Journal of Ethnopharmacology 102 (2005) 170–176 Bioassay-guided isolation of iridoid glucosides with antinociceptive and anti-inflammatory activities from Veronica anagallis-aquatica L. Esra K ¨ upeli a , U. Sebnem Harput b , Mehtap Varel b , Erdem Yesilada a,∗ , Iclal Saracoglu b a Gazi University, Faculty of Pharmacy, Etiler 06330 Ankara,Turkey b Hacettepe University, Faculty of Pharmacy, Sihhiye 06100 Ankara, Turkey Received 24 September 2004; received in revised form 11 May 2005; accepted 26 May 2005 Available online 12 July 2005 Abstract Extracts obtained from the herbs of various Veronica species are used as folk remedy worldwide for the treatment of various inflammatory ailments including rheumatism. In vivo anti-inflammatory and antinociceptive activities of Veronica anagallis-aquatica L. aerial parts were investigated. Methanolic extract of the plant was shown to possess significant inhibitory activity against carrageenan-induced hind paw edema model and of p-benzoquinone-induced writhings in mice. Through bioassay-guided fractionation and isolation procedures eight compounds, aquaticoside A (1), aquaticoside B (2), aquaticoside C (3), veronicoside (4), catalposide (5), verproside (6), verminoside (7) and martynoside (8) were isolated and their structures were elucidated by spectral techniques. Catapol derivative iridoid glucosides, verproside (6) and catalposide (5), were found to possess potent antinociceptive and anti-inflammatory activities, per os without inducing any apparent acute toxicity as well as gastric damage. Results of the present study supported the utilization of the plant in Turkish folk medicine. © 2005 Elsevier Ireland Ltd. All rights reserved. Keywords: Veronica anagallis-aquatica; Scrophulariaceae; Antinociceptive; Anti-inflammatory; Iridoid glucosides; Catalpol derivatives; Carrageenan-induced paw edema; p-Benzoquinone-induced writhings 1. Introduction Several Veronica species are reported to possess applica- tion in traditional medicines worldwide for the treatment of a wide range of disorders; in respiratory diseases against cough or as expectorant, as antiscorbutic, as diuretics and for wound healing (Baytop, 1984; Harput et al., 2002). In Chinese tra- ditional medicine, Veronica anagallis-aquatica L. is used for the treatment of influenza, hemoptysis, laryngopharyngitis and hernia (Su et al., 1999). During our field expeditions on Turkish folk medicine, we have recorded that aerial parts of Veronica anagallis-aquatica is boiled in milk to obtain poul- tice and then is applied to abdomen for abdominal pain or its warm aqueous extract without removing the boiled herbs is used as a bath remedy to alleviate rheumatic pain in northwest Anatolia (Fujita et al., 1995). ∗ Corresponding author. Tel.: +90 312 2154468; fax: +90 312 223 5018. E-mail address: yesilada@gazi.edu.tr (E. Yesilada). Previously, in vitro anti-inflammatory activity of five Veronica species (Veronica cymbalaria, Veronica hederifo- lia, Veronica pectinata var. glandulosa, Veronica persica and Veronica polita) were investigated through evalua- tion the inhibitory effects on nitric oxide (NO) pro- duction in lipopolysaccharide-stimulated mouse peritoneal macrophages as well as cytotoxic activity against KB epider- moid carcinoma and B16 melanoma (Harput et al., 2002). MeOH extracts of five plant materials were found to pos- sess inhibitory effects on NO synthesis in varying degrees. MeOH extracts were further partitioned between water and chloroform, and water fractions showed the activity without inducing any cytotoxicity, while chloroform fractions were cytotoxic dose-dependently. Moreover, water fractions were found to possess remarkable effect on 2,2-diphenyl-1-picryl- hydrazyl (DPPH), suggesting that the inhibitory effect on NO production might be due to their radical scavenging activity. Veronica species have been known to be rich in iri- doid glucosides. Mainly aucubin, catalpol, benzoic and 0378-8741/$ – see front matter © 2005 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2005.05.042