer and contains one chiral center. The double bond adjacent to the dithiolane group
is in the E configuration. It is not official in Indian Pharmacopoeia (IP), British Pharmacopoeia
(BP) and United States Pharmacopoeia (USP). Till date no analytical method was reported for
quantitative estimation of LCZ except stability indicating method. LCZ is a novel antifungal
drug launched in India by Ranbaxy Laboratories Ltd. The compound was originally screened
from active compounds related to lanoconazole, a potent antidermatophytic drug. Literature
survey reveals that the simple and rapid stability-indicating liquid chromatographic method
has been developed and validated for LCZ. The present manuscript describes simple, accurate,
precise, reproducible, and economical RP-HPLC method for the estimation of Luliconazole in
marketed formulation.
Material, Machine and Methods
Instruments
Shimadzu (Prominence i LC2030 3D Plus liquid chromatography), Analytical balance
(Mettler Toledo, Model: ML204/01).
~ 104 ~
The Pharma Innovation Journal
Chemicals and Reagents
Methanol (Active Fine Chemicals Ltd, Bangladesh),
Acetonitrile (Active Fine Chemicals Ltd, Bangladesh),
Luliconazole (Viwit Pharmaceuticals Co. Ltd., India).The
Lulizol 1% Cream was procured from the local market.
Lulizol (Eskayef Pharmaceuticals Limited) 1% Cream
contains 1% Luliconazole, an azole antifungal agent, in a
white cream for topical application.
Methodology (By HPLC)
Chromatographic system
Column : 4.6-mm × 150 -cm, L1 (C18)
Wavelength : 296nm
Flow Rate : 2.00ml / min
Inject volume : 20 l
Temperature : 40.0 0C
Run Time : 20.0 minutes
Preparation of mobile phase
Prepare a filtered and degassed mixture of Water, Acetonitrile
(60: 40). Make adjustments if necessary.
Standard preparation
Weigh accurately about 10 mg of working standard of
Luliconazole and take into a 100 ml volumetric flask. Add 60
ml of methanol and sonicate to dissolve. Make volume up to
the mark with the same solvent and mix well. Filter the
solution through Whatman # 1 filter paper and collect the
filtrate discarding first few ml. Then take 2 ml of solution into
another 50 ml volumetric flask and add methanol upto the
mark and mix well. Filter the solution through 0.45
membrane filter.
Sample preparation
Weigh accurately about 1.0 g of test sample equivalent to
about 10 mg of Luliconazole into a 100 ml volumetric flask.
Add about 60 ml of methanol and sonicate in a water bath at
55.00C to 60.00C until the sample is completely dispersed, and
mix. Cool the solution to below room temperature, mix and
dilute with same solvent to volume and mix well. Filter the
solution through Whatman # 1 filter paper and collect the
filtrate discarding first few ml. Then take 2 ml of solution into
another 50 ml volumetric flask and add methanol up to the
mark and mix well. Filter the solution through 0.45
membrane filter.
Procedure
Equilibrate the system for about 60 minutes with mobile
phase flow rate at 2.0 ml/min. Separately inject equal volumes
(about 20 l) of the standard preparation and the sample
preparation in to the chromatograph, record the
chromatograms, and measure the responses for the major
peaks. %RSD of peak area for replicate injections of standard
solution should be less than 2.0%.
Calculate the amount of Luliconazole per g of test sample by
using the following equation:
= -------------------------- Luliconazole (mg/g)
Where,
Pu = Peak area of test sample solution
Ps = Peak area of standard solution
Wu = Sample weight taken in g
Ws = Standard weight taken in mg
P = Potency of standard (as Luliconazole)
Validation parameters
System precision/system suitability
System suitability testing is an integral part of many
analytical procedures. System suitability test parameters
depend on the type of procedure being validated. System
precision is determined by measuring the peak area of
standard solution containing 100% working concentration for
six times and calculates the % RSD. The % RSD should be
less than 2.0%.
Table 1: System precision
Standard
Concentration
(0.2 mg/ml)
No of measurement
Peak area
Theoretical plates (NLT 2000) (NLT 2000)
1.
119206
8668
2.
