A miniaturised method to quantify microbial mineralisation of C-labelled organic compounds in small soil samples

A miniaturised method developed to measure the mineralisation of ¹³C-labelled organic compounds in small soil samples is presented. Soil samples (<0.5 g) were placed in wells of microtiter plates with CO₂ traps (NaOH-soaked glass microfiber filters) and amended with ¹³C-labelled substrate. The microtiter plate was covered with a seal and placed in a microplate clamp system to ensure that each well was airtight. After incubation, the CO₂ traps were transferred to tightly sealed glass phials under CO₂-free atmosphere and the ¹³C-labelled CO₂ was released by addition of H₃PO₄. The CO₂ was measured by micro-GC and its isotopic signature was determined using a GC-IRMS. The qualitative and quantitative efficiency of the microplate system was demonstrated by comparison with direct measurement of CO₂ in the headspace of phials in which similarly treated soil samples had been incubated. The two methods showed similar mineralisation rates for added ¹³C-substrates but the apparent mineralisation of soil organic matter was greater with the microtiter plate method. The microplate system presented here is suitable for studying the mineralisation of different kinds of ¹³C-labelled substrates in small soil samples and allows analysis of functional and molecular characteristics on the same micro-samples.