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Characterization and Classification of Matrix Effects in Biological Samples Analysis 05 Feb 2010 Manish S. Yadav Bioanalyt...
ACKNOWLEDGEMENT All Bioscientists of Veeda
<ul><li>The matrix effect is defined as the combined effect of all components of the sample other than the analyte.  </li>...
<ul><li>Matrix effect is the  difference  between the mass spectrometric response for an  analyte in standard solution  an...
<ul><li>Ion suppression is one form of matrix effect that LC–MS techniques suffer from, regardless of the sensitivity or s...
<ul><li>Absolute Matrix Effect </li></ul><ul><li>Difference between response of equally concentrated analytes (standards) ...
<ul><li>There are 2 types: </li></ul><ul><li>Qualitative      Post Column Infusion ( Enhancement   or  Suppression  in th...
<ul><li>Factors  –  The possible route cause of  Matrix Effect </li></ul><ul><ul><li>Matrix Components </li></ul></ul><ul>...
Reason for Matrix Effect in ESI
Reason for Matrix Effect in ESI
Reason for Matrix Effect in ESI
Reason for Matrix Effect in APCI <ul><li>APCI does experience ion suppression, which has been explained by considering the...
<ul><li>More selective extraction procedure </li></ul><ul><li>Specific retention time of analytes </li></ul><ul><li>Changi...
<ul><li>Process Efficiency (PE) </li></ul><ul><li>Extraction Recovery (RE) </li></ul><ul><li>Matrix Effect (ME) </li></ul>...
Sample Preparation
Matrix Effect (ME)
Extraction Recovery (RE)
Interference Materials in matrix Affect the  MS Ionization Decrease Extraction  During process MATRIX EFFECT EXTRACTION RE...
Relationship: <ul><li>Neat Standard (Aqueous STD) (a) </li></ul><ul><li>Post-Extraction Spiked Standard (b) </li></ul><ul>...
If RE = -1    Evaluate Extraction Yield (EY) RE ≠ +1   <ul><li>ME +    Enhancement </li></ul><ul><li>ME -    Suppress...
Conclusion: <ul><li>Now-a-days, it is generally accepted that matrix effect evaluation should be a  mandatory part of the ...
Thank You…!
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Matrix Effect

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Ion Suppression & Enhancement
Matrix Effect in Bioanalysis

