Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA021775

Local Sample ID:	sample18
Subject ID:	SU000454
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

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Subject:

Subject ID:	SU000454
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
sample18	SA021775	FL005328	Cognitively normal	Group

Collection:

Collection ID:	CO000448
Collection Summary:	All blood samples in the Mayo Clinic ADRC are collected from nonfasting participants in the sitting position in a clinical laboratory. Serum tubes (red-top) are drawn first, followed by EDTA plasma tubes. Blood is drawn from the median cubital vein with a 21 g needle and typically centrifuged at 2000 g for 10 minutes at 4°C. The serum and plasma are aliquoted into 0.5 mL and stored in a −80°C freezer until use. None of the aliquots were thawed before being pulled to measure the sphingolipids and fatty acids. We have previously shown that long-term storage, up to 38 years, in long-term −80°C freezers does not affect sphingolipid levels [26].
Sample Type:	Blood

Treatment:

Treatment ID:	TR000468
Treatment Summary:	All individuals were enrolled in the Mayo Clinic Alzheimer's Disease Research Center (ADRC) and donated their brain to the Neuropathology Core. Eligibility criteria for the proposed study included an autopsy report and available blood at the last study visit before death. Standardized methods for sampling and neuropathologic examination were performed according to the third report of the DLB consortium (CDLB) [20], [21] and the Consortium to Establish a Registry for Alzheimer's Disease guidelines [22]. Braak neurofibrillary tangle (NFT) stage was determined based on the distribution of NFTs assessed with Bielschowsky silver stain [23]. A consensus clinical diagnosis was determined at each study visit by a panel of neurologists, neuropsychologists, and nurses who reviewed all patient information including neuropsychological results, activities of daily living, and the Clinical Dementia Rating scale. The diagnosis of dementia was based on DSM-III-R criteria [24]. We identified the following four groups: (1) Cognitively normal-normal pathology [CN, n = 21]. These individuals had no LBs, had low-likelihood AD according to the National Institute of Aging (NIA)-Reagan Criteria [25], and were cognitively normal as of their last study visit. (2) High-likelihood DLB [n = 13]. These individuals met criteria for high-likelihood DLB according to the CDLB, had Braak NFT stage ≤ IV, low to intermediate-likelihood AD, and a diagnosis of dementia as of the last study visit. Twelve patients had diffuse LB and one had transitional LB. (3) Intermediate-likelihood DLB (n = 17). These individuals had both DLB and AD pathologies. They had transitional (n = 14) or diffuse (n = 3) LBs, met criteria for intermediate-likelihood DLB according to the CDLB, had Braak NFT stage ≥ IV, intermediate to high-likelihood AD, and a diagnosis of dementia as of the last study visit. (4) Alzheimer's disease pathology (AD, n = 18). These individuals had high (n = 16) or intermediate (n = 2) likelihood AD according to NIA-Reagan criteria, had Braak NFT stage ≥ IV, no LBs, and had a diagnosis of probable AD dementia. Given our previous report that plasma ceramides increase with age and are higher in women [26], we frequency matched the four groups by sex and also by age.

Treatment ID:

TR000468

Treatment Summary:

All individuals were enrolled in the Mayo Clinic Alzheimer's Disease Research Center (ADRC) and donated their brain to the Neuropathology Core. Eligibility criteria for the proposed study included an autopsy report and available blood at the last study visit before death. Standardized methods for sampling and neuropathologic examination were performed according to the third report of the DLB consortium (CDLB) [20], [21] and the Consortium to Establish a Registry for Alzheimer's Disease guidelines [22]. Braak neurofibrillary tangle (NFT) stage was determined based on the distribution of NFTs assessed with Bielschowsky silver stain [23]. A consensus clinical diagnosis was determined at each study visit by a panel of neurologists, neuropsychologists, and nurses who reviewed all patient information including neuropsychological results, activities of daily living, and the Clinical Dementia Rating scale. The diagnosis of dementia was based on DSM-III-R criteria [24]. We identified the following four groups: (1) Cognitively normal-normal pathology [CN, n = 21]. These individuals had no LBs, had low-likelihood AD according to the National Institute of Aging (NIA)-Reagan Criteria [25], and were cognitively normal as of their last study visit. (2) High-likelihood DLB [n = 13]. These individuals met criteria for high-likelihood DLB according to the CDLB, had Braak NFT stage ≤ IV, low to intermediate-likelihood AD, and a diagnosis of dementia as of the last study visit. Twelve patients had diffuse LB and one had transitional LB. (3) Intermediate-likelihood DLB (n = 17). These individuals had both DLB and AD pathologies. They had transitional (n = 14) or diffuse (n = 3) LBs, met criteria for intermediate-likelihood DLB according to the CDLB, had Braak NFT stage ≥ IV, intermediate to high-likelihood AD, and a diagnosis of dementia as of the last study visit. (4) Alzheimer's disease pathology (AD, n = 18). These individuals had high (n = 16) or intermediate (n = 2) likelihood AD according to NIA-Reagan criteria, had Braak NFT stage ≥ IV, no LBs, and had a diagnosis of probable AD dementia. Given our previous report that plasma ceramides increase with age and are higher in women [26], we frequency matched the four groups by sex and also by age.

Sample Preparation:

Sampleprep ID:	SP000461
Sampleprep Summary:	The lipids were extracted from 200 μL of plasma after the addition of internal standards.

Combined analysis:

Analysis ID	AN000683
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Thermo TSQ Quantum Ultra
Column	Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	Triple quadrupole
MS instrument name	Thermo Quantum Ultra
Ion Mode	POSITIVE
Units	micromolar

Chromatography:

Chromatography ID:	CH000495
Chromatography Summary:	The targeted sphingolipid and fatty acid analyses were conducted by the Mayo Clinic Metabolomics Core. Electrospray ionization mass spectrometry was used to quantify plasma ceramides, sphinganine, sphingosine, sphingosine-1-phosphate (S1P), monohexosylceramides, and free fatty acids. The lipids were extracted from 200 μL of plasma after the addition of internal standards. The extracts were measured against a standard curve on the Thermo TSQ Quantum Ultra mass spectrometer (West Palm Beach, FL) coupled with a Waters Acquity UPLC system (Milford, MA) and quantified in μM units.
Instrument Name:	Thermo TSQ Quantum Ultra
Column Name:	Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS000609
Analysis ID:	AN000683
Instrument Name:	Thermo Quantum Ultra
Instrument Type:	Triple quadrupole
MS Type:	ESI
Ion Mode:	POSITIVE