{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001880","ANALYSIS_ID":"AN003046","VERSION":"1","CREATED_ON":"July 21, 2021, 1:17 pm"},

"PROJECT":{"PROJECT_TITLE":"Predator Cues Target Signaling Pathways in Toxic Algal Metabolome","PROJECT_TYPE":"Metabolomics to elucidate signaling pathway","PROJECT_SUMMARY":"Early detection of predators is critical to the survival of all living organisms. For phytoplankton, recognition and response to chemical cues from predators, as evidence of predation risk, is particularly crucial. The phytoplankton Alexandrium minutum upregulates its toxicity when exposed to copepodamides, a suite of compounds released by copepod predators. However, how A. minutum perceives these predatory cues and what metabolic pathways are involved in initiating toxin induction remains unknown. In this study LC/MS and NMR-based metabolomics uncovered subtle physiological responses of A. minutum to copepodamides, including dysregulation of valine biosynthesis and enhancement of butanoate metabolism and arginine biosynthesis. While we have yet to identify a chemoreceptor directly activated by copepod cues, based on the results of inhibition experiments detection of copepodamides appears to disrupt the activity of serine/threonine phosphatases leading to increased jasmonic acid biosynthesis and signaling, which leads to amplified gonyautoxin biosynthesis in A. minutum. This study is an important step toward a better understanding of chemosensory ecology of predator-prey interactions in phytoplankton.","INSTITUTE":"Georgia Institute of Technology","DEPARTMENT":"School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience","LABORATORY":"Kubanek Lab","LAST_NAME":"Brown","FIRST_NAME":"Emily","ADDRESS":"950 Atlantic Dr Atlanta, GA, 30332, USA","EMAIL":"julia.kubanek@biosci.gatech.edu","PHONE":"404-894-8424","PROJECT_COMMENTS":"This study has 3 parts: 2 NMR (polar and non-polar metabolites) and MS","CONTRIBUTORS":"Emily R. Brown, Sam G. Moore, David A. Gaul, and Julia Kubanek"},

"STUDY":{"STUDY_TITLE":"NMR Predator Cues Target Signaling Pathways in Toxic Algal Metabolome (Polar metabolites)","STUDY_TYPE":"1H NMR Metabolomics to elucidate signaling pathway","STUDY_SUMMARY":"Metabolomics investigation of the phytoplankton Alexandrium minutum with and without copepod cues in order to explore cell signaling involved in toxin induction.","INSTITUTE":"Georgia Institute of Technology","DEPARTMENT":"School of Biological Sciences, School of Chemistry and Biochemistry, Center for Microbial Dynamics and Infection, Parker H. Petit Institute for Bioengineering and Bioscience","LABORATORY":"Kubanek Lab","LAST_NAME":"Brown","FIRST_NAME":"Emily","ADDRESS":"950 Atlantic Dr Atlanta, Georgia USA 30332","EMAIL":"julia.kubanek@biosci.gatech.edu","PHONE":"404-894-8424","NUM_GROUPS":"2","TOTAL_SUBJECTS":"40","STUDY_COMMENTS":"Part 1 of 3. This part includes NMR analysis of polar metabolites using oPLSDA. Parts 2 and 3 inlcude NMR analysis of nonpolar metabolites and the corresponding mass spectrometry metabolomics for both polar and non-polar metabolites."},

"SUBJECT":{"SUBJECT_TYPE":"Other organism","SUBJECT_SPECIES":"Alexandrium minutum","TAXONOMY_ID":"39455","GENOTYPE_STRAIN":"CCMP 113","GENDER":"Not applicable"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Control 1",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 1","Cellular_toxicity (fmol)":"0.3","Cells_per_mL":"20547","Cells_in_polar_NMR_analysis":"3821742","Polar_NMR_solvent_volume (uL)":"224","Cells_in_polar_MS_analysis":"1273914","Polar_MS_solvent_volume (uL)":"280"}
},
{
