Summary of Study ST002758
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001719. The data can be accessed directly via it's Project DOI: 10.21228/M8HF01 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
Study ID | ST002758 |
Study Title | Metabolic responses of normal rat kidneys to a high salt intake (Plasma) |
Study Type | Time-course metabolomics experiment |
Study Summary | In this study, novel methods were developed which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic profiles before and following the switch from a 0.4% to 4.0% NaCl diet. |
Institute | Medical College of Wisconsin |
Department | Physiology |
Laboratory | Dr. Allen W. Cowley |
Last Name | Cowley |
First Name | Allen |
Address | 8701 W. Watertown Plank Rd, Milwaukee, WI 53226 |
cowley@mcw.edu | |
Phone | 4149558277 |
Submit Date | 2023-06-26 |
Raw Data Available | Yes |
Raw Data File Type(s) | mzML |
Analysis Type Detail | LC-MS |
Release Date | 2023-07-02 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
Project ID: | PR001719 |
Project DOI: | doi: 10.21228/M8HF01 |
Project Title: | SD Rat Metabolomics in Response to Salt |
Project Type: | Untargeted Four-Mode Metabolomics |
Project Summary: | This study analyzed the effects of a high salt (HS; 4.0% NaCl) diet upon the kidney, arterial plasma, and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. |
Institute: | Medical College of Wisconsin |
Department: | Physiology |
Laboratory: | Dr. Allen W. Cowley |
Last Name: | Cowley |
First Name: | Allen |
Address: | 8701 W. Watertown Plank Rd, Milwaukee, WI 53226 |
Email: | cowley@mcw.edu |
Phone: | 414-955-8277 |
Funding Source: | NHLBI |
Contributors: | Satoshi Shimada, Brian R. Hoffmann, Chun Yang, Theresa Kurth, Andrew S. Greene, Mingyu Liang, Ranjan K. Dash, Allen W. Cowley Jr |
Subject:
Subject ID: | SU002865 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Species Group: | Mammals |
Factors:
Subject type: Mammal; Subject species: Rattus norvegicus (Factor headings shown in green)
mb_sample_id | local_sample_id | Treatment | Source |
---|---|---|---|
SA290237 | 20211007_Cowley3_Plasma_22_73-434-HS14-A_C18pos | HS14 | Artery Plasma |
SA290238 | 20211013_Cowley3_Plasma_28_61-429-HS14-A_C18neg_rep | HS14 | Artery Plasma |
SA290239 | 20211020_Cowley3_Plasma_2_49-421-HS14-A_HILICpos_rep | HS14 | Artery Plasma |
SA290240 | 20211020_Cowley3_Plasma_9_85-463-HS14-A_HILICpos_rep | HS14 | Artery Plasma |
SA290241 | 20211020_Cowley3_Plasma_23_73-434-HS14-A_HILICpos | HS14 | Artery Plasma |
SA290242 | 20211013_Cowley3_Plasma_28_61-429-HS14-A_C18neg | HS14 | Artery Plasma |
SA290243 | 20211020_Cowley3_Plasma_9_85-463-HS14-A_HILICpos | HS14 | Artery Plasma |
SA290244 | 20211013_Cowley3_Plasma_27_37-417-HS14-A_C18neg | HS14 | Artery Plasma |
SA290245 | 20211013_Cowley3_Plasma_18_49-421-HS14-A_C18neg | HS14 | Artery Plasma |
SA290246 | 20211013_Cowley3_Plasma_16_85-463-HS14-A_C18neg_rep | HS14 | Artery Plasma |
SA290247 | 20211013_Cowley3_Plasma_18_49-421-HS14-A_C18neg_rep | HS14 | Artery Plasma |
SA290248 | 20211013_Cowley3_Plasma_21_73-434-HS14-A_C18neg | HS14 | Artery Plasma |
