Dr. Graeme Clark, Head of Analytical Sciences, Cyprotex Discovery Ltd.

LC-MS technology is helping contract research organization Cyprotex Discovery Ltd. perform bioanalysis of small molecules, peptides, and other pharmaceuticals, enabling quantification to be performed in complex matrices during in vitro ADME studies.

Cyprotex Discovery Ltd. in Macclesfield, UK, is a DMPK ADME contract research organization engaged in in-vitro ADME studies for the pharmaceutical industry. In addition to investigating the drug’s action on the body, there is also need to screen out undesirable compounds with poor DMPK parameters, which involves in vitro studies of microsomes, cell lines, and hepatocytes, as well as drug transporters. An important part of this process is the accurate quantification of small molecules, peptides and other kinds of pharmaceuticals, which is achieved by LC-MS analysis.

Graeme Clark, Head of Analytical Sciences, explained, “In bioanalysis, we typically receive plasma and tissue samples from pre-clinical studies for extraction and quantification of various molecules and their metabolites, which is underpinned by MS techniques. We use the SCIEX QTRAP^® technology a great deal, as it offers high sensitivity and allows us to perform more sophisticated MS/MS experiments, such as LC-MS/MS/MS analyses.

This technology is also used to support the in vitro ADME work when more sophisticated analytics are required, coupling multi-dimensional chromatography systems to QTRAP 5500 mass spectrometers.”

Graeme continued, “In the past 12 months, we’ve also introduced a couple of TripleTOF^® 6600 Systems into the company, primarily for in vitro ADME screening, where there is a major bottleneck in setting up optimized MRM transitions for every single compound. For small numbers, this is not an issue, but we routinely screen hundreds of compounds a week and have to develop a method for each one before it can be assayed.

The advantage of the TripleTOF 6600, a high-resolution accurate mass system with SWATH^® Acquisition, is that it collects all the data, all the time, eliminating the need for optimization. Compounds can be assayed straight away, saving time and improving efficiency, and we can retrospectively reprocess the data to look at different analytes, for example, a sodium rather than a proton adduct, or potential metabolites, if necessary, without having to repeat the analysis.”

“The TripleTOF 6600s are also being used for the analysis of biotherapeutic-type molecules, for example, next generation peptides, which may be cyclic or have polymers, dendrimers or other non-peptidic material attached, increasing the molecular weight. While this can be done using a triple quad instrument, it is quite difficult to get selective MRM transitions and may be less sensitive. The high-resolution accurate mass capabilities of the TripleTOF 6600 provide the selectivity we need, without spending days developing analytical methods.

Also, we perform intact protein bioanalysis using an Eksigent MicroLC 200 System, investigating the catabolic fate of biotherapeutic drugs; the lower flow rate gives cleaner spectra, making the data easier to interpret.”

“Because of the sheer volume of samples we handle, on average, we collect around 8,000 data points a day, any manual intervention impacts massively on data turnaround times, and so we have fully automated our data analysis. The relevant MS data is extracted, integrated and exported to the LIMS we have developed in-house, where the required calculations are performed using spreadsheets. Simple, basic metrics highlight which samples need attention, allowing scientists just to come in and look at the data.”

“Moving forward, we’re hoping to implement the SCIEX MPX™-2 technology, which enables switching between two LC systems. This increases flexibility and minimizes dead time due to column equilibration. On a two minute run, we could effectively double our efficiency; an 18 to 20 hour analysis time would be reduced to just 9 or 10 hours and could be completed overnight. Data would be available for analysis sooner, improving turnaround times even further.”

“I’ve worked with SCIEX for years and have a high degree of confidence in its systems. As a result, when we were looking to overhaul our high throughput processes and grow alternative businesses, such as bioanalysis, biotransformation, and biotherapeutics, SCIEX was the first choice because of its reputation in the field and our previous experience of working with the company. We have a very good relationship, and are a demo lab for the TripleTOF 6600, giving potential customers the chance to visit our laboratory and see the system in use in a ‘real world’ situation.

We also have a lot of contact with the technical staff at SCIEX and can call on their application chemists if our clients request something a little different to the norm. It’s a relationship that works well for both of us,” Graeme concluded.Find out more about the SCIEX TripleTOF 6600 System >

This article is from the 2017 edition of the SCIEX VISION journal. You can download the full version of the journal which includes over 10 customer research stories. Download now >