Development and Validation of UV-Spectroscopic methods for simultaneous Estimation of Hydrocortisone and Iodoquinol in Tablet Dosage Forms
Ramavath Mohanbabu Naik1, Syed Ahmed2, S. Vijay Kumar3, K. Radhika4,
Dr. K. Vinod Kumar5
1Asst Professor, Department of Pharmaceutical Analysis and Quality Assurance, Gland Institute of Pharmaceutical Science, Kothapet (V), Near Narsapur, Medak (District), Telangana-502334.
2Asst Professor, Department of Pharmaceutics, Gland Institute of Pharmaceutical Science,
Kothapet (V), Near Narsapur, Medak (District), Telangana-502334.
3Asst Professor, Department of Pharmacognosy, Vishnu Institute of Pharmaceutical Education and Research, Narsapur (Mandal), Medak (District), Telangana-502334.
4Asst Professor, Department of Pharmacology, Gland institute of Pharmaceutical Science,
Kothapet (V), Near Narsapur, Medak (District), Telangana-502334.
5Associate Professor, Department of Pharmaceutical Analysis and Quality Assurance, Raghavendra Institute of Pharmaceutical Education and Research, Krishnam Reddy Pally (V), Ananthapuramu- 515721,
Andhra Pradesh, India.
*Corresponding Author E-mail: mohanbabu623@gmail.com
ABSTRACT:
A simple, precise, accurate and economical UV-Spectrophotometric method has been developed and validated for the simultaneous estimation of Hydrocortisone and Iodoquinol in combined pharmaceutical dosage form using simultaneous equation method. Hydrocortisone has absorbance maxima at 245nm and Iodoquinol has absorbance maxima at 250nm in Methanol. The drugs obeyed beer’s law in the concentration range of 10-60µg/ml for Hydrocortisone and 5-30µg/ml for Iodoquinol. The correlation coefficient of Hydrocortisone and Iodoquinol was found to be 0.999 and 0.999 respectively. The method was statistically validated as per the ICH guidelines. The low RSD values indicate good precision and high recovery values indicate accuracy of the proposed method. The developed method was simple, precise, accurate, reproducible and economical which can be efficiently and easily applied to pharmaceutical dosage form.
KEYWORDS: Hydrocortisone, Iodoquinol, UV-Spectroscopic method.
Hydrocortisone (Hydro) is chemically (1S,2R10S,11S,14R,15S,17S)- 14,17- dihyroxy-14 -(2-hydroxyacetyl)- 2,15-dimethyl tetracyclo [8.7.0.0^{2,7} heptadec- 6- en-5- one. HYDRO belongs to Anti-inflammatory Agents. Structure of Hydro was shown in figure 1[1].
Diiodohydroxy quinoline also known as Iodoquinol, is a quinoline derivative that can be used in the treatment of Amoebiasis. Iodoquinol (Iodo) is chemically 5,7-diiodoquinolin-8-ol. Structure of Iodo was shown in Figure 2[2].
Literature studies revealed that there is no method till date developed for the estimation of these two drugs in the combined dosage form and hence a simple, precise and an accurate UV-Spectrophotometric method has been developed in the presentstudy.
Figure 1: Hydrocortisone
Figure 2: Iodoquinol
EXPERIMENTAL:
MATERIAL AND REAGENTS:
Analytically pure Hydrocortisone was kindly provided by Gland Pharma Laboratory, and Iodoquinol was provided by Mylan Laboratory, as gift samples. Analytical grade methanol was purchased from Merck and Co. Glasswares used in each procedure were soaked over night in a mixture of chromic acid and sulphuric acid rinsed thoroughly with double distilled water and dried in hot air oven. Water were purchased from Merck, India. Triple distilled water is used for all purpose.
Instrumentation:
UV- system (Model-1700 with UV Probe Software) Manufacturer is Shimadzu. Shimadzu-1700+ UV-Visible double beam spectrophotometer with a fixed slit width 1nm and 1cm matched quartz cells was used for all the spectral measurements.
Selection of Common Solvent:
The selection of solvent is made after assessing the solubility of both the drugs in various solvents. Methanol is selected as the common solvent and the spectral characteristics were studied out using this solvent.
