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High noise level is undesirable since it makes it difficult for the integration software to adequately measure all of the peak area. Analysts rely on GC-MS and other GC methods to provide highly sensitive, low-level detection. Analysts using MS detection have an additional concern: high column bleed that can interfere with proper mass spectral identification of analytes.
Figures 2 and 3 show mass spectra of benzo(g,h,i)perylene at a 5 ng on-column level and an oven temperature of 325 °C.
The more instrument downtime you experience, the fewer billable samples you will be able to run in a given period of time. Low detection limits, positive mass spectral identifications, and instruments that are not down for maintenance are all important daily goals for a GC analyst. A decrease in peak shape quality in a capillary GC system can typically be traced to the inlet end of the column. To extend the lifetime of capillary GC columns, Supelco recommends using a 3 m long guard column. For use as guard columns to protect your analytical column from damaging sample components. This device consists of a double-tapered ferrule and a stainless steel compression housing with a threaded cap. You must have JavaScript enabled in your browser to utilize the functionality of this website. HP 6890 is a new Agilent GC, which is designed to replace its older version HP 5890 The HP 6890 Series gas chromatograph system delivers high levels of performance. Onboard sensors automatically compensate for ambient temperature changes and barometric pressure differences to routinely achieve more accurate and reproducible results. Competitive sport has been around for thousands of years and there is evidence to suggest that for a large amount of that time there have been athletes who have been looking to gain an advantage over their competitors by taking drugs (Bowers, 1997: 1299). Beta-2 agonists are generally used to treat respiratory illnesses such as asthma in humans and Chronic Obstructive Pulmonary Disease (COPD) in livestock and racing animals, as well as humans.
Table 1: R groups for beta-2 agonists used in this experiment, as well as adrenaline (epinephrine), the compound on which they are based.
Gas chromatography (GC) is an analytical technique which has been around for several decades and can be used to detect volatile substances in a variety of mixtures. The modulation device used in this experiment was a Longitudinally Modulated Cryogenic System (LMCS), which has a moving trap cooled by liquid carbon dioxide. The techniques which are most commonly used to detect beta-2 agonists are GC-MS and liquid chromatography (LC-MS) (Van Eenoo and Delbeke, 2010: 241).
This study aimed to develop a preliminary method of separating beta-2 agonists from a urine matrix in order to potentially lower detection limits for this class of drug. The three beta-2 agonists used in this exercise were clenbuterol, salbutamol and terbutaline and were provided as a gift by Dr John Vine from Racing Analytical Services Limited (RASL). Table 2 shows the total peak areas for each of the drugs at each of the concentrations, while figure 3 represents these results graphically.
Table 2: The average peak areas and relative standard deviations for each of the beta-2 agonists used.
Figure 3 shows that each of the drugs produces a trend line which is linear over the concentrations measured. While these results show that the relationship between the peak area and the concentration of the drugs is linear over the concentrations measured, they do not factor in how much of these signals may have been due to background interference or noise.
Again, the %RSD values for all of these points show that there is a high level of precision associated with them, which shows that the results are highly reproducible across runs. These results do however show that, even with an FID, which is not as sensitive or specific as an MS detector, the limits of detection are much lower for a GC×GC experiment than a traditional GC experiment. This complete separation from the matrix in the second dimension is important because it allows for more accurate identification of compounds of interest, as well as potential storage of data until a later date, which could potentially lead to retrospective analysis and catching of drug cheats from past samples, without having to store and reanalyse the sample. This research showed that beta-2 agonists could be separated completely from a urine sample using GC×GC technology and detected at levels much lower than could possibly be detected with a single GC column and similar detector.
The authors thank Dr Blagoj Mitrevski and Dr Sung Tong Chin, for their expert advice and assistance with aspects of instrumental analysis, as well as Dr John Vine for providing the drugs which were used in the research. Many analysts require natural gas analysis for BTU value calculations or drilling and mudlogging applications.
