Comprehensive Two Dimensional

Trends in Analytical Chemistry, Vol. 25, No.
4, 2006
Trends
Comprehensive two-dimensional gas chromatography for the analysis of organohalogenated micro-contaminants

Peter Korytar, Peter Haglund, Jacob de Boer, Udo A.Th. Brinkman
We explain the principles of comprehensive two-dimensional gas chromatography (GC GC), and discuss key instrumental aspects with emphasis on column combinations and mass spectrometric detection. As the main item of interest, we review the potential of GC GC for the analysis of organohalogenated micro-contaminants, and highlight its superiority over conventional 1D-GC. We present results for 12 compound classes, including polychlorinated biphenyls, dibenzo-p-dioxins and furans, and n-alkanes, toxaphene and polybrominated diphenyl ethers. We draw attention to target analysis as well as within-class and between-class separations. 2006 Elsevier Ltd. All rights reserved.
Keywords: Comprehensive two-dimensional gas chromatography; GC GC; Mass spectrometry; Organohalogenated compound; Separation
1. Introduction In 1966, the Swedish chemist Jensen [1] announced that he had identied a group of organochlorine compounds that was accumulated to toxic concentrations in nature. Unlike the closely related chlorinated pesticides, such as DDT and the various drins which had, so far, attracted most attention of environmental chemists the newly detected compounds, the polychlorinated biphenyls (PCBs), had entered the environment essentially unintentionally. Since that time, these two classes of persistent organic pollutants, which shared high annual production, widespread usage, long persistence and serious toxic effects, have been the subject of a rapidly increasing number of fundamental as well as applied studies. Over the years, many related classes of compounds have been added to the list; Table 1 gives an overview, and also lists the number of theoretically possible congeners, which gives an impression of the degree of complexity that can be expected in technical mixtures and, thus, in environmental and food samples. For the rest, we assume that the general reader will be familiar with the main characteristics and usage of the various classes of organohalogens and, at least in some cases, with the catastrophes or otherwise, which led to their being considered priority pollutants by many governments and international (UN, EU) bodies. During the entire past half-century, gas chromatography with electron-capture 373
Peter Korytar* Netherlands Institute for Fisheries Research, P.O. Box 68, NL-1970 AB IJmuiden, The Netherlands Free University, Department of Analytical Chemistry and Applied Spectroscopy, de Boelelaan 1083, NL-1081 HV Amsterdam, The Netherlands Peter Haglund Umea University, Department of Chemistry, Environmental Chemistry, SE-901 87 Umea, Sweden Jacob de Boer Netherlands Institute for Fisheries Research, P.O. Box 68, NL-1970 AB IJmuiden, The Netherlands Udo A.Th. Brinkman Free University, Department of Analytical Chemistry and Applied Spectroscopy, de Boelelaan 1083, NL-1081 HV Amsterdam, The Netherlands
Corresponding author. E-mail: Peter.Korytar@wur.nl
0165-9936/$ - see front matter 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.trac.2005.12.003
Trends
Trends in Analytical Chemistry, Vol. 25, No. 4, 2006
Table 1. Main classes of polyhalogenated micro-contaminants Name Acronym Maximum number of congenersa 209 75 135 61 696 16 640 12 288 32 768 ca. 300 8149 209 75 Very high 209 209
Polychlorinated biphenyls Polychlorinated dibenzo-p-dioxins Polychlorinated dibenzofurans Toxaphene components bornane congeners camphene congeners dihydrocamphene congeners Organohalogenated pesticides Polychlorinated terphenyls Polychlorinated diphenylethers Polychlorinated naphthalenes Polychlorinated alkanes Polybrominated biphenyls Polybrominated diphenylethers
a
PCBs PCDDs PCDFs Toxaphene
OCPs PCTs PCDEs PCNs PCAs PBBs PBDEs
Enantiomers not included.
detection (GCECD) was the predominant analytical technique for selective and sensitive determination of organohalogens. In the early part of that period, when packed-column GC was the only means of separation available, total-PCB determination (based on comparing an environmental sample with a technical PCB mixture) was all that could be achieved. The introduction of (fused-silica) capillary columns was a real breakthrough, which, all of a sudden, enabled congener-specic determination of the PCBs and, of course, also of the other classes of organohalogens. However, it also became clear, after this major step forward, that capillary GC was not the solution for all, or even most, problems. Admittedly, single-column (or one-dimensional, 1D) GC can provide the required resolution when the number of target analytes is restricted (e.g., with the organochlorine pesticides (OCPs) or polychlorinated naphthalenes (PCNs)). However, even for the PCBs with their still moderate number of 209 congeners, or for the about 140 CB congeners present in a technical mixture, no satisfactory 1D-GC solution could be found. And, not surprisingly, correspondingly more serious problems were encountered with complicated mixtures, such as toxaphene, polychlorinated terphenyls (PCTs) and, specically, polychlorinated alkanes (PCAs); the PCAs typically show up in 1D-GC as an essentially unresolved broad band covering a major part of the baseline. The magnitude of the problem becomes even clearer when one considers that the present discussion is about within-class separations only: interferences caused by other organohalogens and/or matrix constituents or problems due to widely different concentrations of (partly) co-eluting congeners have not been taken into account. Over the years, several approaches have been used to alleviate the problems. One of these was sample fractionation (e.g., by size-exclusion or adsorption liquid 374
chromatography). Another was to lter out interferences by selective mass spectrometric (MS) detection, which is powerful but fails to resolve co-elutants with closely similar mass spectra, as is often the case for congeners of the same compound class, but also of different classes. On the GC side, one way to go was the parallel use of several stationary phases and the combination of information from the GC runs on different columns to solve specic separation problems. It was a step further to combine two different columns in a single set-up: in socalled multi-dimensional GC (MDGC), selected fractions from the rst column are subjected to a second GC run that uses a completely different separation mechanism. Many successful applications have been reported, but one should keep in mind that, in essentially all cases, they are of a heart-cutting nature: only a single, or at best a few, small fractions are transferred to the second column for further separation. That is, it is an excellent solution when information is required about a few target analytes, but not about the entire sample. In the latter instance (i.e. with complex samples and/or when unknowns have to be traced), MDGC becomes much too complicated and timeconsuming. It is in these situations and their number can condently be said to be increasing daily that a socalled comprehensive approach is needed: MDGC, or GC GC, is now replaced by GC GC, with which, instead of a few selected fractions, the entire sample is subjected to separation on two different columns. One immediate advantage is that the information content is much greater, and another is that the GC GC run is ready once the rst-dimension run is nished; that is, GC GC is much more efcient than MDGC. It is the goal of this review briey to explain the principle of GC GC and to discuss key aspects, such as column-to-column interfacing or modulation, detection and detector requirements (including a comparison of time-of-ight MS (TOF-MS) and fast-scanning quadrupole MS (qMS) instruments) and the selection of properly matched column combinations. We will devote most attention to applications in the eld of organohalogenated micro-contaminants, which will also be used to highlight the added value of GC GC compared with 1D-GC. Readers who are interested in a more detailed review of the various aspects of GC GC and in applications to compound classes other than organohalogens, should consult two extensive reviews [2,3].
2. GC GC: general principle In GC GC, the entire sample is subjected to two GC separations that are based on different separation mechanisms. Fig. 1 shows a schematic of a GC GC system. In most instances, the sample is rst separated on a high-resolution capillary GC column typically a 1530 0.250.32 mm ID, 0.11 lm df column
http://www.elsevier.com/locate/trac
Trends
Figure 1. Schematic of a GC GC system with different modulator types.
