Simultaneous quantitation of five Panax notoginseng saponins by multi heart-cutting two-dimensional liquid chromatography: Method development and application to the quality control of eight Notoginseng containing Chinese patent medicines

• A “monomethod-heterotrait matrix” strategy was proposed for TCM quality control.
• A multi heart-cutting 2D-LC (MHC-2D-LC) assay method was developed and validated.
• The quality of eight Chinese patent medicines was assessed by the MHC-2D-LC method.
• The MHC-2D-LC method was proven as superior over the China Pharmacopoeia methods.

Current China Pharmacopoeia (ChP) standards employ diversified and case-dependent assay methods to evaluate the quality of different Chinese patent medicines (CPMs) that contain Panax notoginseng as the monarch drug. These conventional, HPLC-based approaches, utilizing a complex sample preparation procedure, can easily result in low analytical efficiency and possible component loss. Here, a “monomethod-heterotrait matrix” (MHM) strategy is proposed, that is, developing a universal multi heart-cutting two-dimensional liquid chromatography (MHC-2D-LC) approach that facilitates the simultaneous quantitation of five P. notoginseng saponins (noto-R1, Re, Rg1, Rb1, and Rd) in eight different CPMs. The MHC-2D-LC system was constructed on a dual-gradient liquid chromatography instrument equipped with a Poroshell SB C18 column and a Zorbax SB-Aq column for respective 1D and 2D separation. Method validation was performed in terms of specificity, linearity (r2 and F-test), intra-/inter-day precision (0.4–7.9%), stability (1.2–3.9%), and recovery (90.2–108.7%), and the LODs and LOQs (loaded masses) of the five analytes varied between 4.0–11.0 ng and 6.0–33.0 ng, respectively. The validated MHC-2D-LC approach was subsequently applied to quantify the five saponins in thirty batches of different CPMs. The method demonstrated superiority over the current ChP assay methods in respect of specificity (avoiding co-elution), resolution (Rs > 1.5), sample preparation (easy-to-implement ultrasonic extraction without repeated re-extraction), and transfer rate (minimum component loss). This is the first application of an MHC-2D-LC method for the quantitative assessment of the constituents of CPMs. The MHM approach represents a new, strategically significant methodology for the quality control of CPMs that involve complex chemical matrix.