SA290249 | 20211020_Cowley3_Plasma_23_73-434-HS14-A_HILICpos_rep | HS14 | Artery Plasma |
SA290250 | 20211013_Cowley3_Plasma_21_73-434-HS14-A_C18neg_rep | HS14 | Artery Plasma |
SA290251 | 20211013_Cowley3_Plasma_27_37-417-HS14-A_C18neg_rep | HS14 | Artery Plasma |
SA290252 | 20211020_Cowley3_Plasma_27_37-417-HS14-A_HILICpos | HS14 | Artery Plasma |
SA290253 | 20211029_Cowley3_Plasma_32_37-417-HS14-A_HILICneg_rep | HS14 | Artery Plasma |
SA290254 | 20211029_Cowley3_Plasma_32_37-417-HS14-A_HILICneg | HS14 | Artery Plasma |
SA290255 | 20211029_Cowley3_Plasma_34_73-434-HS14-A_HILICneg | HS14 | Artery Plasma |
SA290256 | 20211029_Cowley3_Plasma_34_73-434-HS14-A_HILICneg_rep | HS14 | Artery Plasma |
SA290257 | 20211029_Cowley3_Plasma_39_49-421-HS14-A_HILICneg_rep | HS14 | Artery Plasma |
SA290258 | 20211029_Cowley3_Plasma_39_49-421-HS14-A_HILICneg | HS14 | Artery Plasma |
SA290259 | 20211029_Cowley3_Plasma_28_61-429-HS14-A_HILICneg_rep | HS14 | Artery Plasma |
SA290260 | 20211029_Cowley3_Plasma_28_61-429-HS14-A_HILICneg | HS14 | Artery Plasma |
SA290261 | 20211020_Cowley3_Plasma_39_61-429-HS14-A_HILICpos | HS14 | Artery Plasma |
SA290262 | 20211020_Cowley3_Plasma_27_37-417-HS14-A_HILICpos_rep | HS14 | Artery Plasma |
SA290263 | 20211020_Cowley3_Plasma_39_61-429-HS14-A_HILICpos_rep | HS14 | Artery Plasma |
SA290264 | 20211029_Cowley3_Plasma_10_85-463-HS14-A_HILICneg | HS14 | Artery Plasma |
SA290265 | 20211029_Cowley3_Plasma_10_85-463-HS14-A_HILICneg_rep | HS14 | Artery Plasma |
SA290266 | 20211013_Cowley3_Plasma_16_85-463-HS14-A_C18neg | HS14 | Artery Plasma |
SA290267 | 20211020_Cowley3_Plasma_2_49-421-HS14-A_HILICpos | HS14 | Artery Plasma |
SA290268 | 20211007_Cowley3_Plasma_29_37-417-HS14-A_C18pos | HS14 | Artery Plasma |
SA290269 | 20211007_Cowley3_Plasma_29_37-417_HS14-A_C18pos_rep | HS14 | Artery Plasma |
SA290270 | 20211007_Cowley3_Plasma_31_85-463-HS14-A_C18pos | HS14 | Artery Plasma |
SA290271 | 20211007_Cowley3_Plasma_31_85-463-HS14-A_C18pos_rep | HS14 | Artery Plasma |
SA290272 | 20211007_Cowley3_Plasma_37_49-421-HS14-A_C18pos_rep | HS14 | Artery Plasma |
SA290273 | 20211007_Cowley3_Plasma_37_49-421-HS14-A_C18pos | HS14 | Artery Plasma |
SA290274 | 20211007_Cowley3_Plasma_28_61-429-HS14-A_C18pos | HS14 | Artery Plasma |
SA290275 | 20211007_Cowley3_Plasma_28_61-429-HS14-A_C18pos_rep | HS14 | Artery Plasma |
SA290276 | 20211007_Cowley3_Plasma_22_73-434-HS14-A_C18pos_rep | HS14 | Artery Plasma |
SA290277 | 20211007_Cowley3_Plasma_20_86-463-HS14-RV_C18pos | HS14 | Vein Plasma |
SA290278 | 20211007_Cowley3_Plasma_20_86-463-HS14-RV_C18pos_rep | HS14 | Vein Plasma |
SA290279 | 20211007_Cowley3_Plasma_24_50-421-HS14-RV_C18pos | HS14 | Vein Plasma |
SA290280 | 20211007_Cowley3_Plasma_11_62-429-HS14-RV_C18pos_rep | HS14 | Vein Plasma |
SA290281 | 20211007_Cowley3_Plasma_11_62-429-HS14-RV_C18pos | HS14 | Vein Plasma |
SA290282 | 20211020_Cowley3_Plasma_7_74-434-HS14-RV_HILICpos | HS14 | Vein Plasma |
SA290283 | 20211007_Cowley3_Plasma_7_38-417-HS14-RV_C18pos | HS14 | Vein Plasma |
SA290284 | 20211007_Cowley3_Plasma_7_38-417-HS14-RV_C18pos_rep | HS14 | Vein Plasma |