"Subject ID":"-",
"Sample ID":"Control 2",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 2","Cellular_toxicity (fmol)":"0.36","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Control 3",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 3","Cellular_toxicity (fmol)":"0.4","Cells_per_mL":"20670","Cells_in_polar_NMR_analysis":"3844620","Polar_NMR_solvent_volume (uL)":"226","Cells_in_polar_MS_analysis":"1281540","Polar_MS_solvent_volume (uL)":"282"}
},
{
"Subject ID":"-",
"Sample ID":"Control 4",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 4","Cellular_toxicity (fmol)":"0.45","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Control 5",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 5","Cellular_toxicity (fmol)":"0.29","Cells_per_mL":"20670","Cells_in_polar_NMR_analysis":"3844620","Polar_NMR_solvent_volume (uL)":"226","Cells_in_polar_MS_analysis":"1281540","Polar_MS_solvent_volume (uL)":"282"}
},
{
"Subject ID":"-",
"Sample ID":"Control 6",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 6","Cellular_toxicity (fmol)":"0.46","Cells_per_mL":"21165","Cells_in_polar_NMR_analysis":"3936690","Polar_NMR_solvent_volume (uL)":"231","Cells_in_polar_MS_analysis":"1312230","Polar_MS_solvent_volume (uL)":"289"}
},
{
"Subject ID":"-",
"Sample ID":"Control 7",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 7","Cellular_toxicity (fmol)":"0.43","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Control 8",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 8","Cellular_toxicity (fmol)":"0.41","Cells_per_mL":"20794","Cells_in_polar_NMR_analysis":"3867684","Polar_NMR_solvent_volume (uL)":"227","Cells_in_polar_MS_analysis":"1289228","Polar_MS_solvent_volume (uL)":"284"}
},
{
"Subject ID":"-",
"Sample ID":"Control 9",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 9","Cellular_toxicity (fmol)":"0.34","Cells_per_mL":"20423","Cells_in_polar_NMR_analysis":"3798678","Polar_NMR_solvent_volume (uL)":"223","Cells_in_polar_MS_analysis":"1266226","Polar_MS_solvent_volume (uL)":"279"}
},
{
"Subject ID":"-",
"Sample ID":"Control 10",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 10","Cellular_toxicity (fmol)":"0.34","Cells_per_mL":"20670","Cells_in_polar_NMR_analysis":"3844620","Polar_NMR_solvent_volume (uL)":"226","Cells_in_polar_MS_analysis":"1281540","Polar_MS_solvent_volume (uL)":"282"}
},
{
"Subject ID":"-",
"Sample ID":"Control 11",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 11","Cellular_toxicity (fmol)":"0.4","Cells_per_mL":"19433","Cells_in_polar_NMR_analysis":"3614538","Polar_NMR_solvent_volume (uL)":"212","Cells_in_polar_MS_analysis":"1204846","Polar_MS_solvent_volume (uL)":"265"}
},
{
"Subject ID":"-",
"Sample ID":"Control 12",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 12","Cellular_toxicity (fmol)":"0.43","Cells_per_mL":"20423","Cells_in_polar_NMR_analysis":"3798678","Polar_NMR_solvent_volume (uL)":"223","Cells_in_polar_MS_analysis":"1266226","Polar_MS_solvent_volume (uL)":"279"}
},
{
"Subject ID":"-",
"Sample ID":"Control 13",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"N/A","Cellular_toxicity (fmol)":"0.31","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Control 14",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 14","Cellular_toxicity (fmol)":"0.36","Cells_per_mL":"20175","Cells_in_polar_NMR_analysis":"3752550","Polar_NMR_solvent_volume (uL)":"220","Cells_in_polar_MS_analysis":"1250850","Polar_MS_solvent_volume (uL)":"275"}
},
{
"Subject ID":"-",
