Incognito believes that taking some time to step back and look at the broader picture could make your laboratory perform more efficiently and effectively.

Articles

Separating the Wood from the Trees

 has identified micro-particles within human urine that could be used for the non‑invasive diagnosis of bladder cancer. The occurrence of bladder cancer is rising every year, with the American Cancer Society 

 estimating that 73, 510 new cases of bladder cancer will be diagnosed by the end of 2012. The main methods of diagnosis are urine cytology and cytoscopy. Cytology is of low sensitivity when used as a diagnostic tool because it is poor at detecting low-grade bladder cancers, and cytoscopy is a highly invasive diagnostic procedure.

The team took urine samples from hernia patients (as a control) and bladder cancer patients and observed the differential expression of proteins within urine micro‑particles found in each of the samples. To do this, they utilized isotopic dimethylation labeling coupled with liquid chromatography–multiple reaction monitoring-mass spectrometry(LC–MRM-MS) to identify biomarkers within urinary micro-particles isolated from urine samples collected from hernia control patients and bladder cancer patients. 2,964 proteins were identified to be differentially expressed, present within the samples of either the controls or test subjects but not both, with 2058 novel proteins not previously reported. A total of 29 proteins were determined by statistical analysis to be differentially expressed at a level that made them potentially suitable to be biomarkers in bladder cancer diagnosis. The study quantified tumour-associated calcium transducer 2 (TACSTD2) within the raw urine specimens, confirming its potential use as a biomarker in diagnosis.

The identification of biomarkers has great potential in a clinical setting to reduce the method invasion and the discomfort to the patient in addition to reducing the cost of surveillance. 

 (Accessed 30th of October 2012, www.pubs.acs.org).

2.	American Cancer Society (Accessed 1st of November 2012,http://www.cancer.org/).

News

A group of scientists in Taiwan1 has identified micro-particles within human urine that could be used for the non?invasive diagnosis of bladder cancer.

Bladder cancer

Two novel peptides have been identified within the crude extract of Asian scorpion Buthus martensi Karsch venom using two-dimensional high perfomance liquid chromatography (2D-HPLC) and tandem mass spectrometry (MS–MS). Historically used in Chinese medicine to treat heart disease, the venom has potential applications in the development and identification of new drugs. Earlier this year, a study was published showing that peptides isolated from scorpion venom had anti-bacterial properties, specifically effective against established antibiotic‑resistant bacteria.

Scorpion venom is a complex mixture of proteins, including a range of neurotoxic peptides that selectively block cell-to-cell signalling by selectively binding to ion-channels. These peptides can be used in pharmocological studies to block specific channels and signalling, potentially revealing new drug targets. 

Based on genetic data, within the Buthus martensi Karsch venom it is estimated that there are 86 toxins, but only the presence of 46 peptides have been confirmed.

 The previous methods have been regarded as too low a sensitivity to detect all the petides present.

 has identified two novel peptides, previously predicted to be present in scorpion venom-based on genetic data, but otherwise not found. Offline orthogonal two‑dimensional reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography (2D-RPLC/HILIC) purification and collision-induced dissociation tandem mass spectrometry (CID-MS–MS) technique was used. Using bioinformatics the two novel peptides were predicted to be selective inhibitors of the potassium channel toxin α-KTx17 subfamily.

This research therefore has the potential to advance drug development through the discovery of new drug targets.

 DOI:10.1371/journal.pone.0040135 (2012).

Two novel peptides have been identified within the crude extract of Asian scorpion Buthus martensi Karsch venom using two-dimensional high perfomance liquid chromatography (2D-HPLC) and tandem mass spectrometry (MS?MS).

A sting in the tale

Since its introduction in 1975, ion chromatography (IC) has been used in most areas of analytical chemistry and has become a versatile and powerful technique for the analysis of a vast number of ions present in the environment. This article is a review of possible uses of IC in combination with MS detection for environmental research.

IC-MS and IC-ICP-MS in Environmental Research

The world’s population is predicted by the UN to reach seven billion by 2013.

