Development and Validation of Stability Indicating RP-HPLC Method for Quantitative Estimation of Tofacitinib in Tofacitinib Tablets Dosage Form

Aim: The main intention of the research work is to develop an effective, sensitive, economical and uncomplicated reverse phase HPLC technique for the estimation of Tofacitinib in Tofacitinib tablets dosage form.

Study Design: HPLC based quantification studies.

Place and Duration of Study: Department of Chemistry, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur, Andhra Pradesh, between August 2022 and November 2022.

Methodology: Estimation of Tofacitinib in Tofacitinib tablets dosage form. The separation was achieved by using a stationary phase Kromasil C18 (150 x 4.6 mm, 5µ) and the mobile phase consisted of pH 4.0 phosphate buffer and acetonitrile in the ratio of (80:20 volume/volume). The flow rate was 1.5 mL/min. Tofacitinib was detected using UV detector at the wavelength of 215 nm. Column temperature 25°C and sample temperature 25°C and injection volume 20µL, run time was 20 minutes.

Results: As there is no meddling between blank and placebo at the retention time of Tofacitinib. Degradation study results were shown significant degradation was observed in alkali (base) stress condition. Hence it can be concluded that Tofacitinib is sensitive to alkali. To obtain system exactitude, a study was conducted with six replicate injections. %RSD was estimated from the peak areas of Tofacitinib found to be 0.16% respectively. The relative standard deviation for method exactitude was found to be 5 mg and 10 mg is 0.26% and 0.75%. The proposed HPLC technique was linear over the range of 24.88-74.64 µg/mL, the correlation coefficient was found to be 1.0000. The accuracy studies were shown as % recovery for Tofacitinib 50% to150% level. The limit of % recovered shown is in the range of 98 and 102% and the results obtained were found to be within the limits. Hence the method was found to be accurate. The solution stability of the standard and samples are stable upto 48 hrs on a bench top and refrigerator (2-8°C). The technique is robust for changes like flow rate, column oven temperature, pH variation and the organic phase of the mobile phase. Performed the filter validation for sample solution 0.45 µm PVDF filterers are suitable for filtration.

The technique has authenticated as per ICH instructions and all the validation parameters are satisfy the ICH Q2 specification acceptance limits.

Conclusion: The urbanized technique was validated for various parameters as per ICH guidelines like accuracy, precision, linearity, specificity, system suitability, solution stability and robustness. The results obtained were within the acceptance criteria. So, it can be concluded that the developed method is simple, precise, cost-effective, eco-friendly, and safe and can be successfully employed for the routine analysis of Tofacitinib in bulk and pharmaceutical dosage forms.