PharmPK Discussion - Separation of dextrorphan from gluconoride conjugate

• On 18 Nov 2007 at 08:25:16, RAKESH DADHANIA (receptor20.at.yahoo.com) sent the message
  

  
  Dear All,
  We are conducting one study in volunteers wherein they are being
  screened for their status on poor or fast metabolizer of CYP 2D6. In
  order to check the same, it is decided to use Dextromethorphan as the
  substrate.
  
  We have also planned to analyse the substrate Dextromethorphan and its
  metabolite dextrorphan in the urine. From the literature, we found
  that dextromethorphan undergoes phase-II metabolism and its main
  metabolite is dextrorphan-gluconoride. As we want to measure the
  concentration of dextromethorphan and dextrorphan and determine its
  ratio.
  
  How can we measure the concentration of dextrorphan in its conjugated
  state in urine. If we cannot measure, it in dextrorphan-gluconoride
  then what method we should use to separate the dextrorphan from its
  conjugated form.
  
  Can I request any members to help us by giving your suggestions,
  ideas, thoughts or guidance with or without literature please.
  
  Many thanks,
  
  Regards,
  Rakesh Dadhania
  
  

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• On 19 Nov 2007 at 08:43:47, "Satya Jadhav" (satya.jadhav.-a-.gmail.com) sent the message
  

  
  Dear Rakesh,
  The conjugated dextrorphan can be separated from the free dextrophan
  using preparative HPLC. Then you can cleave it (enzyme hydrolysis) and
  measure the parent dextrophan. For  details, please refer to  J.
  Chrom. B. vol. 813 (1-2), 217-225.
  
  

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• On 20 Nov 2007 at 16:04:28, millin karthik (millinkarthik.at.yahoo.co.in) sent the message
  

  
  Dear Rakesh
  
  In order to quantify the Dextrorphan glucuronide , you have to use
  glucuronidase enzyme during the Processing of urine sample. This
  enzyme will cleave the drug and conjugate.
  
  Here you need to have replicate of urine sample. One sample you can
  process as such and estimate the drug concentration and another sample
  you could use beta glucuronidase enzyme( Some IU/ml). Then the ratio
  can be calculated easily.
  
  you can find many literature  this type of method.
  
  Thanks.
  
  Best wishes
  A.Karthik
  Research scholar
  MCOPS
  Manipal
  
  

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