INTRODUCTION:

Tamoxifen is an antineoplastic nonsteroidal selective estrogen receptor modulator (SERM). Tamoxifen competitively inhibits the binding of estradiol to estrogen receptors, thereby preventing the receptor from binding to the estrogen-response element on DNA. The result is a reduction in DNA synthesis and cellular response to estrogen. In addition, tamoxifen up-regulates the production of transforming growth factor B (TGFb), a factor that inhibits tumor cell growth, and down-regulates insulin-like growth factor 1 (IGF-1), a factor that stimulates breast cancer cell growth. It is official in IP²⁶, B.P²³, Merck Index²⁴ USP²⁵. Tamoxifen citrate is rapidly and almost completely absorbed after oral administration. The Method of Validation parameters follows by ICH Guidelines. The literature suggests that there are many analytical techniques like UV spectroscopy^1-10 HPLC^12-13, RP-HPLC^14-17, HPTLC^20-21, Biodegradability, TLC, Electrophoresis and Ion Pair Chromatography for Tamoxifen citrate.

Multilinear Regression (MLR) relies upon statistical guidelines of multilinear measurements, which comprises examination and perception examination of more than one factual result variable at once. In structure and investigation, the strategy is utilized to execute exchange examines various concentrations while in consideration of the impacts of all factors on the reactions of premiums.

Figure 1: Chemical structure for Tamoxifen citrate

METHODOLOGY:

Chemicals:

Tamoxifen citrate in local pharmaceutical companies

Distilled water

The marketed tablet formulation (label claim – 50mg)

Solubility:

Rapidly soluble in Distilled water.

Instrumentation:

The multilinear regression was executed in 1.0cm quartz cells using UV 7600 UV- Visible Spectrometer (UV Lamda., Eng.) with a fixed 1nm spectral bandwidth and UV Spectrum Rx v1.04 software was utilize for all absorbance measurements.

Preparation of Standard Stock Solutions:

Standard stock solution for Tamoxifen citrate was prepared by taking 25mg in 100ml of distilled and the final volume makeup with Distilled water in 100mL Volumetric flask and 250µg/ml was determined as final concentration. The 80µg/ml was taken from the working standard solution were examined in the entire Ultraviolet range between 400-200nm to obtain overlay spectra and absorbance spectra.

Determination of λ max:

The standard stock solution of Tamoxifen citrate was dilute with the solvent to get a concentration of 80µg/mL. This scanned under the UV region from 400-200nm. The maximum absorbance was found to be 236 nm.⁵

Preparation of sample solution:

For evaluation of synthetic mixtures. 250mg of the bulk drug weigh accurately and transfer into a 100mL volumetric flask containing the distilled water. The above solutions were filtered through Watmann filter paper and the solution was transferred into a standard flask and makeup to the fixation with a solvent system to obtain a final concentration of 20µg/ml. All determinations were conducted with six variates. The solutions were sonicated for 15 min. and finally makeup to the volume with solvent, filtered and diluted for further analysis.

Method of Validation:

This technique was evaluated as per the ICH Q2 R1 guidelines to examine validation parameters such as accuracy, sensitivity, precision, and linearity.

Linearity:

The prepared standard stock solution 250µg/ml of Tamoxifen citrate was dilute with a solvent to get a concentration in the range 20-120µg/ml. To develop an interactive and underrate instrumental fluctuation absorbance of the solution was calibrated in certain wavelengths around the λmax of the drug 236nm i.e., 267, 269, 271, 273, 275nm. Then the overlay spectra show linearity of the absorbance were recorded at six various concentrations. The overlay spectra were shown in Figure 3

Precision:

Intra-day and Inter-day Precision was performed by examining the absorbance of the solution of concentration 80µg/ml at a selected wavelength. The concentration was scanned two times within a day (intraday precision) and for two days (interday precision). The absorbance value recorded at the selected wavelength for intraday and interday precision. The SD and %RSD values obtained at different wavelengths were calculated.

Assay:

The absorbance of the extracted sample solution was recorded at 236nm and the quantity of drug present in the formulation was determined

Recovery studies:

This method was executed to justify 0.5ml of standard formulation (100µg/mL) were taken in three 10ml standard flask and it 40%, 60%, 80% (i.e, 4mL, 6mL, 8mL) of working standard stock solution (100µg/mL) and make the volume up to the mark. The corresponding absorbance at 236nm was observed against the blank. The quantity of drug added concentration was determined from the obtained percentage recovery and absorbance values for each dosage form.