SA290285 | 20211013_Cowley3_Plasma_10_86-463-HS14-RV_C18neg_rep | HS14 | Vein Plasma |
SA290286 | 20211013_Cowley3_Plasma_11_38-417-HS14-RV_C18neg | HS14 | Vein Plasma |
SA290287 | 20211007_Cowley3_Plasma_24_50-421-HS14-RV_C18pos_rep | HS14 | Vein Plasma |
SA290288 | 20211013_Cowley3_Plasma_34_74-434-HS14-RV_C18neg_rep | HS14 | Vein Plasma |
SA290289 | 20211013_Cowley3_Plasma_34_74-434-HS14-RV_C18neg | HS14 | Vein Plasma |
SA290290 | 20211013_Cowley3_Plasma_31_50-421-HS14-RV_C18neg | HS14 | Vein Plasma |
SA290291 | 20211020_Cowley3_Plasma_7_74-434-HS14-RV_HILICpos_rep | HS14 | Vein Plasma |
SA290292 | 20211013_Cowley3_Plasma_11_38-417-HS14-RV_C18neg_rep | HS14 | Vein Plasma |
SA290293 | 20211013_Cowley3_Plasma_13_62-429-HS14-RV_C18neg | HS14 | Vein Plasma |
SA290294 | 20211013_Cowley3_Plasma_13_62-429-HS14-RV_C18neg_rep | HS14 | Vein Plasma |
SA290295 | 20211013_Cowley3_Plasma_31_50-421-HS14-RV_C18neg_rep | HS14 | Vein Plasma |
SA290296 | 20211020_Cowley3_Plasma_33_38-417-HS14-RV_HILICpos | HS14 | Vein Plasma |
SA290297 | 20211029_Cowley3_Plasma_11_62-429-HS14-RV_HILICneg | HS14 | Vein Plasma |
SA290298 | 20211029_Cowley3_Plasma_8_50-421-HS14-RV_HILICneg_rep | HS14 | Vein Plasma |
SA290299 | 20211029_Cowley3_Plasma_8_50-421-HS14-RV_HILICneg | HS14 | Vein Plasma |
SA290300 | 20211029_Cowley3_Plasma_11_62-429-HS14-RV_HILICneg_rep | HS14 | Vein Plasma |
SA290301 | 20211029_Cowley3_Plasma_16_86-463-HS14-RV_HILICneg | HS14 | Vein Plasma |
SA290302 | 20211029_Cowley3_Plasma_31_38-417-HS14-RV_HILICneg_rep | HS14 | Vein Plasma |
SA290303 | 20211029_Cowley3_Plasma_31_38-417-HS14-RV_HILICneg | HS14 | Vein Plasma |
SA290304 | 20211029_Cowley3_Plasma_16_86-463-HS14-RV_HILICneg_rep | HS14 | Vein Plasma |
SA290305 | 20211029_Cowley3_Plasma_5_74-434-HS14-RV_HILICneg_rep | HS14 | Vein Plasma |
SA290306 | 20211029_Cowley3_Plasma_5_74-434-HS14-RV_HILICneg | HS14 | Vein Plasma |
SA290307 | 20211020_Cowley3_Plasma_25_50-421-HS14-RV_HILICpos_rep | HS14 | Vein Plasma |
SA290308 | 20211020_Cowley3_Plasma_25_50-421-HS14-RV_HILICpos | HS14 | Vein Plasma |
SA290309 | 20211020_Cowley3_Plasma_22_62-429-HS14-RV_HILICpos_rep | HS14 | Vein Plasma |
SA290310 | 20211020_Cowley3_Plasma_30_86-463-HS14-RV_HILICpos_rep | HS14 | Vein Plasma |
SA290311 | 20211013_Cowley3_Plasma_10_86-463-HS14-RV_C18neg | HS14 | Vein Plasma |
SA290312 | 20211007_Cowley3_Plasma_1_74-434-HS14-RV_C18pos | HS14 | Vein Plasma |
SA290313 | 20211007_Cowley3_Plasma_1_74-434-HS14-RV_C18pos_rep | HS14 | Vein Plasma |
SA290314 | 20211020_Cowley3_Plasma_33_38-417-HS14-RV_HILICpos_rep | HS14 | Vein Plasma |
SA290315 | 20211020_Cowley3_Plasma_22_62-429-HS14-RV_HILICpos | HS14 | Vein Plasma |
SA290316 | 20211020_Cowley3_Plasma_30_86-463-HS14-RV_HILICpos | HS14 | Vein Plasma |
SA290317 | 20211029_Cowley3_Plasma_24_40-417-HS21-A_HILICneg_rep | HS21 | Artery Plasma |
SA290318 | 20211029_Cowley3_Plasma_25_52-421-HS21-A_HILICneg | HS21 | Artery Plasma |
SA290319 | 20211029_Cowley3_Plasma_24_40-417-HS21-A_HILICneg | HS21 | Artery Plasma |
SA290320 | 20211029_Cowley3_Plasma_23_76-434-HS21-A_HILICneg_rep | HS21 | Artery Plasma |
SA290321 | 20211029_Cowley3_Plasma_23_76-434-HS21-A_HILICneg | HS21 | Artery Plasma |
