ABSTRACT The main purpose of this study was to improve the accuracy and reliability of nutrient analysis on soilless substrates by comparing two different saturated medium extract (SME) methods, using either water (SMEW) or DTPA (diethylenetriamine-pentaacetic acid, SMED) as extractants. A series of analyses were performed to compare ion levels quantified from either SMEW or SMED. Substrate pH, electrical conductivity (EC) and nutrient data were analyzed from a survey of substrate components (coconut coir, peat, bark, perlite, vermiculite), and blended greenhouse propagation substrates using the SMEW and SMED methods. In addition, a series of SMEW and SMED extracted samples were sent to three commercial soil testing laboratories for nutrient analysis. The objectives of these analyses were to (1) validate that published relationships between SMEW and SMED methods were consistent with correlation curves for pH, EC and nutrient levels across a wide range of soilless substrates, (2) compare statistical variability of results from SMEW and SMED methods and (3) compare variability in results between different laboratories. The results validated that previously published standards for interpreting SMEW extractions can be used for SMED method for macronutrients and EC due to an approximately 1:1 relationship for test results between extraction methods. Up to 2 orders of magnitude higher levels of micronutrients were extracted for iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) using SMED compared with SMEW, with low correlation between extractants. Concentrations of ions from SMED extractions were less variable than SMEW for micronutrient analysis but were comparable for macronutrient analysis. Results from this study were combined with published research to provide suggested ranges using SMED for soilless substrates.