Glomalin, a glycoprotein produced by arbuscular mycorrhizal (AM) fungi, is a major component of the humus fraction of soil organic matter. Glomalin is extracted from soil and hyphae of AM fungi by using sodium citrate at 121 degrees C in multiple 1-h cycles, but extensive extraction does not solubilize all glomalin in all soils. Efficacies of 100 mM sodium salts of citrate, borate or pyrophosphate (pH 9.0, 121 degrees C) were tested for two 1-h cycles for hyphae from four AM fungal isolates and four 1-h cycles for seven soils from four US geographic regions. Residual soil glomalin was examined by pyrophosphate extraction of soils previously extracted with citrate or borate followed by extraction of all soils after treatment with NaOH. Hyphal extracts were compared using Bradford-reactive total protein (BRTP) values, and extracts from soils were compared using BRTP, percentage C and C weight. No difference among extractants was detected for AM fungal isolates or across soils. The residual glomalin across soils for extractants contained the following percentages of the total BRTP: pyrophosphate, 14%; borate, 17%; and citrate, 22%. Comparisons among individual soils indicated that pyrophosphate extracted significantly more BRTP (10-53%) than borate or citrate in six soils and borate was equal to pyrophosphate in one soil. Extraction with borate should be compared with pyrophosphate before initiating an experiment. For routine extractions of ca. 85% of the glomalin across a variety of soils, sodium pyrophosphate appears to be equal to or better than borate and better than citrate.