Food Archive
  
 
 SEPARATION SCIENCE LATEST EDITION 
 
Analysis of Propionic and Acrylic Acids by Reversed Phase HPLC with Anion Suppression and Conductivity Detection
A liquid chromatographic method for the determination of propionic and acrylic acids in aquatic mixtures was developed by combining three techniques: HPLC reversed-phase separation, anion suppression and conductivity detection. This combination yields a similar response for both acids allowing their determination at ppm level and was found to be applicable to samples containing other organic compounds. These substances were chosen because of their environmental relevance and because of the increasing interest related to their involvement in different chemical and biological processes. Reproducibility, accuracy, sensitivity and linearity of this method were evaluated under optimized conditions, showing that suppressed conductivity detection combined with a reversed-phase column improves the separation and detection of both carboxylic acids when compared with non-suppressed UV detection.

 
Optimization of Analytical Methods Using Factorial Designs
Part 1: An Introduction
The conventional approach for optimizing analytical methods in the laboratory is the one-factor-at-a-time approach, where each experimental factor or parameter is optimized separately and independently of other factors. In contrast, factorial designs involve simultaneous optimization of all factors at once. Factorial designs offer a simple, efficient, and statistically valid method for optimizing analytical methods. In this article, the first of a three-article series, we will take an introductory look at factorial designs and its benefits to the analytical laboratory.
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Implementing Robustness Testing for HPLC Methods
This is the second article in a series about robustness testing for HPLC analytical methods. Previously, an overview of the steps involved in investigation of robustness was described. The first step is to decide which factors are going to be studied. In this article the selection of appropriate robustness factors and factor levels relating to the mobile phase will be discussed and in particular, the volume fraction of the organic solvent in the mobile phase for reversed phase HPLC.

 
Method Transfer and Routine-Use Study of the USP Impurities Method for Donepezil Tablets
In this article we investigate some of the sample-matrix related issues encountered when transferring a long drug final formulation separation method to a faster, more sustainable LC method. The geometrically-scaled gradient method presented in this article is 80% faster than the original compendial HPLC method and consumes 92% less mobile phase.
 
 
How to Analyse Unknown Samples by Pyrolysis GC/MS?
Although GC/MS and LC/MS are very powerful analytical techniques, they generally lack the ability to analyse and characterize insoluble materials, such as polymers and other high molecular weight products. An elegant approach to cope with this type of samples involves a combination of thermal decomposition and online chromatographic analysis of the released breakdown products. This approach is called analytical pyrolysis.
 
 
How Fittings Can Impact Chromatographic Results
Anyone who has been involved in chromatography has likely struggled with things like band broadening, split peaks, carryover, and loss of sample. Typically, when these challenges are encountered, it’s normal for chromatographers to suspect degradation of the column’s stationary phase or issues with the mobile phase, or even mechanical problems with primary system components (e.g., the injection valve, the pump, etc.). However, after investing hours and precious financial resources towards “fixing” or replacing the suspected components, it’s entirely possible that the initial challenges may still be present, leaving many a chromatographer puzzled and frustrated.
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 HPLC SOLUTIONS LATEST EDITIONS 
 
How to Calculate k*
In the previous HPLC Solutions (#93) looked at a problem gradient method where peaks in the second and subsequent chromatograms were eluted too early. Our conclusion was that the most likely cause was insufficient equilibration time between runs. However, I also suggested that we make a quick check to see if the gradient conditions are reasonable, and this is even more important now that we’ve already discovered one problem with the method.
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Poor Equilibration
A reader emailed in the following question: I am trying to analyze a peptide (2675 Da) on a 150 x 2.1 mm, 3 μm particle column by LC-MS using an ion trap MS in the positive electrospray ionization (ESI) mode. The first injection is OK, with the peptide eluting at ≈ 15 min with good peak shape, but in subsequent runs the retention time is ≈5 min. If I try to run the method the next day, the same pattern occurs. The A-solvent is 10% acetonitrile (ACN) in 0.1% trifluoroacetic acid (TFA); B is 100% ACN with 0.1% TFA added. The gradient runs from 0-100% B over 30 min, then steps back to 100% A for 20 min prior to the next injection. Can you spot any obvious problems here?