119646
8765
3.
119423
8782
4.
119780
8758
5.
119473
8666
6.
119804
8611
RSD % (Limit NMT 2.0%)
0.193 %
Limit:≥2000
The relative standard deviation of six replicate measurement
of standard solution found 0.193 % (limit NMT 2.0%), which indicates that the system is precise to analyze the sample.
~ 105 ~
The Pharma Innovation Journal
Fig 1: Chromatogram of luliconazole standard & sample
Precision
Repeatability/ method precision
Repeatability was established by analyzing six separate
samples at 100% of the working concentration from a same
batch Lulizol (Eskayef Pharmaceuticals Limited) 1% Cream
Result was calculated against label claim.
Table 2: Data for method precision
S. No.
Sample taken ( in g )
Peak area
Results (mg/g)
RSD %
1
1.06
117736
10.07
1.10
(Limit NMT 2.0%)
2
1.08
118095
9.91
3
1.04
116417
10.15
4
1.06
116159
9.93
5
1.05
116453
10.05
6
1.04
116861
10.18
The relative standard deviation of assay result of six separate
samples from a single batch found 1.10 % (limit NMT 2.0%)
which indicates that the method is precise to analyze
Luliconazole.
Intermediate precision
Intermediate precision was established by analyzing six
separate samples at 100% of the working concentration from
a same batch of in different day by different analyst using
different machine. Result was calculated against label claim.
Table 3: Data for precision in day 2
S. No
Sample taken (in g)
Peak Area
Results (mg/ g )
RSD %
1.
1.03
116336
9.98
1.34
(Limit NMT 2.0%)
2.
1.02
116555
10.10
3.
1.03
116536
10.00
4.
1.05
116396
9.80
5.
1.01
116521
10.20
6.
1.03
116388
9.99
Table 4: Data for intermediate precision
Sample No.
Results ( mg/ g)
Day-1 (Analyst-01)
Day-2 ((Analyst-02)
1.
10.07
9.98
2.
9.91
10.10
3.
10.15
10.00
4.
9.93
9.80
5.
10.05
10.20
6.
10.18
9.99
Mean
10.05
10.01
Overall mean
10.03
% RSD
1.10
1.34
RSD % of 12 units
1.22
The relative standard deviation of assay result of six separate
samples from a single batch found 1.10 % in day 1, 1.34 % in
day-2 and 1.22 % for 12 results (limit NMT 2.0%) which
indicate that the method is precise to analyze the
Luliconazole.
Accuracy
Accuracy was established by analyzing nine sample solution
of Luliconazole spiked with placebo at 80%, 100% and 120%
of the working concentration (Three replicates for each
concentration) and the percent recovery was calculated. The
percent recovery at each level should be within 97.0% to
103.0%. A linear curve was prepared by plotting amount
added Vs amount recovered and correlation co-efficient was
calculated.
Standard preparation
10.9 mg working standard of Luliconazole was weighed and
taken into 100ml volumetric flask. About 60 ml of methanol
was added and Sonicated to dissolve. Volume was made with
diluents and mixed well. The solution was filtered through
~ 106 ~
The Pharma Innovation Journal
whatman # 1 filter paper and the filtrate was collected
discarding first few ml. 2 ml of this filtered solution was taken
into a 50 ml volumetric flask and volume was made up to
mark with diluents and mixed well. Again the solution was
filtered through 0.45 membrane filter.
Placebo stock preparation
1 g of placebo was weighed and taken into 100 ml volumetric
flask.60 ml of methanol was added and sonicated to dissolve.
Made volume up to the mark with same methanol and mix
well. Filtered the solution through Whatman # 1 filter paper
and collected the filtrate discarding first few ml.
Sample preparation for 80%
Sample 01
1.6 ml Standard stock preparation + 2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 02
1.6 ml Standard stock preparation +2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 03
1.6 ml Standard stock preparation +2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample preparation for 100%
Sample 01
2 ml Standard stock preparation + 2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 02
2 ml Standard stock preparation + 2 ml Placebo stock
preparation →500 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 03
2 ml Standard stock preparation + 2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample preparation for 120%
Sample 01
2.4ml Standard stock preparation + 2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 02
2.4 ml Standard stock preparation +2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Sample 03
2.4ml Standard stock preparation + 2 ml Placebo stock
preparation →50 ml with methanol. Again the solution was
filtered through 0.45 membrane filter.
