Testosterone analysis on UPLC/MSMS

SernaJim
SernaJim
in Instrumentation and Mass Spectrometry
<p>Dear all,</p><p></p><p>I have an Acquity UPLC system in tandem with a Xevo TQ, and I'm using a C18 2.1x100mm. I'm having lot of problems with the repeatability of the analysis of testosterone, even in the standards solutions in methanol and formic acid. If someone have been working with testosterone any help will be appreciated. Wrt the equipment, I ran all the leak tests and the static decay tests and they passed. All the solvents used were HPLC grade.</p><p></p><p>Thanks in advance.</p><p></p><p>Regards</p>

Answers

  • RD
    RD

    I worked on an app for hormones in drinking water which included testoterone.  IClass/Xevo TQD.  I did not specifically look at precision, but the linearity of the testosterone response curve implies that precision was good.  Some data and conditions are attached.

    Contact me directly if you would like to discuss my experiences/observations

    rich_demuro@waters.com

  • schaeferb
    schaeferb

    I have had some experience in repeatablitiy for hormones in the past on HPLC, but I narrowed it down to a sample preparation technique.  Have you tried using a generic hormone (ie cortisone) as an internal standard to see if you are getting results to pass the precision/RSD test?

  • gofish
    gofish

    I've found that slightly modifying the gradient can have a big effect.  Make sure your compound is not eluting right at a gradient change, if it is try changing the gradient so your testosterone elutes during a part of the gradient that has a consistent organic concentration.

    As a test you can run it isocratic (say at 50/50) to see if that is the problem.  If you try this make your standard up in the same buffer (50/50), so that you are injecting roughly the same. 

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