{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000612","ANALYSIS_ID":"AN000937","VERSION":"1","CREATED_ON":"May 28, 2017, 1:05 pm"},

"PROJECT":{"PROJECT_TITLE":"Sphingolipid Profiling of Human Aqueous Humor and Trabecular Meshwork in Glaucomatous and Control eyes","PROJECT_SUMMARY":"Using a triple quadrupole mass spectrometer and direct infusion lipidomics we identified various sphingolipids present in both the aqueous humor and trabecular meshwork of the human eye and compared them between glaucomatous and control groups.","INSTITUTE":"University of Miami","LAST_NAME":"Bhattacharya","FIRST_NAME":"Sanjoy","ADDRESS":"1638 NW 10th Avenue, Miami, Florida, 33136, USA","EMAIL":"sbhattacharya@med.miami.edu","PHONE":"3054824103"},

"STUDY":{"STUDY_TITLE":"Sphingolipid Analysis of Human Aqueous Humor in Glaucomatous and Control eyes","STUDY_SUMMARY":"This study examined the profiles of sphin¬golipids and ceramides present in the aqueous humor (AH) of human control and POAG donors. Furthermore, we quantitatively compared distinct differences between glaucomatous and age-matched control eyes, identifying potential molecules for further experimentation to determine their biological role in modulating cell behavior. Lipids were identified and ratiometrically quantified in a two-step process using precursor ion scan (PIS) or neutral loss scan (NLS) with appropriate class-specific lipid standards on a TSQ Quantum Access Max mass spectrometer following established procedures. We identified several species of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramides that were common between control and glaucomatous AQH. Some unique lipid species in these classes were also identified in controls but not in glaucoma and vice versa.","INSTITUTE":"University of Miami","LAST_NAME":"Bhattacharya","FIRST_NAME":"Sanjoy","ADDRESS":"1638 NW 10th Avenue, Miami, Florida, 33136, USA","EMAIL":"sbhattacharya@med.miami.edu","PHONE":"3054824103"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"UGS039",
"Sample ID":"NAH1BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"65","Gender":"F","Race":"H","Medications":"Lisinorpil, Amlodipine"}
},
{
"Subject ID":"UGS040",
"Sample ID":"NAH2BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"80","Gender":"M","Race":"W","Medications":"Aspirin, Zocar, Vasotec, Persantine, Enalapril, Proscar, Glucosamine, MSM, Mycaterourt (sp? Illegible)"}
},
{
"Subject ID":"UGS041",
"Sample ID":"NAH3BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"58","Gender":"F","Race":"H","Medications":"Vitamins, Aspirin, Glucosamine, Mylanta, Promethazine Codeine"}
},
{
"Subject ID":"UGS042",
"Sample ID":"NAH4BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"64","Gender":"M","Race":"N/A","Medications":"N/A"}
},
{
"Subject ID":"UGS043",
"Sample ID":"NAH5BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"82","Gender":"F","Race":"W","Medications":"Nexium, Lipitor, Synthroid"}
},
{
"Subject ID":"UGS044",
"Sample ID":"NAH6BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"63","Gender":"M","Race":"HW","Medications":"Micardis, Corgard, Crestor, Clonazepam, Zyprexa, Lithium"}
},
{
"Subject ID":"UGS045",
"Sample ID":"NAH7BL4",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"78","Gender":"F","Race":"HB","Medications":"Norvasc, Furosemide, Monopril, Lanoxin, Lipitor, Captopril, Celebrex, Centurium, Lisinopril, Atenolol, Baby Aspirin"}
},
{
"Subject ID":"UGS046",
