Simultaneous Quantitative and Chemical Fingerprint Analysis of Receptaculum Nelumbinis Based on HPLC-DAD-MS Combined with Chemometrics

Abstract

A rapid and sensitive method based on HPLC-DAD-MS was developed for quantitative analysis of two flavonoids and chemical fingerprint analysis to evaluate the quality of Receptaculum Nelumbinis. The analysis was conducted on a Poroshell 120 C18 column (100 × 4.6 mm, 2.7 μm) with 0.2% formic acid buffer solution and methanol as mobile phases with gradient elution. This method displayed good linearity with R(2) at >0.9999 and limits of quantity <0.37 μg mL(-1). Relative standard deviation values for intra- and interday precision were <0.82 and 1.03%, respectively. The mean recovery of hyperoside was 95.54% and of isoquercitrin was 92.10%. Hyperoside and isoquercitrin were determined simultaneously, and 12 peaks in the chemical fingerprint were identified. The chemometric methods, including similarity analysis, hierarchical clustering analysis and principal component analysis, were applied to distinguish 11 batches of Receptaculum Nelumbinis samples. The above results could validate each other and successfully divide these samples into two groups. Moreover, hyperoside and isoquercitrin could be selected as chemical markers to evaluate the quality of Receptaculum Nelumbinis from different localities. This study demonstrated that the developed method was a powerful and beneficial tool to carry out the quality control of Receptaculum Nelumbinis.

Figure 1.

Typical chromatograms for chemical analysis of mixed standards (A) and Receptaculum Nelumbinis (B). This figure is available in black and white in print and in color at JCS online.

Figure 2.

Chemical structures and MS data of 12 flavonoids identified in Receptaculum Nelumbinis.

Figure 3.

Chemometric analysis of 11 batches of Receptaculum Nelumbinis samples. (A) Dendrogram for hierarchical clustering by PASW Statistics. (B) Scores plot of PCA by the Unscrambler. (C) Loadings plot of PCA for 12 peaks in their HPLC profiles by the Unscrambler. This figure is available in black and white in print and in color at JCS online.