Int J Pharm Pharm Sci, Vol 12, Issue 11, 19-25Original Article
NEW DEVELOPED AND VALIDATED SPECTROSCOPIC METHOD FOR THE SIMULTANEOUS ESTIMATION OF TERBINAFINE HYDROCHLORIDE AND FLUCONAZOLE
BHUMIKA THAKUR1*, INDER KUMAR2
1*Shiva Institute of Pharmacy, Bilaspur, HP, India, 2School of Pharmacy, Abhilashi University, Mandi, HP, India
Email: bhumi.thakur26@gmail.com
Received: 09 Aug 2020, Revised and Accepted: 10 Sep 2020
ABSTRACT
Objective: A new, simple, precise, accurate, and reproducible method or simultaneous equation method was developed and validated for the simultaneous estimation of terbinafine hydrochloride (TH) and fluconazole (FLZ) in pure form.
Methods: Simultaneous estimation of terbinafine hydrochloride and fluconazole was estimated by the ultraviolet (UV) spectrophotometry method. The method was based on the measurement of absorbance at two wavelengths 222 nm and 239 nm, of terbinafine hydrochloride and fluconazole in 0.1N HCl respectively.
Results: Calibration curves terbinafine hydrochloride and fluconazole were found to be linear in the concentration ranges of 0.5-3.0 μg/ml and 80-400 μg/ml, respectively, with their correlation coefficient values (R2) 0.999 and 0.998. LOD and LOQ of TH were found to be 0.067, 0.203 at 222 nm and 0.175, 0.531 at 239 nm, similarly for FLZ; 31.089, 94.210 at 222 nm and 94.380, 286.00 at 239 nm respectively. In the precision study, the % RSD value was found within limits (%). The percentage recovery at various concentration levels varied from 98.50 % to 103.96 % for TH and 97.27 % to 103.83 % for FLZ confirming that the expected method is accurate.
Conclusion: It could be concluded from the results obtained in the present study that this method for simultaneous estimation of TH and FLZ in pure is simple, precise, and economical. The proposed method can be applied successfully for the simultaneous estimation of TH and FLZ in the pure and pharmaceutical dosage form.
Keywords: Fluconazole, Terbinafine hydrochloride, UV-visible spectroscopy, Simultaneous estimation
© 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijpps.2020v12i11.39344. Journal homepage: https://innovareacademics.in/journals/index.php/ijpps.
INTRODUCTION
Terbinafine hydrochloride (TH) is (E)-N,6,6-trimethyl-N-(naphthalen-1-ylmethyl) hept-2-en-4-yn-1-amine; hydrochloride. It is an allylamine, which has a wide spectrum of antifungal activity in fungal infections of the hair and skin such as Pityriasis Versicolor. Terbinafine, a synthetic allylamine antifungal agent, inhibits ergosterol biosynthesis via squalene epoxidase. When exposed to terbinafine, fungi accumulate squalene while becoming deficient in ergosterol, an essential component of fungal cell membranes [1, 2]. The inhibition of squalene epoxidase leads to fungistatic and fungicidal action. Terbinafine is an orally and topically active drug with primarily fungicidal activity against a broad spectrum of fungi, including dermatophytes, filamentous, dimorphic organisms, and some yeasts such as Candida parapsilosis [3, 4].
Fluconazole (FLZ) is 2-(2,4-difluorophenyl)-1,3-bis(lH-1,2,4-triazo1-yl)propan-2-ol. Fluconazole is a synthetic triazole with antifungal activity. It is a well-established first-line management option for both localized and systemic C. albicans infections. Fluconazole is used against superficial and systemic candidiasis and in the treatment of cryptococcal infections for patients with the acquired immunodeficiency syndrome [5, 6]. Fluconazole main targets the heme protein, which co-catalyzes cytochrome-P450-dependent 14 α-demethylation of lanosterol in the last stage of ergosterol biosynthesis in fungus. Fluconazole is effective against C. albicans infections at a wide range of body sites and tissues, irrespective of the patient's immune status [7, 8]. Therefore, the present study aims to develop and validate the simultaneous estimation method of terbinafine hydrochloride and fluconazole in pure form.
(b)
Fig. 1: Structure of (a) terbinafine hydrochloride (b) fluconazole
MATERIALS AND METHODS
Fluconazole and terbinafine hydrochloride were procured from Lark Pharmaceuticals, Bhiwandi, India as a gift sample. Hydrochloric acid, methanol, and other chemicals and solvents were procured from CDH Chemicals, Mumbai, India. All chemicals and solvents were used of analytical grade.