Table 5: Data for accuracy
Concentration level
Sample No.
Amount added in (mg/ml)
Amount recovered in (mg/ml )
Peak area
% Recovery
80%
Sample-1
0.00349
0.00339
92515
97.1
Sample-2
0.00349
0.00339
92727
97.1
Sample-3
0.00349
0.00339
92857
97.1
100%
Sample-1
0.00436
0.00424
116022
97.2
Sample-2
0.00436
0.00423
115862
97.0
Sample-3
0.00439
0.00425
116492
97.5
120%
Sample-1
0.00523
0.00511
140090
97.7
Sample-2
0.00523
0.00512
140259
97.9
Sample-3
0.00523
0.00511
139839
97.7
The percent recovery was calculated for nine determinations
and found within limit. A graphical presentation between
amount added VS amount recovered also shows linearity.
Thus it has been concluded that the method is accurate to
analyze the Luliconazole.
Specificity
The retention time of the sample preparation for assay is
concordant with the retention time of standard sample from
assay preparation.
Blank effect: To identify the blank effect.
Placebo effect: To identify the placebo effect.
Wt. of placebo: 1 g →100 ml →2 ml→50 ml
Peak area
Blank
No interference
Placebo
No interference
There is no significant interference of blank and placebo. So
the method is specific to analyze Luliconazole.
Linearity
Five different standard solutions were prepared covering a
concentration of 60% to 140 % of the working concentration
of Luliconazole and all peak area was recorded. A linear
curve was prepared by plotting percentage of nominal
concentration Vs peak area and correlation co-efficient was
calculated. The results obtained correlate with the
concentrations resulting in the following calibration curve.
Standard stock solution
10 mg working standard of Luliconazole was weighed and
taken into 50 ml volumetric flask. About 60 ml of methanol
was added and sonicated to dissolve. Volume was made with
methanol and mixed well. The solution was filtered through
whatman # 1 filter paper and the filtrate was collected
discarding first few ml.
~ 107 ~
The Pharma Innovation Journal
Sample preparation for 60%
1.20 ml Standard stock preparation →50 ml with Methanol
.Again the solution was filtered through 0.45 membrane
filter.
Sample preparation for 80%
1.60 ml Standard stock preparation →50 ml with Methanol.
Again the solution was filtered through 0.45 membrane
filter.
Sample preparation for 100%
2.0 ml Standard stock preparation →50 ml with Methanol.
Again the solution was filtered through 0.45 membrane
filter.
Sample preparation for 120%
2.40 ml Standard stock preparation →50 ml with Methanol.
Again the solution was filtered through 0.45 membrane
filter.
Sample preparation for 140%
2.80 ml Standard stock preparation →50 ml with Methanol.
Again the solution was filtered through 0.45 membrane
filter.
Table 6: Data for linearity
S. No.
Concentration of Luliconazole (mcg/ml)
Percentage of nominal concentration
Peak area
1
2.4
60%
70433
2
3.2
80%
95627
3
4.0
100%
119919
4
4.8
120%
144097
5
5.6
140%
168212
Correlation coefficient : 0.999
Fig 2: Graphical presentation of linearity
The correlation co-efficient found 0.999. Thus the graph
confirms the linearity of the method in the range of 60% to
140%.
Limit of detection and limit of quantification
ICH guideline describes several approaches to determine the
detection and quantification limits. These include visual
evaluation, signal-to-noise ratio and the use of standard
deviation of the response and the slope of the calibration
curve. In the present study, the LOD and LOQ were based on
the third approach and were calculated according to the 3.3 ×
(SD/Slope) and 10 × (SD/Slope) criteria, respectively; where
SD is the standard deviation of y-intercept of regression line
and S is the slop of the calibration curve.
Robustness
Robustness of this method was determined by analyzing the
same batch of Lulizol (Eskayef Pharmaceuticals Limited) 1%
Cream by different flow rate and different wavelength.
