#METABOLOMICS WORKBENCH ebrammerrobbins_20231210_185900 DATATRACK_ID:4498 STUDY_ID:ST003017 ANALYSIS_ID:AN004951 PROJECT_ID:PR001878
VERSION             	1
CREATED_ON             	December 18, 2023, 5:58 pm
#PROJECT
PR:PROJECT_TITLE                 	Lipidomics and plasma hormone reveal indicators of reproductive status in
PR:PROJECT_TITLE                 	Florida manatees (Trichechus manatus latirostris)
PR:PROJECT_SUMMARY               	Florida manatees (Trichechus manatus latirostris) are protected as a threatened
PR:PROJECT_SUMMARY               	species, and data are lacking regarding their reproductive physiology. This
PR:PROJECT_SUMMARY               	study aimed to (1) quantify plasma steroid hormones in Florida manatees from two
PR:PROJECT_SUMMARY               	field sites, Crystal River and Indian River Lagoon, at different gestational
PR:PROJECT_SUMMARY               	stages and to (2) determine the relationship between plasma progesterone
PR:PROJECT_SUMMARY               	concentrations and lipid biochemistry in relation to pregnancy status.
PR:PROJECT_SUMMARY               	Ultra-high performance liquid chromatography-tandem mass spectrometric analysis
PR:PROJECT_SUMMARY               	was used to measure plasma steroid hormones and lipids. Pregnant female manatees
PR:PROJECT_SUMMARY               	were morphometrically distinct from male and non-pregnant female manatees,
PR:PROJECT_SUMMARY               	characterized by larger body weight and maximal girth. Progesterone
PR:PROJECT_SUMMARY               	concentrations in manatees were also elevated during early gestation versus late
PR:PROJECT_SUMMARY               	gestation. Cholesterol, an important metabolic lipid and precursor for
PR:PROJECT_SUMMARY               	reproductive steroids, was not different between groups. Lipidomics quantified
PR:PROJECT_SUMMARY               	949 lipids and plasma concentrations of a sphingolipid, ceramide non-hydroxy
PR:PROJECT_SUMMARY               	fatty acid-sphingosine and several glycerophospholipids, including
PR:PROJECT_SUMMARY               	lysophosphatidylcholine, phosphatidylethanolamines,
PR:PROJECT_SUMMARY               	plasmenyl-phosphatidylserines and monomethyl phosphatidylethanolamines, were
PR:PROJECT_SUMMARY               	associated with pregnancy status in the Florida manatee. This research
PR:PROJECT_SUMMARY               	contributes to improving knowledge of manatee reproductive physiology by
PR:PROJECT_SUMMARY               	providing data on plasma steroid hormones relative to reproductive status and by
PR:PROJECT_SUMMARY               	assessing how plasma lipids in healthy Florida manatees correspond to
PR:PROJECT_SUMMARY               	progesterone levels. This lipid panel has potential as a diagnostic approach to
PR:PROJECT_SUMMARY               	identify pregnant individuals in fresh and archived samples. These biochemical
PR:PROJECT_SUMMARY               	and morphometric indicators of reproductive status advance the understanding of
PR:PROJECT_SUMMARY               	manatee physiology.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Physiological Sciences
PR:LABORATORY                    	Dr. Martyniuk
PR:LAST_NAME                     	Brammer-Robbins
PR:FIRST_NAME                    	Elizabeth
PR:ADDRESS                       	2187 Mowry Rd., Bldg 471
PR:EMAIL                         	e.brammerrobbins@ufl.edu
PR:PHONE                         	9104652899
PR:FUNDING_SOURCE                	National Science Foundation Graduate Research Fellowship Program under Grant No.
PR:FUNDING_SOURCE                	(2019285699 to EB-R).
