Abstract: Insamyangpye decoction (ISYPD) is an oriental herbal prescription used in Korea to treat lung-related diseases such as chronic obstructive pulmonary disease. ISYPD is a complex prescription consisting of 13 herbal medicines, and ISYPD sample was obtained by adding 50 L of distilled water to a mixture (5 kg) of 13 herbal medicines, extracting at 100 ◦C for 2 h using an electric extractor, and freeze-drying. In this study, an accurate and sensitive liquid chromatography tandem mass spectrometry (LC–MS/MS) method based on multiple reaction monitoring (MRM) was developed and verified for quality assessment of ISYPD using 10 marker components: mulberroside A (1), amygdalin (2), liquiritin apioside (3), naringin (4), poncirin (5), platycodin D (6), ginsenoside Rb1 (7), glycyrrhizin (8), saikosaponin A (9), and schizandrin (10). These marker compounds were separated using an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 µm) maintained at 30 ◦C with a mobile phase elution gradient of acetonitrile in distilled water, both containing 0.1% (v/v) trifluoroacetic acid. Marker components were quantified using the LC–MS/MS MRM method developed and validated, and found at 0.09–7.47 mg/g.
Authentic reference standards (Figure S1) for LC–MS/MS quantitative analysis of ISYPD samples were supplied by manufacturers of phytochemical standards: compound 1 (C26H32O14, 98.1%) from Ensol Biosciences (Daejeon, Korea); compounds 2 (C20H27NO11, ≥99.0%) and 4 (C27H32O14, 95.0%) from Merck (Seoul, Korea); compounds 3 (C26H30O13, ≥98.0%), 5 (C28H34O14, 98.9%), and 7 (C54H92O23, 98.5%) from Shanghai Sunny Biotech Co. (Shanghai, China); compounds 8 (C42H62O16, 99.4%) and 9 (C42H68O13, 99.4%) from Fujifilm Wako Pure Chemicals (Osaka, Japan); compounds 6 (C57H92O28, 99.9%) and 10 (
C24H32O7, 99.3%) from ChemFaces Biochemical Co. (Wuhan, China) and Chengdu
Biopurify Phytochemicals (Chengdu, China), respectively. LC–MS-grade solvents (methanol, acetonitrile, and distilled water) were purchased from ThermoFisher Scientific (Seoul, Korea) and LC–MS-grade reagent (trifluoroacetic acid; TFA, ≥99.0%) from Merck (Seoul, Korea).