RESULTS AND DISCUSSION:

Tamoxifen citrate was predicted by the prospective multivariate UV spectroscopic method in tablets. The bulk drug of Tamoxifen citrate was rapidly soluble in distilled H₂O then distilled H₂O was fixed as a solvent for Tamoxifen citrate to determine the UV spectrum in the range of 400-200 nm. After the validation of the spectrum, the λ max of Tamoxifen citrate present at 236 nm (Figure 2)

Figure 2: λ max of Tamoxifen citrate

The calibration curves of Tamoxifen citrate at different wavelengths is shown in Figure 3. The technique was evaluated through ICH Q2 B guidelines for accuracy, linearity, precision, and sensitivity.

Linearity:

The concentration range 20-120µg/ml were scanned for the linearity at the selected wavelength of about 240, 238, 236, 234 and 232nm which was shown in Figure 3 and their calibration graphs were shown in Figure 4-8.

Figure 3: Overlay spectra of Tamoxifen citrate

Figure 4: Calibration curve at 232 nm

Figure 5: Calibration curve at 234 nm

Figure 6: Calibration curve at 236 nm

Figure 7: Calibration curve at 238 nm

Figure 8: Calibration curve at 240 nm

Precision:

Inter-day and Intra-day precision was done through scanning the absorbance of 80 µg/ml solution at five selected wavelengths. The %RSD for Inter- day and Intra-day were found lie within range 0.828-1.274 and 0.814- 1.275 and it was within well limited of less than 2% at all the wavelength. The Prepared solvent in 80µg/ml was scanned three times per day and for 3 days at the same intervals time for Inter day and Intraday precision. The least value of % Relative standard deviation shows that the destined technique was accurate and precise.

Assay

The sample solutions were recorded for absorbance at 236nm and the amount of drug present in the dosage form was evaluated. The assay percentage of Tamoxifen citrate was found to be 99.44% w/w.

Recovery:

The percentage recovery of the drug was calculated and was found to be within the range of 99.35-101.15% w/w it shows a percentage recovery belongs to within the limits of 97 – 103 % w/w as per the ICH regulations.

Table 1: Contents of Calibration Parameters:

PARAMETERS	WAVELENGTHS (nm)
PARAMETERS	232	234	236	238	240
Linearity (µg/ml)	20-120	20-120	20-120	20-120	20-120
Correlation coefficient R²	0.999	0.999	0.9999	0.9999	0.9994
Slope (m)	0.0046	0.0053	0.0056	0.0053	0.0053
Intercept (c)	0.0054	0.0043	0.0040	0.0071	0.0020
LOD (µg/ml)	2.91	2.90	2.91	2.90	2.91
LOQ (µg/ml)	8.81	8.82	8.83	8.84	8.84
Precision Inter Day (%RSD)	1.2763	0.8554	0.8281	1.1175	1.1374
Precision Intra Day (%RSD)	0.9993	0.8491	0.8154	1.0029	1.1240
Assay (%w/w)	99.67-101.52	99.05-100.01	99.33-100.11	99.54-100.91	99.31-100.02
Percent Recovery(%w/w)	99.64-101.10	99.10-100.15	99.34-100.98	99.44-101.86	99.40-100.15

CONCLUSION:

UV spectrophotometric of the Multivariate calibration technique was performed as per the ICH protocol shows that lie within the limit. The method was developed in the present studies were establish to be accurate, precise, sensitive and reproducible for the assessment of Tamoxifen citrate and its component formulation. In this way, a basic and fast strategy utilizing scientific substance was created, which was discovered more unsurprising than the other spectroscopic strategies and is unequivocally suggested for the normal quality control analysis of Tamoxifen citrate in pharmaceutical formulation.

ACKNOWLEDGEMENT:

The Authors are Thankful to Respective Management of SRM Institute of Science and Technology and our sincere Thanks to our Dean and Respective Staffs of SRM College of Pharmacy.

CONFLICT OF INTEREST:

The Authors declares no conflicts of Interest

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Received on 10.02.2020 Modified on 31.03.2020

Research J. Pharm. and Tech. 2020; 13(12):6022-6026.

DOI: 10.5958/0974-360X.2020.01050.1