SA290322 | 20211029_Cowley3_Plasma_25_52-421-HS21-A_HILICneg_rep | HS21 | Artery Plasma |
SA290323 | 20211029_Cowley3_Plasma_26_88-463-HS21-A_HILICneg | HS21 | Artery Plasma |
SA290324 | 20211007_Cowley3_Plasma_30_52-421-HS21-A_C18pos | HS21 | Artery Plasma |
SA290325 | 20211007_Cowley3_Plasma_30_52-421-HS21-A_C18pos_rep | HS21 | Artery Plasma |
SA290326 | 20211007_Cowley3_Plasma_12_40-417-HS21-A_C18pos | HS21 | Artery Plasma |
SA290327 | 20211007_Cowley3_Plasma_10_88-463-HS21-A_C18pos_rep | HS21 | Artery Plasma |
SA290328 | 20211007_Cowley3_Plasma_10_88-463-HS21-A_C18pos | HS21 | Artery Plasma |
SA290329 | 20211029_Cowley3_Plasma_12_64-429-HS21-A_HILICneg_rep | HS21 | Artery Plasma |
SA290330 | 20211029_Cowley3_Plasma_12_64-429-HS21-A_HILICneg | HS21 | Artery Plasma |
SA290331 | 20211013_Cowley3_Plasma_37_52-421-HS21-A_C18neg_rep | HS21 | Artery Plasma |
SA290332 | 20211013_Cowley3_Plasma_37_52-421-HS21-A_C18neg | HS21 | Artery Plasma |
SA290333 | 20211013_Cowley3_Plasma_39_76-434-HS21-A_C18neg | HS21 | Artery Plasma |
SA290334 | 20211013_Cowley3_Plasma_39_76-434-HS21-A_C18neg_rep | HS21 | Artery Plasma |
SA290335 | 20211007_Cowley3_Plasma_5_76-434-HS21-A_C18pos_rep | HS21 | Artery Plasma |
SA290336 | 20211013_Cowley3_Plasma_32_64-429-HS21-A_C18neg_rep | HS21 | Artery Plasma |
Collection:
Collection ID: | CO002858 |
Collection Summary: | Plasma was collected through an arterial and renal venous catheter throughout the study (200 µL of arterial and renal venous blood were sampled at the day 7, 14, and 21). Overnight urine (18 hours) from the day before the blood draw was collected on ice. The kidneys were collected either at 14 days of HS (HS14) or 21 days of HS (HS21). The kidneys of only LS fed SD rats were also collected for comparison. The collected kidneys (n=5 for each group for metabolomics) were dissected to cortex and outer medulla and snap frozen with liquid nitrogen. Plasma, urine and tissue were stored in -80°C until further analysis. |
Sample Type: | Arterial plasma; Venous plasma |
Storage Conditions: | -80? |
Treatment:
Treatment ID: | TR002874 |
Treatment Summary: | Rats (n=7, 10-11 weeks of age) were performed renal blood flow (RBF) probe implantation and femoral arterial catheterization5. Briefly, rats were anesthetized with isoflurane and arterial catheter was inserted. Following an abdominal incision, RBF probe was implanted on left renal artery and the cable was exposed at nape of the neck via the subcutaneous route. In addition to the RBF probe implantation, renal venous catheter was inserted through the femoral vein and placed in the left renal vein and secured to the luminal wall with 10-0 nylon. RBF and BP via arterial line were measured by conscious freely moving rats and recorded on average of every minute for 24 h/day. After 7-10 days of recovery period, 200 µL of arterial and renal venous blood were sampled and that blood was replaced from donor rats before and following 7, 14 and 21 days after the switch in diet from 0.4% (LS) to 4.0% (HS) salt diet (Dyets Inc, Bethlehem, PA). Overnight urine (18 hours) from the day before the blood draw was collected on ice. The kidneys were collected either at 14 days of HS (HS14) or 21 days of HS (HS21). The kidneys of only LS fed SD rats were also collected for comparison. The collected kidneys (n=5 for each group for metabolomics and mRNAseq analysis) were dissected to cortex and outer medulla and snap frozen with liquid nitrogen. Plasma, urine and tissue were stored in -80°C until further analysis. |
