Benchmarking GC and HPLC for amino sugar analyses across soils: A comprehensive evaluation

Amino sugars are key tracers of microbial necromass in soil organic carbon, yet the lack of direct methodological comparison between gas chromatography (GC) and high-performance liquid chromatography (HPLC) limits cross-study integration. Here, we provide the first systematic evaluation of both methods using 395 field samples and 1900 published observations across global ecosystems. GC and HPLC showed strong analytical agreement (R 2 > 0.92; RSD <5 %), but GC yielded more accurate measurements in C- and N-rich soils due to superior cleanup and purification, whereas HPLC offered higher throughput and operational simplicity. These results establish a quantitative benchmark for harmonizing amino-sugar datasets and highlight that method choice should align with soil matrix complexity and analytical goals. • GC and HPLC were firstly benchmarked for amino sugar analysis in 395 soil samples. • Both methods exhibited high consistency in quantifying soil microbial necromass. • GC has higher accuracy in C/N-rich soils due to better purification specificity. • HPLC provides efficient, high-throughput quantification for ecosystem-scale studies. • Global meta-analysis (n = 1900) validated this methodological framework across biomes.