Table 7: Data for robustness
S. No.
Changed Flow rate
(ml/min)
Peak area
Assay (%)
Changed
mobile phase
Peak area
Assay (%)
RSD
(%)
1.
Flow rate actual 2.00 ml/min
Wavelength:296 nm
0.49
Standard
120626
99.4
Standard
120626
99.4
Sample
116350
Sample
116350
2.
Flow rate changed to 1.8 ml/min
Wavelength:298 nm
Standard
133443
99.6
Standard
120991
99.6
Sample
128890
Sample
116880
3.
Flow rate changed to 2.20 ml/min
Wavelength:294 nm
Standard
109673
100.6
Standard
119133
100.2
Sample
105967
Sample
114721
~ 108 ~
The Pharma Innovation Journal
Table 8: Summary results
S. No.
Validation parameters
Results
1.
System precision/System suitability
RSD 0.193%
2.
Precision
Repeatability/ Method Precision
RSD 1.10 %
Intermediate Precision
RSD 1.22 %
3.
Accuracy
97.1-97.9 %
4.
Linearity
Correlation co-efficient 0.999
5.
LOD(µg/ml)
0.38
6.
LOQ(µg/ml)
1.06
7.
Robustness
RSD 0.49 %
8.
Specificity
No peak Area detected
Conclusion
The results of our study indicate that the proposed RP-HPLC
method is simple, rapid, precise and accurate. The developed
RP-HPLC method was found suitable for determination of
Luliconazole in marketed formulation without any
interference from the excipients. Statistical analysis proves
that the method is repeatable and selective for the analysis of
Luliconazole. It can therefore be conclude that use of the
method can save time and money and it can be used in small
laboratories with accurate and wide linear range. From the
above data it was observed that all validation parameters (like
system suitability, precision, accuracy, specificity, linearity,
robustness) meet the predetermined acceptance criteria. Thus
it has been concluded that the method is validated for the
analysis of Luliconazole in Lulizol (Eskayef Pharmaceuticals
Limited) 1% Cream.
Acknowledgements
Authors are highly thankful to Department of Pharmacy, State
University of Bangladesh and Department of Pharmacy,
Primeasia University for providing facilities to complete the
work.
References
1. Validation of compendial procedures, USP-42, Chapter-
1225.
2. ICH, Q2 (R1) Validation of Analytical Procedure
Methodology, International Conference on
Harmonization.
3. Tripathi KD, Essential Medical pharmacology, Jaypee
Brother Medical Publisher, Six edition pp 684-689.
4. Mohan H. Textbook of Pathophysiology, Jaypee
brother’s Medical Publishers, Six Edition, 773.
5. Rang HP, Dale MM, Ritter JM, Flower RJ. Rang and
Dales Pharmacology, Churchill Livingstone Elsevier
Publication, Six Edition, 692.
6. Hardman G, Limbird L. Goodman and Gilman’s-The
Pharmacological basis of Therapeutics, Mc Grow Hill
Medical Publishing Division, Tenth Edition, 1181-1182.
7. Chatwal GA. Insrtumental Methods of Chemical
Analysis, Himalaya Publishing House, First edition,
2.167-2.171.
8. Sharma BK. Instrumental methods of chemical analysis
in introduction to analytical chemistry, Nineteenth
Edition, Goel publishing House, 2003, 1-4:71
9. Willard HH, Merritt LL, Dean JA, Frank AS.
Instrumental method of analysis, New Delhi: CBS
publishers and Distributors, Seventh Edition, 1986, 101-
105.
10. Skoog DA, Crouch SR, Holler FJ. Analytical Methods.
Principles of Instrumental Analysis. California Cengage
Learning Inc., sixth Edition, 2006, 6-19.
11. Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC
Method Development, John Wiley and Sons, New York,
A Wiley- Inter science Publication, Second Edition,
1997, 21-56,266-288.
12. Sethi PD. HPLC, Quantitative analysis of pharmaceutical
formulations, CBS Publishers and Distributors, New
Delhi,1996, 3-3.