#STUDY
ST:STUDY_TITLE                   	Lipidomics and plasma hormone reveal indicators of reproductive status in
ST:STUDY_TITLE                   	Florida manatees (Trichechus manatus latirostris)
ST:STUDY_SUMMARY                 	Florida manatees (Trichechus manatus latirostris) are protected as a threatened
ST:STUDY_SUMMARY                 	species, and data are lacking regarding their reproductive physiology. This
ST:STUDY_SUMMARY                 	study aimed to (1) quantify plasma steroid hormones in Florida manatees from two
ST:STUDY_SUMMARY                 	field sites, Crystal River and Indian River Lagoon, at different gestational
ST:STUDY_SUMMARY                 	stages and to (2) determine the relationship between plasma progesterone
ST:STUDY_SUMMARY                 	concentrations and lipid biochemistry in relation to pregnancy status.
ST:STUDY_SUMMARY                 	Ultra-high performance liquid chromatography-tandem mass spectrometric analysis
ST:STUDY_SUMMARY                 	was used to measure plasma steroid hormones and lipids. Pregnant female manatees
ST:STUDY_SUMMARY                 	were morphometrically distinct from male and non-pregnant female manatees,
ST:STUDY_SUMMARY                 	characterized by larger body weight and maximal girth. Progesterone
ST:STUDY_SUMMARY                 	concentrations in manatees were also elevated during early gestation versus late
ST:STUDY_SUMMARY                 	gestation. Cholesterol, an important metabolic lipid and precursor for
ST:STUDY_SUMMARY                 	reproductive steroids, was not different between groups. Lipidomics quantified
ST:STUDY_SUMMARY                 	949 lipids and plasma concentrations of a sphingolipid, ceramide non-hydroxy
ST:STUDY_SUMMARY                 	fatty acid-sphingosine and several glycerophospholipids, including
ST:STUDY_SUMMARY                 	lysophosphatidylcholine, phosphatidylethanolamines,
ST:STUDY_SUMMARY                 	plasmenyl-phosphatidylserines and monomethyl phosphatidylethanolamines, were
ST:STUDY_SUMMARY                 	associated with pregnancy status in the Florida manatee. This research
ST:STUDY_SUMMARY                 	contributes to improving knowledge of manatee reproductive physiology by
ST:STUDY_SUMMARY                 	providing data on plasma steroid hormones relative to reproductive status and by
ST:STUDY_SUMMARY                 	assessing how plasma lipids in healthy Florida manatees correspond to
ST:STUDY_SUMMARY                 	progesterone levels. This lipid panel has potential as a diagnostic approach to
ST:STUDY_SUMMARY                 	identify pregnant individuals in fresh and archived samples. These biochemical
ST:STUDY_SUMMARY                 	and morphometric indicators of reproductive status advance the understanding of
ST:STUDY_SUMMARY                 	manatee physiology.
ST:INSTITUTE                     	University of Florida
ST:LAST_NAME                     	Brammer-Robbins
ST:FIRST_NAME                    	Elizabeth
ST:ADDRESS                       	2187 Mowry Rd., Bldg 471
ST:EMAIL                         	e.brammerrobbins@ufl.edu
ST:PHONE                         	9104652899
ST:TOTAL_SUBJECTS                	57
ST:NUM_MALES                     	31
ST:NUM_FEMALES                   	26
ST:PUBLICATIONS                  	https://doi.org/10.1016/j.ygcen.2023.114250
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Florida manatee (Trichechus manatus latirostris)
SU:AGE_OR_AGE_RANGE              	Adult
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	Quality Control 1	QC01	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=-; RAW_FILE_NAME=FullScan_QC01_neg.raw; RAW_FILE_NAME=FullScan_QC01_pos.raw
SUBJECT_SAMPLE_FACTORS           	Quality Control 2	QC02	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=-; RAW_FILE_NAME=FullScan_QC02_neg.raw; RAW_FILE_NAME=FullScan_QC02_pos.raw
SUBJECT_SAMPLE_FACTORS           	Quality Control 3	QC03	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=-; RAW_FILE_NAME=FullScan_QC03_neg.raw; RAW_FILE_NAME=FullScan_QC03_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1906	Sp01	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp01_neg.raw; RAW_FILE_NAME=FullScan_Sp01_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1832	Sp02	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp02_neg.raw; RAW_FILE_NAME=FullScan_Sp02_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1819	Sp03	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp03_neg.raw; RAW_FILE_NAME=FullScan_Sp03_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1810	Sp04	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp04_neg.raw; RAW_FILE_NAME=FullScan_Sp04_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1714	Sp05	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp05_neg.raw; RAW_FILE_NAME=FullScan_Sp05_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1632	Sp06	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp06_neg.raw; RAW_FILE_NAME=FullScan_Sp06_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1611	Sp07	