 A proposed solution to increase food production has been the introduction of genetically modified (GM) crops, which has faced strong opposition. A group of investigators in the USA

 recently published a study indicating that environmental changes have a greater impact than genetic modifications on the metabolome of crop plants

: “As the population continues to grow, we need to use all the tools available to ensure there is enough food to meet global demand. GM crops are expected to play a critical role in meeting this need. GM crops have been examined for safety for more than 20 years now using targeted assays. Before metabolomics can be used for safety assessments of crops developed using biotechnology, it is necessary to understand the biological variation of metabolites in non-GM crops as a result of a different environment and genetic background.”

The investigators collected 654 grain samples and 695 forage samples from several sites in the USA and Canada, and measured the levels of 156 metabolites in grain and 185 in forage using gas chromatography (GC) coupled to time-of-flight mass spectrometry (TOF-MS). Multivariate statistics were used to identify a 2% change in the metabolome in non-genetically modified maize, whereas a change in location introduced a 50% change in the metabolome. This indicated that genetic modification had a minor effect on the metabolome when compared to the changes induced by factors of the natural environment.

1.	United Nations Information Service, (Accessed: 5th of November: www.unis.unvienna.org/unis/pressrels/2005/pop925.html).

et al., Journal of Agricultural and Food Chemistry,

A group of investigators in the USA2 recently published a study indicating that environmental changes have a greater impact than genetic modifications on the metabolome of crop plants.

GM crop safety

, (London, UK) have developed and optimized an assay for the simultaneous detection of eight forms of vitamin D. Vitamin D is produced within the body on sunlight stimulation in the skin (photo-activated D3 analog) in addition to a supply from dietary sources (vitamin D2) making analysis complex.

There are a range of reports suggesting that a vitamin D deficiency may have links to a number of disorders from neurological disorders through to hypertension. Hypovitaminosis D, a condition describing a deficiency of vitamin D from either dietary or sunlight sources, is now widely recognized to be an affliction of the western world as a result of low sunlight exposure andpoor diet.

Lead author of the study Dr Iltaf Shah told 

: “We observed that there was considerable debate about the veracity of vitamin D assays with many methods suffering from interferences from epimers, and in the case of mass spectrometric methods from isobars. Given that vitamin D deficiency is widespread and linked to a number of diseases, we set about developing a new assay with the capability of quantifying all eight forms of vitamin D. The assay uses two chromatographic columns used in tandem and mass spectrometer set to run three periods sequentially with optimized conditions for each analyte. Certified reference materials were also analysed to meet the standards outlined by National Institute of Standards and Technology (NIST).”

The investigators found that LC–MS–MS could be used to analyse serum samples under low‑light conditions; furthermore, the analysis of the multiple forms of the vitamin did not impact on the analysis of the commonly used 25OHD protein biomarker that is used in research and clinical applications.Shar went on to comment further: “The new method is sensitive, selective and accurate and it is efficient in extraction and chromatographic separation. The method is capable of accounting for the misleading measures due to the epimers and isobars. The results demonstrate the feasibility of applying the assay in research and clinical practice and to facilitate further in vivo investigation into the role of ubiquitous epimers.”

This new method holds promise for the improvement of current vitamin D analytical methods, and for the further development of 

 research looking into the role of different vitamin D epimers.

Scientists from Kingston University1, (London, UK) have developed and optimized an assay for the simultaneous detection of eight forms of vitamin D.

Vitamin D detection

The International Symposium on High Performance Liquid Phase Separations and Related Techniques - HPLC2013 Amsterdam - will take place at the Amsterdam RAI Conference Centre, Amsterdam from 16–20 June 2013.

The International Symposium on High Performance Liquid Phase Separations and Related Techniques - HPLC2013 Amsterdam - will take place at the Amsterdam RAI Conference Centre, Amsterdam from 16?20 June 2013.

HPLC2013 Amsterdam Preview

In recent years the advent of ultra high performance liquid chromatography (UHPLC) has broadened the scope of laboratory applications, making it possible to attain extremely low detection limits. However, there are certain requirements when working with this sensitive analytical technique:high purity water, solvents and reagents are necessary to take full advantage of UHPLC and achieve representative analytical data. Here a series of experiments demonstrates that freshly produced ultrapure water is suitable for UHPLC.

In recent years the advent of ultra high performance liquid chromatography (UHPLC) has broadened the scope of laboratory applications, making it possible to attain extremely low detection limits.

The Column-11-21-2012