title: Analysis of phytochemical variations in dioecious Tinospora cordifolia stems using HPLC/QTOF MS/MS and UPLC/QqQLIT -MS/MS.
abstract: BACKGROUND
  The stem of dioecious Tinospora cordifolia (Menispermaceae) is a commonly used traditional Ayurvedic medicine in India having several therapeutic properties.
  
  
  OBJECTIVE
  To develop and validate LC-MS methods for the identification and simultaneous quantitation of various secondary metabolites and to study metabolomic variations in the stem of male and female plants.
  
  
  METHODS
  Ethanolic extract of stems were analysed by HPLC/ESI-QTOF-MS/MS for rapid screening of bioactive phytochemicals. High resolution MS and MS/MS in positive ESI mode were used for structural investigation of secondary metabolites. An UPLC/ESI-QqQ(LIT) -MS/MS method in MRM mode was developed and validated for the simultaneous quantitation of five bioactive alkaloids.
  
  
  RESULTS
  Identification and characterisation of 36 metabolites including alkaloids, sesquiterpenes and phytoecdysteroids were performed using LC-MS and MS/MS techniques. The bioactive alkaloids such as jatrorrhizine, magnoflorine, isocorydine, palmatine and tetrahydropalmatine were successfully quantified in male and female plants. The mean abundances of magnoflorine jatrorrhizine, and oblongine were significantly (P < 0.05) higher in male plants while mean abundances of tetrahydropalmatine, norcoclaurine, and reticuline were significantly (P < 0.05) higher in female plants.
  
  
  CONCLUSIONS
  Phytochemicals in the stem of male and female Tinospora cordifolia showed significant qualitative and quantitative variations. LC-MS and MS/MS methods can be used to differentiate between male and female plants based on their chemical profiles and quantities of the marker bioactive alkaloids. This chemical composition difference was also evident during vegetative stage when there were no male and female flowers.
authors.affiliation: Sophisticated Analytical Instrument Facility Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.|Academy of Scientific and Innovative Research, New Delhi, 110001, India.
authors.forename: Vikas
authors.initials: V
authors.lastname: Bajpai
authors.affiliation: Sophisticated Analytical Instrument Facility Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.|Academy of Scientific and Innovative Research, New Delhi, 110001, India.
authors.forename: Awantika
authors.initials: A
authors.lastname: Singh
authors.affiliation: Sophisticated Analytical Instrument Facility Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.|Academy of Scientific and Innovative Research, New Delhi, 110001, India.
authors.forename: Preeti
authors.initials: P
authors.lastname: Chandra
authors.affiliation: Biometry and Statistics Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.
authors.forename: M P S
authors.initials: MP
authors.lastname: Negi
authors.affiliation: Betelvine Laboratory, CSIR-National Botanical Research Institute, Lucknow, 226001, India.
authors.forename: Nikhil
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Sophisticated Analytical Instrument Facility Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India.|Academy of Scientific and Innovative Research, New Delhi, 110001, India.
authors.forename: Brijesh
authors.initials: B
authors.lastname: Kumar
mesh_terms: D000470:Alkaloids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004441:Ecdysterone / Q000032:analysis; D064209:Phytochemicals / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D012717:Sesquiterpenes / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D031610:Tinospora / Q000737:chemistry*
chemical_list: D000470:Alkaloids; D064209:Phytochemicals; D012717:Sesquiterpenes; D004441:Ecdysterone
keywords: Alkaloid; LC-MS; MRM; Tinospora cordifolia; simultaneous quantitation