HPLC Methods for the Quantification of Cyproheptadine Hydrochloride, Vitamins Using RP-HPLC with UV Detection

Abstract

For the purposes of this investigation, we have created straightforward RP-HPLC techniques for the analysis of cyproheptadine hydrochloride, thiamine mononitrate, calcium pantothenate, and pyridoxine hydrochloride. The International Conference on Harmonization's (ICH) Q2(R1) 2005 criteria were followed in the conduct of this study. The quality control of several APIs in single dose forms depends heavily on the availability of trustworthy, high-throughput analytical technologies. Here, we present the development and validation of two techniques for the identification and quantification of calcium pantothenate, cyproheptadine hydrochloride (CH), thiamine mononitrate (Vit. B1), and pyridoxine hydrochloride (Vit. B6) in uncoated tablets. For the vitamin test, a mobile phase mixture of phosphate buffer pH 3.5 and methanol in the ratio 93:7 was used. The stationary phase used was Eurospher ODS (150 x 4.5mm), and the flow rate was set at 1.0 mL/min (ambient temperature settings). For thiamine mononitrate and pyridoxine hydrochloride, the wavelength of detection was 270 nm with a run time of 0.00 to 5.50 minutes, and for calcium pantothenate, it was 205 nm with a run time of 5.50 to 11.00 minutes. The injection volume was 20 µL. 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 minutes respectively.

The mobile phase composition for cyproheptadine HCl was methanol and an ion-pairing solution (70:30), set at a flow rate of 1.0 mL/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, with linear dynamic 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, accurate, and robust for the estimation of cyproheptadine hydrochloride, vitamins B₁, B₅ and B_6.