"Sample ID":"NAH8BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"86","Gender":"M","Race":"W","Medications":"Detrol, One-A-Day men, Zocor, Avandia, Coreg, Tylenol 3,"}
},
{
"Subject ID":"UGS047",
"Sample ID":"NAH9BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"71","Gender":"F","Race":"N/A","Medications":"N/A"}
},
{
"Subject ID":"UGS048",
"Sample ID":"NAH10BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"70","Gender":"F","Race":"N/A","Medications":"Zyrtec, Diltiazem, AcipHex (sp?), Premarin, Trazodone, Vytorin, Pomperidone, Zelnorm, Ester-C"}
},
{
"Subject ID":"UGS049",
"Sample ID":"NAH11BL4",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"62","Gender":"M","Race":"B","Medications":"Insulin, Cardizem, Plavix, Prevacid, Protonix, Enalapril, Crestor, Glucophage, Vasotec, Prilosec, Amoxicillin, Loperamide, Novolin, Lantus, Neurotin, Albuterol"}
},
{
"Subject ID":"UGS050",
"Sample ID":"NAH12BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"44","Gender":"F","Race":"N/A","Medications":"N/A"}
},
{
"Subject ID":"UGS051",
"Sample ID":"NAH13BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"52","Gender":"M","Race":"W","Medications":"N/A"}
},
{
"Subject ID":"UGS052",
"Sample ID":"NAH14BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"81","Gender":"F","Race":"W","Medications":"N/A"}
},
{
"Subject ID":"UGS053",
"Sample ID":"NAH15BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"72","Gender":"F","Race":"W","Medications":"PF, Vigamox, Acular Pamelor, CQ10, Lipitor, Lunesta, Calcium, PF, Aspirin, Fish Oil"}
},
{
"Subject ID":"UGS054",
"Sample ID":"NAH16BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"79","Gender":"M","Race":"HW","Medications":"Acular, Pred Forte, Vigamox, Synthroid, Metropolol, Norvasc, Digoxin"}
},
{
"Subject ID":"UGS055",
"Sample ID":"NAH17BL2",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"70","Gender":"F","Race":"H","Medications":"Vigamox, Acular, Prednisolone, Cardizem, Glipizide, Simvastatin, Avapro, Labetalol"}
},
{
"Subject ID":"UGS056",
"Sample ID":"NAH18BL2",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"74","Gender":"M","Race":"N/A","Medications":"Acular, Pred Forte, Vigamox, PF, Stalevo, Mirapex, Amantadine, Sanctura, Seroquel"}
},
{
"Subject ID":"UGS(V)080",
"Sample ID":"NAH19BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"56","Gender":"F","Race":"W","Medications":"Atrovastatin, Cardizem, Glipizide"}
},
{
"Subject ID":"UGS(V)081",
"Sample ID":"NAH20BL3",
"Factors":{"Condition":"Control"},
"Additional sample data":{"Age":"70","Gender":"M","Race":"B","Medications":"Not Known"}
},
{
"Subject ID":"UGS057",
"Sample ID":"GAH1BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"82","Gender":"F","Race":"HW","Medications":"Travatan, Cosopt, Alphagan"}
},
{
"Subject ID":"UGS058",
"Sample ID":"GAH2BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"81","Gender":"F","Race":"N/A","Medications":"Cosopt, Xalatan"}
},
{
"Subject ID":"UGS059",
"Sample ID":"GAH3BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"77","Gender":"F","Race":"H","Medications":"Alphagan, Xalatan, Procardia, Mycardis, Singulair, Lipitor, Advair"}
},
{
"Subject ID":"UGS060",
"Sample ID":"GAH4BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"80","Gender":"F","Race":"HW","Medications":"Travatan, Cosopt, Avapro, Meprobamato, ASA, Clarinex"}
},
{
"Subject ID":"UGS061",