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp07_neg.raw; RAW_FILE_NAME=FullScan_Sp07_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1501	Sp08	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp08_neg.raw; RAW_FILE_NAME=FullScan_Sp08_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1905	Sp09	Capture Location:Crystal River | Reproductive Status:P | Sex:F	Pool #=1; RAW_FILE_NAME=FullScan_Sp09_neg.raw; RAW_FILE_NAME=FullScan_Sp09_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1902	Sp10	Capture Location:Crystal River | Reproductive Status:P | Sex:F	Pool #=1; RAW_FILE_NAME=FullScan_Sp10_neg.raw; RAW_FILE_NAME=FullScan_Sp10_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1901	Sp11	Capture Location:Crystal River | Reproductive Status:P | Sex:F	Pool #=1; RAW_FILE_NAME=FullScan_Sp11_neg.raw; RAW_FILE_NAME=FullScan_Sp11_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1705	Sp12	Capture Location:Crystal River | Reproductive Status:P | Sex:F	Pool #=1; RAW_FILE_NAME=FullScan_Sp12_neg.raw; RAW_FILE_NAME=FullScan_Sp12_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1528	Sp13	Capture Location:Crystal River | Reproductive Status:P | Sex:F	Pool #=1; RAW_FILE_NAME=FullScan_Sp13_neg.raw; RAW_FILE_NAME=FullScan_Sp13_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1510	Sp15	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp15_neg.raw; RAW_FILE_NAME=FullScan_Sp15_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1507	Sp16	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp16_neg.raw; RAW_FILE_NAME=FullScan_Sp16_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1419	Sp17	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp17_neg.raw; RAW_FILE_NAME=FullScan_Sp17_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1408	Sp18	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp18_neg.raw; RAW_FILE_NAME=FullScan_Sp18_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1310	Sp19	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp19_neg.raw; RAW_FILE_NAME=FullScan_Sp19_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1305	Sp20	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp20_neg.raw; RAW_FILE_NAME=FullScan_Sp20_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1213	Sp21	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp21_neg.raw; RAW_FILE_NAME=FullScan_Sp21_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1205	Sp22	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp22_neg.raw; RAW_FILE_NAME=FullScan_Sp22_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1409	Sp23	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp23_neg.raw; RAW_FILE_NAME=FullScan_Sp23_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1311	Sp24	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp24_neg.raw; RAW_FILE_NAME=FullScan_Sp24_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1309	Sp25	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp25_neg.raw; RAW_FILE_NAME=FullScan_Sp25_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1212	Sp26	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp26_neg.raw; RAW_FILE_NAME=FullScan_Sp26_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1209	Sp27	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp27_neg.raw; RAW_FILE_NAME=FullScan_Sp27_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1308	Sp28	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp28_neg.raw; RAW_FILE_NAME=FullScan_Sp28_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1306	Sp29	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp29_neg.raw; RAW_FILE_NAME=FullScan_Sp29_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1305	Sp30	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp30_neg.raw; RAW_FILE_NAME=FullScan_Sp30_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1304	Sp31	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp31_neg.raw; RAW_FILE_NAME=FullScan_Sp31_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1303	Sp32	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp32_neg.raw; RAW_FILE_NAME=FullScan_Sp32_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1302	Sp33	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp33_neg.raw; RAW_FILE_NAME=FullScan_Sp33_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1301	Sp34	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp34_neg.raw; RAW_FILE_NAME=FullScan_Sp34_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1207	Sp35	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp35_neg.raw; RAW_FILE_NAME=FullScan_Sp35_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1823	Sp36	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp36_neg.raw; RAW_FILE_NAME=FullScan_Sp36_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1402	Sp39	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp39_neg.raw; RAW_FILE_NAME=FullScan_Sp39_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1317	Sp40	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp40_neg.raw; RAW_FILE_NAME=FullScan_Sp40_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1208	Sp41	Capture Location:Crystal River | Reproductive Status:NP | Sex:F	Pool #=2; RAW_FILE_NAME=FullScan_Sp41_neg.raw; RAW_FILE_NAME=FullScan_Sp41_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1008	Sp52	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp52_neg.raw; RAW_FILE_NAME=FullScan_Sp52_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1007	Sp53	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp53_neg.raw; RAW_FILE_NAME=FullScan_Sp53_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1006	Sp54	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp54_neg.raw; RAW_FILE_NAME=FullScan_Sp54_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1312	Sp57	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp57_neg.raw; RAW_FILE_NAME=FullScan_Sp57_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1307	Sp58	Capture Location:Brevard County | Reproductive Status:NP | Sex:F	Pool #=6; RAW_FILE_NAME=FullScan_Sp58_neg.raw; RAW_FILE_NAME=FullScan_Sp58_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1907	Sp60	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp60_neg.raw; RAW_FILE_NAME=FullScan_Sp60_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1904	Sp61	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp61_neg.raw; RAW_FILE_NAME=FullScan_Sp61_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1833	Sp62	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp62_neg.raw; RAW_FILE_NAME=FullScan_Sp62_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1825	Sp63	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp63_neg.raw; RAW_FILE_NAME=FullScan_Sp63_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1805	Sp64	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp64_neg.raw; RAW_FILE_NAME=FullScan_Sp64_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1709	Sp65	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp65_neg.raw; RAW_FILE_NAME=FullScan_Sp65_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1706	Sp66	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp66_neg.raw; RAW_FILE_NAME=FullScan_Sp66_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1625	Sp67	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp67_neg.raw; RAW_FILE_NAME=FullScan_Sp67_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1623	Sp68	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp68_neg.raw; RAW_FILE_NAME=FullScan_Sp68_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CCR1523	Sp69	Capture Location:Crystal River | Reproductive Status:NA | Sex:M	Pool #=5; RAW_FILE_NAME=FullScan_Sp69_neg.raw; RAW_FILE_NAME=FullScan_Sp69_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1203	Sp70	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp70_neg.raw; RAW_FILE_NAME=FullScan_Sp70_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1202	Sp71	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp71_neg.raw; RAW_FILE_NAME=FullScan_Sp71_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1106	Sp72	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp72_neg.raw; RAW_FILE_NAME=FullScan_Sp72_pos.raw
SUBJECT_SAMPLE_FACTORS           	Plasma-CBC1104	Sp73	Capture Location:Brevard County | Reproductive Status:NA | Sex:M	Pool #=7; RAW_FILE_NAME=FullScan_Sp73_neg.raw; RAW_FILE_NAME=FullScan_Sp73_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target01	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=1; RAW_FILE_NAME=FullScan_Target01_neg.raw; RAW_FILE_NAME=FullScan_Target01_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target02	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=2; RAW_FILE_NAME=FullScan_Target02_neg.raw; RAW_FILE_NAME=FullScan_Target02_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target03	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=3; RAW_FILE_NAME=FullScan_Target03_neg.raw; RAW_FILE_NAME=FullScan_Target03_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target04	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=4; RAW_FILE_NAME=FullScan_Target04_neg.raw; RAW_FILE_NAME=FullScan_Target04_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target05	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=5; RAW_FILE_NAME=FullScan_Target05_neg.raw; RAW_FILE_NAME=FullScan_Target05_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target06	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=6; RAW_FILE_NAME=FullScan_Target06_neg.raw; RAW_FILE_NAME=FullScan_Target06_pos.raw
SUBJECT_SAMPLE_FACTORS           	-	Target07	Capture Location:- | Reproductive Status:- | Sex:-	Pool #=7; RAW_FILE_NAME=FullScan_Target07_neg.raw; RAW_FILE_NAME=FullScan_Target07_pos.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Blood samples were collected from wild FL manatees at manatee health
CO:COLLECTION_SUMMARY            	assessments. Blood was collected in 7-mL lithium heparinized vacutainers and
CO:COLLECTION_SUMMARY            	centrifuged at 3200 rpm for 10 min onsite at atmospheric temperature. The plasma
CO:COLLECTION_SUMMARY            	supernatant was transferred to cryovials in 1 mL aliquots. Plasma samples were
CO:COLLECTION_SUMMARY            	kept on dry ice during transport and stored at -80 °C at USGS and University of
CO:COLLECTION_SUMMARY            	Florida (UF) until analysis.
