Xevo TQ-XS ****************************************************************************************** * ACQUITY UUPLC I-Class System (SM-FTN)? ****************************************************************************************** Supplier Waters Year of acquisition 2018 Price 12.5 milion CZK Financing OP VVV CORE FACILITIES [ URL "LFHKEN-192.html "] CZ.02.1.01/0.0/0.0/16_017/0002515 Responsible Person doc. Ing. Miloš Hroch, Ph.D. The last decade has seen a huge boom in pharmaceutical and biomedical research, but this h lot of pressure on instrumentation and big data processing. In the field of separation tec mass spectrometry, we can observe a trend of continuous innovation, modernisation and opti instrumentation to meet these demands, especially in terms of miniaturisation, efficiency analysis. Our analytical chemistry laboratory is engaged in the development of new methods for the d of xenobiotics and endogenous substances in biological material with a relatively wide sco these are compounds from the category of drugs, metabolites, or endogenous substances as m pathological conditions. However, with the technique available to us so far, we have quite to encounter physical limits of the instrumentation used and some projects could not be sa addressed at the level of current knowledge. Nevertheless, after many years of great effort, it has been possible to obtain the financi to ensure the purchase of instrumentation up to the standards of today that can be found i laboratories of developed countries. The instrument we have been waiting for is a modern Mass Detector (QQQ MS) and UHPLC syste This instrumentation will form the instrumental basis of our Analytical Laboratory and wil primarily for targeted, highly sensitive quantification of substances. The cell consists o modules. The Acquity ultra performance liquid chromatograph (UPLC) module and the Waters X mass spectrometer (MS). The UPLC has several basic components found in most contemporary liquid chromatographs. It pump capable of forming a high pressure gradient up to 1300 Bar at a flow rate of 0.010 - Furthermore, it is a versatile autosampler allowing to work not only with classical chroma (2x 48 positions), but also, for example, with 96 and 384 well microtiter plates or other formats. The volume of the injected sample can be selected from 0.1 - 10.0 ?L. Samples can temperature range of 4 - 40 °C. Another part of the UPLC is a column thermostat with space up to 150 mm in length. This allows independent switching and heating over a temperature r °C. The column thermostat also contains an in-line mobile phase preheater to prevent tempe during separation. The detector part of the UPLC/MS set-up consists of the Xevo TQ-XS mass spectrometer, whic advanced triple quadrupole tandem mass spectrometers. At the time of acquisition (2018), t was one of the most sensitive instruments of its kind available on the market. The spectro with a combined ESI/APCI ion source in a "Z-spray" arrangement that limits the entry of un into the vacuum section of the instrument. The ion optics in the inlet section uses "StepW to further limit the passage of uncharged particles to the mass analyzer while optimally f beam arriving at the first quadrupole. The instrument's greatest advantages are undoubtedly its very high sensitivity and, thanks UPLC technology in the separation, the significantly shorter analysis times compared to co instrumentation. Other advantages worth mentioning include the ability to quickly switch between ESI and AP having to change the ion source probe. Furthermore, due to the appropriate design of the i the input of uncharged particles to the mass analyzer is minimized. This actively reduces noise on the detector. The use of a photomultiplier in the incident ion detector provides a longer instrument lifetime compared to a conventional electro-multiplier. Finally, we would like to demonstrate the advantages of the new instrumentation with a con 2017, a method for the determination of 17 bile acids in rat and human bile was developed in our laboratory. The analysis time per sample was 40 minutes. Due to this time, only abo could be analyzed per day, including a 7-point calibration curve. This is not really much. quantification of this method was around 200 nM, which unfortunately did not allow the use for the analysis of individual bile acids in plasma, where we find concentrations much low Using the new instrumentation, not only was the time per analysis reduced to 11 minutes, b sensitivity of the instrument also allowed us to move the limit of quantification by two o magnitude compared to the original method. Thus, we can now reach low plasma concentration bile acids with sensitivity. For illustration, a comparison of the separation using the or instrumentation is shown below. In order to make the chromatograms comparable, the time ax both cases from 0 to 40 min. Author: doc. Ing. Miloš Hroch, Ph.D *========================================================================================= * Methods *=========================================================================================