See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/275018881 Phytochemical analysis of isoflavonoids using liquid chromatography coupled with tandem mass spectrometry Article in Phytochemistry Reviews · March 2015 CITATIONS READS 0 587 4 authors, including: Kanumuri Siva Rama Raju Isha Taneja University of Florida Central Drug Research Institute 72 PUBLICATIONS 835 CITATIONS 50 PUBLICATIONS 576 CITATIONS SEE PROFILE SEE PROFILE Muhammad Wahajuddin Central Drug Research Institute 130 PUBLICATIONS 2,243 CITATIONS SEE PROFILE Some of the authors of this publication are also working on these related projects: Targeting GPCRs for the development novel therapeutics for obesity View project Identification & evaluation of candidate drug target from natural plant resources, chemical synthetic routes, chemical modification for their anti - diabetic. View project All content following this page was uploaded by Kanumuri Siva Rama Raju on 17 April 2015. The user has requested enhancement of the downloaded file. Phytochem Rev DOI 10.1007/s11101-015-9400-x Phytochemical analysis of isoflavonoids using liquid chromatography coupled with tandem mass spectrometry Kanumuri Siva Rama Raju • Naveen Kadian • Isha Taneja • M. Wahajuddin Received: 31 December 2014 / Accepted: 12 March 2015 Ó Springer Science+Business Media Dordrecht 2015 Abstract Isoflavonoids are the biologically active made it possible to analyse and characterize several secondary metabolites of plants that are being used for constituents and their metabolites in a single run along several health promoting and restoring effects medi- with high selectivity and sensitivity. In this review, we ated through different pathways. Isoflavonoids are have summarised the application of LC–MS/MS for structurally similar to estrogens due to which also the identification and quantification of isoflavonoids known as phytoestrogens and have shown potent reported for various plant extracts and food products estrogenic and anti-estrogenic activity. Association along with their general extraction procedures and with large biological activity lead to the need of rapid, factors affecting extraction providing a view towards sensitive and precise quantitation of different iso- the conditions used for their analysis. The most flavonoids in different plant extracts, food materials suitable and widely acceptable extraction solvent and biological matrices as the biological activities system for the isoflavonoids is methanol or ethanol mainly depends on the quantities and nature of in combination with water ranging from 40 to 60 % isoflavonoids present in them. The characterisation, organic solvent based on the type of tissues and the standardisation and quantification of herbal extracts or isoflavonoids to be extracted by different extraction food products require techniques that are highly techniques. ESI ionization with Q-TOF MS was the selective, sensitive and also provide mass measure- most useful detection system for the characterisation ment precisions and structural information. Liquid and quantification of the diverse isoflavonoids with chromatography with tandem mass spectroscopy has molecular insights. Keywords LC–MS/MS Á Isoflavonoids Á Kanumuri Siva Rama Raju, Naveen Kadian and Isha Taneja Phytoestrogens Á Extraction Á Polyphenols have contributed equally to this work. CDRI Communication No. 8948. Abbreviations APCI Atmospheric pressure chemical K. S. R. Raju Á N. Kadian Á I. Taneja Á M. Wahajuddin Pharmacokinetics and Metabolism Division, CSIR- ionization Central Drug Research Institute, Lucknow, India CHS Chalcone synthase CID Collision induced dissociation & K. S. R. Raju Á I. Taneja Á M. Wahajuddin ( ) DAD Diode array detector Academy of Scientific and Innovative Research, New Delhi, India DHB 2,5-Dihydroxybenzoic acid e-mail: [email protected]; [email protected] ECD Electron capture dissociation 123 Phytochem Rev ESI Electro spray ionization Lissin and Cooke 2000). Along with their estrogen EST Expressed sequence tag receptor modulatory effects, other pharmacological FWHM Full peak width at one-half maximum activities such as antifungal, antibacterial, antioxidant, HPLC High performance liquid anti-inflammatory, anti-diabetic, anticancer, cardio- chromatography vascular diseases, menopause, osteoporosis and anti- HPLC– High performance liquid obesity activity have been reported for various MS chromatography coupled with mass isoflavonoids (Barnes 1998; Hsu et al. 2000; Davis spectrometer et al. 1998; Hillman et al. 2001; Davis et al. 2000; Hsu IFS Isoflavone synthase et al. 1999; Kenny and Prestwood 2000; Anthony ILUAE Ionic liquid based ultrasonic assisted 2000; Hsu et al. 2000; Van der Schouw et al. 2000; extraction Kenny and Prestwood 2000; Rostagno et al. 