#METABOLOMICS WORKBENCH NReisdorphLab_20190922_152446 DATATRACK_ID:1819 STUDY_ID:ST001257 ANALYSIS_ID:AN002086 PROJECT_ID:PR000843
VERSION             	1
CREATED_ON             	September 30, 2019, 8:14 pm
#PROJECT
PR:PROJECT_TITLE                 	Foodomics Dash Diet
PR:PROJECT_SUMMARY               	Untargeted metabolomics was used to analyze urine and a variety of foods from
PR:PROJECT_SUMMARY               	the Dietary Approaches to Stop Hypertension (DASH) diet in order to understand
PR:PROJECT_SUMMARY               	which food compounds result in health benefits.
PR:INSTITUTE                     	University of Colorado, Denver
PR:DEPARTMENT                    	Anschutz Medical Campus
PR:LAST_NAME                     	Reisdorph
PR:FIRST_NAME                    	Nichole
PR:ADDRESS                       	12850 East Montview Blvd, Aurora, CO, 80045, USA
PR:EMAIL                         	nichole.reisdorph@ucdenver.edu
PR:PHONE                         	3037249234
PR:FUNDING_SOURCE                	NIH (NIDDK, NCATS, NCRR), National Pork Board, NIH Indiana CCTSI, USDA
#STUDY
ST:STUDY_TITLE                   	Nutrimetabolomics and DASH diet
ST:STUDY_SUMMARY                 	Although health benefits of the Dietary Approaches to Stop Hypertension (DASH)
ST:STUDY_SUMMARY                 	diet are established, it is not understood which food compounds result in these
ST:STUDY_SUMMARY                 	benefits. We used a step-wise approach to identify unique compounds from
ST:STUDY_SUMMARY                 	individual foods of a DASH-style diet, determined if these Food-Specific
ST:STUDY_SUMMARY                 	Compounds (FSC) are detectable in urine, and then examined relationships between
ST:STUDY_SUMMARY                 	urinary FSC and blood pressure (BP). Nineteen subjects were randomized into
ST:STUDY_SUMMARY                 	6-week controlled DASH-style diet interventions. Untargeted, LC/MS-based
ST:STUDY_SUMMARY                 	metabolomics was performed on 24-hour urine samples collected before and after
ST:STUDY_SUMMARY                 	each intervention and on 12 representative DASH-style foods.
ST:INSTITUTE                     	University of Colorado, Denver
ST:DEPARTMENT                    	Anschutz Medical Campus
ST:LAST_NAME                     	Reisdorph
ST:FIRST_NAME                    	Nichole
ST:ADDRESS                       	12850 East Montview Blvd, Aurora, CO, 80045, USA
ST:EMAIL                         	nichole.reisdorph@ucdenver.edu
ST:PHONE                         	3037249234
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Male and female
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	01 Grapefruit	Sample type:Food | Food type:Grapefruit | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	02 Apple Juice	Sample type:Food | Food type:Apple Juice | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	03 Blueberries	Sample type:Food | Food type:Blueberries | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	04 Broccoli	Sample type:Food | Food type:Broccoli | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	05 Apple	Sample type:Food | Food type:Apple | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	06 Pork Loin	Sample type:Food | Food type:Pork Loin | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	07 Cucumber	Sample type:Food | Food type:Cucumber | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	08 Peanut Butter 8 ul	Sample type:Food | Food type:Peanut Butter | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	09 Chicken Tenderloin	Sample type:Food | Food type:Chicken | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	10 Tilapia	Sample type:Food | Food type:Tilapia | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	11 coffee	Sample type:Food | Food type:Coffee | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	12 Beef Tenderloin	Sample type:Food | Food type:Beef | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	13 Coffee diluted	Sample type:Food | Food type:Coffee diluted | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	14 Coffee Neat	Sample type:Food | Food type:Coffee neat | Pre or Post and food:na	
SUBJECT_SAMPLE_FACTORS           	-	S31-101 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-101 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-101 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-101 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-102 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-102 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-102 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-102 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-105 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-105 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-105 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-105 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-106 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-106 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-106 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-106 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-107 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-107 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-107 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-107 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-108 Post Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-108 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-108 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-108 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-109 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-109 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-109 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-109 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-111 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-111 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-111 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-111 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-114 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-114 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-114 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-114 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-115 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-115 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-115 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-115 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-116 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-116 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-116 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-116 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-119 Post Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-119 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-119 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-119 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-120 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-120 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-120 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-120 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-141 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-141 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-141 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-141 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-142 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-142 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-142 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-142 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-144 Post Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-144 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-144 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-144 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-145 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-145 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-145 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-145 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-146 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-146 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-146 Pre Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-146 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-148 Post-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Post Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-148 Post-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Post Pork	
SUBJECT_SAMPLE_FACTORS           	-	S31-148 Pre-Ch	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Chicken	
SUBJECT_SAMPLE_FACTORS           	-	S31-148 Pre-Po	Sample type:Urine | Food type:urine | Pre or Post and food:Pre Pork	
#COLLECTION
CO:COLLECTION_SUMMARY            	Twenty-four-hour urine collections were obtained before and near the end of each
CO:COLLECTION_SUMMARY            	of the two DASH-style diet periods. A total of 76 urine samples were used for
CO:COLLECTION_SUMMARY            	untargeted metabolomics analysis, including 38 collected while the 19
CO:COLLECTION_SUMMARY            	participants consumed their typical diets (19 before each intervention period)
CO:COLLECTION_SUMMARY            	and 38 while the DASH-style diet was consumed (19 during each intervention
CO:COLLECTION_SUMMARY            	period). For simplicity, these are referred to as pre- and post-diet urine
CO:COLLECTION_SUMMARY            	samples. Food was purchased from a local grocery store for metabolomic analyses.