"Sample ID":"GAH5BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"78","Gender":"F","Race":"B","Medications":"Cosopt, Xalatan, Cozaar, Calcium, Baby Aspirin, Fosamax, Norvasc, Celebrax, Lipitor, Tylenol"}
},
{
"Subject ID":"UGS062",
"Sample ID":"GAH6BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"46","Gender":"F","Race":"B","Medications":"Cosopt, Lumigan, AIDs meds per pt. Everything else is illegible"}
},
{
"Subject ID":"UGS063",
"Sample ID":"GAH7BL4",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"78","Gender":"M","Race":"W","Medications":"Xalatan, Coumadin, Prednisone, Maxzide, Folic Acid, Vitamin E, Vitamin B6"}
},
{
"Subject ID":"UGS064",
"Sample ID":"GAH8BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"81","Gender":"F","Race":"W","Medications":"Alphagan, Trusopt, Xalatan, Synthroid, Tamoxifen, Altace, Vit. C - Beta Carotene, Vit. E, Aspirin, Calcium, Avapro, Lipitor, Lumigan, Alphagan P, BID"}
},
{
"Subject ID":"UGS065",
"Sample ID":"GAH9BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"83","Gender":"M","Race":"N/A","Medications":"Cosopt, Travatan Namenda, Nitro Patch, Atenolol, Coumadin/Warfarin, Lipitor, Triamterene, Water pill, Maxide, Meloxicam"}
},
{
"Subject ID":"UGS066",
"Sample ID":"GAH10BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"60","Gender":"M","Race":"B","Medications":"Cosopt, Synthroid, Glucotrol, Zocor"}
},
{
"Subject ID":"UGS067",
"Sample ID":"GAH11BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"62","Gender":"M","Race":"HB","Medications":"Xalatan, Timolol"}
},
{
"Subject ID":"UGS068",
"Sample ID":"GAH12BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"76","Gender":"M","Race":"N/A","Medications":"Cosopt, Diovan, Protonix"}
},
{
"Subject ID":"UGS069",
"Sample ID":"GAH13BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"78","Gender":"F","Race":"H","Medications":"Alphagan"}
},
{
"Subject ID":"UGS070",
"Sample ID":"GAH14BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"72","Gender":"F","Race":"B","Medications":"Xalatan, Cosopt, Aricept: 5mg, Micardis HCT:N/A, Bayer: N/A, Vytorin: 10/40,Norvasc: 5 mg, Novolin Insulin: 70/30, HCT2: 25 mg, Tramadol: 50mg, Catapres: 3 patches, Ecotrin: 81mg, Nifedipine CR: 60mg, Nexium: N/A, Atenolol: 25mg, Lisinopril: 10mg, Oxycodone/APAP: N"}
},
{
"Subject ID":"UGS071",
"Sample ID":"GAH15BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"82","Gender":"F","Race":"HW","Medications":"Travatan: 1 drop (OD) - bedtime, Alphagan: 1 drop (OD) - 2x/day, Dorzolamide: 1 drop (OD) - 2x/day, Ofloxacin: 1 drop (OS ) - 4x/day, Prednisolone: 1 drop (OS) - 4x/day, Vigamox: (OS), Paraoxetine: 1 tab. daily, Namenda: 1 tab. daily, Avandia: 1"}
},
{
"Subject ID":"UGS072",
"Sample ID":"GAH16BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"69","Gender":"F","Race":"W","Medications":"Travatan, Alphagan, Metformin: 2x/day, Aricept: N/A, Ibuprofen: N/A, Abilify: N/A, Lexapro: N/A, Zanax: N/A"}
},
{
"Subject ID":"UGS073",
"Sample ID":"GAH17BL4",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"78","Gender":"M","Race":"N/A","Medications":"Timolol, Xalatan"}
},
{
"Subject ID":"UGS074",
"Sample ID":"GAH18BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"66","Gender":"M","Race":"N/A","Medications":"Cosopt, Xalatan"}
},
{
"Subject ID":"UGS075",
"Sample ID":"GAH19BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"78","Gender":"F","Race":"H","Medications":"Timolol, Xalatan, Synthroid, Calcium, ASA"}
},
{
"Subject ID":"UGS076",
"Sample ID":"GAH20BL3",
"Factors":{"Condition":"Glaucomatous"},
