{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002283","ANALYSIS_ID":"AN003731","VERSION":"1","CREATED_ON":"September 21, 2022, 12:19 pm"},

"PROJECT":{"PROJECT_TITLE":"ForensOMICS","PROJECT_TYPE":"Untargeted metabolomics","PROJECT_SUMMARY":"Analysis of bone biomolecules for post mortem and age at death estimation in forensic contexts","INSTITUTE":"University of Central Lancashire","LAST_NAME":"Bonicelli","FIRST_NAME":"Andrea","ADDRESS":"Fylde Rd, Preston, Lancashire, PR1 2HE, United Kingdom","EMAIL":"abonicelli@uclan.ac.uk","PHONE":"07383974949","PROJECT_COMMENTS":"raw data ID DataTrackID3449","CONTRIBUTORS":"Andrea Bonicelli"},

"STUDY":{"STUDY_TITLE":"The “ForensOMICS” approach to forensic post-mortem interval estimation: combining metabolomics, lipidomics and proteomics for the analysis human skeletal remains","STUDY_SUMMARY":"The combined use of multiple omics methods to answer complex system biology questions is growing in biological and medical sciences, as the importance of studying interrelated biological processes in their entirety is increasingly recognized. We applied a combination of metabolomics, lipidomics and proteomics to human bone to investigate the potential of this multi-omics approach to estimate the time elapsed since death (i.e., the post-mortem interval, PMI). This “ForensOMICS” approach has the potential to improve accuracy and precision of PMI estimation of skeletonized human remains, thereby helping forensic investigators to establish the timeline of events surrounding death. Anterior midshaft tibial bone was collected from four female body donors in a fresh stage of decomposition before placement of the bodies to decompose outdoors at the human taphonomy facility managed by the Forensic Anthropological Center at Texas State (FACTS). Bone samples were again collected at selected PMIs (219, 790, 834 and 872 days). Liquid chromatography mass spectrometry (LC-MS) was used to obtain untargeted metabolomic, lipidomic and proteomic profiles from the pre- and post-placement bone samples. Multivariate analysis was used to investigate the three omics blocks by means of Data Integration Analysis for Biomarker discovery using Latent variable approaches for Omics studies (DIABLO), to identify the reduced number of markers that could effectively describe post-mortem changes and classify the individuals based on their PMI. The resulting model showed that pre-placement bone metabolome, lipidome and proteome profiles were clearly distinguishable from post-placement profiles. Metabolites associated with the pre-placement samples, suggested an extinction of the energetic metabolism and a switch towards another source of fuelling (e.g., structural proteins). We were able to identify certain biomolecules from the three groups that show excellent potential for estimation of the PMI, predominantly the biomolecules from the metabolomics block. Our findings suggest that, by targeting a combination of compounds with different post-mortem stability, in future studies we could be able to estimate both short PMIs, by using metabolites and lipids, and longer PMIs, by including more stable proteins.","INSTITUTE":"University of Central Lancashire","LAST_NAME":"Bonicelli","FIRST_NAME":"Andrea","ADDRESS":"Fylde Rd, Preston PR1 2HE","EMAIL":"abonicelli@uclan.ac.uk","PHONE":"07383974949","NUM_GROUPS":"5","TOTAL_SUBJECTS":"4","NUM_FEMALES":"4"},

