Because routine analytical methods cannot differentiate D- from L-arabinitol, a combined microbiological and gas chromatographic method was developed to study the stereoisomeric configuration of the arabinitol in humans and rats with invasive candidiasis. D-Arabinitol was defined as the difference between arabinitol concentrations measured with and without incubation with 5.0 X 10(5) blastospores of Candida tropicalis strain CT 12 at 37 C for 24 hr. The yeast consumed at least 95% of the D-arabinitol and none of the L-arabinitol added to normal serum and urine. D-Arabinitol as a fraction of D,L-arabinitol was 0.43 +/- 0.15 (mean +/- SD) in the urine of 10 normal humans, 0.82 +/- 0.12 in the serum or urine of five patients with cancer and invasive candidiasis (P less than .001), and 1.0 in the kidneys of rats with candidiasis. Because most or all of the excess arabinitol in body fluids or tissues in candidiasis was the D isomer, which is produced by fungal metabolism, stereospecific quantitation of arabinitol should improve the sensitivity of this approach to diagnosis of candidiasis.