Preformulation studies
Various preformulation studies were carried out to identify the purity of the drug i.e. physical characterization, melting point, partition coefficients, solubility, UV, and FTIR studies [9, 10].
Determination of absorption maxima (λmax) and preparation of calibration curve
Absorption maxima (λmax) of TH in 0.1N HCl
Standard known concentration solution (3.0 µg/ml in HCl) was scanned in the range of 400-200 nm using a UV-visible spectrophotometer to get the absorption maxima.
Preparation of calibration curve of TH in 0.1N HCl
100 mg of TH was dissolved in 0.1N HCl in the 100 ml volumetric flask and a series of concentrations 0.5 µg/ml, 1.0 µg/ml, 1.5 µg/ml, 2.0 µg/ml, 2.5 µg/ml, 3.0 µg/ml of TH were prepared, and absorbance was measured at 222 nm in UV-visible spectrophotometer.
Determination of absorption maxima (λmax) of FLZ in 0.1N HCl
Absorption maxima (λmax) of FLZ was determined in 0.1N HCl by scanning the 400 µg/ml solution in a range of 200-400 nm using a UV-visible spectrophotometer.
Preparation of calibration curve of FLZ in 0.1N HCl
100 mg of FLZ was dissolved in 0.1N HCl in the 100 ml volumetric flask a series of concentration, 80 µg/ml, 120 µg/ml, 160 µg/ml, 200 µg/ml, 240 µg/ml, 280 µg/ml, 320 µg/ml, 360 µg/ml and 400 µg/ml of FLZ were prepared and absorbance was measured at 260 nm.
Compatibility studies
Both the drugs were uniformly mixed with dry powdered KBr in the ratio of 1:100 and the mixture was then compressed into the transparent disc under high pressure using special dies. The disc was placed in the FTIR spectrophotometer and the spectrum was recorded [11, 12].
Development and validation of simultaneous estimation method for TH and FLZ by UV-visible spectrophotometer
Q-Analysis method was developed and validated for the simultaneous estimation of TH and FLZ by UV-visible spectrophotometer
Standard stock solutions of TH and FLZ
Accurately weighed 100 mg of TH was dissolved in 0.1N HCl in 100 ml volumetric flask. The solution was sonicated for 10 min and the final volume was made up with the same solvent. From the standard stock solution, pipetting out 1 ml and dissolved it with 0.1N HCl to prepare the working standard solution of 100 µg/ml. Then, a series of concentrations were prepared using 0.5 ml, 1.0 ml, 1.5 ml, 2.0 ml, 2.5 ml, and 3.0 ml working standard solution in 10 ml volumetric flasks.
The FLZ standard solution was prepared by weighed 100 mg of drug and dissolved in 0.1N HCl in 100 ml volumetric flask for 10 min sonication and make up the final volume. From the above solution 0.8 ml, 1.2 ml, 1.6 ml, 2.0 ml, 2.4 ml, 2.8 ml, 3.2 ml, 3.6 ml, and 4.0 ml were pipetted out in series of 10 ml volumetric flask and volume was made up with 0.1N HCl.
Selection of wavelength of maximum absorbance (λmax)
3.0 µg/ml solution was prepared from the working stock solution of TH in a 10 ml volumetric flask. For FLZ, 400 µg/ml solution was prepared by a standard stock solution of FLZ in a 10 ml volumetric flask. Both the solutions were scanned in 400-200 nm against 0.1N HCl as blank using a UV-visible spectrophotometer.
Determination of iso-absorptive point
1.5 µg/ml solution of TH and 200 µg/ml of FLZ and scanning in spectrum mode in the range of 400-200 nm. An overlain spectrum of TH and FLZ was found and an iso-absorptive point was determined [13, 14].
Linearity range
Linearity of TH and FLZ was determined by preparing different concentrations i.e. 0.5 µg/ml, 1.0 µg/ml, 1.5 µg/ml, 2.0 µg/ml, 2.5 µg/ml and 3.0 µg/ml for TH and 160 µg/ml, 200 µg/ml, 240 µg/ml, 300 µg/ml, 360 µg/ml and 400 µg/ml for FLZ from their working standard solution and absorbance was measured at 222 nm and 239 nm [15].