containing a non-polar stationary phase. As will be explained in Section 3, an interface called a modulator is used to separate the rst-column eluate into a very large number of adjacent small fractions. To maintain the rst-column separation, these fractions should be no larger than one quarter of the peak width, or r, in that dimension. In order to meet this so-called modulation criterion, temperature programming in GC GC is slower than in 1D-GC, and typically occurs at a rate of 13C/min. Each individual fraction is trapped, refocused and, next, launched into the second GC column, which is much shorter and narrower than the rst one typical dimensions are 12 m 0.1 mm ID 0.1 lm df. The second-column separation generally is of a polar or shape-selective nature. That is, the separation mechanisms are indeed different or, in other words,
orthogonal separation conditions have been created. The separation in the second column is extremely fast and usually takes only 28 s as against 45120 min for the rst-dimension separation. Consequently, it is performed under essentially isothermal conditions. The fast separation in the second dimension causes the analyte peaks to be very narrow with widths of, typically, 100600 ms at the baseline. These narrow peaks require fast detectors with a small internal volume and a short rise time in order to achieve a proper reconstruction of the (seconddimension) chromatograms. In some systems, the second column is housed in a separate oven to allow more exible, independent temperature programming. The outcome of a GC GC run is a large series of highspeed, second-dimension chromatograms, which are usually stacked side by side to form a two-dimensional 375
Trends
(2D) chromatogram with one dimension representing the retention time on the rst column and the other the retention time on the second column. The most convenient way to visualize these chromatograms is as contour plots, where peaks are displayed as spots in a 2D plane using colors and/or shading to indicate signal intensities. Apex plots, which use specic symbols to indicate the position of the peak apexes, are another frequently used means to display GC GC chromatograms; the overall presentation becomes much simpler, and that is especially advantageous when ordered structures are studied (in this review, apex plots are used (e.g., Figs. 4 and 8) and are combined with contour plots (e.g., Figs. 9 and 10); contour plots are also displayed (e.g., Figs. 2 and 5). There is general agreement regarding the main advantages of GC GC over 1D-GC. Most strikingly, the peak capacity is much higher, and that yields a dramatically improved separation of the analytes of interest from each other but also and this is often even more important from interfering matrix constituents. In addition, a main benet of the trapping-plus-refocusing occurring during modulation is, typically, a 310-fold improved
signal-to-noise ratio compared with 1D-GC. Finally, compound identication is more reliable in GC GC because each substance now has two identifying retention values rather than one. Specically, when orthogonal conditions are used, chemically related compounds show up as so-called ordered structures (i.e. as clusters or bands). This phenomenon greatly facilitates group-type analysis, ngerprinting studies and the provisional classication of unknowns. It is particularly important in the study of classes of organohalogens, as will become clear from most of the gures included in Sections 6 and 7. In the following sections, we will discuss in some detail three topics of general analytical interest: modulation; detection and analytical performance; and, column selection.
3. Modulation The key component of a GC GC instrument is the modulator that joins the two columns. It serves three main goals:
Figure 2. GC GClECD chromatograms of PCA-60 technical mixture on DB-1 in the rst dimension and each of the six columns indicated in the second dimension [24].
376
Trends
(i) collecting and focusing of each of the fractions eluting from the rst-dimension column; (ii) re-injecting/launching of each collected fraction into the second-dimension column; and, (iii) trapping of the next eluent fraction from the rst column during the launch of the preceding fraction. Over the years, different ways of modulating the rstcolumn efuent have been reported (Fig. 1). Initially, heating was preferred, with a rotating slotted heater, the sweeper, rapidly moving over a thick-lm modulation capillary, heating it locally. Despite its frequent use in the early years of GC GC, because of the vulnerability of the set-up, the time-consuming optimization of the multi-parameter design and the restricted application range, the sweeper and related modulator types are obsolete today. The rst major step forward was the introduction of the longitudinally modulating cryogenic system (LMCS). This modulator uses expanding CO2 (liquid) for trapping of the analytes at the top of the second-dimension column. By subsequently moving the trap rapidly to an upstream position, the re-focused zone is exposed to the GC oven air and instantaneously volatilized and launched. Today, jet-based modulators with no moving parts at all with either CO2 or liquid N2 for cooling are generally preferred. Single-, dual- and quad-jet modulators have been introduced, and several of these have been extensively compared in a recent study [4]. The main conclusion was that all cryogenic modulators, if properly optimized, can satisfactorily be used for most applications, and certainly for those in the eld of organohalogen analysis. One caveat should be added. In order to ensure proper modulation of high-boiling compounds (i.e. highly substituted congeners), cooling should be just enough to trap the target compounds safely. Cooling that is too strong can cause remobilization to be inefcient and may lead to distorted and/or tailing peaks [5]. In addition, severe cooling will lead to more bleed from the rst-dimension column being accumulated and that, in turn, will yield noisier seconddimension chromatograms. Some studies have indicated that modulation of high-boiling compounds can easily be achieved by air cooling [6]. If found to be true in further studies, this will make GC GC of organohalogens less expensive and simpler to operate.
4. Detection As briey mentioned in Section 2, detectors with a high data-acquisition rate and a negligible internal volume are required to describe properly the very narrow peaks that are the outcome of a GC GC run. Flame ionization detectors (FIDs) have data-acquisition rates up to 200 Hz and dead volumes that are effectively zero. It does not
therefore come as a surprise that virtually all early GC GC studies were carried out with an FID, including those dealing with organohalogens [7]. However, as soon as real-life studies and analyte detectability became an issue of interest, it became clear that FID would have to be replaced by ECD to obtain sufcient selectivity and sensitivity. Conventional ECDs have data-acquisition rates up to 50 Hz, but the main problem is their 1.5-ml cell volume, which causes severe peak broadening [8]. Kristenson et al. [4] showed that, from amongst the miniaturized ECDs marketed in recent years, only the Agilent micro-ECD (lECD) with an internal volume of 150 ll gives acceptable peak widths. The best results were obtained when working at the maximum ow of make-up gas (150 ml/min) and at temperatures above 300C. However, as is to be expected, even under optimum conditions, the lECD delivers about 2-fold broader peaks than an FID [9]. All recent GC GClECD studies use an Agilent detector. The main problem of element-selective detection in general and, therefore, also of ECD is that no structural information is provided. In other words, MS is indispensable for identication/conrmation of the numerous separated compounds, whether target analytes or unknowns. Today, the preferred choice is a TOFMS that can, typically, acquire up to 50500 mass spectra per second (with unit mass resolution). The coupling of GC GC separation and TOF-MS detection presents no difculties, and there is no additional peak broadening. This is true for both conventional electron impact (EI) ionization and the recently introduced electron-capture negative ionization (ECNI) mode. An additional advantage of a TOF-MS is that the sensitivity is higher than that of the full-scan mode of conventional scanning MS detectors, while the high acquisition rate prevents spectral skewing, and deconvolution consequently becomes a very powerful tool. Unfortunately, TOF-MS instruments are very expensive. From the start therefore, the potential and limitations of qMS instruments, which are available in essentially all GC laboratories, were investigated by several groups of workers [1013]. Briey, the outcome of these studies was that, for restricted mass ranges of 150200 Da, acquisition rates of 2030 Hz can be obtained, and that tentative identication is then indeed often possible, but that proper quantication cannot be achieved on the basis of the 35 data points registered across a peak. Recently, the situation has improved (i.e. when two rapid-scanning qMSs were marketed, the Shimadzu QP 2010 and the Perkin Elmer Clarus 500, which allow scanning up to 10,000 Da/s [14,15]). Table 2 provides an illustrative comparison of these two instruments and a state-of-the-art conventional qMS. A clear indication of the improved performance is that, if the lower limit of the peak width at the baseline is set at 300 ms, the conventional qMS is restricted to a mass
377
Trends
Table 2. Comparison of performance of two rapid-scanning and a conventional qMS in GC GC [11,1315,62] Mass range (Da) or number of ions monitored Perkin Elmer Clarus 500 maximum acquisition (Hz) 17 23 31 63 minimum peak widtha (ms) 410 300 230 110 Shimadzu QP 2010 maximum acquisition (Hz) 20 25 33 50 minimum peak widtha(ms) 350 280 210 140 Agilent MSD HP 5973 maximum acquisition (Hz) 12 15 20 n.a. minimum peak widtha(ms) 540 470 350 n.a.
Full scan 400 300 200 100 SIM 1 ion 2 ions 3 ions
91 45 30
80 160 230
n.a. n.a. n.a.
33 18 12
210 390 580
n.a., not available. a Calculated for seven points per peak from: minimum peak width = 7/maximum acquisition.
range of about 100 Da, whilst 23-fold larger ranges can be handled by rapid scanning detectors. This is fully satisfactory for many, and certainly for most, organohalogen applications. In the selected ion monitoring (SIM) mode, acquisition rates of up to 90 Hz can be used when only one ion is monitored (i.e. seven data points can be collected even for peaks having a baseline width of only 80 ms). While the present conclusions are gratifying, they do not imply that rapid-scanning qMS instruments can replace TOF-MS in all instances, as, particularly when a wide mass range has to be covered (e.g., in searching for unknowns), TOF-MS has to be used. 4.1. Analytical performance data Analyte detectability and linearity are inuenced by quite a number of parameters such as GC conditions
and set-up, and the number of modulations but the key factor is the detector used. Table 3 shows instrumental limits of detection (iLODs) and dynamic ranges for various detectors and compound classes quantied so far by GC GC. The data can be called fully satisfactory, specically the iLODs, which are 35-fold lower than in 1D-GC due to peak re-focusing in the modulator.
