The carbon and nitrogen concentration of lichen samples were analysed by an elemental analyser (EA 1108, Carlo-Erba-Milano, Italy) with an analytical precision of 0.1%. The stable C isotopic ratio was measured with an isotope ratio mass spectrometer (CONFLO interface, Thermo, MAT Bermen, Germany) operating in continuous flow mode after the combustion of the samples in an elemental analyser (EA 1108, Carlo-Erba-Milano, Italy). Samples were weighted by using a high precision Ultra Micro Balance and the percentage composition were calculated based on Carlo Erba Elemental Standards B2005, B2035, and B2036, with an error of <1%. Standards of ammonium sulphate (IAEA-N1 and IAEA-N2) for nitrogen, and sugar (IAEA-CH6) and graphite (EIL-32) for carbon were used for calibration.
Ea 1108
Manufactured by Thermo Fisher Scientific
Sourced in Italy
The EA 1108 is an elemental analyzer designed for the determination of carbon, hydrogen, nitrogen, and sulfur content in a wide range of organic and inorganic materials. It is a versatile instrument capable of analyzing solid, liquid, and gaseous samples with high precision and accuracy.
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4 protocols using ea 1108
1
Lichen Elemental and Isotopic Analysis
2
Elemental Analysis of Plant Samples
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The oven dried samples of each replicate pot and harvest were powdered. Subsamples of 50 mg were digested on 10 mL tubes with 1 mL of 67–70% HNO3 and allowed to rest overnight prior to boiling for about 4 h to almost dryness, then diluted to 10 mL and analysed for elements by ICP-OES. Internal reference material was used to verify analytical accuracy. Total N was measured by dry combustion with an elemental analyzer (Thermo Scientific, EA1108) via combustion at 900 °C and subsequent analysis of CO2 and N2 with gas chromatography.
3
Measuring N2 Fixation in Cyanobacteria
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The PON and POC content were measured for Nodularia cultures and Nostoc punctiforme sp. 73.1 grown in N-free medium on day 14. Due to budget constraints, the remaining Nostoc cultures were not studied. Filters containing culture samples were trimmed, sectioned, then loaded into tin capsules and palletised for isotopic analysis. Measurement was done by means of flash combustion in a Carlo Erba EA 1108 at 1020 °C in a Thermo Finnigan Delta S mass-spectrometer. Calibration material for N and C analysis was acetanilide (Merck). N 2 fixation activity was determined by incubating cultures in two replicates per treatment with bubble addition of 15 N-N 2 enriched gas (99% 15 N 2 ) for 24 h, guaranteeing sufficient dissolution of the 15 N gas in the incubation bottle (Wannicke et al. 2018a ). Tracer incubations were terminated by gentle vacuum filtration (< 200 mbar) of the culture material through pre-combusted GF/F filters (Whatman) that were then dried at 60 °C, analysed and the N 2 fixation rates calculated (Montoya et al. 1996) . Two technical replicates were conducted per bottle.
4
Elemental Carbon Quantification by Combustion
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Elemental quantification of total carbon content was performed by bulk combustion using a Thermal combustion element analyzer, Thermo EA 1108(TC/EA), working in standard conditions (Helium flow at 120 mL/min, combustion furnace at 1000°C, chromatographic column oven at 60°C, oxygen loop 10 mL at 100 kPa). The amount of carbonate was calculated according to the following equation:
where, is the molecular weight of carbonate ion, is the molecular weight of carbon, and is the percentage of carbon given by the measurement. Prior to analysis, samples were crushed using an agate mortar, and the powders were dried at 120°C overnight.
where, is the molecular weight of carbonate ion, is the molecular weight of carbon, and is the percentage of carbon given by the measurement. Prior to analysis, samples were crushed using an agate mortar, and the powders were dried at 120°C overnight.
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