Accuracy
2.0 µg/ml, 2.5 µg/ml, 3.0 µg/ml and 240 µg/ml, 300 µg/ml, 360 µg/ml solutions were prepared for TH and FLZ, respectively. These were the three different levels 80 %, 100 %, and 120 % by considering 2.5 µg/ml and 300 µg/ml as 100 % for TH and FLZ, respectively. The recovery study was performed three times at each level [16].
Intra-day and inter-day precision
The intra-day and the inter-day precision study was performed by preparing the dilution of 2.0 µg/ml, 2.5 µg/ml, 3.0 µg/ml for TH and 240 µg/ml, 300 µg/ml and 360 µg/ml for FLZ and from the respective stock solutions for both the drugs, respectively and recording their absorbance at 222 nm and 239 nm against 0.1N HCl as a blank. The absorbance of these concentrations was measured three times a day at regular intervals of one hour for intra-day and day1, day 2, and day 3 for inter-day study [17, 18].
Repeatability
3.0 µg/ml and 240 µg/ml of concentrations were prepared from the stock solution of TH and FLZ, and absorbance was measured at 222 nm and 239 nm against 0.1N HCl as a blank. The standard deviation and relative standard deviation was calculated [19].
Reproducibility
2.0 µg/ml and 300 µg/ml solutions were prepared separately from stock solutions of TH and FLZ and analyzed by one analyst (analyst1) at 222 nm and 239 nm. The same solutions were prepared by another analyst (analyst2) and values obtained were evaluated using a t-test to verify their reproducibility [20, 21].
Limit of detection (LOD)
The limit of detection was calculated from the data obtained from the linearity studies. The slope of the linearity plot was determined and calculated by the following formula.
Where S is the standard deviation of the residuals around the regression line.
b is the slope of the regression line.
Limit of quantitation (LOQ)
The LOQ is the lowest concentration of an analyte in a sample that can be determined with acceptable precision and accuracy under the stated conditions of the test [22-24]. The limit of quantitation can be calculated from the standard deviation of the response and the slope.
Where S is the standard deviation of the residuals around the regression line.
b is the slope of the regression line.
RESULTS AND DISCUSSION
Preformulation studies
Determination of λmax and preparation of calibration curve
Determination of absorption maxima (λ max) of TH and FLZ in 0.1N HCl
The absorption maxima of TH and FLZ were found at 222 nm and 260 nm respectively (fig. 2). The correlation coefficient, intercept, and slope for the calibration data for TH and FLZ also were calculated (table 1).
Compatibility study of TH and FLZ
For respective functional groups, the important peaks of TH and FLZ were present in the spectrum (fig. 4 and table 2). It concludes that the TH and FLZ are compatible with each other.
Development and validation of simultaneous estimation method of TH and FLZ by UV-visible spectrophotometer
Selection of wavelength of maximum absorbance (λmax) wavelength
TH and FLZ showed wavelength maximums at 222 nm and 260 nm in 0.1N HCl respectively as shown in fig. 2.
Determination of iso-absorptive point
1.5 µg/ml solution of TH and 200 µg/ml of FLZ were scanned in spectrum mode 200-400 nm and the overlain spectrum was found and it shows an iso-absorptive point at 239 nm as shown in fig. 5.
Results of validation parameters of simultaneous estimation method of TH and FLZ by UV-visible spectrophotometer
Linearity curve
The linearity curve for TH was constructed by plotting concentration versus absorbance (fig. 6). The linearity of TH was found in a concentration range of 0.5-3.0 µg/ml at 222 nm and 239 nm. The linearity of FLZ was found in the concentration range 160-400 µg/ml at 222 nm and 239 nm. A linear graph was obtained between absorbance and concentration in the range given in table 3.