CO:COLLECTION_SUMMARY            	All four types of meat included in the diet (pork, chicken, fish, and beef loin)
CO:COLLECTION_SUMMARY            	were analyzed. Fruits and vegetables, if not pre-washed, were washed with tap
CO:COLLECTION_SUMMARY            	water and prepared with inedible parts (i.e. leaves or peels) removed. Meat and
CO:COLLECTION_SUMMARY            	eggs were cooked as instructed to match prescribed cooked weight.
CO:SAMPLE_TYPE                   	Urine
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	During the 18-week study period, participants initially consumed their
TR:TREATMENT_SUMMARY             	self-chosen, unrestricted (typical) diets for two weeks. Participants were then
TR:TREATMENT_SUMMARY             	randomly assigned to consume a DASH-style diet with either lean pork or chicken
TR:TREATMENT_SUMMARY             	and fish as the predominant sources of dietary protein for six weeks. This time
TR:TREATMENT_SUMMARY             	period was followed by a four-week washout period during which participants
TR:TREATMENT_SUMMARY             	again consumed their typical diets prior to consuming the DASH-style diet
TR:TREATMENT_SUMMARY             	containing the other predominant protein sources. Samples utilized for the
TR:TREATMENT_SUMMARY             	current research consisted of 12 individual foods consumed during the DASH-style
TR:TREATMENT_SUMMARY             	diet and urine samples collected prior to each intervention and during the final
TR:TREATMENT_SUMMARY             	two weeks of each controlled feeding period.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Individual food and urine samples were stored at -80 °C prior to sample
SP:SAMPLEPREP_SUMMARY            	preparation. Individual foods (including apple juice) were individually dried
SP:SAMPLEPREP_SUMMARY            	using a FreeZone 2.5 plus lyophilizer (Labconco, Kansas City, MO, USA). Dried
SP:SAMPLEPREP_SUMMARY            	samples were then divided and macerated using a chilled bio-pulverizer (BioSpec
SP:SAMPLEPREP_SUMMARY            	Products, Bartlesville, OK). Approximately 50 mg of each sample was measured
SP:SAMPLEPREP_SUMMARY            	into chilled microcentrifuge tubes and stored at -80o C prior to further
SP:SAMPLEPREP_SUMMARY            	processes and analysis. 10 µL of coffee was diluted with 90 µL of water,
SP:SAMPLEPREP_SUMMARY            	vortexed, aliquotted and stored at -80° C until analysis. An aliquot of coffee
SP:SAMPLEPREP_SUMMARY            	was also reserved for neat (i.e. unprocessed) analysis, and stored at -80° C
SP:SAMPLEPREP_SUMMARY            	until analysis. Freeze-dried foods were removed from -80° C and allowed to thaw
SP:SAMPLEPREP_SUMMARY            	on ice. To each sample, 480 μl of chilled methanol, 10 µl each of a
SP:SAMPLEPREP_SUMMARY            	hydrophilic spike mix, hydrophilic positive control mix, hydrophobic spike mix
SP:SAMPLEPREP_SUMMARY            	and hydrophobic positive control mix were added (see Supplemental Table S1).
SP:SAMPLEPREP_SUMMARY            	Samples were gently vortexed for 10 seconds, placed at -80o C for 60 minutes,
SP:SAMPLEPREP_SUMMARY            	and then centrifuged 0o C at 18,000 x g for 15 minutes to facilitate protein
SP:SAMPLEPREP_SUMMARY            	precipitation. Supernatants were transferred to new microcentrifuge tubes and
SP:SAMPLEPREP_SUMMARY            	dried using vacuum centrifugation at 45o C for approximately 60 minutes. Each
SP:SAMPLEPREP_SUMMARY            	sample was suspended in 50 µl of 95:5 LC/MS grade water-acetonitrile and gently
SP:SAMPLEPREP_SUMMARY            	vortexed for 30 seconds. Following a quick centrifugation, each sample was then
SP:SAMPLEPREP_SUMMARY            	divided into replicate 20 µl aliquots, while 10 µl was removed from each
SP:SAMPLEPREP_SUMMARY            	sample to generate a pooled QC sample. Samples were stored at -80° C until
SP:SAMPLEPREP_SUMMARY            	analysis. Urine samples were analyzed neat (i.e. without any preparation) except
SP:SAMPLEPREP_SUMMARY            	that authentic standards were spiked into urine samples to monitor variability
SP:SAMPLEPREP_SUMMARY            	in instrument response or batch effects.
SP:PROCESSING_STORAGE_CONDITIONS 	Described in summary
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 6520
CH:COLUMN_NAME                   	Agilent Zorbax Rapid Resolution HT (RRHT) SB-AQ, 1.8 micron (2.1 x 100 mm)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6520 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Agilent 6520 Time-of-Flight (TOF-MS) with dual electrospray ionization (ESI)
MS:MS_COMMENTS                   	source, scan rate 2.21 spectra/second, mass range 50-1700 m/z, gas temperature
MS:MS_COMMENTS                   	325 °C, gas flow 12.0 L/min, nebulizer 30 psi, skimmer 60 V, capillary voltage
MS:MS_COMMENTS                   	4000 V, fragmentor 120 V, reference masses 121.050873 and 922.009798 (Reference
MS:MS_COMMENTS                   	mix, Agilent Technologies,).
MS:MS_RESULTS_FILE               	ST001257_AN002086_Results.txt	UNITS:Abundance (Counts Log2 Transformed)	Has m/z:Neutral masses	Has RT:Yes	RT units:Minutes
#END