5. Columns and column combinations To summarize, and simultaneously extend, what has so far been said about columns or, more appropriately column combinations to be used in GC GC, in essentially all early studies, a conventional 1530 m non-polar rst-dimension column was combined with a
Table 3. Analytical performance data for various detectors and classes of organohalogens Detector (mode) Class/compound iLOD [pg] Linearity* Range [pg] lECD PCBs PCDD/Fs 2,3,7,8-TCDD PCBs PCDD/Fs 2,3,7,8-TCDD PBDEs OCPs PCBs PBDEs PCDD/Fs 2,3,7,8-TCDD 0.010.07 0.040.15 0.09 0.110 n.a. 0.20.5 5 510 12 1040 10710 710 1400 0.1200 0.140 0.51000 0.2500 0.5200 0.22000 51000 101000 n.a. n.a. n.a. R2 >0.999 >0.998 >0.998 >0.993 >0.996 >0.996 >0.994 >0.991 >0.997 n.a. n.a. n.a. [9,19,20] [9,19] [9,19] [22,37] [36,38] [36,38,63] [37] [37] [15] [14] [14] [14] Reference
TOF-MS (EI)
qMS (EI, scan 50 Da) qMS (ECNI, SIM)
n.a., not available.
378
Trends
much shorter, narrower (medium-)polar or shapeselective second-dimension column. Using such a non-polar more polar column set has three distinct advantages: A wealth of information on the 1D-GC separation of the compound classes of interest is available in the literature and can be used for the rst-dimension separation. The non-polar columns also have a high thermal stability and, consequently, cause little bleeding. As was shown in one study [19], use of some polar columns in the rst dimension (e.g., an LC-50 or a DB-Dioxin stationary phase) can cause LODs to be 10 times higher. While in the rst dimension there is a truly volatilitybased boiling-point separation, for all other, more selective columns, separation will indeed be mainly governed by specic analyte/stationary phase interactions with, however, also a volatility-based contribution. Fortunately, the extremely rapid second-dimension separation will be essentially isothermal. This implies that, for sample constituents present in each individual rst-dimension efuent fraction (which will have closely similar boiling points), there will be no volatility contribution. In other words, the two separation mechanisms are indeed independent, and the conditions are orthogonal. In the early literature on GC GC, most of the above was already clearly understood and, possibly because of
this and also because of a rather limited availability of short and narrow-bore second-dimension columns, not too much attention was devoted to optimizing the selection of column combinations. In recent years, this attitude has changed and, today, several interesting studies on, specically, second-dimension column selection are available notably in the context of organohalogen analysis [1923]. We include illustrative examples in Sections 6 and 7 below. The present discussion is therefore limited to two comments: (i) when aspects such as nature of substituents (Cl vs. Br), molecular shape (planar vs. non-planar) or parent-compound nature (aromatic vs. nonaromatic) play a role, even a brief study of second-column characteristics can be most rewarding; and, (ii) the column combination providing the best structural ordering (often considered the Number One criterion) does not always offer the best overall resolution of a class of compounds (highly relevant when unraveling the composition of relatively unknown mixtures such as toxaphene or PCAs). As an example, Fig. 2 shows what is found when a non-polar DB-1 rst-dimension column is combined with six different second-dimension columns for the study of PCAs [24]. In this case, using 007-65HT as the second column yielded optimal structural ordering (less band
Table 4. First- and second-dimension columns used for GC GC of organohalogens Code First-dimension columns DB-1, HP-1, VF-1ms Rtx-Dioxin 2 Rtx-500 HT-5 HT-8 DB-XLB DB-Dioxin LC-50 Chirasil-Dex BGB-172 BGB-176SE Second-dimension columns BPX-5 HT-8 Rtx-500 DB-17 BPX-50 007-65HT OV 1701, DB-1701 BPX-70 SP-2340 VF-23ms 007-210 LC-50 SupelcoWax-10, CP-WAX-52CB Phase 100% methylpolysiloxane Proprietary Proprietary (carborane) 5% phenyl-methylpolysiloxane (carborane) 8% phenyl-methylpolysiloxane (carborane) Proprietary Proprietary (44% methyl, 28% phenyl, 20% cyanopropyl, 8% polyoxyethylene-polysiloxane) 50% liquid crystalline-methylpolysiloxane 2,3,6-tri-O-methyl-b-cyclodextrin 25% 2,3,6-tert.-butyldimethylsilyl-b-cyclodextrin 20% 2,3-di-O-methyl-6-O-tert.-butyldimethyl-b-cyclodextrin References [19,20,2224,33,35,37,54] [36] [38] [19] [22] [9,19,21,22] [19] [7,19] [2527] [28] [28]
5% phenyl-methylsilphenylene 8% phenyl-methylpolysiloxane (carborane) Proprietary (carborane) 50% phenyl-methylpolysiloxane 50% phenyl-methylpolysiloxane (silphenylene) 65% phenyl-methylpolysiloxane 14% cyanopropyl-phenyl-methylpolysiloxane 70% cyanopropyl polysilphenylene-siloxane 100% biscyanopropyl polysiloxane Proprietary (7090% cyano-containing polymer) 50% triuoropropyl-methylpolysiloxane 50% liquid crystalline-methylpolysiloxane Polyethylene glycol
[7] [19,20,2224,35,37,54] [36] [35] [19,22,38] [19,23,24] [19,35] [21] [21] [19,23,24,26,27] [19,23,24] [9,19,21,2326] [19,23,24,27,33,35]
379
Trends
overlap than with SupelcoWax-10 and LC-50), but, with VF-23ms, a much larger part of the 2D-separation space was used and overall resolution was, consequently, better. The only exception to what was said above is in the area of chiral separations. Since separations such as those of the pairs of atropisomeric PCBs require long columns and long run times for these applications, the more discriminating (i.e. in this case shape-selective) column is used in the rst dimension [2528]. As can be seen in Table 4, substituted b-cyclodextrins were the preferred stationary phases in these studies. Relevant examples are included in Section 6 below.