CO:COLLECTION_PROTOCOL_FILENAME  	Collection Summary.txt
CO:SAMPLE_TYPE                   	Blood (plasma)
CO:STORAGE_CONDITIONS            	-80℃
CO:COLLECTION_VIALS              	lithium heparinized vacutainers
CO:STORAGE_VIALS                 	Cryovials
#TREATMENT
TR:TREATMENT_SUMMARY             	Plasma was not treated. Lipids were measured and compared between pregnant and
TR:TREATMENT_SUMMARY             	non-pregnant manatees.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Lithium heparinized manatee plasma samples, quality control samples, extraction
SP:SAMPLEPREP_SUMMARY            	and solvent blanks, and pooled group samples were extracted via the Folch
SP:SAMPLEPREP_SUMMARY            	extraction method (2:1 chloroform: methanol) (Folch et al., 1957). The plasma
SP:SAMPLEPREP_SUMMARY            	samples were thawed on ice and briefly homogenized. Next, 50 µL of each sample
SP:SAMPLEPREP_SUMMARY            	was added to a glass test tube. Three quality control samples were prepared the
SP:SAMPLEPREP_SUMMARY            	same manner using 50 µL of SRM 1950. All samples, except the solvent blanks,
SP:SAMPLEPREP_SUMMARY            	were spiked with 50 µL of deuterium-labeled internal standards from 10
SP:SAMPLEPREP_SUMMARY            	different lipid classes at known concentrations (diluted 1:10) (Splash
SP:SAMPLEPREP_SUMMARY            	Lipidomix, Avanti Polar Lipids, Alabaster, AL). Biphasic extraction was
SP:SAMPLEPREP_SUMMARY            	conducted by adding 3 mL of 2:1 chloroform:methanol solution to each test tube
SP:SAMPLEPREP_SUMMARY            	including the quality control samples and extraction blanks. The samples were
SP:SAMPLEPREP_SUMMARY            	then vortexed for 10 s, 500 µL of Optima-grade water was added, and the tubes
SP:SAMPLEPREP_SUMMARY            	were inverted 5 times. All samples were centrifuged at 2500 g for 5 min at 4
SP:SAMPLEPREP_SUMMARY            	°C. The resulting bottom lipid containing layer was then removed and
SP:SAMPLEPREP_SUMMARY            	transferred to a new glass test tube using glass Pasteur pipettes. Samples were
SP:SAMPLEPREP_SUMMARY            	divided into sub-groups determined by study site, sex, and reproductive status.
SP:SAMPLEPREP_SUMMARY            	Extract pools were created by equal contributions from each sample within a
SP:SAMPLEPREP_SUMMARY            	group. Finally, the samples, controls, and pools were evaporated under a steady
SP:SAMPLEPREP_SUMMARY            	stream of nitrogen at 25 °C for about 25 min. The samples were then
SP:SAMPLEPREP_SUMMARY            	reconstituted with 100 µL of isopropanol and vortexed for 10 s. The
SP:SAMPLEPREP_SUMMARY            	reconstituted solutions were then transferred to autosampler vial inserts inside
SP:SAMPLEPREP_SUMMARY            	glass autosampler vials for UHPLC-MS/MS analysis.