Capillary gas chromatography with flame ionisation detection (FID) or mass spectrometry are the techniques most frequently used for analysing the 16 PAH's targeted by the US EPA.
Excessive levels of column bleed which interfere with peak identification and quantitiation. For rapid analysis of PAH’s not requiring the separation of the benzo (b) and (k) fluoranthene isomers the HT8 range of columns are the best choice.
A specially designed palladium membrane is included in the design to optimize the purity of hydrogen (99.99999+ %). When a new gas tank is required, the analyst must transport a tank from a secure storage area to the laboratory.
A standard tank is quite heavy and can become a guided missile if the valve is compromised during transportation.
The hydrogen generator is a self contained unit that requires the user to simply add water on a periodic basis. 2.1The Licensed Material may not be used in any final materials distributed inside of your company or any materials distributed outside of your company or to the public, including, but not limited to, advertising and marketing materials or in any online or other electronic distribution system (except that you may transmit comps digitally or electronically to your clients for their review) and may not be distributed, sublicensed or made available for use or distribution separately or individually and no rights may be granted to the Licensed Material.
2.2One copy of the Licensed Material may be made for backup purposes only but may only be used if the original Licensed Material becomes defective, destroyed or otherwise irretrievably lost.


The better the signal-to-noise ratio, the more area counts are obtained, resulting in the ability to achieve lower detection limits.
When measurement is required at the ppb or even ppt level, extreme care must be taken to ensure that nothing interferes with the analysis. In addition to the primary, or quant ion < in a mass spectrum, MS programs are set to measure the abundance of other ions that are characteristic to the analyte of interest. When the fouling becomes severe enough to warrant action, the detector must be dismantled and cleaned, a procedure that may remove an instrument from service for one or more days. When you have a backlog of analyses to perform, you simply cannot afford unnecessary instrument downtime. Using a capillary column, such as Supelco SLB-5ms, that has a low bleed characteristic is an important step in achieving these goals.
Over time, the inlet end of the column becomes contaminated from an accumulation of non-volatile material. A guard column is a short piece of uncoated deactivated fused silica tubing which is placed in-line between the GC injection port and the analytical column. The HP 6890 series GC features electronic pneumatics control of all gas pressures and flows. By providing stable results, EPC reduces recalibration frequency and improves laboratory productivity. There has been a large amount of research which focused on detecting these and similar compounds in doping situations (Van Eenoo and Delbeke, 2010).
The most common methods which are thought of when sports doping is mentioned include steroids, erythropoietin (EPO) and, more recently, peptides and blood doping. Table 1 below lists the side (R) groups for each of the drugs used in this experiment, as well as adrenaline, on which their structures are based. This essentially leads to animals which become heavier much faster, consuming less feed and thus saving farmers money on the feed, while increasing profits when the animals are sold. The compounds in the mixture are heated up and vaporised, before passing through a column which is also heated by the instrument oven. The trap cools a section of the column, before moving up and releasing that section on to the short 2D column, moving back down and trapping the next section of the 1D effluent.
The instrument used was an Agilent 7890A GC-FID, with a 7683B autosampler, retrofitted with an LMCS modulation device. The other columns mentioned above were tested as 2D columns in experiments not described here; however, the separation of the components was not as complete, as well as there being a large amount of 2D tailing, so this column set was chosen for all experiments regarding the calibration curve.
The organic layer was then collected and treated in the same manner as the methanol standards previously, in that it was refrigerated when not in use and derivatised as described prior to analysis. The first measurement only takes into account the peak area of the compound, without factoring in any interference which may have been caused by either the electronic noise caused by the instrument or the chemical noise caused by the matrix from which the compound is being separated.
The %RSD statistic is a representation of how much of the mean peak area can be accounted for by the standard deviation. The highlighted point for clenbuterol was not used in the calibration curve for reasons discussed below.
Table 3 has the equations and coefficients of determination (R2 values) for each of these lines, which is essentially a measurement of how well each of the points is fitted to the trend line.
