Overground parts of Centaurium spicatum (L.) FRITSCH (Gentianaceae) from two different habitats have been investigated on their content of Secoiridoid glucosides. Sweroside (I), Swertiamarin (II) and gentiopicrin (III) (= gentiopicroside) have been identified. In plant material of C. spicatum from both habitats sweroside is the major compound. In plant material from one habitat gentiopicrin was found in almost equal amounts as sweroside. Swertiamarin is only present as a minor component. For the isolation of sweroside and gentiopicrin a preparative column chromatographic method using silica gel as adsorbent is described. The chromatographic analysis of these Secoiridoid glucosides has been carried out by means of newly elaborated TLC and HPLC separation systems. The composition of sweroside (I), swertiamarin (II) and gentiopicrin (III) in C. spicatum is compared with that occurring in C. pulchellum (SW.) DRUCE and C. erytkraea RAFN. (= C. minus MOENCH). On the basis of chemical derivation, chromatographic (TLC and HPLC) and spectroscopical (IR, Mass) data it is shown that sweroside is very likely identical with the compound known as „kantaurin“ in literature. 1 Part 1: v. d. SLUIS, W. G. and R. P. LABADIE, Pharm. Weekblad 113, 21 (1978) Part of these studies were presented at the 25 th annual meeting of the Gesellschaft für Arzneipflanzenforschung in Zürich, Switzerland, September 12-16, 1977 (see Planta Medica 32A, 52 (1977)