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Sample Prep.txt
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	The extracted lipid samples, pooled groups, quality controls, and blanks were
CH:CHROMATOGRAPHY_SUMMARY        	analyzed by UHPLC-MS/MS on a Thermo Vanquish UHPLC system coupled to a Thermo
CH:CHROMATOGRAPHY_SUMMARY        	Q-Exactive Orbitrap mass spectrometer. The column used was a Waters Acquity BEH,
CH:CHROMATOGRAPHY_SUMMARY        	100 mm x 2.1 mm x 1.7 µm. Mobile phase A and B contained 60:40
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile/water (v/v) and 90:10 isopropanol/acetonitrile (v/v) respectively,
CH:CHROMATOGRAPHY_SUMMARY        	with both phases containing 5mM ammonium formate and 0.1% formic acid. The
CH:CHROMATOGRAPHY_SUMMARY        	mobile phase gradient was as follows: a linear increase of solvent B from 40% to
CH:CHROMATOGRAPHY_SUMMARY        	55% over 7 min, then increased to 65% by min 8, held at 65% for 4 min, linear
CH:CHROMATOGRAPHY_SUMMARY        	increase to 95% over 8 min, increased to 100% over 2 min, solvent B was held at
CH:CHROMATOGRAPHY_SUMMARY        	100% for 5 min before decreasing to 40% in a tenth of a min. Finally, solvent B
CH:CHROMATOGRAPHY_SUMMARY        	was held at 100% for another 3 min. The sample injection volume was 10 µL at a
CH:CHROMATOGRAPHY_SUMMARY        	flow rate of 0.25 mL/min, the column compartment was 45 °C, and the autosampler
CH:CHROMATOGRAPHY_SUMMARY        	temperature was 10 °C.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um)
CH:SOLVENT_A                     	60:40 acetonitrile/water (v/v), 5mM ammonium formate and 0.1% formic acid
CH:SOLVENT_B                     	90:10 isopropanol/acetonitrile (v/v), 5mM ammonium formate and 0.1% formic acid
CH:FLOW_GRADIENT                 	a linear increase of solvent B from 40% to 55% over 7 min, then increased to 65%
CH:FLOW_GRADIENT                 	by min 8, held at 65% for 4 min, linear increase to 95% over 8 min, increased to
CH:FLOW_GRADIENT                 	100% over 2 min, solvent B was held at 100% for 5 min before decreasing to 40%
CH:FLOW_GRADIENT                 	in a tenth of a min. Finally, solvent B was held at 100% for another 3 min.
CH:FLOW_RATE                     	0.25 mL/min
CH:COLUMN_TEMPERATURE            	45
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	MS Metadata.txt
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	All samples were analyzed in full scan positive and negative ion modes with
MS:MS_COMMENTS                   	spray voltages +3.0kV and -3.0kV, respectively, at 70,000 resolution at m/z 200,
MS:MS_COMMENTS                   	the scan range was m/z 100-1500 and source temperature at 300 °C. Tandem mass
MS:MS_COMMENTS                   	spectra were collected via criterion based on data-dependent acquisition for the
MS:MS_COMMENTS                   	top-10 ions with stepped normalized collision energy 20, 25, and 30, an
MS:MS_COMMENTS                   	isolation window of 1Da, dynamic exclusion of 6 s, and using IE-Omics for the
MS:MS_COMMENTS                   	generation of exclusion lists in iterative-exclusion experiments (Koelmel,
MS:MS_COMMENTS                   	Kroeger, Gill, et al., 2017). Extracted Lipid Data Analysis LipidMatch software
MS:MS_COMMENTS                   	was implemented to identify and integrate the detected lipids (Koelmel, Kroeger,
MS:MS_COMMENTS                   	Ulmer, et al., 2017). The peak areas were normalized relative to the internal
MS:MS_COMMENTS                   	standards of the same subclass as the lipid of interest. For lipids that did not
MS:MS_COMMENTS                   	have an internal standard of the same subclass, the internal standard with the
MS:MS_COMMENTS                   	closest chemical structure was used (i.e., internal standard with the same head
MS:MS_COMMENTS                   	group). Otherwise, the internal standard with the closest chromatographic
MS:MS_COMMENTS                   	retention time (same ionization polarity) was used for analytes without a
MS:MS_COMMENTS                   	corresponding internal standard of the same lipid subclass. The plasma lipid
MS:MS_COMMENTS                   	concentrations were reported as µg/mL.
MS:MS_RESULTS_FILE               	ST003017_AN004951_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Seconds
#END