(a) (b)
Fig. 2: UV spectra of (a) TH (b) FLZ in 0.1N HCl
Table 1: Calibration curve data of TH and FLZ in 0.1N HCl
| S. No. | Concentration (µg/ml) | Absorbance | Concentration (µg/ml) | Absorbance |
| TH | FLZ | |||
| 1 | 0 | 0 | 0 | 0 |
| 2 | 0.5 | 0.178±0.0005 | 80 | 0.165±0.0024 |
| 3 | 1.0 | 0.345±0.0005 | 120 | 0.243±0.0010 |
| 4 | 1.5 | 0.498±0.0008 | 160 | 0.318±0.0014 |
| 5 | 2.0 | 0.661±0.0005 | 200 | 0.396±0.0013 |
| 6 | 2.5 | 0.802±0.0006 | 240 | 0.475±0.0018 |
| 7 | 3.0 | 0.972±0.0008 | 280 | 0.569±0.0007 |
| 8 | 320 | 0.650±0.0010 | ||
| 9 | 360 | 0.720±0.0008 | ||
| 10 | 400 | 0.778±0.0017 |
*mean±SD; n=3, SD= standard deviation, TH= terbinafine hydrochloride, FLZ= fluconazole
Fig. 3: Calibration curve of TH (left) and FLZ (right) in 0.1N HCl
Fig. 4: FTIR spectra of a mixture of TH and FLZ
Table 2: Interpretation of FTIR spectra of a mixture of TH and FLZ
| Functional groups | Standard values (cm-1) | Observed vales (cm-1) |
| -OH stretching | 3200-2800 | 3119.99 |
| -SH stretching | 2800-2400 | 2446.81 |
| -CN stretching | 2400-2000 | 2227.88 |
| -C=O stretching | 1800-1600 | 1621.24 |
| -CH bending | 1600-1400 | 1465.96 |
| -COOH stretching | 1400-1200 | 1365.66 |
| -S=O stretching | 1200-1000 | 1076.33 |
| -CH bending | 1000-800 | 808.21 |
| -C-Cl stretching | 800-600 | 773.21 |
| -OH stretching | 3320.96 | 2964.72 |
| -C-F stretching | 1209.42 | 1210.38 |
| -C=N stretching | 1620.00 | 1621.24 |
Fig. 5: Overlain spectra of TH and FLZ in 0.1N HCl
Table 3: Linearity data of TH and FLZ at 222 nm, 239 nm, and in 0.1N HCl
| Conc. (µg/ml) | Conc. (µg/ml) | Absorbance at 222 nm | Absorbance at 239 nm | Absorbance at 222 nm | Absorbance at 239 nm |
| TH | FLZ | TH | FLZ | ||
| 0.5 | 160 | 0.178±0.0020 | 0.035±0.0010 | 0.265±0.0005 | 0.065±0.0012 |
| 1.0 | 200 | 0.345±0.0010 | 0.077±0.0015 | 0.334±0.0012 | 0.082±0.0012 |
| 1.5 | 240 | 0.498±0.0010 | 0.102±0.0006 | 0.431±.0010 | 0.105±0.0010 |
| 2.0 | 300 | 0.661±0.0020 | 0.140±0.0012 | 0.524±0.0005 | 0.126±0.0005 |
| 2.5 | 360 | 0.802±0.0010 | 0.178±0.0010 | 0.628±0.0015 | 0.154±0.0011 |
| 3.0 | 400 | 0.972±0.0010 | 0.217±0.0012 | 0.717±0.0012 | 0.176±0.0015 |
*mean±SD; n=3, SD= standard deviation, TH= terbinafine hydrochloride, FLZ= fluconazole
(a) (b)
(c) (d)
Fig. 6: Linearity curve of TH and FLZ at 222 nm and 239 nm in 0.1N HCl by Q-Analysis method, (a) TH at 222 nm (b) TH at 239 nm (c) FLZ at 222 nm (d) FLZ at 239 nm
Accuracy
Accuracy results obtained in table 4 showed excellent mean recovery percent values, close to 100 %, and low standard deviation values (SD<1.0) which indicate high accuracy of the proposed analytical methods. Percent recovery of TH was found in between of 98.50 %-103.96 % and FLZ it was found between 97.27 %-103.83 % in all recovery level and both wavelengths.