Preparation of Standard Drug Solution:
Standard Hydrocortisone of 10mg and Iodoquinol of 10mg was accurately weighed and transfered in to a 100 ml volumetric flask. Add small amount of methanol in volumetric flask and dissolve the drug in to it and then adjust the volume up to 100ml. This solution from 100ppm of concentration and then prepare other dilution.
Selection of Analytical concentration Range:
From the standard stock solution of Iodoquinol (100µg/ml), appropriate dilutions of 5ppm, 10ppm up to 30ppm are prepared.
From the standard stock solution of Hydrocortisone (100µg/ml), appropriate dilutions of 10ppm, 20ppm up to 60ppm are prepared.
Method I (Simultaneous Equation Method):
Two wavelengths selected for the method are 245nm and 250nm that are absorption maxima of HC and Iodo respectively in Methanol. The stock solutions of both the drugs were further diluted separately with methanol to get a series of standard solutions of 5-60µg/mL concentrations.
The absorbances were measured at the selected wavelengths and absorptivities for the drugs were determined. Concentrations in the sample were obtained by using following equations-
A1= ax1Cx + ay1 Cy A2= ax2Cx + ay2 Cy
Where, A1 and A2 are absorbance’s of mixture at 245nm and 250 nm respectively, ax1 and ax2 are absorptivities of HC at λ1 and λ2 respectively and ay1 and ay2 are absorptivities of SP at λ1 and λ2 respectively. Cx and Cy are concentrations of HC and Iodo respectively.
0.392 = 464.64 Cx + 143.95 Cy
1.331= 27.385 Cx + 635.832 Cy
Method II (Multicomponent Mode):
Multicomponent mode was employed for the estimation of these drugs in the combined dosage form. For this method of analysis 245and 250 nm were selected as the sampling wavelengths. The drugs showed linearity in the range of 5- 30µg/mL for HC and 10-60µg/mL for Iodo. Three mixed standard solutions containing both the drugs were prepared in the ratio of 1:2 and the spectral characteristics were studied after the data is fed into the instrument. The instrument then gave the concentrations of individual drugs in the test by an inbuilt microprocessor.
Method Validation:
The method was validated according to ICH Q2B guidelines 8 for linearity, accuracy and precision.
1. Linearity:
Linearity solutions for the method were prepared from the stock solutions of both the drugs and spectral runs were carried out.
2. Precision:
This is studied to find out the intra and the inter day variations in the developed method. The %RSD of the three assay values at three different concentrations on same day (intraday) and on three different days (interday) values were calculated.
3.Accuracy:
The accuracy of this method was evaluated at three concentration levels and the recovery studies were performed by standard addition method. Percent recovery for both the methods was then calculated.
RESULTS AND DISCUSSION:
The overlain spectra of Hydrocortisone and Iodoquinol exhibit λmax of 245nm and 250nm respectively. These wavelengths were selected for both the methods. Standard calibration curves for the drugs were described with equations Y=0.021x+0.005 for Hydro and Y=0.0428x+0.0002 for for simultaneous equation method and Y=0.021x+0.005 and Y=0.0428x + 0.0002 for Hydro and Iodo respectively for Multicomponent mode. The correlation coefficient for both the drugs was found to be 0.999 in both the methods.
The beer Lambert’s linearity range of these drugs include 10-60µg/mL for Hydro and 5-30µg/mL of Iodo. The results were tabulated in table 1. The %RSD for the intraday was found to be 0.130 for Hydro and 0.113 for Iodo. Similarly 0.286 and 0.467 are the %RSD of HC and Iod respectively for inter day studies for simultaneous equation method.
Table 1: Precision Studies
|
Drug |
Concentration(µg/ml) (n=3 ) |
%RSD |
|
|
Intraday |
Interday |
||
|
Iodoquinol |
4 |
0.113 |
0.467 |
|
5 |
0.203 |
0.320 |
|
|
6 |
0.205 |
0.304 |
|
|
Hydrocortisone |
8 |
0.130 |
0.286 |
|
10 |
0.080 |
0.155 |
|
In the case of Multicomponent mode the %RSD for intraday was found to be 0.33 for both the drugs and %RSD for inter day studies were found to be 0.82 and 0.76 for HC and Iodo respectively.
