Validation of a NIR quantification method for the determination of chlorogenic acid in Lonicera japonica solution in ethanol precipitation process

► The feasibility study of NIR for low dose API in water extract of Lonicera japonica was verified. ► The PLS model based on Fourier Transform NIR and Holographic Grating NIR was built. ► The combination of conventional criteria (R2, RMSEC, RMSEP) and accuracy profile was applied. ► The LLOQ was about 1.5mg/mL for two types of NIR instruments. The feasibility of near-infrared spectroscopy (NIRS) for chlorogenic acid content analysis in ethanol precipitation process of water extract of Lonicera japonica was verified in this work. A calibration and validation set was designed for the conception and evaluation of the method adequacy. An experimental protocol was then followed, involving two different NIR instruments for data acquisition. On the basis of this protocol, the model was developed based on partial least squares regression (PLS) and the determination coefficient (R2cal and R2val), standard error of calibration and prediction (SEC and SEP) were 0.9962, 0.9955, 111.1μg/mL and 107.1μg/mL for Holographic Grating NIR instrument, and 0.9984, 0.9971, 53.6μg/mL and 83.3μg/mL for Fourier Transform NIR instrument. However, such above criteria did not clearly demonstrate the model's prediction error over each analyzed content range. Consequently, a novel approach based on accuracy profile which allowed the acquisition of the lower limit of quantification (LLOQ) was used to validate the robustness and accuracy of PLS model. The resulting accuracy profile showed that PLS model was able to determine chlorogenic acid content by two NIR systems, whose LLOQ was about 1550μg/mL. It was concluded that the two NIR systems were suitable for use as Process Analytical Technology (PAT) to understand ethanol precipitation process of water extract of Lonicera japonica.