6. Within-class separations 6.1. Polychlorinated biphenyls (PCBs) PCBs are products of anthropogenic activity and comprise a class of chlorinated aromatic compounds with 110 chlorine atoms attached to a biphenyl backbone. In total, there are 209 CB congeners; the presence of about 140 of these has been conrmed in technical formulations, such as Aroclors and Clophens, and also in environmental samples. For regulatory and monitoring purposes, seven CBs (CBs 28, 52, 101, 118, 138, 153 and 180) have been selected because of their abundance in humans and in foodstuff of animal origin, and they are often called the EU indicator CBs. In the past two decades, much attention has been paid to the toxicology of PCBs, particularly to the congeners that show the same type of toxicity as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), the so-called dioxin-like or WHO CBs. They include four non-ortho (CBs 77, 81, 126 and 169) and eight monoortho (CBs 105, 114, 118, 123, 156, 157, 167 and 189) substituted CBs. Due to restricted rotation around the central CC bond of biphenyl, some CB congeners occur as enantiomers. Separation of such enantiomer pairs is interesting, since it enables the study of enantioselective bioaccumulation and biodegradation. There are 19 such pairs (CBs 45, 84, 88, 91, 95, 131, 132, 135, 136, 139, 144, 149, 171, 174, 175, 176, 183, 196 and 197), which exist as stable atropisomers at ambient or physiological temperatures. The separation of any of the groups of CBs listed above and, specically, of all CB congeners from each other and from the plethora of matrix constituents is a challenging task. Even though 1D-GC (with ECD or MS detection) can separate some 100150 CB congeners, there still is no unambiguous chromatographic separation of the 12 WHO CBs, or even the seven EU indicator CBs, from the 209 congeners or, even, the 140 present in technical formulations [29]. To achieve the intended goal, it is 380
necessary to use sample fractionation, multiple injections on different stationary phases, heart-cut multidimensional GC or columns coupled in series. It should, therefore, come as no surprise that, since the introduction of GC GC, there have been many attempts to use this technique for improving separation of PCBs. There are over 20 papers [4,5,79,15,1923,25 28,3038] that discuss PCB separation by GC GC. However, most of these use PCBs merely as test analytes to optimize and/or demonstrate modulator tuning, proper modulator or detector performance, and instrument set-up. These studies have clearly contributed to the development of robust GC GC procedures but provide little information on the separation itself. The present review therefore mainly discusses papers aimed at improving separation and/or detection of CB congeners [7,2022,32] and the atropisomeric CB pairs [2528]. The rst study on the GC GC separation of PCBs was presented by Phillips and Xu [32], who used the retention database of all 209 CBs published for 20 stationary phases [39] to construct 2D gas chromatograms. The authors did not discuss the separation of individual congeners, but they predicted structured chromatograms for the non-polar semi-polar DB-1 CNBP column combination. The CBs in the 2D plane were grouped together according to the number of chlorine substituents, and the position of the congeners within a homologue group was determined by the number of ortho chlorines, with the retention sequence being: 4 < 3 < 2 < 1 < 0. In 2001, Haglund et al. [7] analyzed PCBs in technical Clophen A50. The authors used a rather unconventional column set-up, with a highly shape-selective liquid crystal LC-50 (10 m 0.15 mm 0.1 lm) column in the rst dimension and a non-polar BPX-5 (0.25 m 0.1 mm 0.1 lm) column in the second dimension. The non-ortho CBs were most strongly retained by the LC-50 column, followed by the mono-, di- and multi-ortho CBs. As a consequence, the non- and mono-ortho congeners eluted at the highest temperature within each homologue group, and the toxic non-ortho CBs 77, 126 and 169 and the mono-ortho CBs 105, 118 and 156 all showed up in the lower right-hand corner of the contour plot. In order to achieve reasonable second-dimension elution times (<10 s) for the multi-ortho CBs, which were least retained and eluted at very low temperatures, a very short second column (0.25 m) and an unusually fast temperature gradient (>10C/min) had to be used. Under these conditions, all six planar CBs quoted above were successfully separated from other CBs present in the mixture within a run time of only 17 min. In addition, the seven EU indicator CBs were also adequately resolved. In our opinion, despite the excellent separation of the non-ortho CBs from the other congeners, their quantication may be problematic, because the excessive
Trends
bleed of the LC-50 phase at the elevated temperatures at which non-ortho CBs elute will also be modulated and dramatically increase the noise level and, thus, increase the LODs. In addition, the column combination does not allow the determination of the many other CBs, because, due to the fast temperature programme required, the separation power of the rst-dimension column cannot be used fully and many CBs co-elute. In 2002, Korytar et al. [20] tested three column combinations for the separation of 90 CBs with emphasis on the separation of the 12 WHO CBs. They preferred a classical set-up, with a non-polar HP-1 (30 m
0.25 mm 0.25 lm) separating solely on the basis of volatility in the rst dimension, and three more polar phases, BPX-50, HT-8 and SupelcoWax-10 (all 1 m 0.1 mm 0.1 lm), in the second dimension. A complete separation of the WHO CBs from each other and from the other CBs was obtained with the latter two columns. With HP-1 SupelcoWax-10, only six congeners were involved in co-elutions, as against 12 congeners for HP-1 HT-8. However, the latter combination provided the highest information content, because structured chromatograms were obtained. The CBs were found to be grouped together according to the number of chlorine
Table 5. Co-eluting PCBs on six column combinations [21,22]* ** DB-1 HT-8 Non-ortho CB congeners Mono-ortho CB congeners Marker CB congeners 52/69 101/90 138/163/164 23/54a 16/32 20/21/33 43/49 48/75 42/59 41/64 61/70 56/60 98/102 88/91 83/112 115/116 108/107 139/149 134/143 146/165 182/187 196/203 163 HT-8 BPX-50 DB-XLB BPX-50 77/144 153/168 DB-XLB SP-2340 118/131a 101/90 153/168 DB-XLB LC-50 DB-XLB BPX-70
Other CB congeners
132/179a 160/175a 20/33 47/48 93/95/98 112/119 97/117 108/107 163/164 182/187
38/47/62a 20/21/33 66/155 84/89 90/101 107/123
21/33 47/62/65 42/59 37/40a 57/94a 58/67 63/76 86/125 107/134a 160/163 175/182 201/204 196/203
4/10 20/33 43/69 62/65 58/67 63/76 88/95 84/89 83/119 86/125 160/163 175/182a 201/204 196/203
31/53a 47/62/65 42/59 57/94a 86/112 106/107 175/182
No. of CBs resolved by GC GC No. of CBs resolved by GC GCMS Temperature ramp (C/min) Modulation time (s) Run time (min)
188
191
176
181
194
165
192
192/194
184
183
198
1 3 140
1 3 146
1 3 144est.
1.5 4 100est.
1.5 5 90
0.5 5 240
, all congeners separated. a Resolved by MS. est. Run time not explicitly mentioned in paper; estimated from available information. * CB congener classication according to their presence in any of the Aroclors 1242, 1254 or 1260: Bold, >1.0 wt.%; Bold, 0.051.0 wt.%; italics, trace or undetected. ** IUPAC numbering is used. Numbering of [21] was corrected accordingly.
381
Trends
substituents and, within a group, the number of ortho chlorines. This is the structure predicted by Phillips and Xu [32], but now veried experimentally. The ordered structure is very useful because it helps to predict the position of other CB congeners in the 2D plane. This enabled the prediction, on the basis of published 1D-GC retention data, that the WHO CBs will be separated from all other congeners present in Aroclor mixtures. In addition, a large number of unknown peaks detected in a cod-liver extract could be provisionally identied as CB congeners. The effort to analyze PCBs by means of GC GC culminated in two recent studies [21,22], in which attempts were made to separate all 209 CBs. In these studies, seven column combinations were evaluated. The separation for six of these is visualized in Table 5. The DB-XLB HT-8 set-up is not included because it provided a very limited improvement over 1D-GC due to the very similar separation mechanisms applied in both dimensions. When comparing the data of Table 5, one should keep in mind that the results also depend on run time or, more precisely, the temperature ramp during elution, and on modulation time. In general, slower elution and a shorter modulation time yield more efcient separation. Table 5 therefore includes these two parameters. As for the DB-1 HT-8 column combination, the ordered structure quoted above has been conrmed for all CBs (Fig. 3). The separation of the homologue groups was so clear cut that only one co-elution of two congeners, CBs 23 and 54, was caused
by the overlapping of two (tri- and tetra-substituted) homologue series. All other co-elutions occurred within these series. For the rest, this column set performed much poorer, in terms of number of congeners resolved, than the other column combinations. Because most co-eluting compounds have closely similar mass spectra, additional separation by means of MS detection is possible in a limited number of cases only such as here for CBs 23 and 54. As regards the other column sets, the best result was found for DB-XLB BPX-70, which separated 198 congeners. However, the run time was as high as 4 h, or almost double that of most other procedures and it is questionable whether, in several instances, a limited overall increase of resolution justies such an excessive demand of time. In other words, from a practical point of view, HT-8 or DB-XLB combined with BPX-50 or DB-XLB LC-50 are to be preferred with the rst two sets separating 192 congeners in 2.5 h, and the latter a lower number (183) of analytes, but in a very short time (1.5 h). Two of these combinations separate all WHO CBs and EU indicator CBs. The HT-8 BPX-50 set has the added advantage of yielding structured chromatograms. For the penta-CBs, this is shown in the apex plot of Fig. 4, which also clearly visualizes the non-orthonumber-based sub-division. One should consider that the above separations were studied with the congener concentrations in the standard mixture being essentially the same and with RS > 0.5 as the separation criterion. As is well known,
Figure 3. GC GCTOF-MS chromatogram of all CB congeners on DB-1 HT-8 column set [22].
382
Trends
1.9 1.6
tR (s)
104
1.3 1 0.7 4200
82 105 115 98 126 89 116 85 114 95 122 110 93 91 86 97,117 106 83 88 94 124 118 99 109 87 102 125 90 123 127 112,119 103 100 108 92 113 101 107 111 120 121 96 84
4700 5200 0
1
5700 5
6200
6700
tR (s)
Figure 4. GC GCTOF-MS apex plot of the penta-CBs on HT-8 BPX-50 column set. Pink, tetra-ortho CBs; green, tri-ortho CBs; red, di-ortho CBs; light blue, mono-ortho CBs; dark blue, non-ortho CBs. Boxes represent co-eluting congeners [22].
this resolution will not be satisfactory for quantifying partly co-eluting congeners with widely different concentrations, as is often true for PCBs and other classes of organohalogens in real-life samples. Even if RS > 1, the determination of the WHO CBs in one run together with predominant congeners may be difcult. Another problem was encountered when the target analytes had to be determined in a seal-blubber sample [21]. After the usual sample treatment, some of them were found to be present in concentrations too low to be detected by GC GClECD. One solution would be to concentrate the extract. However, some of the major congeners would then overload the second-dimension column and, in addition, it would be difcult to keep all congeners within the working range of the detector. Therefore, two separate injections will be required. 6.1.1. Chiral analysis. With atropisomeric CBs, the preferred method is heart-cut MDGC [29]. Most reported methods use two-oven systems and comprise an achiral precolumn and a chiral main column. Heart cuts (from the precolumn) containing the atropisomers of interest and, inevitably, other PCBs or interferences are directed towards the chiral column. With typical precolumns, such as DB-5 or DB-XLB, three or four out of the 11 atropisomeric pairs present in technical formulations co-elute with some of the 140 CBs [29] and must be separated in the second dimension. ChirasilDex (permethylated 2,3,6-tri-O-methyl-b-cyclodextrin) is the most commonly used chiral column that can resolve nine out of the 19 stable, and seven out of the 11 interesting pairs of chiral congeners [29]. The column sequence used in heart-cut MDGC has to be changed for GC GC, because the second-dimension column must be short and, if a short chiral column is used, no separation of the enantiomers would be achieved (i.e. the chiral column is used in the rst dimension, and a polar or shape-selective column in the second dimension).
