Identification and Characterization of Terpenoid Lactones and Flavonoids from Ethanolic Extract ofAndrographis Paniculata(Burm.f.) Nees Using Liquid Chromatography/Tandem Mass Spectrometry

Abstract

Andrographis paniculata is traditionally used for the prevention and treatment of infectious diseases due to its bioactive terpenoid lactones and flavonoids. So, there is an urgent need to develop a liquid chromatography with tandem mass spectrometry method for identification and characterization of terpenoid lactones and flavonoids. The chromatographic separation of terpenoid lactones and flavonoids from crude extract was carried on Thermo Betasil C8 column (250 mm × 4.5 mm, 5 μm) using 0.1% formic acid in water and 0.1% formic acid in acetonitrile as a mobile phase by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry. Terpenoid lactones showed characteristic consecutive losses of H 2 O, HCHO and CO whereas flavonoids gave successive losses of CH 3 , CH 4 , CO and retro Diels Alder fragment ions. In total 68 compounds including 28 terpenoid lactones and 40 flavonoids were tentatively identified and characterized on the basis of their chromatographic and mass spectrometric features (molecular formula, exact mass measurements and tandem mass spectrometry). Andrographolide and wogonin were unambiguously identified by comparison with their authentic standards. The present method can be used for identification and characterization of terpenoid lactones and flavonoids. K E Y W O R D SAndrographis paniculata, flavonoids, tandem mass spectrometry, terpenoid lactones 762

Secondary metabolite profiling and characterization of diterpenes and flavones from the methanolic extract of Andrographis paniculata using HPLC-LC-MS/MS

“…One of the most important techniques is the combination of liquid chromatography and mass spectrometry that symbolizes a robust, sensitive, quick, and reproducible analytical method for the identification and characterization of secondary metabolites present in bioactive extracts. Several studies have reported the presence of diverse secondary metabolites from leaves, root, stem, or from whole plants of A. paniculata extracts, such as andrographolide (major compound), neoandrographolide, 14-deoxy-11, 12-didehydroandrographide and isoandrographolide [8,25,26], 14-deoxy-17-hydroxyandrographolide, 12S-hydroxyandrographolide, andrographatoside, 14-deoxy-11, 12-didehydroandrographolide, andrographolide, andrographic acid, 14-deoxyandrographolide, andrographolide derivatives, andrograpanin, bisandrographolide, skullcapflavone I, 5-Hydroxy-7,8,2′,5′-tetra-methoxyflavone 5-O-glucoside, and 7-O-methylwogon [27,28].…”

“…The positive ion mode was much more efficient for the analysis of these kinds of compounds. Kumar et al (2018) also reported fragmentation of Andrographolide from the methanol extract of aerial parts of A. paniculata, but the fragmentation pattern differed at m/z 303 and 275 due to loss of CO in reverse Diels-Alder (RDA) [27]. The biological function of a specific diterpenoid cannot be representative for the whole class of molecules due to their diverse nature and often restricted distribution [29].…”

Diterpenoids as potential anti-malarial compounds from Andrographis paniculata

“…Natural products with medicinal activities can be directly isolated from plants and used in therapies or can act as a basic structure for the development of several derivatives for the development of more active and less toxic analogs [45]. Several diterpenoids including 14-deoxy-11, 12-di-dehydroandrographide, andrographolide, neoandrographolide, and isoandrographolide have been earlier reported from A. paniculata [12,[46][47][48]. Kumar et al (2018) have reported Skullcapflavone I, 5-Hydroxy-7,8,2′,5′-tetra-Methoxyflavone 5-O-glucoside and 7-O-Methylwogonin to be present in the methanolic extract of aerial parts of A. paniculata.…”

“…Several diterpenoids including 14-deoxy-11, 12-di-dehydroandrographide, andrographolide, neoandrographolide, and isoandrographolide have been earlier reported from A. paniculata [12,[46][47][48]. Kumar et al (2018) have reported Skullcapflavone I, 5-Hydroxy-7,8,2′,5′-tetra-Methoxyflavone 5-O-glucoside and 7-O-Methylwogonin to be present in the methanolic extract of aerial parts of A. paniculata.…”

Simultaneous Quantification of Major Bio‐Active Diterpenoid Lactones and Flavonoids in Andrographis paniculata (Burm. F.) Nees: LC‐ESI‐MS/MS Method Validation and Uncertainty Determination

“…The combination of LC‐MS/MS with positive electrospray ionization (ESI) and multiple reactions monitoring (MRM) has proven to be an effective method for co‐analyzing multiple complex compounds in medicinal herbs [18–20] . Various chromatographic methods for the quantification of single or multiple diterpenoid lactones in A. paniculata had been developed [21–22] . However, these methods lack the ability to identify isomers and have low sensitivity and selectivity.…”