Sample Preparation:
Sampleprep ID: | SP002871 |
Sampleprep Summary: | Plasma/Urine Metabolite Extraction. Metabolites were extracted from 20 µL of plasma and 20 µL of urine from each SD rat in the study according to standard operating procedures in the Mass Spectrometry and Protein Chemistry Service at The Jackson Laboratory34. Metabolites were extracted using 500 µL of an ice cold 2:2:1 methanol:acetonitrile:water (MeOH:ACN:H2O) buffer; the sample was part of the water fraction. Caffeine, 1-napthylamine, and 9-anthracene carboxylic acid were all added at 0.5 ng/ µL in the extraction buffer as internal standards. Each sample was then vortexed for 30 seconds on the highest setting, subject to one minute of mixing with the Tissue Lyser II in pre-chilled cassettes, and then sonicated at 30 Hz for 5 minutes of 30 seconds on 30 seconds off in an ice water bath. Samples were then placed in the -20°C freezer overnight (16 hours) for extraction. Following the extraction, samples were centrifuged at 21,000 x g at 4°C and supernatant from each metabolite extract was equally divided into five 2 mL microcentrifuge tubes. Each sample supernatant was divided into five equal volume aliquots, one for each of the four modes and the rest to create equal representation pools of all samples, one for each mode. Each aliquot was then dried down using a vacuum centrifuge for storage at -80°C until further use. Tissue Metabolite Extraction. Metabolites were extracted from 20 mg of kidney cortex and medulla from each SD rat in the study according to standard operating procedures in the Mass Spectrometry and Protein Chemistry Service at The Jackson Laboratory34 as described for the plasma and urine samples with slight modification. Metabolites were extracted using 1000 µL of an ice cold 2:2:1 methanol:acetonitrile:water (MeOH:ACN:H2O) buffer containing internal standards as above per 20 mg of sample to ensure the extraction equivalents were normalized. Each sample had a 5 mm stainless steel bead added, then were pulverized in extraction buffer for two minutes usingTissue Lyser II. Samples were then placed in the -20°C freezer overnight (16 hours) for extraction and the supernatant was collected as with the urine/plasma samples. Each sample supernatant was divided into five equal volume aliquots, one for each of the four modes and the rest to create equal representation pools of all samples, one for each mode. Each aliquot was then dried down using a vacuum centrifuge for storage at -80°C until further use. |
Combined analysis:
Analysis ID | AN004475 | AN004476 | AN004477 | AN004478 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | Reversed phase | Reversed phase | HILIC | HILIC |
Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
Column | Agilent InfinityLab Poroshell 120 EC-C18 (2.1 x 50 mm; 2.7-Micron) | Agilent InfinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) | Agilent InfinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) | Agilent InfiinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
Units | Area | Area | Area | Area |
Chromatography:
Chromatography ID: | CH003360 |
Chromatography Summary: | This chromatography method was utilized for all C18 positive polarity runs in this study. |
Instrument Name: | Thermo Vanquish |
Column Name: | Agilent InfinityLab Poroshell 120 EC-C18 (2.1 x 50 mm; 2.7-Micron) |
Column Temperature: | 25C |
Flow Gradient: | 0-1 minutes at 98% A1/2% B1, 1-13 minutes from 98% A1/2% B1 to 10% A1/90% B1, 13-15 minutes at 10% A1/90% B1, 15-16 minutes from 10% A1/90% B1 to 98% A1/2% B1, and was re-equilibrated from 16-25 minutes at 98% A1/2% B1 |
Flow Rate: | 0.1 mL/minute |
Internal Standard: | Caffeine, 1-napthylamine, and 9-anthracene carboxylic acid were all added at 0.5 ng/ µL in the extraction buffer as internal standards |
Solvent A: | 100% water, 0.2% acetic acid |
Solvent B: | 100% acetonitrile, 0.2% acetic acid |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH003361 |
Chromatography Summary: | This chromatography method was utilized for all C18 negative polarity runs in this study. |
Instrument Name: | Thermo Vanquish |
Column Name: | Agilent InfinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) |
Column Temperature: | 25C |
Flow Gradient: | 0-1 minutes at 2% A/98% B, 1-11 minutes from 2% A/98% B to 30% A/70% B, 11-12 minutes from 30% A/70% B to 40% A/60% B, 12-16 minutes from 40% A/60% B to 95% A/5% B, was held at 95% A/5% B from 16-18 minutes, 18-20 minutes from 95% A/5% B to 2% A/98% B, and was re-equilibrated from 20-25 minutes at 2% A/98% B |
Flow Rate: | 0.1 mL/minute |
Internal Standard: | Caffeine, 1-napthylamine, and 9-anthracene carboxylic acid were all added at 0.5 ng/ µL in the extraction buffer as internal standards |
Solvent A: | 100% water, 0.2% acetic acid |
Solvent B: | 100% acetonitrile, 0.2% acetic acid |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH003362 |
Chromatography Summary: | This chromatography method was utilized for all HILIC positive polarity runs in this study. |
Instrument Name: | Thermo Vanquish |
Column Name: | Agilent InfinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) |
Column Temperature: | 25C |
Flow Gradient: | 0-1 minutes at 2% A/98% B, 1-11 minutes from 2% A/98% B to 30% A/70% B, 11-12 minutes from 30% A/70% B to 40% A/60% B, 12-16 minutes from 40% A/60% B to 95% A/5% B, was held at 95% A/5% B from 16-18 minutes, 18-20 minutes from 95% A/5% B to 2% A/98% B, and was re-equilibrated from 20-25 minutes at 2% A/98% B |
Flow Rate: | 0.1 mL/minute |
Internal Standard: | Caffeine, 1-napthylamine, and 9-anthracene carboxylic acid were all added at 0.5 ng/ µL in the extraction buffer as internal standards |
Solvent A: | 10 mM ammonium formate in H2O with 0.1% formic acid (Solvent A2) |
Solvent B: | 90% ACN with 10 mM ammonium formate in H2O with 0.1% formic acid (Solvent B2) |