The %RSD for intraday and inter day precision were found to be less than 2 indicating that the method is precise. The results of these were presented in table 2.
The accuracy was confirmed by the recovery studies and the percentage recovery was found to be in the range of 96-102% thus justifying the method. From accuracy studies it was found that the RSD was less than 2% indicating that the method is accurate. The results were presented in table 3and 4.
Table 2: Accuracy of Iodoquinol
|
Concentration of standard Iodoquinol (µg/ml) |
Recovery level |
Amount of drug added (µg/ml) |
Total amount of drug added |
Amount of drug found mean± SD (n=3 ) |
%Recovery |
%RSD |
|
5 |
80% |
4 |
9 |
8.57±0.02 |
95.2% |
0.113 |
|
100% |
5 |
10 |
9.95±0.04 |
99.5% |
0.203 |
|
|
120% |
6 |
11 |
10.97±0.045 |
99.7% |
0.205 |
Table 3: Accuracy of Hydrocortisone
|
Concentration of standard Hydrocortisone (µg/ml) |
Recovery level |
Amount of drug added (µg/ml) |
Total amount of drug added |
Amount of drug found mean± SD (n=3 ) |
%Recovery |
%RSD |
|
10 |
80% |
8 |
18 |
17.57±0.02 |
97.6% |
0.130 |
|
100% |
10 |
20 |
19.95±0.04 |
99.7% |
0.080 |
|
|
120% |
12 |
22 |
21.98±0.045 |
99.9% |
0.074 |
Table 4: LOD and LOQ Studies
|
Drug |
LOD |
LOQ |
|
Iodoquinol |
0.23µg/ml |
1.91µg/ml |
|
Hydrocortisone |
0.47µg/ml |
2.33µg/ml |
Table 5: Analysis data of Formulation
|
Drug |
Labeled claim(mg) |
Amount found(mg) Mean± SD (n=3 ) |
Assay value (%) |
%RSD |
|
Iodoquinol |
10 |
9.67±0.02 |
96.7 |
0.1033 |
|
Hydrocortisone |
20 |
20.67±0.01 |
103.3 |
0.0322 |
CONCLUSION:
The proposed UV Spectrophotometric method was found to be simple, precise, accurate and economical for the simultaneous estimation of Hydrocortisone and Iodoquinol in combined dosage forms. Hence, this method can be easily used for routine quality control analysis of Hydrocortisone and Iodoquinol in pure and its combined dosage form.
REFERENCE:
1. Ramavath Mohanbabu Naik, Syed Ahmed , S. Vijaykumar, Dr. D. Ramakrishna, K. Venkatalakshmi. Simultaneous Estimation of hydrocortisone and ketoconazole in tablet dosage form by UV-Spectroscopy. Journal of Pharma Research. 2019;8(5):323-326.N: 2319-5622.
2. Ramavath Mohanbabu Naik, Swapna Gumpula and Prajakta Shamsundar Mhaskar. Simultaneous Estimation of hydrocortisone and ketoconazole in pharmaceutical dosage form by RP-HPLC method. International Journal of Pharmaceutical Sciences and Research. 2018; 9(11): 4805-4811.
3. Sunil Kumar, R.K. Nanda, Pritam Kuttepali, S.K. Sharma. Development and Validation of RP-HPLC Method for Estimation of Hamycin and Ketoconazole in Pharmaceutical Cream. Pharm Analysis and Quality Assurance. 2014; 5(1): 263-268.
4. Ramavath Mohanbabu Naik, Syed Ahmed, G. Naveen Kumar Reddy. RP-HPLC Method Development and Validation for Simultaneous Estimation of Hydrocortisone and Acyclovir in Pharmaceutical dosage forms. American Journal of Pharmacy and Health Research. 2018; 6(6):5-18.
Received on 24.09.2019 Modified on 27.11.2019
Accepted on 21.01.2020 © RJPT All right reserved
Research J. Pharm. and Tech. 2020; 13(8):3777-3780.
DOI: 10.5958/0974-360X.2020.00668.X