title: Reverse-phase HPLC method for the quantification of two antihyperglycemic glycolipids in Oplismenus burmannii.
abstract: Glycolipids and sphingolipids are well known for their diverse biological activities like anticancer, anti-inflammatory, antistress, anti-HIV, hepatoprotective and antimicrobial. The present study deals with the activity-guided isolation and characterization of two antihyperglycemic glycolipids, (2S)-1,2-di-O-octadecanoyl-3-O-[α-d-galctopyranosyl-(1'' → 6')-O-β-d-galactopyranosyl] glycerol (1) and 1-O-β-d-glucopyranosyl-(2S,3S,4R,8E)-2-[(2R)-2-hydroxy-tetracosanoylamino]-2,3,4-octadecanetriol-8-ene (2) from Oplismenus burmannii and the development of a simple and validated reverse-phase HPLC analytical method for their quantification in the methanolic extracts of O. burmannii. The marker compounds 1 and 2 were isolated from the methanolic extract of O. burmannii and characterized on the basis of their spectroscopic data. Their antihyperglycemic potential was evaluated by determining their glucose uptake-stimulating potential in L6-GLUT4myc myotube cells. Finally, these analytes were separated on a Waters Spherisorb ODS 2 column with a binary gradient of methanol and water at a constant flow rate of 0.8 mL/min and detected using a photodiode array detector at 230 nm. The calibration curve was linear (r(2)  > 0.999) over 1.2 orders of magnitude with acceptable accuracy, reproducibility and recovery (98.16-100.50%). The limits of detection and quantification for 1 and 2 were 1.36, 4.11 and 1.11, 3.35 µg/mL respectively. The method is simple, accurate, precise and selective and may be routinely used for the quality control analysis of whole plant extract of O. burmannii for these two glycolipids.
authors.affiliation: Medicinal Chemistry Department, CSIR - Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow, 226 015, India.
authors.forename: Surjeet
authors.initials: S
authors.lastname: Verma
authors.affiliation: Analytical Chemistry Department, CSIR - Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow, 226 015, India.
authors.forename: Ram Kishor
authors.initials: RK
authors.lastname: Verma
authors.affiliation: Medicinal Chemistry Department, CSIR - Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow, 226 015, India.
authors.forename: Dibyaranjan
authors.initials: D
authors.lastname: Sahoo
authors.affiliation: Medicinal Chemistry Department, CSIR - Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow, 226 015, India.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Srivastava
mesh_terms: D066241:Carbon-13 Magnetic Resonance Spectroscopy; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006017:Glycolipids / Q000032:analysis*; D007004:Hypoglycemic Agents / Q000032:analysis*; D057230:Limit of Detection; D006109:Poaceae / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D006017:Glycolipids; D007004:Hypoglycemic Agents
keywords: Oplismenus burmannii; antihyperglycemic; cerebroside; glycoglycerolipid; method validation; quantitative determination