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Matrix Effect

  1. 1. Characterization and Classification of Matrix Effects in Biological Samples Analysis 05 Feb 2010 Manish S. Yadav Bioanalytical Research, Veeda CR, Ahmedabad Dinesh Kumar & Amit Patel Bioanalytical Research, Veeda CR, Ahmedabad Presented by: Conceptualized by
  2. 2. ACKNOWLEDGEMENT All Bioscientists of Veeda
  3. 3. <ul><li>The matrix effect is defined as the combined effect of all components of the sample other than the analyte. </li></ul><ul><li>or </li></ul><ul><li>Components originating from the sample matrix that co-elute with the compound(s) of interest can interfere with the ionization process in the mass spectrometer, causing ionization Suppression or Enhancement that Negatively affect the measurement of quantity. This phenomenon, so-called matrix effect. </li></ul><ul><li>Described by Kebarle and Tang in 1993. </li></ul><ul><li>Questionable in Quantitative Analysis…!!! </li></ul>Matrix Effect
  4. 4. <ul><li>Matrix effect is the difference between the mass spectrometric response for an analyte in standard solution and the response for the same analyte at the same concentration in a biological matrix (plasma, urine, etc.). </li></ul><ul><li>A major source of matrix effect is phospholipids, but there are other matrix components causing matrix effect. You can also get ion suppression from another analyte, a metabolite, concomitant medications, or even from the internal standard. </li></ul>Matrix Effect
  5. 5. <ul><li>Ion suppression is one form of matrix effect that LC–MS techniques suffer from, regardless of the sensitivity or selectivity of the mass analyzer used. </li></ul><ul><li>Ion suppression negatively affects several analytical figures of merit, such as detection capability, precision, and accuracy. </li></ul><ul><li>- introduced originally by Buhrman and coworkers. </li></ul>Ion Suppression
  6. 6. <ul><li>Absolute Matrix Effect </li></ul><ul><li>Difference between response of equally concentrated analytes (standards) in solvent and in matrix extracts. </li></ul><ul><ul><ul><li>Signal Enhancement </li></ul></ul></ul><ul><ul><ul><li>Signal Suppression </li></ul></ul></ul>Types of Matrix Effect <ul><li>Relative Matrix Effect </li></ul><ul><li>Variation of absolute matrix effects between several lots of the same matrix. </li></ul>
  7. 7. <ul><li>There are 2 types: </li></ul><ul><li>Qualitative  Post Column Infusion ( Enhancement or Suppression in the signal) </li></ul><ul><li>- Proposed in 1999 by Bongfiglio et. al. </li></ul><ul><li>Quantitative  Matrix Effect, Extraction Recovery and Process Efficiency </li></ul><ul><li>- Proposed in 2003 by Matuszewski et. al. </li></ul><ul><li>The quantitative measure of matrix effects is the Matrix Factor (MF) </li></ul>Matrix Effect Evaluation
  8. 8. <ul><li>Factors – The possible route cause of Matrix Effect </li></ul><ul><ul><li>Matrix Components </li></ul></ul><ul><ul><li>Residue released from materials used during extraction process </li></ul></ul><ul><ul><li>Inter-conversion during extraction and analysis (drug-drug, drug-meta, meta to drug, meta to meta or isomer to isomer) </li></ul></ul><ul><ul><li>Purity & Assay of Reference / working standards </li></ul></ul><ul><ul><li>Stability of analytes during extraction & analysis </li></ul></ul><ul><ul><li>Presence of lipids – phospholipids (separate session is required), fats, detergents, anticoagulants, plasticizers etc in matrix </li></ul></ul><ul><ul><li>Reagents used in the extraction process and mobile phase </li></ul></ul><ul><ul><li>Elution flow rate </li></ul></ul><ul><ul><li>Hydrophobicity of Analyte </li></ul></ul><ul><ul><li>Analyte concentration </li></ul></ul><ul><ul><li>Mass and Charge of Analyte </li></ul></ul><ul><ul><li>Ion Source </li></ul></ul>
  9. 9. Reason for Matrix Effect in ESI
  10. 10. Reason for Matrix Effect in ESI
  11. 11. Reason for Matrix Effect in ESI
  12. 12. Reason for Matrix Effect in APCI <ul><li>APCI does experience ion suppression, which has been explained by considering the effect of sample composition on the efficiency of charge transfer from the corona discharge needle. </li></ul><ul><li>In addition, because there is very little chance for analytes to pass through the vaporization region and remain in solution, another mechanism of ion suppression in APCI is solid formation, either as pure analyte or as a solid co-precipitate with other nonvolatile sample components. </li></ul>
  13. 13. <ul><li>More selective extraction procedure </li></ul><ul><li>Specific retention time of analytes </li></ul><ul><li>Changing buffer, strength and pH </li></ul><ul><li>Different Ionization Process </li></ul><ul><li>ESI source design </li></ul>Approches – Our target…!!! To mininize Matrix Effect / Ion Suppression…
  14. 14. <ul><li>Process Efficiency (PE) </li></ul><ul><li>Extraction Recovery (RE) </li></ul><ul><li>Matrix Effect (ME) </li></ul><ul><li>Extraction Yield (EY) </li></ul>Terminology
  15. 15. Sample Preparation
  16. 16. Matrix Effect (ME)
  17. 17. Extraction Recovery (RE)
  18. 18. Interference Materials in matrix Affect the MS Ionization Decrease Extraction During process MATRIX EFFECT EXTRACTION RECOVERY
  19. 19. Relationship: <ul><li>Neat Standard (Aqueous STD) (a) </li></ul><ul><li>Post-Extraction Spiked Standard (b) </li></ul><ul><li>Pre-Extraction Spiked Standard (c) </li></ul><ul><li>Neat Extraction Standard (d) </li></ul>
  20. 20. If RE = -1  Evaluate Extraction Yield (EY) RE ≠ +1   <ul><li>ME +  Enhancement </li></ul><ul><li>ME -  Suppression </li></ul><ul><li>ME 0  No Effect </li></ul><ul><li>RE 0  No Effect </li></ul><ul><li>RE -  Low Recovery </li></ul>-1 PE ME ME ME RE RE RE RE RE RE -1 0 -1 -1 0 -1 -1 0 +1 0 -1 0 +1 +1 +1 0
  21. 21. Conclusion: <ul><li>Now-a-days, it is generally accepted that matrix effect evaluation should be a mandatory part of the validation procedure . </li></ul><ul><li>Use of an ( stable labeled isotope ) internal standard is highly recommended. </li></ul><ul><li>Prefer APCI Ion Source instead of ESI, if possible. </li></ul><ul><li>Specific Extraction Procedure. </li></ul><ul><li>Use mobile phase pH to move the analyte away from the interference and improve retention, resolution and sensitivity. </li></ul>
  22. 22. Thank You…!

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