title: Immunoaffinity Knockout of Saponin Glycosides from Asparagus racemosus to Assess Anti-lipid Peroxidation.
abstract: BACKGROUND
  Asparagus racemosus Willd (Asparagaceae family), known as Shatavari, is important in Ayurveda and traditional Thai medicines. The saponin glycosides, shatavarin I and IV are major constituents in its roots and may be responsible for their actions including protection against lipid peroxidation and carcinogenesis.
  
  
  OBJECTIVE
  To develop an immunoaffinity column for isolating compounds with structures related to shatavarin IV from crude extracts of A. racemosus root.
  
  
  METHODS
  The monoclonal antibody recognising shatavarin IV (mAbShavIV) was coupled to an Affi-Gel Hz gel to isolate compounds with structures related to shatavarin IV from the other components of crude extracts of A. racemosus root. The saponin glycosides in each fraction were analysed by mAbShavIV ELISA and LC-MS/MS.
  
  
  RESULTS
  The pooled wash-through fractions contained 3% of loaded mAbShavIV reactive saponin glycosides, while eluted fractions released ~ 90% of shatavarin saponin glycosides in a single step. Using thiobarbiturate (TBARs) to measure lipid-peroxidation, the extract, and the pooled wash-through fractions showed moderate protection against Cu+-induced oxidation of human low density lipoprotein (LDL) (IC5011.3 ± 1.4 and 12.6 ± 0.9 μg/mL, respectively). In contrast, the saponin glycosides eluted from the mAbShavIV-column had weaker protectant (IC5029.7 ± 1.8 μg/mL) suggesting that A. racemosus shatavarins do not inhibit carcinogenesis through preventing lipid peroxidation.
  
  
  CONCLUSIONS
  The strategy described here demonstrates its utility for isolating a group of related compounds from the rest of the extract with selectivity and recovery rate. Pharmacological efficacy and synergistic effects of the components obtained can be further investigated. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Bioscreening Unit, Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Centre of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok, 65000, Thailand.
authors.forename: Churanya
authors.initials: C
authors.lastname: Onlom
authors.affiliation: Faculty of Public Health, Naresuan University, Phitsanulok, 65000, Thailand.
authors.forename: Watoo
authors.initials: W
authors.lastname: Phrompittayarat
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
authors.affiliation: Cosmetic and Natural Product Research Centre, Faculty of Pharmaceutical Sciences, Naresuan University, Phitsanulok, 65000, Thailand.
authors.forename: Neti
authors.initials: N
authors.lastname: Waranuch
authors.affiliation: Bioscreening Unit, Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Centre of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok, 65000, Thailand.
authors.forename: Kornkanok
authors.identifier: http://orcid.org/0000-0002-4415-8489
authors.initials: K
authors.lastname: Ingkaninan
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D027761:Asparagus Plant / Q000737:chemistry*; D002846:Chromatography, Affinity; D006027:Glycosides / Q000302:isolation & purification*; D006801:Humans; D015227:Lipid Peroxidation; D008077:Lipoproteins, LDL / Q000737:chemistry; D010936:Plant Extracts / Q000032:analysis*; D012503:Saponins / Q000302:isolation & purification*; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D006027:Glycosides; D008077:Lipoproteins, LDL; D010936:Plant Extracts; D012503:Saponins
keywords: Asparagus racemosus; anti-lipid peroxidation; immunoaffinity column; monoclonal antibody; saponin glycoside