13. Chung HCC, Lee YC. Analytical method validation and
instrument performance verification, Wiley- Inter science
Publication, 1999, 51-67.
... Mixtures in each bottle were transferred into Eppendorf vials, and then centrifuged at 12,000 rpm for 15 min to precipitate particles of drug. The amounts in the supernatants were assayed using HPLC, according to a previously reported method [36]. Briefly, the quantitative supernatants were diluted with methanol, and the solubility of LCZ in oils, surfactants and co-surfactants was assayed via the HPLC method equipped with an ultraviolet detector (Waters 2487) and an octadecyl bonded silica column (C18, Waters XBridge ® , φ 4.6 mm × 150 mm, particle size: 3.5 µm) at a detection wavelength of 290 nm. ...
Article
Full-text available
Luliconazole (LCZ), a novel imidazole drug, has broad-spectrum and potential antifungal effects, which makes it a possible cure for fungal keratitis; nevertheless, its medical use in ocular infections is hindered by its poor solubility. The purpose of this study was to design and optimize LCZ nanoemulsion (LCZ-NE) formulations using the central composite design-response surface methodology, and to investigate its potential in improving bioavailability following ocular topical administration. The LCZ-NE formulation was composed of Capryol 90, ethoxylated hydrogenated castor oil, Transcutol® P and water. The shape of LCZ-NE was spherical and uniform, with a droplet size of 18.43 ± 0.05 nm and a low polydispersity index (0.070 ± 0.008). The results of an in vitro release of LCZ study demonstrated that the LCZ-NE released more drug than an LCZ suspension (LCZ-Susp). Increases in the inhibition zone indicated that the in vitro antifungal activity of the LCZ-NE was significantly improved. An ocular irritation evaluation in rabbits showed that the LCZ-NE had a good tolerance in rabbit eyes. Ocular pharmacokinetics analysis revealed improved bioavailability in whole eye tissues that were treated with LCZ-NE, compared with those treated with LCZ-Susp. In conclusion, the optimized LCZ-NE formulation exhibited excellent physicochemical properties, good tolerance, enhanced antifungal activity and bioavailability in eyes. This formulation would be safe, and shows promise in effectively treating ocular fungal infections.
... Chemical structures of both the drugs are shown in Figure 1. Literature survey reveals various analytical methods for the estimation of LCZ using UV spectrophotometry, [12,13] high performance liquid chromatography (HPLC), [14][15][16][17] and high performance thin layer chromatography (HPTLC). [18] Various method for the estimation of GLA using UV spectrophotometry, [19,20] HPLC, [21][22][23] and HPTLC. ...
Article
Full-text available
Introduction: Three new UV spectrophotometric approaches specifically, simultaneous equation (SME), absorbance ratio (ASR), and first derivative (zero crossing) spectroscopic techniques were developed and validated for simultaneous estimation of luliconazole (LCZ) and gallic acid (GLA) in emulgel formulation which were simple, sensitive, precise, and accurate. Materials and Methods: In SME method, absorbance was measured at 299 and 259 nm for both the drugs. LCZ and GLA were estimated at 299 and 266 nm in the ASR method. First derivative (zero crossing) method depended on the change of UV spectra in to first derivative spectra followed by measurement of first derivative signal at 249 and 259 nm for LCZ and GLA, respectively, using 4 nm as wavelength interval (Δλ) and 4 as scaling factor. Developed methods were validated according to ICH guidelines including parameters, namely, specificity, linearity, range, precision and accuracy, limit of detection, and limit of quantification. Results and Discussion: All three techniques showed direct relation of absorbance in the concentration range of 1-30 µg/ml for both the drugs. Good repeatability, low intra and inter-day variability, indicate that precise agreement within the value. Recovery studies for both drugs ranged from 97 to 102 percent, recommending that the methods are effective. Results of method validation parameters such as linearity and range, precision, and accuracy adhere to ICH guideline acceptable limit. Conclusion: All the developed methods were found to be quick, profoundly accurate and financially effective; and henceforth can be valuable for simultaneous estimation of LCZ and GLA in emulgel formulation for routine quality control analysis.