"Sample ID":"Control 15",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 15","Cellular_toxicity (fmol)":"0.37","Cells_per_mL":"20052","Cells_in_polar_NMR_analysis":"3729672","Polar_NMR_solvent_volume (uL)":"219","Cells_in_polar_MS_analysis":"1243224","Polar_MS_solvent_volume (uL)":"274"}
},
{
"Subject ID":"-",
"Sample ID":"Control 16",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 16","Cellular_toxicity (fmol)":"0.43","Cells_per_mL":"20670","Cells_in_polar_NMR_analysis":"3844620","Polar_NMR_solvent_volume (uL)":"226","Cells_in_polar_MS_analysis":"1281540","Polar_MS_solvent_volume (uL)":"282"}
},
{
"Subject ID":"-",
"Sample ID":"Control 17",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 17","Cellular_toxicity (fmol)":"0.43","Cells_per_mL":"21165","Cells_in_polar_NMR_analysis":"3936690","Polar_NMR_solvent_volume (uL)":"231","Cells_in_polar_MS_analysis":"1312230","Polar_MS_solvent_volume (uL)":"289"}
},
{
"Subject ID":"-",
"Sample ID":"Control 18",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 18","Cellular_toxicity (fmol)":"0.34","Cells_per_mL":"21165","Cells_in_polar_NMR_analysis":"3936690","Polar_NMR_solvent_volume (uL)":"231","Cells_in_polar_MS_analysis":"1312230","Polar_MS_solvent_volume (uL)":"289"}
},
{
"Subject ID":"-",
"Sample ID":"Control 19",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 19","Cellular_toxicity (fmol)":"0.41","Cells_per_mL":"20547","Cells_in_polar_NMR_analysis":"3821742","Polar_NMR_solvent_volume (uL)":"224","Cells_in_polar_MS_analysis":"1273914","Polar_MS_solvent_volume (uL)":"280"}
},
{
"Subject ID":"-",
"Sample ID":"Control 20",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"Control 20","Cellular_toxicity (fmol)":"0.35","Cells_per_mL":"19309","Cells_in_polar_NMR_analysis":"3591474","Polar_NMR_solvent_volume (uL)":"211","Cells_in_polar_MS_analysis":"1197158","Polar_MS_solvent_volume (uL)":"264"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 1",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 1","Cellular_toxicity (fmol)":"0.95","Cells_per_mL":"19804","Cells_in_polar_NMR_analysis":"3683544","Polar_NMR_solvent_volume (uL)":"216","Cells_in_polar_MS_analysis":"1227848","Polar_MS_solvent_volume (uL)":"270"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 2",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 2","Cellular_toxicity (fmol)":"0.78","Cells_per_mL":"19309","Cells_in_polar_NMR_analysis":"3591474","Polar_NMR_solvent_volume (uL)":"211","Cells_in_polar_MS_analysis":"1197158","Polar_MS_solvent_volume (uL)":"264"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 3",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 3","Cellular_toxicity (fmol)":"0.64","Cells_per_mL":"21165","Cells_in_polar_NMR_analysis":"3936690","Polar_NMR_solvent_volume (uL)":"231","Cells_in_polar_MS_analysis":"1312230","Polar_MS_solvent_volume (uL)":"289"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 4",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 4","Cellular_toxicity (fmol)":"1.02","Cells_per_mL":"18938","Cells_in_polar_NMR_analysis":"3522468","Polar_NMR_solvent_volume (uL)":"207","Cells_in_polar_MS_analysis":"1174156","Polar_MS_solvent_volume (uL)":"228"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 5",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 5","Cellular_toxicity (fmol)":"0.69","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 6",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 6","Cellular_toxicity (fmol)":"0.74","Cells_per_mL":"18938","Cells_in_polar_NMR_analysis":"3522468","Polar_NMR_solvent_volume (uL)":"207","Cells_in_polar_MS_analysis":"1174156","Polar_MS_solvent_volume (uL)":"258"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 