Diatoms are widely distributed in many temperate areas and some species frequently form extensive blooms in spring. Hence, monitoring the variations of specific genera or species of diatoms is necessary for studying phytoplankton population dynamics in marine ecosystems. To test whether pigment ratios can be used to identify diatoms at a below-class taxonomic level, we analyzed 14 species/strains of diatoms isolated from Chinese seas using high performance liquid chromatography (HPLC). We normalized all pigment concentrations to total chlorophyll a to calculate the ratios of pigment to chlorophyll a, and calculated the ratios between accessory pigments (or pigment sums). Cluster analysis indicated that these diatoms could be classified into four clusters in terms of three accessory pigment ratios: chlorophyll c2: chlorophyll Cl, fucoxanthin:total chlorophyll c and diadinoxanthin:diatoxanthin. The classification results matched well with those of biological taxonomy. To test the stability of the classification, pigment data from one species, cultured under different light intensities, and five new species/strains were calculated and used for discriminant analysis. The results show that the classification of diatom species using pigment ratio suites was stable for the variations of pigment ratios of species cultured in different light intensities. The introduction of new species, however, may confuse the classification within the current scheme. Classification of marine diatoms using pigment ratio suites is potentially valuable for the fine chemotaxonomy of phytoplankton at taxonomic levels below class and would advance studies on phytoplankton population dynamics and marine ecology.