Stability-Indicating HPLC Methods for the Quantification of Cyproheptadine Hydrochloride, Vitamins B1, B5 And B6 in Tablets using RP-HPLC with UV Detection

Abstract

The availability of reliable and fast analytical method is crucial for the quality control of multi-API in a single dosage form. Herein, two methods have been developed and validated for the determination of Cyproheptadine hydrochloride (CH), Thiamine mononitrate (Vit. B₁), Calcium pantothenate (Vit. B₅) and Pyridoxine hydrochloride (Vit. B₆) in uncoated tablets. To assay the vitamins, a mobile phase combination of Phosphate buffer pH 3.5 and Methanol in the ratio 93:7, using Eurospher ODS (150 x 4.5mm) as the stationary phase with a flow rate set at 1.0 mL/min at ambient temperature conditions was adopted. Wavelength of detection was 270 nm with a run time of 0.00 to 5.50 minutes for Thiamine mononitrate and Pyridoxine hydrochloride, and 205 nm from 5.50 to 11.00 minutes for Calcium pantothenate. A good resolution and a short run time of 11 minutes were achieved with the validated conditions. The retention times of Thiamine mononitrate, Pyridoxine hydrochloride and Calcium pantothenate were 2.823±0.020, 4.184±0.007 and 10.025±0.015 respectively.

The mobile phase for Cyproheptadine HCl was methanol and an ion-pairing solution (70:30), set at a flow rate of 1mL/min, a runtime of 8 minutes, an injection volume of 20 µL and a wavelength of 285 nm using Eurospher ODS (150 x 4.5 mm) as the stationary phase. The retention time for Cyproheptadine HCl was 4.961±0.006. Both methods were found to be specific, robust, accurate and precise over the concentration ranges of 0.0192 mg/mL–0.0288 mg/mL for Thiamine mononitrate, 0.0128–0.0192 mg/mL for Pyridoxine hydrochloride, 0.032–0.048 mg/mL for Calcium pantothenate and 0.032-0.048 mg/mL for Cyproheptadine hydrochloride. The Correlation Coefficient (r²) for Cyproheptadine hydrochloride, Thiamine mononitrate, Pyridoxine hydrochloride and Calcium pantothenate were greater than 0.999.

The purpose of this study was to develop and validate simple HPLC methods for the estimation of Cyproheptadine hydrochloride, Vitamins B₁, B₅ and B₆ in combined dosage forms. The proposed methods were found to be precise, specific, accurate and robust for the estimation of Cyproheptadine hydrochloride, Vitamins B₁, B₅ and B_6.

Keywords: Cyproheptadine hydrochloride, Thiamine mononitrate, Pyridoxine hydrochloride, Calcium pantothenate, RP-HPLC, Validation

DOI: 10.7176/CMR/12-6-06

Publication date: August 31^st 2020