In all published papers [2528], Chirasil-Dex was used as the rst-dimension column; in one study [28], BGB-172 (25% 2,3,6-tert.-butyldimethylsilyl-b-cyclodextrin) and BGB-176SE (20% 2,3-di-O-methyl-6-Otert.-butyldimethyl-b-cyclodextrin) were also evaluated. As second-dimension columns, Bordajandi et al. tested HT-8, VF-23ms and SupelcoWax-10 for 65 [28] or 90 [27] CBs, whilst Harju et al. [25,26] used VF-23ms and LC-50 for all 140 Aroclor CBs. The results of the latter, more comprehensive study are displayed in Table 6. It shows that all nine atropisomeric pairs, which are resolved on Chirasil-Dex, partly or completely co-elute with one or more CBs in 1D-GC, while all but two or three of the co-elutions are resolved in the second dimension. As an application, grey-seal tissue was analyzed on both column combinations. Seven out of the nine atropisomeric pairs were detected, and enantiomeric fractions could be determined for six of these (CBs 91, 95, 132, 135, 149 and 174) by using lECD and/or TOF-MS detection; the second eluting enantiomer of CB 84 co-eluted with CB 56. The most abundant
Table 6. Co-elutions for nine atropisomeric CB pairs in 1D-GC and GC GC with 140 Aroclor CBs [26]* Atropisomeric CBs 84 91 95 132 135 136 149 174 176
a *
Co-elutants on Chirasil-Dex (CD) 56, 90, 99, 101 63 93 176, 141 110, 82 115 77, 124 202 132, 141 CD VF-23ms 56 141 CD LC-50 99 93 141 (0.7)a
RS in second dimension. For classication code, see Table 5.
383
Trends
atropisomers, CBs 135, 149 and 174, had enantiomeric fractions (EFs) close to the racemic value, while the EFs for CBs 91, 95 and 132 invariably deviated signicantly from that value. 6.2. Polychlorinated dibenzo-p-dioxins/furans / (PCDD/Fs) / PCDD/Fs are highly toxic compounds formed as by-products during a variety of chemical and combustion processes [40]. Notorious examples are their (tracelevel) presence in technical PCB mixtures, chlorinated phenoxyalkanoic acid pesticides and y ash. The number of chlorine substituents can vary from one to eight to produce up to 75 CDD and 135 CDF positional isomers. There is a pronounced difference in toxic and biological effects amongst these CDD/F congeners and toxicities vary 1,00010,000-fold. Seven 2,3,7,8-substituted CDDs and 10 CDFs are generally considered the most toxic, since they have toxic properties similar to 2,3,7,8tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), which is the most toxic congener of the group. Consequently, the analytical challenge is to detect, and quantify, these 17 priority congeners and, also, the 12 WHO CBs discussed above (which also show a dioxin-like effect). Such ultratrace analyses are expensive, both because of the timeconsuming pre-treatment and clean-up, and the need to use GChigh-resolution MS (GCHRMS) with its unrivalled sensitivity and selectivity [18]. In other words, sample throughput is low and cost is high. Not surprisingly, therefore, the potential of GC GC with its improved selectivity is eagerly explored to nd less demanding but equally rewarding analytical approaches. The rst attempt to analyze dioxins and dioxin-like CBs by GC GC was made by Grainger et al. [16,17]. They used an early sweeper for modulation and HRMS (resolution power 3000) for detection of a 24-compound mixture containing the non-ortho CBs 77, 81, 126 and 169 and a suite of PCDD/Fs. Rather unconventional column dimensions were used: a very short rstdimension DB-5 column (2 m 0.25 mm 0.25 lm) and a rather long second-dimension OV-1701 column (3 m 0.1 mm 0.05 lm). With this set-up, the separation took only 18 min, with an impressive iLOD of 335 attog (S/N 9) for 2,3,7,8-TCDD. Unfortunately, no further information was provided. r As a part of their PCB study quoted above, Koryta et al. [20] tried to separate the priority CDD/Fs and WHO CBs from each other and the bulk of 90 CBs on HP-1 HT-8. The outcome was successful for all except one pair of target analytes, 1,2,3,7,8-PeCDD/CB 169. This stimulated a further search for a column combination that would separate all target compounds from each other and, as importantly, from matrix co-extractants. In a subsequent study [19], therefore, seven rst-dimension and eight second-dimension 384
columns were tested. With a 100% methylpolysiloxane stationary phase (DB-1) in the rst dimension to create orthogonal conditions, all congeners with different toxic equivalency factor (TEF) values could be separated if VF-23ms or LC-50 were used in the second dimension. When other types of rst-dimension column were used (and orthogonality was partly sacriced), a DB-XLB column combined with 007-65HT, VF-23ms or LC-50 gave a complete separation of all 29 priority congeners. With a spiked, fractionated milk extract, DB-XLB LC50 was found to be the most powerful column combination, because of the good separation of all priority congeners from each other as well as matrix constituents. Analytical performance was satisfactory with a close to 3-order linearity, and iLODs of 30150 fg injected mass. Two of the quoted column sets the preferred DB-XLB LC-50 and also VF-1 LC-50 were used by Danielsson et al. [9,41], who studied the quantication of dioxins and dioxin-like CBs by GC GClECD. Fish oil from herring, spiked cows milk, vegetable oil and an eel extract were analyzed by two GC GC and four GCHRMS laboratories, with the latter serving as references. Fig. 5 shows typical GC GClECD chromatograms of the mono-ortho-CB, and non-ortho-CB and CDD/F fraction of herring oil on DB-XLB LC-50, and clearly demonstrates the efciency of the LC-50 phase to separate the target analytes from matrix co-extractants. The quantication data for WHO CBs obtained on both GC GClECD systems were closely similar and agreed very well with the GCHRMS data. As for the dioxins, data produced on DB-XLB LC-50 also showed good agreement, but this was not true for VF-1 LC-50 due to interferences co-eluting with some congeners. For the rest, the CVs were somewhat higher for GC GC than for HRMS (540% vs. 227%). The total toxic equivalent (TEQ) data obtained on the preferred GC GC system compared well with those obtained by GCHRMS, with CVs for both techniques being below 10%. These results show that the proposed method meets the EC requirements for a WHO CDD/F-plus-CB screening method, which should have a false negative rate of less than 1% and a TEQ CV of less than 30%. However, more samples need to be analyzed to conrm fully the criterion for the false negative rate. For the rest, as Danielsson et al. noted [9,41], before GC GClECD can be considered a conrmatory method, the EC Directives will have to be revised; currently, MS is the only mode of detection allowed for this purpose. Focant et al. [36,38] devoted two studies to GC GC TOF-MS. In their rst report [36], Rtx-Dioxin 2 was employed as the rst-dimension column because it is known for its excellent separation of the priority CDD/Fs from each other and also from non-2,3,7,8-substituted congeners. For the second dimension, they selected a relatively non-polar Rtx-500 column (similar to HT-8),
Trends
Figure 5. GC GClECD of the (A) mono-ortho-CB and (B) non-ortho-CB and CDD/F fractions of a sh oil on DB-XLB LC-50. Assignment for (B): 1, CB 77; 2, CB 126; 3, CB 169; IS1, 1,2,3,4-TCDD; 4F1, 2,3,7,8-TCDF; 4D1, 2,3,7,8-TCDD; 5F1, 1,2,3,7,8-PeCDF; 5F2, 2,3,4,7,8-PeCDF; 5D1, 1,2,3,7,8-PeCDD; 6D3, 1,2,3,7,8,9-HxCDD; IS2, 1,2,3,4,6,7,9-HpCDD; 7D1, 1,2,3,4,6,7,8-HpCDD; 8D1, OCDD [9].