2002). IT Ion trap Along with the above mentioned pharmacological LC Liquid chromatography activities, various anticancer mechanisms (i.e. asso- MALDI Matrix-assisted laser desorption ciation with lower incidences of hormonally dependent MS Mass spectroscopy cancers) were detected by in vitro experiments, using MS/MS Tandem mass spectroscopy cell cultures, and immune-suppressed mice carrying NMR Nuclear magnetic resonance xenotransplants of human cancer cells (Klejdus et al. spectroscopy 2005a). They are believed to have some beneficial PDA Photodiode array effects in humans upon epidemiological studies indi- PLE Pressurized liquid extraction cating that the Asian people are less prone to incidence Q-TOF Quardrupole time-of flight mass of carcinoma of the breast, prostate, large intestine, and spectrometer cardiovascular diseases compared to western people as rDA retro-Diels–Alder the Asian diets contain higher amounts of the RP-HPLC Reverse-phase liquid chromatography isoflavonoids (Adlercreutz et al. 2000). But there are SFE Supercritical fluid extraction no experimental evidences to prove the same till now. SPE Solid phase extraction Isoflavonoids occur predominantly in their precursor SPME Solid-phase micro extraction forms such as acetate, malonyl and glycoside e.g. SRM Selected reaction monitoring daidzin, the precursor of diadzein (the glycosidic form TLC Thin layer chromatography that contains a carbohydrate moiety) (Sharma and UAE Ultrasound assisted extraction Ramawat 2013). UHPLC Ultra high performance liquid The basic pathways for the synthesis of core chromatography isoflavonoid skeletons have been established both UV Ultraviolet–Visible spectroscopy enzymatically and genetically. Many flavonoids and isoflavonoids are derived from the flavonoid biosyn- thesis pathway using liquiritigenin or naringenin as precursors (Database; Ralston et al. 2005). The other Introduction precursors of flavonoid biosynthesis include shikimic acid, phenylalanine, cinnamic acid, and p-coumaric Isoflavonoids are heterocyclic phenolic secondary acid. Shikimic acid acts as an intermediate in the metabolites present in plants which are biogenetically biosynthesis of aromatic acid (Sharma and Ramawat related to flavonoids but constitute a distinctly separate 2013). The entry point enzymes are the polyketide class containing rearranged C15 skeleton and may be synthase, chalcone synthase (CHS) and isoflavone regarded as derivatives of 3-phenylchroman while synthase (IFS), more correctly termed as 2-hy- flavonoids are 2-phenylchromans (Cheng et al. 1997). droxyisoflavanone synthase, a cytochrome P450 that Most of the isoflavonoids structurally resemble en- catalyzes the aryl migration reaction that converts a dogenous 17b-estradiol and has the ability to modulate 2-phenylchroman to a 3-phenylchroman (Database). the estrogen receptors, showing both estrogenic and The structural diversity of isoflavonoids is derived by anti-estrogenic effects, due to which they are com- substitution of these basic carbon skeletons through monly known as phytoestrogens (Casanova et al. 1999; further hydroxylation, glycosylation, methylation, 123 Phytochem Rev acylation and prenylation as well as, in the case of procedure according to the type of plant tissue from proanthocynadins (also known as condensed tannins) which they are to be sought i.e. heartwood, seeds, and phalobaphenes, by polymerization (Dixon and leaves, rhizomes, roots etc. The need for sample Pasinetti 2010; Dixon and Steele 1999). The enzymes preparation depends strongly on the sample type and that catalyses the substitution reactions are often the analysis techniques to be employed (Bajer et al. encoded by large gene families, which can be recog- 2007). Extraction technique, extraction solvent, pH of nized in expressed sequence tag (EST) and genome the solvent, extraction time, temperature, sample to data sets through family-specific conserved sequence solvent ratio, sample characteristics are the defining motifs (Dixon and Pasinetti 2010). In comparison to factors that affect extraction efficiency, isoflavonoids the flavonoids (2-phenylchromans), which are found profile and recovery (Rostagno et al. 2010). The main widely in higher plants, the isoflavonoids appear in a objective of sample treatment is to eliminate the limited occurrence of taxon, probably owing to the undesired matrix components and the enrichment of limited occurrence of IFS enzyme, being confined samples. The extraction procedures must allow quan- essentially to the subfamily Papilionoideae (Lo- titative recovery of isoflavonoids in their native form, toideae) of the leguminosae (Klejdus et al. 2005a; whilst avoiding any chemical modification or degra- Veitch 2007, 2009). There are however, occasional dation