Current testing methods rely on target tests for compounds of interest, which may overlook other performance-enhancing or illegal drugs which may be present. This project was presented as a poster at both the 20th annual RACI Research and Development Conference in December 2012 and the 1st Monash-Warwick alliance International Conference for Undergraduate Research in May 2013, where it was awarded runner-up for best poster. He undertook postdoc research at the University of Bristol, then commenced his academic career at National University of Singapore. For more information on what data is contained in the cookies, please see our Privacy Policy page. Although there are only 16 compounds in this separation, many problems are encountered with this analysis. Figure 1 shows the separation of all 16 EPA nominated PAH's (with the exception of the two benzofluoranthene isomers) in only 18 minutes. In contrast, hydrogen is readily available via the electrolysis of water or as a high-pressure bottled gas. Peter Froehlich has over 30 years of experience in the analytical instrumentation industry. Except as specifically provided in this Agreement, the Licensed Material may not be shared or copied for example by including it in a disc library, image storage jukebox, network configuration or other similar arrangement. System noise refers to everything else, other than the analyte, producing a response in the detector. Figure 1 shows a graphic illustration of a peak when column bleed (a significant source of system noise) is low vs. For this reason, today’s chemists require capillary columns, such as Supelco SLB-5ms, that exhibit a very low level of bleed in addition to being inert towards the various analytes in the method.
For the software to assign a high probability of positive identification, these so-called secondary, or qualifier ions must be within specific ratio ranges relative to the primary ion when the mass spectrum from a peak in a realworld sample extract is compared to a mass spectral library entry. In order for the software to assign a high probability of positive identification, the mass spectrum from the sample extract must compare well with the mass spectral library entry. For TICs, the situation would be less desirable since retention time data would not be available to assist with identification. The phase can also be damaged from the continuous condensation and vaporisation of solvent and analytes. This study aimed to develop a preliminary method of separating beta-2 agonists from a urine sample, which could be used as a starting point for more complete identification, through multidimensional gas chromatography (MDGC), a technique which had not been applied to this class of drugs previously.


Clenbuterol is another member of this class, which is no longer used as an asthma treatment due to potentially harmful side effects. Beta-2 agonists act on the beta-2 adrenergic receptors in the body, which has several desirable effects, such as relaxing of the smooth muscles of the airways, which allows for easier breathing in patients suffering from asthma, as well as some potentially lethal side effects such as increased heart rate and tremors, although these are associated with stimulation of the beta-1 receptors, rather than the desired beta-2 stimulation (Van Eenoo and Delbeke, 2010: 229, 237).
The stationary phase of the column separates the components depending on their properties such as polarity, boiling point and chirality.
The advantage which GC×GC offers is that it can potentially separate compounds completely from a sample matrix, which allows for full mass spectra of each of the components to be obtained and a more complete representation of what is present in a sample found.
For the present work, an FID was used, but mass spectrometry offers better sensitivity and specificity. Although the detector being used was a Flame Ionisation Detector (FID) rather than a more sensitive and specific detector such as a Mass Spectrometer (MS) which has been previously used, if the technique was shown to separate the compounds of interest completely from the sample matrix then it could be reasonably assumed that more complete mass spectra of the compounds could be obtained, which would lead to lower detection limits. The LMCS modulator was programmed to come on at 6 min into the run and switch off at 17.5 min into the run.
Although the chemical noise is generally much lower in GC×GC experiments due to the improved separation offered by the second dimension, the electronic noise is generally higher due to the increased data acquisition rates which are required. For example, a %RSD of 50% would mean that the standard deviation is half the magnitude of the mean and there is too much variation between the data points to consider the data precise.
The point was also further than three standard deviations away from the expected result following the trend line obtained from using only the highest three concentrations and, as such, was considered an outlier and not used in any further measurements or discussions of the results. By allowing for full mass spectra of the entire mixture to be obtained, every compound could be analysed, rather than only specific drugs which would be looked for in a standard drug testing run.