because they expected the second dimension to bring more in terms of signal enhancement after modulation than added resolution retention on that column should, consequently, be rather weak. An iLOD of 0.5 pg was obtained for 2,3,7,8-TCDD. As for the separation of the 17 CDD/Fs and the four non-ortho CBs analyzed, one co-elution persisted in both dimensions (viz between CB 126 and 2,3,7,8-TCDD). This co-elution is especially problematic because CB 126 is usually present in concentrations one or two orders of magnitude higher than 2,3,7,8-TCDD, causing a serious over-estimation of the sample TEQ due to the much higher toxicity of 2,3,7,8TCDD. The problem was solved by deconvoluting the masses involved. The congener-specic data found by GC GCTOF-MS and GCHRMS for various environmental (sediment, y ash) and biological (vegetation, sh) samples showed good agreement for 2,3,7,8-TCDD and most other congeners. However, in their next study [38], the authors used another column combination, Rtx-500 BPX-50. All priority CDD/Fs and four non-ortho CBs were then separated and, in a next run, all EU indicator and mono-ortho CBs were also separated from each other and other CBs. The iLOD of 2,3,7,8-TCDD improved to an impressive 0.2 pg. The system was used to analyze
sh, pork, and milk samples and the results were compared with conventional GCHRMS. Not unexpectedly, the earlier conclusions on CBs were conrmed. For the CDD/Fs, the results were strongly concentration dependent. When the average congener concentrations were rather high (above 0.4 pg/g fat weight, which corresponds to 1.1 pg of compound injected), as for sh, and a large sample intake was used (15 g), the two techniques showed good agreement. But, again, the CVs for GCHRMS were 714%, as against 1060% for GC GCTOF-MS. With pork and milk, for which the CDD/F concentrations were much lower (above 0.030.1 pg/g fat weight), there were many over-estimations with CVs up to 90%. However, the congener distribution was still well dened in all instances and can be used (e.g., for tracking sources of contamination). Despite the problems, the TEQ results for the CDD/Fs and CBs compared favorably with those of GCHRMS because a rather good description of the main TEQ contributors (2,3,7,8-TCDD, 1,2,3,7,8PeCDD and 2,3,4,7,8-PeCDF) was achieved. 6.3. Toxaphene Toxaphene is an organochlorine pesticide mixture of complex composition. The major constituents are 385
Trends
chlorobornanes (ca. 75 wt.%), with chlorocamphenes in second place, while chlorodihydrocamphenes are present as minor components [42]. Individual members of these compound classes have been isolated from the technical mixture, and they are likely products of synthesis. The presence of chlorobornenes, which has also been reported [42] as a possible class of constituents, is discussed at the end of this section. The number of congeners that can be found theoretically exceeds 60,000 (cf. Table 1). 1D-GC, which is the method of choice to study the composition of technical toxaphene, cannot create a satisfactory separation. Typically, about 100 peaks show up in a GCECD chromatogram. Better results were obtained by combining GC with other separation techniques, such as adsorption chromatography on silica [43] or active carbon [44], normalphase LC [45], or heart-cut MDGC [46]. With these techniques, the number of compounds in technical toxaphene was estimated to be at least 177 [43], 246 [45], 300 [46], and 675 [44]; all quoted methods were very time-consuming (e.g., the 675-peak experiment required pre-fractionation into no less than 160 fractions with a subsequent 30-min GC analysis of each fraction). In the environment and particularly in higher organisms, many toxaphene constituents are degraded, and only a few are bio-accumulated. This leads to a signicantly simpler toxaphene-residue pattern compared to the technical mixture. One problem with the quantication of individual congeners is the limited availability of the standards. Today, only 23 standards are commercially available (Table 7). Since various
nomenclature rules were proposed for the toxaphene components [4753], each congener in Table 7 has several names. In view of the separation problems outlined above, expectations were high when GC GC became available as a tool to unravel the composition of complex mixtures [54]. Indeed, the use of an HP-1 HT-8 column combination yielded highly structured chromatograms and easily revealed a complex mixture of over 1000 compounds in a run of less than 3 h. Subsequent analysis of the 23-standard mixture and EI-TOF-MS evaluation of technical toxaphene showed that the 2D chromatogram is structured according to the number of chlorine substituents in a molecule (see Fig. 6), with little, if any, dependence on the class of compounds (bornanes or camphenes). The range of chlorination found with GC GCEI-TOF-MS was 511 substituents per molecule. Using home-made software to calculate the total area for each iso-substitution band, hexa- to nonachlorinated compounds were found to be the major components of toxaphene and represented some 97% of the total toxaphene mass. In a more recent study [23], six column combinations were tested for the separation of 12 classes of organohalogenated compounds (see Section 7); one of these was technical toxaphene. Ordered structures were then also found for DB-1 combined with 007-210, 007-65HT or LC-50. Somewhat surprisingly, another column combination, DB-1 VF-23ms, did not deliver ordered structures but offered the best overall separation; the entire GC GC plane was used and visual
Table 7. List of 23 commercially available standards for toxaphene and their various codes IUPAC name 2,2,3-exo,8,9,10-hexachlorocamphene 2-exo,3-endo,8,8,9,10-hexachlorocamphene 2-exo,3-endo,7,8,9,10-hexachlorocamphene 2,2,5,5,9,10,10-heptachlorobornane 2,2,3-exo,8,8,9,10-heptachlorocamphene 2-endo,3-exo,5-endo,6-exo,8,8,10,10-octachlorobornane 2,2,3-exo,8,8,9,9,10-octachlorocamphene 2,2,5-endo,6-exo,8,9,10-heptachlorobornane 2,2,5,5,9,9,10,10-octachlorobornane 2,2,3-exo,5-endo,6-exo,8,9,10-octachlorobornane 2-endo,3-exo,5-endo,6-exo,8,9,10,10-octachlorobornane 2-exo,3-endo,5-exo,8,9,9,10,10-octachlorobornane 2,2,5-endo,6-exo,8,8,9,10- octachlorobornane 2,2,5-endo,6-exo,8,9,9,10-octachlorobornane 2-exo,5,5,8,9,9,10,10-octachlorobornane 2-endo,3-exo,5-endo,6-exo,8,8,9,10,10-nonachlorobornane 2,2,5,5,8,9,10,10-octachlorobornane 2,2,5-endo,6-exo,8,8,9,10,10-nonachlorobornane 2,2,3-exo,5,5,8,9,10,10-nonachlorobornane 2,2,5-endo,6-exo,8,9,9,10,10-nonachlorobornane 2,2,5,5,8,9,9,10,10-nonachlorobornane 2-exo,3-endo,5-exo,6-exo,8,8,9,10,10-nonachlorobornane 2,2,5,5,6-exo,8,9,9,10,10-decachlorobornane Parlar [47,48] 11 12 15 21 25 26 31 32 38 39 40 41 42a 42b 44 50 51 56 58 59 62 63 69 Nikiforov [49] Wester et al. [50,51] C[032001]-(11) C[021001]-(21) C[021011]-(11) B[30030]-(012) C[032001]-(21) B[12012]-(202) C[032001]-(22) B[30012]-(111) B[30030]-(022) B[32012]-(111) B[12012]-(112) B[21020]-(122) B[30012]-(211) B[30012]-(121) B[20030]-(122) B[12012]-(212) B[30030]-(112) B[30012]-(212) B[32030]-(112) B[30012]-(122) B[30030]-(122) B[21022]-(212) B[30032]-(122) AV-code [52] OK-code [53]
HpCB-6533 OCB-4921 HpCB-6452 OCB-6535 OCB-6964 OCB-4917 OCB-3223 OCB-6460 OCB-6454 OCB-2455 NCB-4925 OCB-6549 NCB-6461 NCB-7061 NCB-6455 NCB-6551 NCB-3261 DCB-6583
B7-499 B8-1413 B7-515 B8-789 B8-531 B8-1414 B8-1945 B8-806 B8-809 B8-2229 B9-1679 B8-786 B9-1046 B9-715 B9-1049 B9-1025 B9-2206 B10-1110
99-013 198-303 195-111 99-033 199-111 198-113 41-133 195-311 195-131 97-033 198-313 99-113 195-313 103-113 195-133 99-033 169-313 227-133
386
Trends
Figure 6. Total-ion GC GCECNI-TOF-MS chromatogram of technical toxaphene on DB-1 HT-8, with the mass spectra of two peaks as indicated. Lines indicate position of iso-substitution bands [23].
inspection showed that, here, the highest number of congeners was observed. The column sets that yielded ordered structures indicated the presence of a cluster of unknown compounds (Fig. 6). This group of compounds was not observed in the earlier study [54], because the temperature programme then used was much slower. This caused much more spreading of the toxaphene congeners in the 2D plane; wrap-around occurred and led to co-elution with the unknown compounds. Combined information on the peak shapes and the ECNI-TOF mass spectra added as inserts demonstrated that the unknowns were decomposition products formed during the rst-dimension GC run (i.e. chlorinated bornenes, probably formed by HCl elimination). So far, not a single polychlorinated bornene has been isolated from technical mixtures and the assumption about their presence was based only on GCMS and GCFTIR data [42]. The observations cited created serious doubts about the presence of bornenes in technical toxaphene.
