title: Spermicidal activity of bivittoside D from Bohadschia vitiensis.
abstract: BACKGROUND
  A recent revelation about increased susceptibility to HIV by use of nonoxynol-9 (N-9) has called for identification of novel molecules with potent sperm-attenuating activity and lower side-effect profile, as suitable alternatives. The present study was designed to investigate spermicidal activity in Bohadschia vitiensis whole-body extracts followed by isolation and characterization of bioactive molecule.
  
  
  METHODS
  Bohadschia vitiensis (Semper) was collected from the Southern Andaman coast of India. Freshly collected marine animals were extracted with methanol. A portion of the crude extract was fractionated into four fractions by macerating with hexane, chloroform, and n-butanol successively. All fractions were evaluated for spermicidal activity. Because maximum activity was localized in the n-butanol soluble fraction, it was chromatographed over a silica gel column, and elution with chloroform-methanol-water (35:10:2, v/v) yielded the major compound bivittoside D (400 mg). Bivittoside D [molecular weight (MW) 1426] is a lanostane triterpenoid with six monosaccharide units. The structure of the compound was established on the basis of physicochemical data, acid hydrolysis of saponin, identification of sugar units and aglycon, melting point, and by comparison with data reported in the literature.
  
  
  RESULTS
  The aqueous methanol extract of the Bohadschia vitiensis caused 100% mortality of human sperm at 0.01% concentration in vitro, whereas N-9 (reference control) exhibited an equivalent activity at 0.05%. On further fractionation, activity was localized in n-butanol soluble fraction from which the major compound purified was a lanostane triterpenoid called bivittoside D. Bivittoside D was found to be a more potent spermicide (approximately 2.3 times) than N-9 and killed 100% human sperm at the concentration of 350 muM in approximately 20 sec in vitro. Supravital staining and hypoosmotic swelling test revealed sperm membrane permeabilization by bivittoside D as the major mode of spermicidal action. However, bivittoside D was much safer than N-9 towards normal vaginal flora (Lactobacillus) in vitro, although it affected the viability of HeLa cells like other surfactants.
  
  
  CONCLUSIONS
  Bivittoside D from B. vitiensis can adequately replace N-9 in vaginal contraceptives to make them more vaginally safe and ecofriendly.
authors.affiliation: Medicinal and Process Chemistry, Central Drug Research Institute, Lucknow, India. vijlakshmius@yahoo.com
authors.forename: Vijai
authors.initials: V
authors.lastname: Lakshmi
authors.forename: Ajet
authors.initials: A
authors.lastname: Saxena
authors.forename: Sunil Kumar
authors.initials: SK
authors.lastname: Mishra
authors.forename: Ram
authors.initials: R
authors.lastname: Raghubir
authors.forename: Mahendra N
authors.initials: MN
authors.lastname: Srivastava
authors.forename: Rajeev K
authors.initials: RK
authors.lastname: Jain
authors.forename: Jagdamba P
authors.initials: JP
authors.lastname: Maikhuri
authors.forename: Gopal
authors.initials: G
authors.lastname: Gupta