A green sustainable approach for the determination of a complex quintet hepatoprotective mixture using dual wavelength HPLC coupled with diode array detection: Compliance with greenness and whiteness metrics

Abstract

The wide spread of chronic liver disease is increasing constituting a worldwide health burden. Accordingly, mixed combinations of silymarin, N-acetyl cysteine, with antioxidant vitamins have been introduced. However, the great diversity between the analyzed components makes their simultaneous determination a rather difficult task. The water-soluble vitamins, vitamin C, and N-acetyl cysteine are highly hydrophilic with poor retention while the lipophilic vitamins, vitamin E and B-carotene, in addition to silymarin are strongly retained on the column. In this study, a novel and selective HPLC with diode array detection method has been proposed for the simultaneous determination of the five compounds using a gradient elution system of methanol and 0.025 M KH2PO4 adjusted to pH 2.5 with different flow rates. The method was fully validated as per ICH guidelines. Low limits of detections in the range 0.35–133.5 and of quantitation in the range of 1.07–404.7 μgmL−1 were obtained. The proposed method enabled the successful determination of the cited compounds in their combined pharmaceuticals with high accuracy and precision. Finally, method’s eco-friendliness and sustainability were investigated using greenness and whiteness assessment tools. The results revealed that the proposed method showed high degree of greenness and whiteness and is better than selected reference methods.