title: UPLC-TOF-MS analysis of Galium spurium towards its neuroprotective and anticonvulsant activities.
abstract: BACKGROUND
  Galium species have been reported to be used against epilepsy in traditional Turkish folk medicine.
  
  
  OBJECTIVE
  The present work was undertaken to evaluate the in vivo anticonvulsant and in vitro neuroprotective effects of Galium spurium L. and to determine the major constituents by UPLC-TOF-MS.
  
  
  METHODS
  Anticonvulsant activity of the aerial parts of Galium spurium was investigated using pentylenetetrazole, picrotoxin, and maximal electroshock-induced seizure animal models. In order to evaluate the safety, neurotoxicity (Rota rod test) of the ethanol extract was also determined. In vitro neuroprotective effect of the ethanol extract of Galium spurium was assessed by acetylcholinesterase and butrylcholinesterase inhibitions. Ultra Performance Liquid Chromatography-Time of Flight Mass Spectrometer (UPLC-TOF-MS) was used to identify the major compounds in the extract.
  
  
  RESULTS
  In pentylenetetrazole-induced seizure, the ethanol extract at doses of 250 and 1000mg/kg prolonged the onset of seizures. Similarly, Galium spurium (250 and 500mg/kg) significantly delayed the onset of picrotoxin-induced seizures in mice and these doses also exhibited 12.5% and 17% protection, respectively, against picrotoxin-induced seizures. Furthermore, Galium spurium extract showed a significant protective effect against maximal electroshock-induced seizures at doses of 250 and 1000mg/kg (50% and 37.5%, respectively) and also all tested doses prolonged the onset of seizures. No motor co-ordination was observed with intraperitoneal injection of Galium spurium extract at doses of 500 and 1000mg/kg. The extract exhibited 16.2% inhibition against butrylcholinesterase at 200μg/mL concentration, whereas it did not inhibit acetylcholinesterase. Phytochemical analysis of the extract based on the MS data by UPLC-TOF-MS, ten major compounds (phenolic and triterpenic acids, flavonoids and iridoids) were determined.
  
  
  CONCLUSIONS
  The results indicate that Galium spurium may have anticonvulsant activity against picrotoxin and maximal electroshock-induced seizures in mice. Phenolic acids, flavonoids and iridoids might be responsible for anticonvulsant activity. The results offer possible beneficial effects by the plant's aerial parts and may suggest a realistic explanation for its traditonal usage in epilepsy.
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Hipodrom, Ankara, Turkey.
authors.forename: Nilüfer
authors.initials: N
authors.lastname: Orhan
authors.forename: Didem
authors.initials: D
authors.lastname: Deliorman Orhan
authors.forename: Mustafa
authors.initials: M
authors.lastname: Aslan
authors.forename: Murat
authors.initials: M
authors.lastname: Süküroğlu
authors.forename: Ilkay Erdogan
authors.initials: IE
authors.lastname: Orhan
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D000927:Anticonvulsants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D002091:Butyrylcholinesterase / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002853:Chromatography, Liquid; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004597:Electroshock; D000431:Ethanol / Q000737:chemistry; D005260:Female; D032061:Galium / Q000737:chemistry; D008297:Male; D051379:Mice; D009043:Motor Activity / Q000187:drug effects; D018696:Neuroprotective Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010433:Pentylenetetrazole; D008517:Phytotherapy; D010852:Picrotoxin; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D011930:Reaction Time / Q000187:drug effects; D012640:Seizures / Q000209:etiology / Q000517:prevention & control*; D012997:Solvents / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
chemical_list: D000110:Acetylcholinesterase; D000927:Anticonvulsants; D002091:Butyrylcholinesterase; D002800:Cholinesterase Inhibitors; D000431:Ethanol; D018696:Neuroprotective Agents; D010433:Pentylenetetrazole; D010852:Picrotoxin; D010936:Plant Extracts; D012997:Solvents