Protocol detail

UHPLC-DAD-MS/MS Analysis of Seawater

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Liquid chromatography mass spectrometry (LC-MS) grade H₂O, acetonitrile (ACN) and formic acid for UHPLC-DAD-MS/MS analysis and HPLC grade methanol (MeOH), reagent grade diethyl ether, hydrochloric acid (HCl, 37%), and NaCl used for sample preparation were all purchased from VWR Chemicals (Oslo, Norway). Milli-Q H₂O (Merck; Darmstadt, Germany) was used in sample and buffer preparation. Purine and HP-0921, Agilent part number: G1969-85003, were used as internal standards for MS spectra calibration, purchased from Matriks AS (Oslo, Norway). In this study, calcium and magnesium (Ca²⁺/Mg²⁺) free seawater (CMFSW)^{58 (link)} and calcium and magnesium free modified anticoagulant buffer containing 462 mM NaCl, 10.7 mM KCl, 7 mM Na₂SO₄, 2.1 mM NaHCO₃, and 20 mM EGTA (CMFSW-E), pH 7.8^{59 (link)} were used.

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Hira J., Wolfson D., Andersen A.J., Haug T, & Stensvåg K. (2020). Autofluorescence mediated red spherulocyte sorting provides insights into the source of spinochromes in sea urchins. Scientific Reports, 10, 1149.

Publication 2020

LC-MS) grade H2O acetonitrile (ACN) formic acid HPLC grade methanol (MeOH), Milli-Q H2O Purine HP-0921

Acetonitrile Anticoagulant Buffer Calcium Diethyl ether Egta Formic acid Hplc Hydrochloric acid Liquid chromatography Magnesium Mass spectrometry Methanol Ms ms Nacl Nahco3 Purine

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Variable analysis

independent variables

Liquid chromatography mass spectrometry (LC-MS) grade H2O, acetonitrile (ACN) and formic acid
HPLC grade methanol (MeOH), reagent grade diethyl ether, hydrochloric acid (HCl, 37%), and NaCl
Calcium and magnesium (Ca2+/Mg2+) free seawater (CMFSW)
Calcium and magnesium free modified anticoagulant buffer containing 462 mM NaCl, 10.7 mM KCl, 7 mM Na2SO4, 2.1 mM NaHCO3, and 20 mM EGTA (CMFSW-E), pH 7.8

dependent variables

Not explicitly mentioned

control variables

Milli-Q H2O (Merck; Darmstadt, Germany) used in sample and buffer preparation
Purine and HP-0921, Agilent part number: G1969-85003, used as internal standards for MS spectra calibration

positive controls

Not mentioned

negative controls

Not mentioned

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