7",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 7","Cellular_toxicity (fmol)":"0.7","Cells_per_mL":"19061","Cells_in_polar_NMR_analysis":"3545346","Polar_NMR_solvent_volume (uL)":"208","Cells_in_polar_MS_analysis":"1181782","Polar_MS_solvent_volume (uL)":"260"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 8",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 8","Cellular_toxicity (fmol)":"0.74","Cells_per_mL":"19433","Cells_in_polar_NMR_analysis":"3614538","Polar_NMR_solvent_volume (uL)":"212","Cells_in_polar_MS_analysis":"1204846","Polar_MS_solvent_volume (uL)":"265"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 9",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 9","Cellular_toxicity (fmol)":"0.89","Cells_per_mL":"20299","Cells_in_polar_NMR_analysis":"3775614","Polar_NMR_solvent_volume (uL)":"222","Cells_in_polar_MS_analysis":"1258538","Polar_MS_solvent_volume (uL)":"277"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 10",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 10","Cellular_toxicity (fmol)":"0.85","Cells_per_mL":"19928","Cells_in_polar_NMR_analysis":"3706608","Polar_NMR_solvent_volume (uL)":"218","Cells_in_polar_MS_analysis":"1235536","Polar_MS_solvent_volume (uL)":"272"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 11",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 11","Cellular_toxicity (fmol)":"0.86","Cells_per_mL":"19928","Cells_in_polar_NMR_analysis":"3706608","Polar_NMR_solvent_volume (uL)":"218","Cells_in_polar_MS_analysis":"1235536","Polar_MS_solvent_volume (uL)":"272"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 12",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"N/A","Cellular_toxicity (fmol)":"0.82","Cells_per_mL":"19061","Cells_in_polar_NMR_analysis":"3545346","Polar_NMR_solvent_volume (uL)":"208","Cells_in_polar_MS_analysis":"1181782","Polar_MS_solvent_volume (uL)":"260"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 13",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 13","Cellular_toxicity (fmol)":"0.83","Cells_per_mL":"18566","Cells_in_polar_NMR_analysis":"3453276","Polar_NMR_solvent_volume (uL)":"203","Cells_in_polar_MS_analysis":"1151092","Polar_MS_solvent_volume (uL)":"253"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 14",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"N/A","Cellular_toxicity (fmol)":"0.71","Cells_per_mL":"18319","Cells_in_polar_NMR_analysis":"3407334","Polar_NMR_solvent_volume (uL)":"200","Cells_in_polar_MS_analysis":"1135778","Polar_MS_solvent_volume (uL)":"250"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 15",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 15","Cellular_toxicity (fmol)":"0.87","Cells_per_mL":"19433","Cells_in_polar_NMR_analysis":"3614538","Polar_NMR_solvent_volume (uL)":"212","Cells_in_polar_MS_analysis":"1204846","Polar_MS_solvent_volume (uL)":"265"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 16",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 16","Cellular_toxicity (fmol)":"0.59","Cells_per_mL":"19433","Cells_in_polar_NMR_analysis":"3614538","Polar_NMR_solvent_volume (uL)":"212","Cells_in_polar_MS_analysis":"1204846","Polar_MS_solvent_volume (uL)":"265"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 17",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 17","Cellular_toxicity (fmol)":"0.71","Cells_per_mL":"19185","Cells_in_polar_NMR_analysis":"3568410","Polar_NMR_solvent_volume (uL)":"209","Cells_in_polar_MS_analysis":"1189470","Polar_MS_solvent_volume (uL)":"262"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 18",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 