Inter-laboratory validation of an ISO test method for measuring enzyme activities in soil samples using colorimetric substrates
2022
Cheviron, Nathalie | Grondin, Virginie | Marrauld, Christelle | Poiroux, Françoise | Bertrand, Isabelle | Abadie, Josiane | Pandard, Pascal | Riah-Anglet, Wassila | Dubois, Caroline | Malý, Stanislav | Marques, Catarina | Asenjo, Inmaculada Valverde | Alonso, Alejandro | Díaz, Domingo Marquina | Mougin, Christian | Ecologie fonctionnelle et écotoxicologie des agroécosystèmes (ECOSYS) ; AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Plateforme BIOCHEM-ENV ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Ecologie fonctionnelle et biogéochimie des sols et des agro-écosystèmes (UMR Eco&Sols) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut de Recherche pour le Développement (IRD)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Montpellier ; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro) | Institut National de l'Environnement Industriel et des Risques (INERIS) | Agro-écologie, Hydrogéochimie, Milieux et Ressources (AGHYLE) ; UniLaSalle | UniLaSalle | Central Institute for Supervising and Testing in Agriculture [Brno] (UKZUZ) | Centre for Environmental and Marine Studies [Aveiro] (CESAM) ; Universidade de Aveiro = University of Aveiro | Universidad Complutense de Madrid = Complutense University of Madrid [Madrid] (UCM) | ADEME (Agence de l'Environnement et de la Maitrise de l'Energie, grant No1960C0096) in France | national funds (OE), through FCT -Fundacao para a Ciencia e a Tecnologia -I.P., in the scope of the framework contract foreseen in the numbers 4, 5, and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017 of July 19
International audience
Show more [+] Less [-]English. The evaluation of soil quality requires the use of robust methods to assess biologically based indicators. Among them, enzyme activities are used for several decades, but there is a clear need to update their measurement methods for routine use, in combining feasibility, accuracy, and reliability. To this end, the platform Biochem-Env optimized a miniaturized method to measure enzyme activities in soils using colorimetric substrates in micro-well plates. The standardization of the method was carried out within the framework of ISO/TC 190/SC 4/WG 4 "Soil quality - Biological methods" workgroup, recommending an inter-laboratory evaluation for the publication of a full ISO standard. That evaluation, managed by the platform, was based on the measurement, in six soils of contrasted physicochemical properties, of the ten soil enzyme activities described in the standard. Eight laboratories were involved in the validation study. Only 2.7% of outliers were identified from the analyses of the whole dataset. The repeatability and reproducibility of the method were determined by computing, respectively, the intra-laboratory (CVr,) and inter-laboratory (CVR) coefficients of variation for each soil and enzyme. The mean CVr ranged from 4.5% (unbuffered phosphatase) to 9.9% (alpha-glucosidase), illustrating a reduced variability of enzyme activities within laboratories. The mean CVR ranged from 13.8% (alkaline phosphatase) to 30.9% (unbuffered phosphatase). Despite this large CVR noticed for unbuffered phosphatase, the method was repeatable, reproducible, and sensitive. It also proved to be applicable for measuring enzyme activities in different types of soils. These results have been found successful by ISO/TC 190/SC4 and resulted in the publication of ISO 20130:2018 standard.
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