In our recent studies on bioactive saponins, two Chinese medicinal plants: Mussaenda pubescens Ait.f (Rubiaceae) and Clematis chinensis Osbeck (Ranunculaceae), were investigated. Of the two medicinal plants, M. pubescens is a Chinese folk medicine which has been used as a diuretic, antiphlogistic, diaphoretic and antipyretic agent, and has also been used to detoxify mushroom poisons and to terminate early pregnancy. Clematis chinensis is a Chinese traditional medicine which has been used as an analgesic, diuretic, antitumor, antiinflammatory and insecticidal agent for ages. As a result of our studies, eighteen saponins were identified from M. pubescens, among which seventeen were new compounds, while eleven saponins were isolated from C. chinensis, three of which were new compounds. In the course of our structural studies, mass fragment analysis in FAB-mass spectra and 1H- and 13C-NMR spectra were used to determine the structures of the sapogenin and oligosaccharide moieties. In those more complicated and minor saponins, various 2D-NMR experiments were carried out on 400-, 500- or 600-MHz NMR instruments, which permitted the identification of new sapogenins in glycoside form. In addition, it was also possible to assign all the proton and carbon signals of the sugar units on the basis of 1H-1H DQF COSY, TOCSY and HMQC spectra, which further permitted the establishment of linkage sites and sequences among the sugar units and aglycones by means of NOESY, ROESY and HMBC spectra. When the proton signals of sugar units are overlapped seriously in 1H-NMR spectra, peracetylation is a very helpful technique which can spread proton signals in wider range, thus simplifying their assignment by means of 2D-NMR spectra. When some of the sugar proton signals of a peracetylate are also overlapped coincidentally, alternation of deuterated solvents for measuring NMR spectra can circumvent the difficulties. Pharmacological tests indicated that mussaendoside O, the most abundant saponin from M. pubescens, can inhibit significantly the secretions of the lachrymal and salivary glands induced by galanthamine, and can also inhibit the contraction of the isolated longitudinal muscle strip from guinea pig ileum evoked by an M-Ach receptor agonist (carbachol, 10(-6) M) at concentrations of 10(-4) and 10(-5) M. From these results, the saponin should be an antagonist of the M-Ach receptor, and was presumed to be responsible for its antitoxicity activity toward some mushroom poisons of medicinal plant origin. In addition, mussaendoside O also showed immunopromotive and hemolytic activities.