Quality by Design Approach in Quantitative Determination of the Red Dye from Extract of Onosma Echioides (Boraginaceae) by UV-visible Spectrophotometer

Abstract

Implementation of the Quality by Design (QbD) approach to analytical method development for quantifying red dye from the extract of Onosma echioides was performed. The soxhlet extraction technique was prepared by the dried root bark of O. echioides extract with petroleum ether (60–80°C). A column chromatographic technique used a mobile phase toluene: formic acid (99:1) toluate the major coloring compound from the petroleum ether extract. The extract was standardized using an isolated compound by High-Performance Thin Layer Chromatography (HPTLC). To develop the UV-visible spectrophotometric method, the two-level full factorial model with three variable factors was designed. QbD approach facilitated to calculate the spectrophotometric risk factor of the variable. The QbD approach proved that the selected spectrophotometric condition of each factor was lying in the middle of the design space and has wide boundaries and space. The middle values of the range were selected for method validation. The calibration curve showed a good linear relationship over the concentration range of 10 to 70 ppm. The LOQ and LOD were 0.95 ppm and 2.86 ppm, which indicates adequate method sensitivity. The mean RSD of interday precision, intraday precision and robustness was less than 2, indicating that the method is precise and robust. The accuracy of the method was found to be 105.65%. Thus, an accurate method was developed for the quantification of red dye from extract of O. echioides by using QbD approach. The proposed spectrophotometric method along with QbD approach, can be used as an alternative tool in the drug quality control laboratories for the quantitative determination of red dye in O. echioides in the extract.