This study presents a new HPLC method using evaporative light scattering detection for the simultaneous determination of five major iridoid glucosides, namely 7-epi-loganin, sweroside, loganin, 7-epi-vogeloside, and secoxyloganin in Flos Lonicerae, an important traditional Chinese medicinal herb. The optimal conditions of separation and detection were achieved on a C-18 analytical column with an isocratic mobile phase consisting of methanol-water (30:70, v/v) containing 0.5% acetic acid at the flow-rate of 1.0 ml/min, temperature for the detector drift tube set at 90 degreesC and the nitrogen flow-rate of 2.6 l/nim. The limit of detection (S/N = 3) is less than 35.1 mug/ml and the limit of quantification (S/N= 10) is less than 140.1 mug/ml. All calibration curves show good linear regression (r(2) > 0.996) within test ranges. This method provides good reproducibility for the quantification of the major iridoid glucosides in four Lonicera species with overall intra- and inter-day variation of less than 5% and 9%, respectively. The assay was successfully applied to quantify the main iridoid glucosides in the herb and to identify the botanical origin of Flos Lonicerae. (C) 2003 Elsevier B.V. All rights reserved.