Table 4: Result of accuracy of TH at 222 nm and 239 nm by Q-Analysis method
% recover level |
Conc. of drug added | Abs | Mean | % RSD | % Recov | Abs | Mean | % RSD | % Recov |
| TH | (μg/ml) | TH at 222 nm | TH at 239 nm | ||||||
| 80% | 2.0 | 0.605 0.607 0.612 |
0.608±0.0360 | 0.592 | 103.40% | 0.138 0.139 0.140 |
0.139±0.0010 | 0.719 | 98.50% |
| 100% | 2.5 | 0.733 0.739 0.734 |
0.735±0.0032 | 0.435 | 100% | 0.176 0.177 0.176 |
0.176±0.0006 | 0.341 | 100% |
| 120% | 3.0 | 0.915 0.916 0.920 |
0.917±0.0026 | 0.283 | 103.96% | 0.212 0.212 0.209 |
0.211±0.0017 | 0.805 | 99.90% |
| FLZ | (μg/ml) | FLZ at 222 nm | FLZ at 239 nm | ||||||
| 80% | 240 | 0.429 0.430 0.431 |
0.430±0.0010 | 0.233 | 103.36% | 0.101 0.102 0.102 |
0.102±0.0006 | 0.588 | 101.19% |
| 100% | 300 | 0.519 0.522 0.520 |
0.520±0.0015 | 0.288 | 100% | 0.125 0.126 0.127 |
0.126±0.0010 | 0.769 | 100% |
| 120% | 360 | 0.607 0.605 0.610 |
0.607±0.0025 | 0.412 | 97.27% | 0.159 0.157 0.156 |
0.157±0.0015 | 0.955 | 103.83% |
*mean±SD, n=3, SD= standard deviation, %Recov = percent recovery, Abs= absorbance, conc= concentration, %RSD= relative standard deviation, TH= terbinafine hydrochloride, FLZ= fluconazole
Intermediate precision (Intra-day and inter-day precision)
Intra-day and inter-day precision were assessed using triplicate analysis of three different concentrations. The calculated relative standard deviation values were found to be very small below 2 % indicating good repeatability and reliability of the proposed methods. The results and their statistical analysis are summarized in table 5.
Table 5: Intraday and interday precision of TH and FLZ at 222 nm and 239 nm
Conc. (µg/ml) |
Intra-day | % RSD | Inter-day | % RSD | Intra-day | % RSD | Inter-day | % RSD |
| TH | Absorbance at 222 nm | Absorbance at 239 nm | ||||||
| 2.0 | 0.646±0.0017 | 0.216 | 0.641±0.0031 | 0.483 | 0.146±0.0012 | 0.822 | 0.144±0.0010 | 0.694 |
| 2.5 | 0.790±0.0045 | 0.569 | 0.790±0.0015 | 0.189 | 0.181±0.0012 | 0.662 | 0.182±0.0035 | 0.019 |
| 3.0 | 0.911±0.0064 | 0.702 | 0.917±0.0031 | 0.338 | 0.216±0.0006 | 0.277 | 0.217±0.0010 | 0.555 |
| FLZ | Absorbance at 222 nm | Absorbance at 239 nm | ||||||
| 240 | 0.450±0.0023 | 0.511 | 0.454±0.0021 | 0.465 | 0.104±0.0006 | 0.576 | 0.108±0.0006 | 0.555 |
| 300 | 0.562±0.0015 | 0.267 | 0.556±0.0010 | 0.180 | 0.129±0.00058 | 0.387 | 0.127±0.0012 | 0.945 |
| 360 | 0.658±0.0023 | 0.349 | 0.656±0.0015 | 0.229 | 0.154±0.0012 | 0.779 | 0.152±0.0010 | 0.658 |
*mean±SD; n=3, SD= standard deviation, %RSD= relative standard deviation, conc= concentration, TH= terbinafine hydrochloride, FLZ= fluconazole
Repeatability
The repeatability result indicates the precision under the same operating conditions over a short interval of time and % RSD was found less than 2 % which indicates good repeatability. % RSD was in the range of 0.016-0.0193 %, which were less than the standard value and show significant results (table 6).
Reproducibility
For the proposed method, the t-value found from the t-test indicates good reproducibility, and results are shown in table 7. The T-value of both the drugs was 0.8857, 0.4000 at 222 nm, and 239 nm in TH, wherein FLZ was 0.8000, 0.5066 respectively. Both the drug shows no significant difference.
