LCGC North America-09-01-2015

LCGC North America
LC Troubleshooting

September 01, 2015

I recently returned from a tour of teaching liquid chromatography (LC) classes to users in Minnesota, the United Kingdom, Poland, and Malta. One thing that always impresses me on such trips is that no one group has a corner on the LC problem market. The same problems pop up in most laboratories, no matter where they are located, the role of the laboratory (for example, analytical, forensic, production, research), what industry is involved, or the brands of instrumentation used.

LCGC North America

A systematic approach for formation of sigmoidal gradient for analysis of biopharmaceutical proteins has been reviewed and discussed together with two case studies. The first application involves HPLC separation of the various product related variants of a microbial expressed biotherapeutic, granulocyte colony stimulating factor (GCSF). The second case study involves separation of the charge heterogeneity related variants for a monoclonal antibody (mAb) biotherapeutic product. In all cases, it is observed that the use of sigmoidal gradient successfully reduces the analysis time significantly (from 70 minutes to 15 minutes for GCSF and from 40 minutes to 4 minutes for the mAb) while retaining the selectivity and the resolution.

Ion-mobility spectrometry (IMS) is an anlytical technique that provides for the separation of ions in the gas phase. The separation, occuring on a timescale of milliseconds, is based on the differing mobility of ions according to their charge, shape, and size. These characteristics make IMS suitable for coupling with mass spectrometry (MS), to serve in current MS-based workflows for metabolomics and lipidomics. IM-MS improves peak capacity and signal-to-noise ratios, and it provides more confidence during compound identification or confirmation than conventional analyses. Combining collision-induced dissociation with ion-mobility separation improves the specificity of MS/MS-based approaches. Significantly, ion-mobility-derived information provides an orthogonal, physicochemical parameter-the collision cross section (CCS)-which relates to a metabolite's shape. Novel hardware and software solutions enable analysts to process and exploit IMS-MS data for qualitative and quantitative metabolomics and lipidomics applications.

A simple and accurate method for determining trace level of carcinogenic solvent carbon tetra chloride (CCl4) in Flunixin Meglumine, a drug substance which is white crystalline powder was developed by Gas chromatography .The method was observed to be robust and complete analytical method validation was performed and meets the requirements as per ICH guidelines. The method possesses the lowest detection level when compared with other methods currently available. The LOQ achieved by this method was 0.8 ppm and calibration curves were linear, R >0.998.In general, for analysis of halogenated compounds electron capture detector (ECD) were mostly used and in this proposed method flame ionization detector (FID) with internal standard was used. The method was implemented for various Active pharmaceutical ingredients (API) successfully.

LCGC North America
Column Watch

September 01, 2015

HPLC 2015, chaired by Gérard Hopfgartner was held in Geneva, Switzerland, from June 21–25. This installment covers some of the highlights observed at the symposium including stationary phase developments, particle technology, and areas of growing application of HPLC. In addition, trends and perspectives on future developments in HPLC culled from the conference are presented.

LCGC North America
The Essentials

September 01, 2015

Improve your headspace sampling methods through a deeper understanding of the effects of key method parameters

Issue PDF
LCGC North America

September 01, 2015

Click the title above to open the LCGC North America September 2015 regular issue, Vol 33 No 9, in an interactive PDF format.