Robustness of Sample Rankings by Fluorimetric Enzyme Activities Against Varied Protocol Conditions in Coarse-Textured Soils

Soil enzyme activities are sensitive biochemical indicators that could benefit soil health assessments, especially in coarse-textured soils. Current protocols are inconsistent for fluorimetric assays and an optimized assay would facilitate comparisons of activities across climates and soils. A factorial experiment was conducted to evaluate how assay conditions affect the activity of three enzymes (acid phosphatase, β-glucosidase, and N-acetyl-β-glucosaminidase) across seven Florida mineral soils (>89% sand) by crossing two temperatures, four pH values, and two reaction termination reagents. Results between microplate fluorimetry and benchtop colorimetry and between air-dried and frozen (−80 °C) soils were also compared. For these soils, a pH of 4.5 with sodium hydroxide termination and a temperature of 25 °C were deemed “optimal” for maximizing activities and maintaining consistent trends. Activities measured with benchtop colorimetry and microplate fluorimetry were related for each enzyme (R2 range: 0.58–0.83) and activities from air-dried soils were 50–90% of those from frozen soils (R2 range: 0.75–0.91). Enzyme activities were positively correlated with other indicators (total C, nutrients), supporting their use in soil health assessments. As the rankings of soil samples by highest enzyme activities were similar regardless of protocol variations, this suggests that inherent soil properties were the dominant drivers of enzymatic activity.