On-Line Coupling of Gel Electrophoresis and Inductively Coupled Plasma-Sector Field-Mass Spectrometry for the Determination of dsDNA Fragments

Abstract

The on-line coupling of gel electrophoresis (GE) and inductively coupled plasma-mass spectrometry (ICPMS) is described for the first time. The new method combines the separation power of GE for large biomolecules and the high sensitivity and elemental selectivity of ICPMS. This coupling has been achieved by means of gels housed in glass tubes (length 2.5-20 cm; i.d. 0.5-5 mm). The gel is fixed by glass frits permeable for the analytes. After the electrophoretic separation at voltages up to 500 V, the analytes are transferred to the nebulizer of the ICPMS by an eluent stream, which is separated from the electrode chamber by a membrane. This filter blocks molecules with molecular masses larger than 500 Da. Using double-stranded DNA fragments, commercially available standard solutions were analyzed with on-line GE-ICPMS for the first time by monitoring (31)P(+) with a double-focusing mass spectrometer at a mass resolution of 4000.

Characterization of nanoparticles by capillary electromigration separation techniques

“…With a detection unit consisting of a laser emitting at 266 nm and a DAD recording from 500 to 700 nm coupled to an ITP unit, the recording of emission spectra was possible for migrating CdTe nanoparticles coated with mercaptoacetic acid [99]. Selective and sensitive detection of metal and semiconductor nanoparticles is also possible by ICP-MS. To this end, Brüchert and Bettmer [100] and Helfrich et al [101] developed a device that allows the on-line coupling of microcolumn gel electrophoresis with ICP-MS. The authors demonstrated the size-selective separation of gold nanoparticles (mean diameter 5 and 20 nm, agarose gel, 2.0%) coupled on-line with element-selective detection.…”

Modification of tricine–SDS–PAGE for online and offline analysis of phosphoproteins by ICP-MS

“…The µLC-ICP-MS (pioneering work described by Lehmann and co-workers [32] The recently introduced on-line coupling of GE to ICP-MS is a further development and was first described by Brüchert and Bettmer [35] for the determination of dsDNA fragments. More recently, it has been reported for several applications including: size characterization of gold nano particles [36], detection of iron in metalloproteins [37], determination of the degree of phosphorylation in casein [38], and for iodide and iodate determination in aerosols [39].…”

Inductively Coupled Plasma–Mass Spectrometry (ICP-MS) for Quantitative Analysis in Environmental and Life Sciences: A Review of Challenges, Solutions, and Trends

“…17 Recently Brüchert and Bettmer developed an interface for the online coupling of 1D slab gel electrophoresis and ICP-MS that allows the separation and quantification of DNA strands. 158 Detection limits of 1 ng DNA absolute have been achieved, which corresponds to 96 pg of phosphorus. In comparison to CE, relatively poor separation efficiency can be obtained with a 1D agarose gel, which represents the main limitation of this setup.…”

“…195 They introduced a new interface comparable to those described by Brüchert and Bettmer for on-line isotope-dilution GE-ICP-MS. Iron-sulfur ratios were measured to monitor possible iron loss during the sodium dodecyl sulfatepolyacrylamide gel electrophoresis separation. 158,195,196 In summary it is fair to say that most examples regarding the absolute quantification of a biomolecule via its sulfur content have been carried out with proteins. In consequence, currently only protein samples of low complexity can be quantified with a sulfur-based approach due to the problems related to the chromatographic separation of intact proteins.…”