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The following message was posted to: PharmPK
Dear all,
I want to validate the process of xylanase activity but I want to know
whether I should start the method with enzyme and substrate to produce
reduced sugar from which I can make a calibration curve or I can take
the
reduced sugar (Xylose) directly as a standard for making a calibration
curve. Please let me know if anybody can help in this matter.
Uttam Budhathoki
Department of Pharmacy
Kathmandu University
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Dear Uttam,
If you intend to carry out the validation of your assay lege artis,
you definitely should use the xylose standard (with known purity),
spike the blank matrix to produce calibration standards and then
perform the assay.
You should pay attention to the matrix, however. It should contain
all the substances that will be present in your real samples except
the analyte. Careful matrix preparation is especially important if
you intend to use just UV detection in a microtitter plate reader (or
any assay without chromatographic separation).
Best wishes,
Jurij Trontelj
Faculty of pharmacy
University in Ljubljana, Slovenia
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