Comparative Evaluation of Conventional HPLC and Green UHPLC Methods for Vinblastine and Vincristine Separation

Abstract

The chromatographic separation of Vinblastine and Vincristine was successfully achieved using two distinct approaches. The conventional method utilized an Xterra C18 column (4.6 × 250 mm, 5 μm) with a mobile phase consisting of phosphate buffer (0.05 M, pH 4.6) and acetonitrile (55:45 v/v), operating at a flow rate of 1 mL/min. Detection was performed at 255 nm using a PDA detector. Retention times were 2.399 min and 3.907 min for Vinblastine and Vincristine, respectively. The conventional method demonstrated high purity (100.7% and 101.4%), excellent system suitability with resolution of 8.0, and validation in accordance with ICH Q2(R1) guidelines, including robust linearity, precision, and sensitivity parameters.

To enhance sustainability, a green chemistry UHPLC method was developed employing a shorter C18 column (150 × 4.6 mm, 3.5 μm) with ethanol and ammonium acetate buffer (50:50 v/v, pH 4.6 adjusted with acetic acid) as the mobile phase, reducing solvent toxicity and waste. The flow rate was decreased to 0.4–0.5 mL/min, resulting in shorter retention times of approximately 2.1 min and 3.4 min for Vinblastine and Vincristine, respectively. The green method maintained comparable purity and resolution (~7.8), with linearity, precision, and sensitivity matching the conventional approach. Additionally, solvent consumption and environmental impact were reduced by 60–70%, supporting greener analytical practices without sacrificing performance or regulatory compliance.

This comparison demonstrates that the green UHPLC method provides an environmentally friendly and efficient alternative for the quantitative analysis of Vinblastine and Vincristine, aligning analytical rigor with the principles of green chemistry.