What is the role of LogP in sample prep methods?

The goal of sample preparation is to create cleaner samples, collecting the compounds of interest and eliminating interferences – that “junk” we don’t care about the can cause ion suppression and matrix effects that affect sensitivity, accuracy and precision.  The ideal sample prep method removes all interfering compounds and produces 100% recovery of all analytes of interest.  The problem is that many interfering compounds have properties that are similar to the compounds we need to detect and quantitate.  It takes some skill and knowledge to develop a method that washes interfering compounds away and elutes the analytes of interest in a separate step.  You need to make sure your washes don’t elute the compounds you care about, and you don’t want interfering junk eluting with your analytes. 

Understanding the chemical properties of the compounds in your sample matrix, both the ones you want to detect and the ones you want to eliminate, is necessary for successful method development.  Is the molecule acidic or basic?  What functional groups are present?  Can they be ionized?  Is the molecule hydrophilic or hydrophobic? Polar or non-polar?  In this post, I am going to discuss the octanol-water partition coefficient, or LogP and its role in sample preparation using supported liquid extraction (SLE) and solid phase extraction (SPE).

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Urine hydrolysis: how did I choose which enzyme to use?

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.

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How to calculate percent recovery and matrix effects for your analytical assays

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!)

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Choosing the best ION Exchange Mode for Solid Phase Extraction

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.

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Extracting both acidic and basic analytes in a single supported liquid extraction (SLE) procedure

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.

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When should I choose SLE or SPE for sample cleanup?

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.

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How to develop a sample prep method in 16 easy steps, using supported liquid extraction

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.

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