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MB Sample ID: SA218646
| Local Sample ID: | D1_TF_A_Hilic_Pos |
| Subject ID: | SU002369 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 61-91 |
| Gender: | Female |
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Sample Preparation:
| Sampleprep ID: | SP002375 |
| Sampleprep Summary: | Chloroform (Chl), AnalaR NORMAPUR® ACS was purchased from VWR Chemicals (Lutterworth, UK). Water Optima™ LC/MS Grade, Methanol (MeOH) Optima™ LC/MS Grade, Pierce™ Acetonitrile (ACN), LC-MS Grade and Isopropanol (IPA), Optima™LC/MS Grade were purchased from Thermo Scientific (Hemel Hempstead, United Kingdom). In total two replicates for each of the six specimens were extracted according to a modified Folch et al. [17] as follow: 25 mg of bone powder was placed in tube A and 750μL of 2:1 (v/v) Chl:MeOH were added, vortexed for 30s and sonicated in ice for additional 20 min. 300μL of LC-MS grade water was added to induce phase separation and sonicate for another 15 mins. The sample were then centrifuged at 10°C for 5 mins at 2000 RPM. The respective upper and lower fractions were collected and transferred to fresh Eppendorf tubes and the samples were re-extracted with a second time using 750μL of 2:1 (v/v) Chl:MeOH. The two respective fractions were combined and concentrated. The organic lipid fraction was preconcentrated using a vacuum concentrator at 55oC for 2.5 hours or until all organic solvents has been removed. The aqueous metabolite fractions were flash frozen in liquid nitrogen and preconcentrated using a lyophilizer cold trap -65oC over night to remove all water content. The respective dry fractions were then stored at -80 until analysis. The metabolite fraction was resuspended in 100μL in 95:5 ACN/water (v/v) and sonicated for 15 mins and centrifuged for 15 min at 15K RPM at 4oC and supernatant was then transferred to 1.5mL autosampler vials with 200μL microinsert and caped. 20μL of each sample were collected and pooled to create the pooled QC. The lipid extracts were resuspended in 100μL of 1:1:2 (v/v) water:ACN:IPA and sonicated for sonicated for 15 min and centrifuged for 15 min at 15K RPM at 10oC and supernatant was then transferred to 1.5mL autosampler vials with 200μL microinsert and caped. 20μL of each sample were collected and pooled to create the pooled QC. The sample set was then submitted for analysis |
| Processing Storage Conditions: | On ice |
| Extraction Method: | Chloroform methanol biphasic extraction |
| Extract Storage: | -80℃ |
