Urine hydrolysis is a part of sample preparation that we do to eliminate glucuronides. So first, what are glucuronides? And why do we want to get rid of them? Glucuronides are formed during metabolism. These glucuronide compounds attach to drugs to make them more water-soluble. This allows for easier excretion of the drugs in urine. However, this is not what we want to analyze when looking for our drugs of interest. We would rather look for just morphine or just oxazepam instead of morphine-3-β-D glucuronide or the oxazepam glucuronide. The process of removing these glucuronides from our analytes of interest is called hydrolysis.
In this post, I’ll tell you about my experience with urine hydrolysis and my success (or lack of success) when using different enzymes.
Continue reading Urine hydrolysis: how did I choose which enzyme to use?
For many of us developing an analytical assay requires numerous experiments in addition to lots of data review, and yet despite the feeling of confidence in our success, how reliable is our assay really? How do we measure reliability of our assay? Well, there are many ways but here I’ll explain the most common approach using a theoretical analyte called compound X, which, in this case, is readily prescribed for arthritis. And like many drugs, compound X is excreted in large quantities, unmodified, in human urine. So, for arguments sake, let’s say we’ve already defined our LC/MS-MS method and we’ve ironed out an extraction method using SLE+ (Thanks Bruce!)
Continue reading How to calculate percent recovery and matrix effects for your analytical assays
There is often confusion as to how to choose the proper SPE ion exchange media. How do you determine if you should be looking at anion or cation, weak or strong? In this post, I will give some simple guidelines on making the best choice. You will see that ion exchange is all about acids and bases.
Continue reading Choosing the best ION Exchange Mode for Solid Phase Extraction
If you are frustrated with low recoveries caused by protein binding during supported liquid extraction (SLE) and solid phase extraction (SPE), this blog post will give you a few tips. Proper sample pre-treatment is the key to solving this issue.
Continue reading Techniques for disrupting protein binding in sample prep
Often the question arises asking how can I extract both acids and bases with Supported Liquid Extraction (SLE). In this blog post I will discuss some ways to extract both acidic and basic analytes in the same method.
Continue reading Extracting both acidic and basic analytes in a single supported liquid extraction (SLE) procedure
When it comes to sample cleanup the question occasionally comes up asking what technique is best for my application? Solid Phase Extraction (SPE) or Supported Liquid Extraction (SLE). Well, what’s right for you depends on your application and the characteristics of your analytes. Both techniques offer high recoveries, are reproducible, and are easily automated. In this post I would like to outline some of the advantages, disadvantages and limitations of each technique.
Continue reading When should I choose SLE or SPE for sample cleanup?
Taking the time to develop a Supported Liquid Extraction (SLE) method using these 16 easy steps can save you hours or days of headaches down the road as well as giving you the peace of mind that you’re getting the best recoveries possible.
Continue reading How to develop a sample prep method in 16 easy steps, using supported liquid extraction