PharmPK Discussion - Fatty acid Extraction from Biological matrices

• On 5 Jan 2007 at 08:57:05, "shalini malho" (shalinimalho.aaa.gmail.com) sent the message
  

  
  Dear all,
  
  I wish you a prosperous new year to all.
  I am facing a problem in extracting the fatty acids from the
  biological matrices, please can any one of you can suggest a method
  for extracting fatty acids with good recovery and estimation of the
  same by LCMS-MS.
  
  Note: Preferably LC-MS-MS methods but  not GCMS methods
  
  Thanks regards
  Shallo
  
  

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• On 4 Jan 2007 at 22:00:09, "Ed O'Connor" (Ed.OConnor.at.tandemlabs.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  extraction should be the same. resuspension will differ.
  
  Ed O'Connor, PhD
  Technical Director, Immunoanalytical
  Tandem Laboratories
  115 Silvia Street
  West Trenton, New Jersey
  609-228-0243
  
  

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• On 5 Jan 2007 at 13:41:53, "shalini malho" (shalinimalho.-a-.gmail.com) sent the message
  

  
  Dear, Ed O'Connor
  
      Thanks for ur reply. but can you suggest a method for extraction
  i.e. either with LLE, SPE, or any other. please specify that.
  
  we had tried with LLE and resuspended in Acetonitrile and methanol
  but it has not worked out fine.
  
  

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• On 5 Jan 2007 at 10:43:37, "Ed O'Connor" (Ed.OConnor.-a-.tandemlabs.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  Shalini:  A resource with a modest cost but great benefit is advanced
  chemistry development at http://www.acdlabs.com/ They have an online
  suite ( intro is free) that has a modest cost very well worth it.   you
  can enter the structure and simulate LC/GC and MS results.  You can also
  get solubilities at different pH.
  Having this helps greatly.  Another resource is the TOXI-labs tubes
  offered by Varian.  These are routinely used in forensic tox and can be
  helpful in setting up extractions.
  
  Again though you might try derivatizing the material which should help
  in the extraction and resolution.
  
  Ed O'Connor, PhD
  Technical Director, Immunoanalytical
  Tandem Laboratories
  115 Silvia Street
  West Trenton, New Jersey
  609-228-0243
  
  

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• On 5 Jan 2007 at 13:18:12, "Murad Melhem" (melhemmr.at.gmail.com) sent the message
  

  
  The following message was posted to: PharmPK
  
  Dear Shallo,
  
  A nice article that describes a very efficient method of extraction is
  the one by Patterson and Klein in the Journal of Lipid Research
  (November 1999). However, if I remember correctly, the group used GC
  for final quantitative analysis. recent articles have used
  derivatization or direct HPLC methods to identify, separate and
  analyze diffrent types of fatty acids in various matrices. I am
  alisting a few below.
  
  I hope these are of help.
  
  Murad Melhem, Ph.D.
  Cognigen Corporation
  Buffalo, NY
  
  1) Bravi E et al., J Chromatogr A 2006, 1134, 210
  2) Delmonte P et al., Lipids 2005, 40, 509
  3) Kroumova AB et al., Anal Biochem 1995, 225, 270
  4) Mansour MP, J Chromatogr A, 2005, 1097, 54
  5) Ueno Y et al., Chem Pharm Bull 1999, 47, 1375
  
  

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• On 8 Jan 2007 at 14:52:01, Conti Roberto (Roberto.Conti.at.sigma-tau.it) sent the message
  

  
  The following message was posted to: PharmPK
  
  The best and more reliable way of extracting fatty acid from tissues
  is the ancient Folch method.
  100mg of tissue, homogenated by potter glass/glass in 1ml of methanol
  + 2ml of chloroform + 0.6ml  of saline. Leave overnite in extraction
  and centrifuge 3500rpm for 20 min. The fatty acids will be in the
  chloroform phase on the bottom of the GLASS tube be careful to the
  the cap that may release plastic that may be seen at the CG.
  Ciao
  Dr Roberto Conti
  
  

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