Chromatography Type: | HILIC |
Chromatography ID: | CH003363 |
Chromatography Summary: | This chromatography method was utilized for all HILIC negative polarity runs in this study. |
Instrument Name: | Thermo Vanquish |
Column Name: | Agilent InfiinityLab Poroshell 120 HILIC-Z (2.1 x 50 mm; 2.7 micron; #689775-924) |
Column Temperature: | 25C |
Flow Gradient: | 0-1 minutes at 2% A/98% B, 1-11 minutes from 2% A/98% B to 30% A/70% B, 11-12 minutes from 30% A/70% B to 40% A/60% B, 12-16 minutes from 40% A/60% B to 95% A/5% B, was held at 95% A/5% B from 16-18 minutes, 18-20 minutes from 95% A/5% B to 2% A/98% B, and was re-equilibrated from 20-25 minutes at 2% A/98% B |
Flow Rate: | 0.1 mL/minute |
Internal Standard: | Caffeine, 1-napthylamine, and 9-anthracene carboxylic acid were all added at 0.5 ng/ µL in the extraction buffer as internal standards |
Solvent A: | 10 mM ammonium acetate in H2O, pH 9.0 with 0.1% AffinityLab Deactivator Inhibitor (Agilent, #5191-3940; Solvent A3) |
Solvent B: | 85% ACN with 10 mM ammonium acetate in H2O with 0.1% AffinityLab Deactivator Inhibitor (Solvent B3) |
Chromatography Type: | HILIC |
MS:
MS ID: | MS004222 |
Analysis ID: | AN004475 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | C18 positive plasma data: The tandem mass spectrometry RAW data files (consisting of MS1 and MS2 spectra collected) were analyzed using Thermo Compound Discoverer (v3.2.0.421). The MS1 and MS2 data was searched against the Thermo mzCloud database, ChemSpider database, Metabolika Pathways, and mzLogic predicted composition in the Compound Discoverer workflow. |
Ion Mode: | POSITIVE |
MS ID: | MS004223 |
Analysis ID: | AN004476 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | C18 negative plasma data: The tandem mass spectrometry RAW data files (consisting of MS1 and MS2 spectra collected) were analyzed using Thermo Compound Discoverer (v3.2.0.421). The MS1 and MS2 data was searched against the Thermo mzCloud database, ChemSpider database, Metabolika Pathways, and mzLogic predicted composition in the Compound Discoverer workflow. |
Ion Mode: | NEGATIVE |
MS ID: | MS004224 |
Analysis ID: | AN004477 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | HILIC positive plasma data: The tandem mass spectrometry RAW data files (consisting of MS1 and MS2 spectra collected) were analyzed using Thermo Compound Discoverer (v3.2.0.421). The MS1 and MS2 data was searched against the Thermo mzCloud database, ChemSpider database, Metabolika Pathways, and mzLogic predicted composition in the Compound Discoverer workflow. |
Ion Mode: | POSITIVE |
MS ID: | MS004225 |
Analysis ID: | AN004478 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | HILIC negative plasma data: The tandem mass spectrometry RAW data files (consisting of MS1 and MS2 spectra collected) were analyzed using Thermo Compound Discoverer (v3.2.0.421). The MS1 and MS2 data was searched against the Thermo mzCloud database, ChemSpider database, Metabolika Pathways, and mzLogic predicted composition in the Compound Discoverer workflow. |
Ion Mode: | NEGATIVE |