6.4. Polychlorinated alkanes (PCAs) PCAs are complex mixtures with a degree of chlorination of 3070 wt.%, and carbon chain lengths of C10C13 (short-chain PCAs), C14C17 (medium-chain PCAs) or >C17 (long-chain PCAs). They are used as extreme-pressure additives in industrial cutting uids, plasticizers and ame retardants for polyvinyl chloride (PVC) and other plastics and rubbers, and as additives in paints and sealants. PCAs are persistent and nonbiodegradable, and they accumulate in the food chain. The global production of PCAs has been reduced since the early 1980s [55], but is still in the range of 380,000 tons [56]. Short-chain PCAs cause particular concern due to the high amounts released into the environment, and their toxicity, which is higher than that of other PCAs. Analysis of PCAs is difcult because of the extreme complexity of the mixtures and the lack of quantication standards, and semi-quantication is all that can be achieved. In many environmental studies dealing with 387
Trends
halogenated micro-contaminants, an essentially unresolved band covering a large part of the GC baseline is all that indicates the presence of PCAs. Currently, some 40 individual standards are available, but most of them, as will be demonstrated below, are not present in the technical mixtures. Mixtures of PCAs, with various chlorine contents and carbon-chain lengths, are used as
reference standards for semi-quantication. Analysis is usually carried out by GC coupled to HRMS in ECNI mode. The method is based on monitoring [MCl] ions to determine the concentrations of individual classes (i.e. congeners with the same number of carbon and chlorine atoms). HRMS is highly selective and eliminates interferences caused by other polychlorinated pollutants and
Figure 7. (A) GC GCECNI-TOF-MS extracted ion chromatogram (m/z 7073) of polychlorinated decanes with 55 wt.% Cl. (B) GC GCECNI-TOF-MS chromatograms of C10C13 technical mixture with 55 wt.% Cl. Colored lines indicate the position of apices within the band of polychlorinated (red) decanes, (green) undecanes, (blue) dodecanes and (black) tridecanes. (C) Overlay of GC GCECNI-TOF-MS chromatograms of (red) short-, (green) medium- and (blue) long-chain PCA mixtures with different Cl content. White numbers indicate number of (carbon + chlorine) atoms of the compounds present in the bands [24].
388
Trends
PCAs with the same nominal mass. However, this detection method is not available in many laboratories and is too expensive for routine analysis, so low-resolution MS (LRMS) is also used for quantication. An international inter-comparison study for short-chain PCAs has shown that the two techniques can give comparable quantitative results (e.g., in biota [57,58]). Nevertheless, because of the increased risk of interferences, improved sample clean-up and/or the use of GC GC instead of 1D-GC is urgently required. The potential of GC GC was investigated in a recent paper in which six column combinations were tested [24]. The highest number of separated congeners was found with the DB-1 VF-23ms combination, but, for characterization and quantication, DB-1 007-65HT was preferred because this set provided more information in terms of ordered structures (i.e. group and subgroup separation). As demonstrated for polychlorinated decanes in Fig. 7A, the separation of PCA congeners with the same chain length is based on the number of chlorine substituents. The authors added that the number of congeners in the technical mixtures (formed by uncontrolled synthesis) was relatively rather limited. If a larger number were present, the iso-substitution bands would become broader and neighboring bands would start to overlap. This is demonstrated in Fig. 7A by the outlying positions of several individual congeners, which should be considered unlikely products of uncontrolled synthesis. With mixtures of PCAs of varying chain length, the ordered structures comprise compounds having the same number of carbon-pluschlorine atoms for example, C10Cl8 is on the same diagonal line as C11Cl7, C12Cl6 and C13Cl5. This is elegantly visualized in Fig. 7B. The position of the various compounds in each diagonal band depends on the number of carbon atoms: compounds with longer carbon chains have lower second-dimension retention times. This carbon-chain-length selectivity creates a distinct separation of compounds that differ by at least three carbons; C10 and C13 compounds show no overlap in Fig. 7B. For a mixture of short-, medium- and long-chain PCAs, the (carbon+chlorine)-based ordering is seen to hold over a summed-number range of at least 14 to 26. In addition, the carbon-chain-length selectivity creates a partial separation of the three PCA groups (Fig. 7C). Obviously, using GC GC is a major step forward in PCA analysis, although, simultaneously, it is also clear that additional separation power possibly provided by LC-based sample fractionation will be needed for a satisfactory overall unraveling of the composition of PCA mixtures. Two relevant examples of the added value of GC GC compared to 1D-GC are as follows. In the quoted study [24], two dust samples were analyzed, and signicantly different GC GClECD patterns of the short- and medium-chain PCAs were easily observed. Visual evaluation
of the chromatograms showed that one sample contained more medium- than short-chain PCAs and that these medium-chain PCAs had a relatively low degree of chlorination, while the other sample contained more short- than medium-chain PCAs, with the short-chain PCAs having a higher degree of chlorination. In other words, pattern recognition based on the visual evaluation allowed provisional identication of sample composition. Another advantage of GC GC is the elimination of LRMS interferences amongst the PCAs. Recently, Reth and Oehme [56] discussed three critical limitations of LRMS for the analysis of short- and medium-chain PCAs. [MCl] ions of a specic PCA (e.g., [MCl + 2] of C11H17Cl7 with m/z 360.9) interfere with: (i) the [MCl] ions of a PCA with ve carbon atoms more and two chlorine atoms less (i.e. [MCl] (m/z 361.1) of C16H29Cl5); (ii) the [MCl] ions of a PCA with two carbon atoms more and one chlorine atom less (i.e. [MCl + 8] (m/z 361.0) of C13H22Cl6); and, (iii) with [M + Cl] ions of a PCA with the same number of carbon atoms and two chlorine atoms less (i.e. [M + Cl] (m/z 360.9) of C11H19Cl5). GC GC solves these three problems due to the enhanced chromatographic resolution. 6.5. Polybrominated diphenylethers (PBDEs) PBDEs are widely used as ame retardants (e.g., in polymers, textiles, electronic boards) and, similar to the PCBs, there are 209 BDE congeners. However, with only some 2025 congeners, the composition of the technical PBDE mixtures is rather simple. Most environmental monitoring programmes focus on the analysis of seven congeners (BDEs 28, 47, 99, 100, 153, 154 and 209), which are most abundant in technical mixtures and are conventionally considered a type of reference set, similar to the seven EU indicator CBs. However, in environmental and biota samples, many other BDEs are found than the 2025 technical congeners because of their photolytic and biological debromination and metabolism in higher animals. The analysis of PBDEs is carried out by GCECD or GCMS (EI or ECNI mode) [59]. Today, 125 BDE congeners are commercially available and their retention characteristics on seven stationary phases have been reported [60]. No stationary phase separates all congeners and not even the seven reference BDEs from all others. The best separation was achieved on a DB-XLB column with which 70 congeners were separated, while 55 congeners were involved in 22 co-elutions. In real-life samples, the situation is even worse because other brominated compounds (e.g., polybrominated biphenyls (PBBs), hexabromocyclododecane (HBCD), tetrabromobisphenol-A (TBBP-A) or dimethyl-tetrabromobisphenolA (me-TBBP-A)) may be present in the sample extract 389
Trends
and cause further interferences, which, often, cannot be solved by means of MS. GC GC of BDEs is challenging because of the high boiling points and thermal instability of the higher brominated congeners. In a study to be discussed in more detail in Section 7, Focant et al. [35,37] found that the higher brominated congeners were retained too strongly in the second dimension, with the limited thermal stability of most stationary phases not allowing a sufciently high temperature to speed up the separation. In the end, an HT-8 column was used; it was stable up to 360C. However, even with a DB-1 HT-8 column set, hepta-BDE 183 and deca-BDE could not be included because retention was still too great. Korytar et al. [61] used GC GClECD to separate the 125 BDEs recently marketed. For the rst dimension, DB-XLB was not a good choice because nona- and decasubstituted congeners were decomposed completely on this phase and partial decomposition was observed down to hexa-BDEs. Fortunately, on DB-1, decomposition of nona-BDEs was negligible and only slight decomposition of deca-BDE was observed. As for the second dimension, from amongst six columns tested, 007-65HT was found to add most to the selectivity of the rst-dimension separation. In contrast with the ndings quoted above, no
extreme peak broadening or trapping was observed for the nona-BDEs 206, 207 and 208, and deca-BDE 209. Possibly, the band broadening in [35] was caused by a low temperature of the MS transfer line, which houses a signicant part of the second-dimension column or by too low a temperature of the cooling gas used for modulation. Fig. 8 shows the considerably improved separation on the DB-1 007-65HT column set, with only 17 co-eluting pairs involving 35 congeners. In addition, the seven reference BDEs were separated from all other BDEs. When a dust extract was analyzed (insert in Fig. 8), 18 BDE congeners were identied. Various other brominated ame retardants and a number of BDE metabolites were all found to elute within the BDE band, and that caused several more co-elutions (Fig. 8). Of course, some separations were improved, notably that of TBBP-A and BDE 153; the latter analyte can now be quantied even if a high concentration of TBBP-A is present, as was true for the dust sample. Finally, Fig. 8 shows that second-dimension separation facilitates the use of uorinated BDEs as internal standards, because all F-BDEs that co-elute with the parent compound in the rst dimension are separated in the second dimension, appearing just below that parent.