Calculated Peak Width
In last week’s HPLC Solutions (#91), we looked at a problem where we suspected that the data acquisition rate of the data system was too slow to gather a sufficient number of data points across a peak with a 2-μm particle column. The data of Table 1 were generated to help give us a reality check on the hypothesis. This week we’ll look at those calculations in a little more detail, in case you want to perform a similar exercise for your HPLC system.
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Peaks Too Narrow
I recently had a reader send me an email complaining that the peaks in his chromatogram were too narrow. At first, this seems a bit odd – after all, most people complain because peaks are too wide, so narrow peaks usually are preferred over broader ones. Digging into the question a bit more will highlight the real problem.
     


 GC SOLUTIONS LATEST EDITIONS 


Is it Working?
Is the analytical result you are reporting valid? Maybe the ones from yesterday or last week were, but will the ones from today also be? Will the results from your tray of 70 samples have to be thrown out and the analyses be run again? This month we talk about avoiding problems through routine maintenance and verifying that your instrument is working through the use of a performance verification method.
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What’s New?
As I prepare to attend PittCon 2012, I suspect that the answer to the question “what’s new in gas chromatography?” is “not much”. Is that because there is nothing left for instrument manufacturers and researchers to do in the area or is there something else going on?
  


 MS SOLUTIONS LATEST EDITIONS 

Use of ECD and ETD Fragmentation Mechanisms for Peptide Sequencing
In the previous installment of MS Solutions, I described some of the problems encountered with collisionally-activated dissociation (CAD) of peptides when used in MS/MS sequencing experiments. We noted that the number and type of structurally significant ions produced is dependent on the sequence of amino acid residues. Some sequences, most notably those containing a a C-terminal arginine, result in very little in the way of useful MS/MS spectra.
The Importance of Multiple-Charge Ion Precursors in Peptide MS/MS Sequencing
We typically think of a mass spectrum as consisting of a precursor ion at high m/z and a series of increasingly lower m/z product (fragment) ions. These types of mass spectra are shown in training courses and textbooks as didactic examples because they are easy to interpret “by hand”. 


Determination of Intact Protein Molecular Mass
From Multiple-Charge Electrospray Mass Spectra 
In two previous instalments of MS Solutions, I discussed the 
sequencing of simple, single-charge peptide ions using 
MS/MS. In this and another upcoming edition I am going to discuss 
the use of multiple-charge ions in protein and peptide analysis.
 


CE SOLUTIONS LATEST EDITION 
 
Method Development in CE: Selecting your background electrolyte
The selection of the proper background electrolyte (BGE) is key to a successful CE method. In this second issue of our series on CE Solutions we first look into the choices for BGE based on our analyte properties. Capillary electrophoresis (CE) is based on migration of charged components in an electric field. So when we start our method development, the first thing we need to consider is how to get our analytes to carry charge. We will look into the effects of pH on analyte charge and discuss opportunities to induce charge on neutral analytes. Additionally, we look into those aspects of a BGE that make the peaks sharp and the precision and robustness good.

 
Why CE?
This article is the first in a new series about capillary electrophoresis (CE), aims to be a low-threshold, practical and pragmatic aid for that implementation process by focusing on good CE working practices and troubleshooting.
 


 HPTLC SOLUTIONS LATEST EDITION 
 
Parameters of the HPTLC Process
An inherent advantage of planar chromatography is its enormous flexibility derived from the off-line principle and interaction of many parameters. It is helpful to know these parameters for each step of the process and to understand how they can affect the overall result in order to utilize the advantages for unlocking the full potential of high performance thin-layer chromatography (HPTLC). This issue of HPTLC Solutions is dedicated to the first part of the HPTLC process which includes plate handling, sample application, and chromatogram development. Read on for more information...

 
HPTLC Today
What is High Performance TLC and how can you benefit from it?
In contrast to column chromatography (e.g., GC, HPLC), planar chromatography utilizes a flat (planar) stationary phase for separation. In thin-layer chromatography (TLC) this stationary phase is supported by a glass plate or a foil (plastic or aluminium). The TLC plate constitutes an open system, which during analysis passes through the individual steps (sample application, chromatogram development, detection, etc.) in an off-line mode. In this process many samples can be analysed in parallel on the same plate. The possibilities for adjusting and combining numerous parameters in order to optimize the separation create a flexibility that is unsurpassed by any other chromatographic technique.
 

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