18","Cellular_toxicity (fmol)":"0.62","Cells_per_mL":"19928","Cells_in_polar_NMR_analysis":"3706608","Polar_NMR_solvent_volume (uL)":"218","Cells_in_polar_MS_analysis":"1235536","Polar_MS_solvent_volume (uL)":"272"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 19",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 19","Cellular_toxicity (fmol)":"0.88","Cells_per_mL":"18442","Cells_in_polar_NMR_analysis":"3430212","Polar_NMR_solvent_volume (uL)":"201","Cells_in_polar_MS_analysis":"1143404","Polar_MS_solvent_volume (uL)":"252"}
},
{
"Subject ID":"-",
"Sample ID":"Copepodamide 20",
"Factors":{"Treatment":"Copepodamide"},
"Additional sample data":{"RAW_FILE_NAME":"Copepodamide 20","Cellular_toxicity (fmol)":"0.6","Cells_per_mL":"18814","Cells_in_polar_NMR_analysis":"3499404","Polar_NMR_solvent_volume (uL)":"205","Cells_in_polar_MS_analysis":"1166468","Polar_MS_solvent_volume (uL)":"257"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Alexandrium minutum cells were collected by vacuum filtration onto GF/F filters and quenched with liquid nitrogen. Frozen cells with filters were stored in foil (previously muffled for 3 h at 450 °C) at -80 °C until extraction.","COLLECTION_PROTOCOL_FILENAME":"Predator Cues Target Signaling Pathways in Toxic Algal Metabolome_protocol.pdf","SAMPLE_TYPE":"Algae","COLLECTION_METHOD":"Filtration and Freeze","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Twice, first at the start of the experiment and again 24 h later, 1.5 mL of either 0.2 µM copepodamides (copepod cue) in DMSO (treatment, n=20) or DMSO alone (control, n=20) were added aseptically to 300 mL cultures of Alexandrium minutum resulting in a final concentration of 1 nM copepodamides.","TREATMENT_PROTOCOL_FILENAME":"Predator Cues Target Signaling Pathways in Toxic Algal Metabolome_protocol.pdf"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Polar extracts were dissolved at a concentration equivalent to 1.70 x 10^7 A. minutum cells mL-1 in 90:10 H2O/D2O (99.96% atom D2O; Cambridge Isotope Labs) with 0.2 mM phosphate buffer (pH 7.4) and 0.25 mM 3 (trimethylsilyl)propionic 2,2,3,3d 4 acid (TMSP), as internal standard, for 1H NMR spectroscopy analysis. The media blank extract was prepared in the smallest volume of solvent possible (175 µL). Particulates were excluded by briefly centrifuging the samples and then carefully transferring the extracts into 3 mm NMR tubes. The extracts were analyzed using a Bruker Avance IIIHD 800 MHz NMR spectrometer equipped with a 3 mm triple resonance broadband cryoprobe. Spectra for the polar extracts were acquired using a 1D water presaturation (Bruker zgpr pulse sequence) with a spectral width of 12 kHz centered at 3758.36 Hz, with a relaxation delay 3 s, over 480 scans.","SAMPLEPREP_PROTOCOL_FILENAME":"Predator Cues Target Signaling Pathways in Toxic Algal Metabolome_protocol.pdf","EXTRACT_STORAGE":"-80℃"},

"ANALYSIS":{"ANALYSIS_TYPE":"NMR"},

"NM":{"INSTRUMENT_NAME":"Bruker Avance IIIHD","INSTRUMENT_TYPE":"FT-NMR","NMR_EXPERIMENT_TYPE":"1D-1H","SPECTROMETER_FREQUENCY":"800 MHz","NMR_PROBE":"3 mm triple resonance broadband cryoprobe","NMR_SOLVENT":"90:10 H2O/D2O (99.96% atom D2O; Cambridge Isotope Labs) with 0.2 mM phosphate buffer (pH 7.4) and 0.25 mM 3-(trimethylsilyl)propionic-2,2,3,3d4-acid","NMR_TUBE_SIZE":"3 mm","PULSE_SEQUENCE":"zgpr","NUMBER_OF_SCANS":"480","RELAXATION_DELAY":"3s","SPECTRAL_WIDTH":"12 kHz","BASELINE_CORRECTION_METHOD":"Spline","BINNED_INCREMENT":"0.005 ppm","NMR_RESULTS_FILE":"Preprocessed_binned_polar_NMRmetabolomics.txt UNITS:area under the curve"}

}