Let’s take a look at one of the more irritating issues in LC/MS method development: signal loss. I’m not talking about signal attenuation, which can be much more convoluted to diagnose; but rather a complete loss of signal – nothing, not even a blip on your TIC. So where do we begin?

What's the process behind tuning our analytes to the mass spectrometer’s optics? And why is that so important?

Before we get into our extractions, we need to set up our LC/MS to detect our analytes of interest. The LC will provide the chromatography - or separation of our analytes - and the MS will support detection. So which one do we evaluate first? Well, if you can’t “see” the analytes, what good is perfecting their chromatography? So, first, let’s get the MS focused, or tuned, for optimum analyte detection.

When trying to develop a new method for the detection of a drug panel, you need to refer to reliable sources to learn all you need to know regarding your analytes. For this post, we’ll go in detail through the search process with analytes like naloxone, buprenorphine, norfentanyl and methadone using urine as our matrix of focus.

The sole purpose for method development is to construct a robust and analytically sound method that will not just pass the barriers of validation, but provide physicians and patients with sound and reliable results. But where to start?

When running through the exhaustive process of method development, most of us put the majority of our focus on validation and how to complete our crazy validation checklists. Throughout this process, the last thing we want to see is some random hiccup in our workflow. But a whole validation without a hiccup is just wishful thinking, right.

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?