"Additional sample data":{"Age":"65","Gender":"F","Race":"B","Medications":"Cosopt, Xalatan, Vigamox, Pred forte, Norvasc: 10 mg - 2x/day, Motrin: 800mg - 2x/day, Humalog: 70/30 - 2x/day, Metformin: 5 mg - 2x/day, 3 illegible medications"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Aqueous humor from POAG and control human subjects (n=20 each totaling 40 samples) were procured following institutional review board approved protocols and adhering to the tenets of the Declaration of Helsinki. AH samples were stored at −80 °C until time of use. The mean age of donors was 71.2 +/- 10.2 and both genders were included in these studies."},

"TREATMENT":{"TREATMENT_SUMMARY":"No treatment was applied to the groups."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Aqueous humor was subjected to lipid extraction using a modified Bligh and Dyer method. Prior to extraction, a fixed amount of a standard (1,2-ditridecanoyl-sn-glycero-3-phosphocholine) was added to determine lipid recovery, ensuring >99% and uniform recovery across all samples analyzed. The organic phase with extracted lipids was flushed with argon gas to prevent oxidation and dried in a Speed-Vac (Model 5301, Brinkmann Instruments Inc., Westbury, NY). The aqueous phase containing proteins was stored at −80 °C for protein quantification. All extractions and subsequent handling were carried out using glass vials to a void contamination from plastics. Extracted lipids were dried and resuspended in liquid chromatography–mass spectrometry (LC-MS) grade acetonitrile:isopropanol (1:1). A triple quadrupole electrospray mass spectrometer (TSQ Quantum Access Max; Thermo Fisher Scientific, Pittsburgh, PA) was used for analysis of lipids in infusion mode using the TSQ Tune of Xcalibur 2.3 software package. Samples were infused with a flow rate of 10 μl/ min and analyzed for 1.00 min with a 0.500 s scan. Scans typically ranged from 200 m/z to 1,000 m/z unless specified otherwise. The full width at half maximum peak was set at 0.7 and collision gas pressure was set at 1 mTorr. Sheath gas (nitrogen) was set to 20 arbitrary units. Auxiliary gas (argon) was set to 5 arbitrary units. For analyses of the sphingomyelin, sphingoid base, and ceramide classes, the identifications were performed using neutral loss scan (NLS) for m/z 213.2, 48 and 256.2 with collision energies of 50, 18 and 32 V, respectively; except for ceramide (in negative ion mode), all other scans were carried out in positive mode. For sphingoid base-1-phosphate, PIS was performed for product ion m/z of 79.1 in negative ion mode at 24 V collision energy. The spray voltage, ion mode, and collision energies were based on previous studies. The analytical parameters for sphingolipids and ceramides described here are based on standardized collision energy settings as suggested in the recent literature for automated shotgun lipidomics."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"None (Direct infusion)","INSTRUMENT_NAME":"TSQ Quantum Access Max","COLUMN_NAME":"none"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Thermo Quantiva Access Max","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED"},

"MS_METABOLITE_DATA":{
"Units":"Peak Area",