Although these levels are not lower than what can be obtained with a GC-MS instrument, they were shown to be much lower than what is possible with a single dimension GC-FID and, if an MS were used as the detector, it could be reasonably assumed that the detection levels would significantly drop. His main research activities are in Analytical Chemistry, specifically gas chromatography (multidimensional GC and comprehensive 2D GC) with mass spectrometry. This short analysis results from the ability of HT8 columns to operate routinely at very high temperatures. Upon download of any film Licensed Material, you will be invoiced a non-refundable access service fee of one hundred fifty dollars ($150) USD or such other local currency amount as Getty Images may apply from time to time.
If extraneous ions, such as those column bleed, are present in the mass spectrum, the software will assign a lower probability of positive identification. High column bleed levels can interfere with this comparison, resulting in the reporting of TICs that are either poorly identified or misidentified. The active foil in an ECD can become fouled to the point the entire detector needs to be sent out for refurbishing. By clipping the guard column periodically to restore performance instead of the analytical column, the analytical column remains unaltered. This class of drug has been on the WADA banned list for over a decade and, in that time, has been in the top 6 most detected doping techniques in terms of the number of Adverse Analytical Findings (AAFs) attributed to it (Van Eenoo and Delbeke, 2010: 239). They are also thought to have anabolic properties if taken at higher than prescribed doses and, as such, are banned for use by athletes unless it can be proven that they have a medical condition which requires their use (Polettini, 1996: 28). Multidimensional GC (MDGC) is a more recent technique which involves a second column being coupled to the first either by a switching device which cuts out a section of the original chromatogram to be analysed on the second column, or by a modulation device which traps a small segment of the first dimension (1D) effluent before releasing it into the second dimension (2D) column and trapping the next segment. The GC columns used in this experiment were a VF200ms, DB-5MS UI and DB1701, purchased from Agilent Technologies, as well as an Rxi-17Sil, purchased from Restek Corp. Experiments which are not described here were carried out and the optimum PM was found to be 5 s and TM 40°C. A %RSD value of under 5% is generally considered acceptable in a practical sense, although this number is not an absolute maximum value and values up to 10% are often considered to still be acceptable, particularly when dealing with low concentrations such as those which are seen in this experiment. This means that the points show a very high level of correlation to the trend line obtained.
This means that the signal from the compound of interest is almost impossible to determine due to the background interference.
He has almost 300 refereed International publications, book chapters and patents and has collaborated with researchers from all around the globe. The Licensed Material may only be used in materials for personal, noncommercial use and test or sample use, including comps and layouts.
If Licensed Material featuring a person is used (i) in a manner that implies endorsement, use of or a connection to a product or service by that model; or (ii) in connection with a potentially unflattering or controversial subject, you must print a statement that indicates that the person is a model and is used for illustrative purposes only. Create your slideshowBy using the code above and embedding this image, you consent to Getty Images' Terms of Use. The ratio of the peak height to the baseline variability is termed the signal-to-noise ratio. While this is not as low as is possible using a GC-MS instrument, this technique showed great potential for separating these drugs from the urine matrix and, if coupled to a more sensitive detector such as a Mass Spectrometer (MS), is expected to deliver much greater sensitivity and certainty of identification than traditional GC techniques. Therefore the need to be able to detect low concentrations of beta-2 agonists in a variety of matrices is not only a problem associated with the integrity of sport, but also potentially affects the health of the meat eating public. The time between each of the movements of the modulator is known as the modulation period (PM) and is one of the modulation factors which has to be optimised when conducting a GC×GC experiment, as well as the modulation temperature (TM), which has to be carefully chosen in order to maximise the trapping of the effluent from the first column.
These ratios are generally much higher for GC×GC experiments than single dimension GC due to the almost complete separation offered by the second dimension. This is well illustrated in Figure 1 where effectively no baseline change is observed even when the final temperature is 380 °C.



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