Table 6: Repeatability data for TH and FLZ at 222 nm, 239 nm
Drugs conc S. No. |
TH | FLZ | ||
| 3 µg/ml | 3 µg/ml | 300 µg/ml | 300 µg/ml | |
| Abs at 222 nm | Abs at 239 nm | Abs at 222 nm | Abs at 239 nm | |
| 1 | 0.948 | 0.217 | 0.561 | 0.129 |
| 2 | 0.914 | 0.219 | 0.562 | 0.129 |
| 3 | 0.915 | 0.218 | 0.564 | 0.128 |
| 4 | 0.904 | 0.209 | 0.564 | 0.128 |
| 5 | 0.920 | 0.221 | 0.560 | 0.128 |
| 6 | 0.916 | 0.219 | 0.557 | 0.125 |
| Mean | 0.918±0.015 | 0.217±0.0042 | 0.561±0.0027 | 0.128±0.0015 |
| % RSD | 0.016 | 0.0193 | 0.481 | 0.0171 |
*mean±SD; n= 6, SD= standard deviation, %RSD= relative standard deviation, Conc= concentration, Abs= absorbance, TH= terbinafine hydrochloride, FLZ= fluconazole
Table 7: Reproducibility data for TH at 222 nm by Q-Analysis method
| Analyst 1 | Analyst 2 | t-value | Inference | Analyst 1 | Analyst 2 | t-value | Inference |
| TH at 222 nm (2.0 μg/ml) | TH at 239 nm (2.0 μg/ml) | ||||||
| 0.643±0.0015 | 0.646±0.0030 | 0.8857 | No significant difference | 0.145±0.038 | 0.149±0.0035 | 0.4000 | No significant difference |
| FLZ at 222 nm (300 μg/ml) | FLZ at 239 nm (300 μg/ml) | ||||||
| 0.566±0.0020 | 0.565±0.0040 | 0.8000 | No significant difference | 0.126±0.0025 | 0.128±0.0026 | 0.5066 | No significant difference |
*mean±SD; n=3, SD= standard deviation, TH= terbinafine hydrochloride, FLZ= fluconazole
Table 8: LOD and LOQ for TH and FLZ at 222 nm and 239 nm
| Wavelength | 222 nm | 239 nm | 222 nm | 239 nm |
| Drugs | TH | FLZ | ||
| LOD (µg/ml) | 0.067 | 0.175 | 31.089 | 94.380 |
| LOQ (µg/ml) | 0.203 | 0.531 | 94.210 | 286.00 |
| Mean of Residual values | 0.00020 | 0.000074 | 0.001619 | 0.001034 |
*LOD= limit of detection, LOQ= limit of quantification, TH= terbinafine hydrochloride, FLZ= fluconazole
Limit of detection (LOD) and limit of quantitation (LOQ)
The limit of detection and limit of quantitation was calculated from the linearity curve of TH and FLZ at 222 nm and 239 nm. LOD and LOQ of TH were found to be 0.067, 0.203 at 222 nm and 0.175, 0.531 at 239 nm, similarly for FLZ; 31.089, 94.210 at 222 nm and 94.380, 286.00 at 239 nm respectively (table 8).
DISCUSSION
A new, simple, precise, accurate, and reproducible method or simultaneous equation method was developed and validated for the simultaneous estimation of terbinafine hydrochloride (TH) and fluconazole (FLZ) in pure form. The absorption maxima of TH and FLZ were found at 222 nm and 260 nm respectively. It shows an iso-absorptive point at 239 nm. Compatibility studies show that both the drugs were compatible with each other. The linearity of TH was found in a concentration range of 0.5-3.0 µg/ml at 222 nm and 239 nm and the linearity of FLZ was found in the concentration range 160-400 µg/ml at 222 nm and 239 nm. Percent recovery of TH was found in between of 98.50 %-103.96 % and FLZ it was found between 97.27 %-103.83 % in all recovery level and both wavelengths. Percent recovery of TH and FLZ at all levels were found within the specific limit, which revealed that the method is accurate. The repeatability result indicates the precision under the same operating conditions over a short interval of time and % RSD was found less than 2 % which indicates good repeatability. Interday and intraday precision studies were also within the specific limits, which depicts that method is correct. The T-value of both the drugs was 0.8857, 0.4000 at 222 nm, and 239 nm in TH, wherein FLZ was 0.8000, 0.5066 respectively [23]. Both the drug shows no significant difference. LOD and LOQ of TH were found to be 0.067, 0.203 at 222 nm and 0.175, 0.531 at 239 nm, similarly for FLZ; 31.089, 94.210 at 222 nm and 94.380, 286.00 at 239 nm respectively [25]. All the parameters were done as per the ICH guidelines.
CONCLUSION
Simultaneous estimation of fluconazole and terbinafine hydrochloride was developed and validated by UV-visible spectroscopy. All the statically analyses were within the standard limits. It proves that the method was repeatable and selective for the simultaneous estimation of TH and FLZ in pure and that it can be successfully used for simultaneous estimation of TH and FLZ in pharmaceutical dosage forms. The developed method was found to be simple, precise, specific, and accurate.
FUNDING
Nil
AUTHORS CONTRIBUTIONS
All the author has contributed equally.
CONFLICT OF INTERESTS
Declared none
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