"Data":[{"Metabolite":"Sphingosine","NAH1BL3":"121482.1","NAH2BL3":"","NAH3BL3":"","NAH4BL3":"22268.9","NAH5BL3":"3196.2","NAH6BL3":"3196.2","NAH7BL4":"19738.3","NAH8BL3":"","NAH9BL3":"","NAH10BL3":"33169.3","NAH11BL4":"","NAH12BL3":"51934.5","NAH13BL3":"236020.8","NAH14BL3":"56959.1","NAH15BL3":"519089","NAH16BL3":"53822.9","NAH17BL2":"39013.1","NAH18BL2":"3999.5","NAH19BL3":"34141.3","NAH20BL3":"","GAH1BL3":"56257.7","GAH2BL3":"74360.2","GAH3BL3":"129760.1","GAH4BL3":"74635.2","GAH5BL3":"7227.9","GAH6BL3":"354238.7","GAH7BL4":"211802.8","GAH8BL3":"3401.2","GAH9BL3":"109474.4","GAH10BL3":"283548","GAH11BL3":"","GAH12BL3":"35270.6","GAH13BL3":"83770.4","GAH14BL3":"13414.7","GAH15BL3":"19226","GAH16BL3":"475104","GAH17BL4":"541955.5","GAH18BL3":"8423.2","GAH19BL3":"1033.8","GAH20BL3":""},{"Metabolite":"Penazetidine A","NAH1BL3":"","NAH2BL3":"","NAH3BL3":"","NAH4BL3":"","NAH5BL3":"","NAH6BL3":"","NAH7BL4":"","NAH8BL3":"","NAH9BL3":"","NAH10BL3":"39115.4","NAH11BL4":"","NAH12BL3":"","NAH13BL3":"","NAH14BL3":"","NAH15BL3":"","NAH16BL3":"","NAH17BL2":"","NAH18BL2":"","NAH19BL3":"","NAH20BL3":"","GAH1BL3":"","GAH2BL3":"","GAH3BL3":"92324.3","GAH4BL3":"102644.2","GAH5BL3":"","GAH6BL3":"9863.1","GAH7BL4":"11585","GAH8BL3":"234433.4","GAH9BL3":"128139.1","GAH10BL3":"96331.9","GAH11BL3":"","GAH12BL3":"","GAH13BL3":"","GAH14BL3":"","GAH15BL3":"","GAH16BL3":"","GAH17BL4":"","GAH18BL3":"","GAH19BL3":"","GAH20BL3":""},{"Metabolite":"N,N-dimethylsphingosine","NAH1BL3":"","NAH2BL3":"13334.2","NAH3BL3":"24456.4","NAH4BL3":"42947","NAH5BL3":"","NAH6BL3":"","NAH7BL4":"4268.9","NAH8BL3":"","NAH9BL3":"56324.2","NAH10BL3":"29672","NAH11BL4":"","NAH12BL3":"73040.6","NAH13BL3":"212663.9","NAH14BL3":"342285","NAH15BL3":"444789.6","NAH16BL3":"42481.3","NAH17BL2":"","NAH18BL2":"382044.3","NAH19BL3":"31662.5","NAH20BL3":"53094.6","GAH1BL3":"4033.7","GAH2BL3":"36526.8","GAH3BL3":"69693.5","GAH4BL3":"5807.8","GAH5BL3":"4877","GAH6BL3":"75786.4","GAH7BL4":"36681.8","GAH8BL3":"24931.6","GAH9BL3":"1184.8","GAH10BL3":"93275.2","GAH11BL3":"37848.9","GAH12BL3":"245410","GAH13BL3":"165236.3","GAH14BL3":"216079","GAH15BL3":"262487.2","GAH16BL3":"152788.2","GAH17BL4":"25967.7","GAH18BL3":"390841.5","GAH19BL3":"","GAH20BL3":"16571.8"},{"Metabolite":"Capnine","NAH1BL3":"","NAH2BL3":"16500.4","NAH3BL3":"","NAH4BL3":"","NAH5BL3":"","NAH6BL3":"","NAH7BL4":"","NAH8BL3":"","NAH9BL3":"","NAH10BL3":"","NAH11BL4":"","NAH12BL3":"","NAH13BL3":"","NAH14BL3":"","NAH15BL3":"","NAH16BL3":"","NAH17BL2":"","NAH18BL2":"","NAH19BL3":"","NAH20BL3":"","GAH1BL3":"","GAH2BL3":"37487.6","GAH3BL3":"64653.7","GAH4BL3":"","GAH5BL3":"9027.4","GAH6BL3":"21510.7","GAH7BL4":"4085.6","GAH8BL3":"86601.6","GAH9BL3":"37247.9","GAH10BL3":"59200.4","GAH11BL3":"","GAH12BL3":"","GAH13BL3":"","GAH14BL3":"","GAH15BL3":"","GAH16BL3":"","GAH17BL4":"","GAH18BL3":"","GAH19BL3":"","GAH20BL3":""},{"Metabolite":"(4E,6E,d14:2) sphingosine","NAH1BL3":"25832.2","NAH2BL3":"","NAH3BL3":"","NAH4BL3":"8809.3","NAH5BL3":"21507","NAH6BL3":"21507","NAH7BL4":"","NAH8BL3":"","NAH9BL3":"","NAH10BL3":"","NAH11BL4":"","NAH12BL3":"","NAH13BL3":"8762.5","NAH14BL3":"7438.1","NAH15BL3":"23153.9","NAH16BL3":"25767.1","NAH17BL2":"4702","NAH18BL2":"21702.7","NAH19BL3":"58403.3","NAH20BL3":"","GAH1BL3":"7689.2","GAH2BL3":"56863.4","GAH3BL3":"59369","GAH4BL3":"11631.6","GAH5BL3":"33671.3","GAH6BL3":"3698.1","GAH7BL4":"28071.8","GAH8BL3":"14839.6","GAH9BL3":"331824.2","GAH10BL3":"649405.2","GAH11BL3":"","GAH12BL3":"17893.4","GAH13BL3":"1350.5","GAH14BL3":"","GAH15BL3":"16013.1","GAH16BL3":"","GAH17BL4":"1186.8","GAH18BL3":"","GAH19BL3":"","GAH20BL3":"6279.5"},{"Metabolite":"C16 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