Interview Silvestro
Optimization of analytical set-up, based on this type of equipment, in terms of HPLC conditions (e.g., flow rate and mobile phase composition) is very important to obtain the best results, as well as the evaluation of gas phase modifier to improve specificity.
Q At what stage in your career did you first become involved in bioanalysis & what attracted
you to this area? I should like to discuss shortly how I arrived to bio- analysis but also how, more precisely, to LC-MS. As already discussed I began working as an oncologist and, in this field, it is not uncommon to end up doing, at least part-time, laboratory research. This was also my case and I was interested in cell culture models to study anticancer drugs, in particular new molecules and therapy individualization (cloning assay and simi- lar). Working in this field my curiosity to understand why some molecules are active on some tumors only and to clarify drug resistance mechanisms led me to enter in PK/PD studies as well as biochemistry; HPLC soon became my preferred analytical tool to work with in these interesting areas of research. Then in 1986 I began a cooperation with an Italian
company developing sulfated glycosaminoglycans as drugs. Information on PK was minimal, in large part due to the inadequate analytical tools available at that moment, and also PD studies were difficult. I entered so far in this field with enthusiasm and, after realiz- ing that conventional LC detectors were of poor help, I decided to test MS hyphenated to HPLC. The begin- ning, despite the premises, was quite disappointing and until 1989 when I first employed electrospray, the progress minimal. The appearance on the market of API III from Sciex was a real breakthrough and from then on, impressed by the analytical capabilities of this new instrument, I dedicated my work on applications of LC-MS to a large spectra of molecules: from glycans to polynucleotide, from biomarkers to drug metabo- lites. After so many years I am still not bored and I greatly enjoy working in this field.
Progress & obstacles in 2015
Q What do you think is the biggest regulatory challenge that bioanalysts currently face? I didn’t see a new regulatory challenge coming out in 2015. As always in the last years, that regulatory authorities are continuously growing the stack in terms of quality in bioanalysis. This is a natural evolution in this activity, but a more realistic vision of the situation, avoiding targets sometime beyond the real possibilities, should be considered. We are in fact observing requests
Bioanalysis
of sensitivity, precision, accuracy and specificity that are often approaching the technical limits of the equip- ment. I should like also to see more interaction with
researchers before introduction of new rules, as well as evaluation periods to see the real possibility of imple- menting them. Beside this I am surprised that each new regulation is introduced without clearly consid- ering a transition period and often companies suffer important financial losses from rejections of studies already performed in the past (as it is the case with the missing incurred sample re-analyses in bioequivalence studies).
Q What do you think is the greatest technological obstacle faced by bioanalysts
today? One point that I find really critical in my work as bio- analyst is the gap existing between analytical capabili- ties of injecting in LC-MS extremely small samples and the difficulties to routinely process these kinds of microsamples. We often extract samples of hundred microliters to get extracts in the range of tens of micro- liters, but in reality we need to inject just a few micro- liters! From liquid handler, to SPE systems and autos- amplers, there is a lot of work required to scale down all the technologies preceding the mass spectrometer to avoid huge loss of costly reagents and disposable mate- rials without any return in terms of sensitivity and/or operational ease.
Looking to the future
Q What are you excited about working on in 2016? There are three main areas where I am looking to work with interest, and I can say with excitement these are: first, further applications of ion mobility MS; sec- ond, improvement of sample preparation methods to be faster and more cost effective; and finally, increas- ing the analytical sensitivity by applying microflow or nanoflow HPLC, which besides many other advan- tages are also environment friendly methods (less sol- vent and chemicals in use and waste with the risk to pollute the environment). All three topics are not abso- lute novelties but there is so much to do until robust methods are obtained that serious development work has to be carried out. Efforts are especially needed to select the best equipment and, non negotiable, to train the laboratory team to these new procedures.
Q What new technology would you like to see in the future, & how would this improve your work?
More than new techniques I am fascinated by the future science group
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