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},
{
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"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_C_Hilic_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
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"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_D_Hilic_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_E_Hilic_Neg",
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"Additional sample data":{"RAW_FILE_NAME":"D4_TF_E_Hilic_Neg.mzML"}
},
{
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{
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"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_E_Hilic_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_E_Hilic_Neg.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_A_Lip_Pos",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_B_Lip_Pos",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_C_Lip_Pos",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_D_Lip_Pos",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_E_Lip_Pos",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_A_Lip_Pos",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_B_Lip_Pos",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_C_Lip_Pos",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_D_Lip_Pos",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_E_Lip_Pos",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_A_Lip_Pos",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_B_Lip_Pos",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_C_Lip_Pos",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_D_Lip_Pos",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_E_Lip_Pos",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_A_Lip_Pos",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_B_Lip_Pos",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_C_Lip_Pos",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_D_Lip_Pos",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_E_Lip_Pos",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_A_Lip_Pos",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_B_Lip_Pos",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_C_Lip_Pos",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_D_Lip_Pos",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_E_Lip_Pos",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_A_Lip_Pos",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_B_Lip_Pos",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_C_Lip_Pos",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_D_Lip_Pos",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_E_Lip_Pos",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_A_Lip_Pos",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_B_Lip_Pos",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_C_Lip_Pos",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_D_Lip_Pos",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_E_Lip_Pos",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_A_Lip_Pos",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_A_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_B_Lip_Pos",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_B_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_C_Lip_Pos",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_C_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_D_Lip_Pos",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_D_Lip_Pos.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_E_Lip_Pos",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_E_Lip_Pos.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_A_Lip_Neg",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_B_Lip_Neg",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_C_Lip_Neg",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_D_Lip_Neg",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Fresh",
"Sample ID":"D1_TF_E_Lip_Neg",
"Factors":{"Age":"91","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TF_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_A_Lip_Neg",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_B_Lip_Neg",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_C_Lip_Neg",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_D_Lip_Neg",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D1_Decomposed",
"Sample ID":"D1_TS_E_Lip_Neg",
"Factors":{"Age":"91","PMI":"219","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D1_TS_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_A_Lip_Neg",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_B_Lip_Neg",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_C_Lip_Neg",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_D_Lip_Neg",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Fresh",
"Sample ID":"D2_TF_E_Lip_Neg",
"Factors":{"Age":"67","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TF_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_A_Lip_Neg",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_B_Lip_Neg",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_C_Lip_Neg",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_D_Lip_Neg",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D2_Decomposed",
"Sample ID":"D2_TS_E_Lip_Neg",
"Factors":{"Age":"67","PMI":"790","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D2_TS_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_A_Lip_Neg",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_B_Lip_Neg",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_C_Lip_Neg",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_D_Lip_Neg",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Fresh",
"Sample ID":"D3_TF_E_Lip_Neg",
"Factors":{"Age":"61","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TF_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_A_Lip_Neg",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_B_Lip_Neg",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_C_Lip_Neg",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_D_Lip_Neg",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D3_Decomposed",
"Sample ID":"D3_TS_E_Lip_Neg",
"Factors":{"Age":"61","PMI":"834","Deposition":"Burial","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D3_TS_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_A_Lip_Neg",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_B_Lip_Neg",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_C_Lip_Neg",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_D_Lip_Neg",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Fresh",
"Sample ID":"D4_TF_E_Lip_Neg",
"Factors":{"Age":"77","PMI":"2-10","Deposition":"Pre deposition","State":"Fresh"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TF_E_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_A_Lip_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_A_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_B_Lip_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_B_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_C_Lip_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_C_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_D_Lip_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_D_Lip_Neg.mzML"}
},
{
"Subject ID":"D4_Decomposed",
"Sample ID":"D4_TS_E_Lip_Neg",
"Factors":{"Age":"77","PMI":"872","Deposition":"Open pit","State":"Skeletonised"},
"Additional sample data":{"RAW_FILE_NAME":"D4_TS_E_Lip_Neg.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Bone samples (ca. 1 cm3) of the anterior midshaft tibia (left) were collected prior to placement of the body outdoors, and again upon retrieval of the completely skeletonized remains (right). Each body was in “fresh” stage of decomposition when pre-placement samples were taken, and in “skeletonization” stage when post-placement samples were collected, based on scoring of the gross morphological changes37. The duration of each placement and the deposition context are reported in Table 1. The soft tissue was incised with a disposable scalpel, and a 12 V Dremel cordless lithium-ion drill with a diamond wheel drill bit was used at max. 5000 revolutions to collect ~1 cm3 of bone. Sampling instruments were cleaned with bleach and deionised water between each individual sample collection. A total of eight samples were collected in Ziploc bags, transferred immediately to a -80 °C freezer, and subsequently shipped overnight on dry ice to the Forensic Science Unit at Northumbria University, U.K. The samples were then transferred to a lockable freezer at -20 °C as per UK Human Tissue Act regulations (licence number 12495). The bone samples were defrosted, and fine powder was obtained with a Dremel drill equipped with diamond-tipped drill bits operated at speed 5000 rpms, to avoid heat damage caused by the friction with the bone. The collected powder was homogenised and stored in 2 mL protein LoBind tubes (Eppendorf UK Limited, Stevenage, UK) at -80 °C until extraction and testing. The powder sample was later divided into 25 mg aliquots. The research and bone sample analyses were reviewed and approved by the Ethics committee at Northumbria University (ref. 11623).","SAMPLE_TYPE":"Bone","STORAGE_CONDITIONS":"-20℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Anterior midshaft tibial bone was collected from four female body donors in a fresh stage of decomposition before placement of the bodies to decompose outdoors at the human taphonomy facility managed by the Forensic Anthropological Center at Texas State (FACTS). Bone samples were again collected at selected PMIs (219, 790, 834 and 872 days)"},

"SAMPLEPREP":{"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℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"RP ESI-","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Waters Acquity BEH Amide (150 x 2.1mm, 1.7um)","FLOW_GRADIENT":"60% “B” at T0 hold for 1.5 min, linear increase to 85% “B” at 7 min, increase to 95% “B” at 12.5 min and hold for 4.5 min","FLOW_RATE":"200 μL/min","COLUMN_TEMPERATURE":"45°C","METHODS_FILENAME":"AB_Human_Bone_Chromatography.pdf","SOLVENT_A":"LC/MS grade water:ACN, 40:60 % V/V + containing 10 mM ammonium formate + 0.1% ammonia solution","SOLVENT_B":"IPA:ACN, 90:10 % V/V + containing 10 mM ammonium formate + 0.1% 0.1% ammonia solution","INJECTION_TEMPERATURE":"300°C","SAMPLE_INJECTION":"5μL","PRECONDITIONING":"4.5 mins B"},

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

"MS":{"INSTRUMENT_NAME":"Obitrap ID-X Tribrid","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Mass spectrometry (MS) data were acquired using the AcquieX acquisition workflow (data dependent analysis). The MS operating parameters were as follows: MS1 mass resolution 60K, for MS2 30K, stepped energy (HCD) 20, 25, 50, scan range 100-1000, RF len (%) 35, AGC gain, intensity threshold 2e4, 25% custom injection mode with an injection time of 54 ms. An extraction blank was used to create a background exclusion list and a pooled QC was used to create the inclusion list."},

"MS_METABOLITE_DATA":{
"Units":"Area integration",

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}

}