Standard Analysis of Blood Plasma or Serum Samples

Product Description

Coverage:
This analysis provides a comprehensive coverage of 16 distinct lipid classes:

• DAG – diacylglycerol;
• TAG – triacylglycerol;
• PC – phosphatidylcholine;
• PE – phosphatidylethanolamine;
• PI – phosphatidylinositol;
• LPC – lyso-phosphatidylcholine;
• LPE – lyso-phosphatidylethanolamine;
• PC O- – ether-linked PC;
• PE O- – ether-linked PE;
• LPC O- – ether-linked LPC;
• LPE O- – ether-linked LPE;
• Cer – ceramide;
• HexCer – hexosylceramide;
• SM – sphingomyelin;
• Chol – cholesterol;
• CE – cholesteryl ester;

The exact number of discovered lipids provided in results depends on samples.

Analysis:
Each sample is extracted, acquired, and analyzed once. We recommend including multiple sample replicates to obtain statistically meaningful data.

Approach:
Untargeted high-resolution shotgun lipidomics-generated data analyzed using LipotypeXplorer, our proprietary software for identifying all measurable lipids using the molecular fragmentation query language (MFQL) and thus unimpeded by the traditional reliance on predefined spectral databases.

Results:
Results are fully quantitative and expressed in absolute values – pmol per sample volume and mol%. The absolute quantification is achieved using lipid class-specific internal standards. Lipotype will provide the data on sample volume analyzed and molar amounts of lipid molecules per sample. Please note: Lipotype will not and shall not be obliged to provide any Lipotype background intellectual property (including Lipotype know-how) or any data incorporating such Lipotype background intellectual property, notably no raw data will be provided.

Structural details:
With the advantage of MS2 fragmentation glycerophospholipids and glycerolipids (except TAG) are reported as subspecies, giving additional information on fatty acids – e.g., PI 18:1;0_16:0;0 denotes phosphatidylinositol with octadecenoic (18:1;0) and hexadecanoic (16:0;0) fatty acids, for which the exact position (sn-1 or sn-2) in relation to the glycerol backbone cannot be discriminated. PC O-18:1;0/16:0;0 denotes an ether- phosphatidylcholine, where an alkyl chain with 18 carbon atoms and 1 double bond (O-18:1;0) is ether-bound to sn-1 position of the glycerol and a hexadecanoic acid (16:0;0) is connected via an ester bond to the sn-2 position of the glycerol. The remaining lipid classes are reported as species – e.g. TAG 56:3;0 denotes triacylglycerol with the total length of its fatty acids of 56, 3 double bonds, and 0 hydroxylations.

Report:
The Results are delivered in the form of a Standard Report containing information on methodology, analysis details, used sample amounts, technical variation and quantitative lipidomics results for each sample in tabular format. The Extended Report additionally contains a graphical presentation of: (a) averaged lipid class values with replicate standard deviations, (b) averaged (sub)species, double bond, chain length and hydroxylation profiles for each lipid class, (c) principal component analysis for un-averaged replicates. The optional Extended Report can be viewed via internet browser (as an offline local webpage) or from data files that can be browsed manually. Cohorts defined in the sample list associated with analyzed samples are the basis for averaging, visualizing, comparing or any other replicate-dependent statistical proceeding included in the Standard and Extended Report.

Delivery time:
The Results will be delivered via email within 6 weeks after receiving the samples prepared according to sample preparation guidelines as well as all information regarding the samples and analysis to be performed. The delivery date of samples to Lipotype will be discussed and agreed between the Customer and Lipotype. LIPOTYPE’s obligation to perform under this Quotation shall be suspended as long as and to the extent LIPOTYPE is not able to fully and unrestrainedly use its facilities and personnel due to injunctions, restraints, directions or recommendations (individual or public) by any local, national or international governmental or non-governmental institutions (“Authority Measures”), regardless of whether such Authority Measures affect LIPOTYPE, its directors, employees, agents or subcontractors directly or indirectly.

Quality assurance:
Each analysis includes internal blank and quality control reference samples to assess and improve the technical quality of the analysis. The technical variation of an analytical run assessed from quality control reference samples is provided within the Report. Please note: No further information or data (especially also including individualized data) on blank and quality control reference samples and/or their analysis will be provided to the Customer.

Optional features

• Interactive Data Visualization with LipotypeZoom (available free of charge). LipotypeZoom displays the content of the extended report and is internet-accessible via browser. Lipotype GmbH uploads the results only having the customer’s consent.
• Statistical Report includes an advanced data analysis. It contains results of up to 8 pairwise Cohort Comparisons or Correlation Analyses. Comparisons or correlations are based on experimental conditions e.g., a comparison of treated vs. untreated or a correlation of a drug dose with lipids.
  • A. Cohort Comparison
    This package compares two specified groups each containing multiple samples, and includes:
    • Cohort Comparison of lipid (sub-)species.
      Contains significance tests, graphical presentation of the result of the pairwise comparison (volcano plot) and individual graphical presentation of significant hits (box plots).
    • Cohort Comparison of lipid classes and fatty acids
      Contains sums within classes measured as subspecies. Contains significance tests and individual graphical presentation of significant hits (box plots).
    • Cohort Comparison of lipid features
      (Total length, total double bond and hydroxylation sums within a class.) Contains significance tests and individual graphical presentation of significant hits (box plots).
    • (Optional) Pathway enrichment analysis
      Contains an analysis based on the lipid classes of the significant lipid (sub-)species hits.
  • B. Correlation Analysis
    Within this package a continuous parameter describing samples (e.g., age, a clinical parameter, protein expression level etc.) is correlated with amounts of lipid (sub-)species/classes and other data, and includes:
    • Correlation analysis with lipid (sub-)species.
      Contains histogram of correlation estimates, their p-values and individual graphical presentation of significant correlations (scatter plots).
    • Correlation analysis with lipid classes and fatty acids
      Contains sums within classes measured as subspecies. Contains histogram of correlation estimates, p-values and individual graphical presentation of significant correlations (scatter plots).
    • (Optional) Pathway enrichment analysis
      Contains an analysis based on the lipid classes of significantly correlating lipid (sub-)species.

  The delivery time for the Statistical Report will be 3 weeks after the Standard/Extended Report is delivered to the Customer. There is one revision round of the Statistical Report included which can prolong the delivery time.

• Reduced delivery time for standard and extended reports (3 weeks turbo analysis).