High Speed Analysis: Combining Fast GC with Time-of-Flight Mass Spectrometry for Complex Sample Analysis in Under One Minute
Poster Apr 23, 2018
Jonelle Shiel, Matthew Soyk, Viatcheslav Artaev, Alexander B. Fialkov, Aviv Amirav.
The demand for more efficiency and productivity in the laboratory has led to applications using Fast GC-MS methods for routine analysis in a wide range of fields. Many of these applications can involve complex mixtures with some components at trace levels. The fast analysis time requires some unique capabilities, including a GC with fast column heating and cooling for high throughput, a mass spectrometer with high data acquisition speed, and a fast response ion source in order to correctly characterize narrow peaks generated by Fast GC. To make sense of the resulting compressed data, the software needs the capabilities of automatic peak finding and deconvolution of coeluting peaks to perform correct and meaningful identifications and quantitation. The performance of an instrument capable of combining Fast GC and TOFMS is illustrated in this poster.
To demonstrate capabilities, a separation of the entire Mega Mixture (69 of 76 components), can be completed in under one minute using Fast GC and a pre-production prototype benchtop TOFMS, which, with the FAST option, increases the scan speed maximum from 50 to 100 spectra/second. In addition, a sample containing Synthetic Drug Standards was also separated in under one minute. Forensic laboratories typically experience large workloads with little resources, therefore, fast turnaround times for analyses are valuable for this application. The total analysis time, including time between runs, is about 2.5 minutes for these examples.
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PfABCk2 gene is ligated into pET21a+ vector with His-tag at C-terminus and transformed into BL-21 (DE3) competent cells that are verified through Miniprep and DNA sequencing. Furthermore, this gene construct is utilized to heterologous express this protein with IPTG and afterwards purified the recombinant protein kinase using nickel affinity chromatography as shown on 10% SDS-PAGE with the expected 36 kDa protein band.READ MORE