... Different analytical methods such as spectrophotometry [3][4][5], TLC [6], HPTLC [6,7], RP-HPLC [8,9], RP-UFLC [10] and other liquid chromatographic methods involving the estimation of related [11] substances and experimental design [12] were published for the assay of Luliconazole in pharmaceutical formulations and biological fluids. In the present study we have developed a simple, accu- Figure 1: Chemical structure of Luliconazole. ...
Article
Aim: A new stability-indicating liquid chromatography method was developed and validated for the quantitative determination of luliconazole. Materials and methods: Preliminary forced degradation study demonstrated an additional peak of the degradation product at the same retention time to the drug, due to this, the method was developed optimizing the chromatographic conditions to provide sufficient peak resolution (R ≥ 2). The experimental design was evaluated to assess the robustness and the best chromatographic conditions to be used for the validation. Methodology: Luliconazole solutions were exposed to various stress conditions to evaluate the method indication stability, in which the degradation product (DP-1) formed was isolated, identified, and evaluated in silico to predict degradation pathway and toxicity. The procedure was validated by robustness, selectivity, linearity, precision, and accuracy. Liquid chromatography was performed in a Phenomenex® RP-18 column with a mixture of acetonitrile and 0.3% (v/v) triethylamine solution as a mobile phase in isocratic elution. Results and conclusions: The method demonstrated robustness, good recovery, precision, linear response over a range from 5.0 to 40.0 μg.mL⁻¹- and to be stability indicating. The alkaline stress condition resulted in the formation of DP-1. HRMS studies identified this product as an hydroxyacetamide derivative, and in silico studies did not show toxic potential.
Article
Objective: A new, simple, economical, sensitive, precise and reproducible UV visible spectrophotometric method was developed for the estimation of luliconazole in pure form and pharmaceutical formulation as per ICH guidelines. Method: A UV spectrophotometric method has been developed using methanol and water as solvent to determine the luliconazole in bulk and pharmaceutical dosage formulation. The λmax of luliconazole in methanol and water was found to be 297nm. Results: The drug was proved linear in the range of 3-15µg/ml and exhibited good correlation coefficient (R2= 0.9993) and excellent mean recovery (98-99%). The % RSD for intra-day and inter-day precision was found to be 1.051288 and 1.138658 respectively. The LOD and LOQ of Luliconazole was found to be 1.1168µg/ml and 3.3845µg/ml respectively. This method was successfully applied to luliconazole content in marketed brands and results were in good agreement with the label claims. Conclusion: The method was validated for linearity, precision, repeatability and reproducibility. The obtained results proved that the method can be employed for the routine analysis of luliconazole in bulks as well as in commercial formulations.
Textbook of Pathophysiology, Jaypee brother's Medical Publishers, Six Edition
  • H Mohan
Mohan H. Textbook of Pathophysiology, Jaypee brother's Medical Publishers, Six Edition, 773.
Rang and Dales Pharmacology
  • H P Rang
  • M M Dale
  • J M Ritter
  • R J Flower
Rang HP, Dale MM, Ritter JM, Flower RJ. Rang and Dales Pharmacology, Churchill Livingstone Elsevier Publication, Six Edition, 692.
Goodman and Gilman's-The Pharmacological basis of Therapeutics
  • G Hardman
  • L Limbird
Hardman G, Limbird L. Goodman and Gilman's-The Pharmacological basis of Therapeutics, Mc Grow Hill Medical Publishing Division, Tenth Edition, 1181-1182.
Instrumental methods of chemical analysis in introduction to analytical chemistry, Nineteenth Edition
  • B K Sharma
Sharma BK. Instrumental methods of chemical analysis in introduction to analytical chemistry, Nineteenth Edition, Goel publishing House, 2003, 1-4:71
Quantitative analysis of pharmaceutical formulations
  • P D Sethi
  • Hplc
Sethi PD. HPLC, Quantitative analysis of pharmaceutical formulations, CBS Publishers and Distributors, New Delhi,1996, 3-3.
Analytical method validation and instrument performance verification
  • Hcc Chung
  • Y C Lee
Chung HCC, Lee YC. Analytical method validation and instrument performance verification, Wiley-Inter science Publication, 1999, 51-67.