Comparing Extraction Methods for Biomarker Steroid Characterisation from Soil and Slurry
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Comparing Extraction Methods for Biomarker Steroid Characterisation from Soil and Slurry Amber Manley & Adrian L. Collins & Adrian Joynes & Per-Erik Mellander & Phil Jordan
Received: 2 July 2020 / Accepted: 22 September 2020 # The Author(s) 2020
Abstract Clean water is a precious resource, and policies/programmes are implemented worldwide to protect and/or improve water quality. Faecal pollution can be a key contributor to water quality decline causing eutrophication through nutrient enrichment and pathogenic contamination. The robust sourcing of faecal pollutants is important to be able to target the appropriate sector and to engage managers. Biomarker technology has the potential for source confirmation, by using, for example the biomarker suite of steroids. Steroids have been used in the differentiation of human and animal faeces; however, there is no unequivocal extraction technique. Some of the methods used include (i) Soxhlet extraction, (ii) Bligh and Dyer (BD) extraction, and (iii) accelerated solvent extraction (ASE). The less costly and time intensive technique of ASE is particularly attractive, but a current research gap concerns further comparisons regarding ASE lipid extraction from soils/slurries compared with the more traditional Soxhlet and BD extractions. Accordingly, a randomised complete block experiment was implemented to assess differences between the three A. Manley : A. L. Collins : A. Joynes Sustainable Agriculture Sciences, Rothamsted Research, North Wyke, Okehampton EX20 2SB, UK
extraction methods, differences between the different sample types, and the interactions between these two factors. Following GC-MS, it was found that there was no significant difference between the results of the steroid extraction methods, regardless of the type of sample used, for the quantity of each steroid extracted. It was concluded that ASE could be used confidently instead of the more established steroid extraction methods, thereby delivering time and cost savings. Keywords Lipids . ASE . Bligh and Dyer . Soxhlet . Faecal pollution Abbreviations BD Bligh and Dyer ASE Accelerated solvent extraction N Nitrogen P Phosphorus IMS Industrial methylated spirit DCM Dichloromethane TLE Total lipid extract BDS Bligh and Dyer solvent PCA Principal component analysis PC Principal component CVA Canonical variate analysis
A. Manley : P. 26 mg/L)
Area (ha)
pH value
P (index)
P (mg/L)
K (index)
K (mg/L)
Mg (index)
Mg (mg/L)
0.99
6.9
3
27.4
1
108
2
97
thoroughly and rinsed in acetone before use. The custom-made 2 cm depth soil corer was washed in Virkon disinfectant, MilliQ water, and acetone prior to use. Between samplings in the field, equipment was washed with Virkon disinfectant and MilliQ water to minimise contamination. Sieves and grinders were similarly washed thoroughly with Microsol4 disinfectant, MilliQ water, and grinders also received an industrial
methylated spirit (IMS) wash both prior to, and between, sample use to avoid cross-contamination. For each block, cores were taken and loosely
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