Figure 8. GC GClECD of ( ) BDEs, ( ) uorinated BDEs, ( ) other brominated ame retardants and ( ) BDE metabolites on DB-1 007-65HT. Insert: GC GClECD contour plot of dust extract [61].
390
Trends
7. Between-class separations For obvious reasons, the common denominator of nearly all papers devoted to GC GC analysis of organohalogens is the (much) improved within-class resolution that can be achieved, with the joint addressing of the priority CDD/Fs and CBs as the only, and logical, exception. Because of the marked successes so obtained, we now see a gradual shift of interest to the more intriguing between-class separations [23,35,37]. As a rst attempt, Focant et al. [35] used GC GC TOF-MS for the simultaneous determination of 38
predominant CBs, 11 persistent halogenated pesticides (OCPs), one PBB and eight PBDEs. With the DB-1 HT-8 column combination selected, of 58 test compounds, only one pair of CBs was not resolved. In their next study [37], the authors determined the priority compounds in serum and milk. Single-injection GC GC was compared with a validated GCHRMS procedure, which required three separate injections with three different temperature programmes and was, of course, much more time-consuming. The data shown for human serum corresponded very well between the two techniques. For the milk sample and the more abundant
Figure 9. GC GClECD on DB-1 LC-50 of: (A) PCBs, PBBs, PCDEs, PBDEs, PCDTs, h PCNs, PCDD/Fs, OCPs, individual toxaphene standards; (B) PCAs (PCA-60) as color contour plot and other classes as black dots; (C) PCTs (Aroclors 5442 + 5460) as color contour plot and dioxin-like CBs (black dots) and planar PCTs (white arrows); (D) PCDD/F fraction of a sediment extract [23].
391
Trends
congeners (>1 ng/g of lipid), per-cent deviations between the two methods were below 20%, which is acceptable. However, for some less-abundant congeners (<1 ng/g of lipid), the GC GC-based concentrations were higher than those of the reference method and per-cent deviations reached 100%. These rst attempts were obviously promising, but the results also showed that there still were interfering co-elutions that could not be solved by GC GC separation or MS deconvolution. Not unexpectedly, they caused problems specically for analytes present at the ultra-trace level. Part of the
problem could probably be solved by utilizing a more selective column combination; as will be discussed below, the DB-1 HT-8 set does not yield much betweenclass separation. The between-class separation of 12 groups PCBs, PBBs, PCDD/Fs, PCAs, PBDEs, toxaphene, OCPs, PCDEs, PCNs, PCDTs and PCTs has been studied very recently [23]. Five column combinations were selected DB1 007-210, DB-1 HT-8, DB-1 LC-50, DB-1 00765HT and DB-1 VF-23ms to study between-class, but occasionally also within-class, separation. Both types of
Figure 10. GC GClECD on DB-1 007-65HT of: (A) PCBs, PBBs, PCDEs, PBDEs, PCDTs, h PCNs, PCDD/Fs, OCPs, individual toxaphene standards and PCAs (PCA-60); (B) PCAs (PCA-60), PCTs (Aroclors 5442 + 5460) and toxaphene technical mixture; (C) a dust extract for PCA and PBDE determination [23].
392
Trends
separations were found to strongly depend on the column combination used. We summarize a few relevant conclusions here: On DB-1 HT-8, congener separation on the basis of the number of halogen substituents, well-known for PCBs and toxaphene, was also observed for PCTs and PBDEs. However, there was no group-type separation (i.e. although the DB-1 HT-8 set can be successfully used for within-class separation, there is a high risk of interferences if congeners from other compound classes are present). Group separation based on planarity was obtained with the DB-1 LC-50 column set. Fig. 9AC shows that three groups of analytes could be distinguished. Three-ring planar compounds, such as PCDD/Fs, PCDTs and planar PCTs, were most strongly retained. Next in line were two-ring planar compounds, such as PCNs and planar PCBs. Non-planar analytes showed least retention and did not interfere with the planar compounds. The practicability of this column set is demonstrated in Fig. 9D, which shows a chromatogram of the PCDD/F fraction of a sediment sample after fractionation on a carbon column. Obviously, a properly tuned GC GC system could accommodate a very high number of compounds in the 2D plane, and could separate dioxins (indicated by black acronyms) from co-extractants (white-yellow band along rst-dimension axis). Group-type separation was also delivered by DB-1 007-65HT (Fig. 10A-B). Here, the PBDEs were the strongest retained compounds in the second dimension and the PCAs are the least retained organohalogens. This is a rewarding result because PCAs and PBDEs are usually present in the same fraction after clean-up. As an example, the analysis of a dust sample is shown in Fig. 10C: the group separation is clear-cut. Finally, Fig. 10B displays the overlay of three mixtures toxaphene, PCA-60 and Aroclors 5442 plus 5460. What shows up in the 2D plane can be interpreted in two rather different ways: one is to emphasize that the presence of these three types of mixtures in any real-life sample will virtually obscure all other classes of organohalogens, even in GC GC; but, it is as interesting to note that there is a striking separation of the three compound classes of interest. In addition, clearly ordered structures are observed for toxaphene and PCAs. The last column combination tested, DB-1 VF23ms, yielded excellent within-class separation, especially of non-aromatic compounds OCPs, toxaphene and PCAs but there was no group separation. This column set should, therefore, be preferred when unraveling the composition of, specically, toxaphene and the PCAs, is the main aim of a study.
8. Conclusions and perspectives Accurate, precise congener-specic analysis of organohalogenated compounds has been a main goal of analytical research since the early days of OCP and PCB determination in environmental samples in the 1960s. Three areas of interest can be recognized, viz. to ensure/ improve: (i) the separation of the analytes of interest from other constituents present in a sample to enable their quantication; (ii) the sensitivity of the selected procedures and achieve the required LODs; and, (iii) the reliability of the identication/conrmation of the target analytes and/or unknowns. The common denominator of well-known milestones of analytical chemistry, such as the introduction of capillary GC columns, the coupling of GC with selective detectors, such as the ECD and low- or highresolution MS, and the introduction of heart-cut MDGC, is that they all simultaneously improved these three aspects. The present review shows that GC GC is effecting a considerable further improvement and that this technique will no doubt become another milestone in improving the analysis of the various classes of organohalogenated compounds of interest at present. It is worthwhile adding that the relatively brief introductory sections on instrumental aspects show that GC GC can be considered a mature technique (with the exception of sufciently rapid and (semi-)automated data handling and interpretation) and that proper instrumentation is commercially available. The degree to which the overall resolution of organohalogens can be improved by means of GC GC depends on differences in the physicochemical properties of the target analytes but, of course, also on the (non)complexity of the technical mixtures. The best results for within-class separations were observed for very complex mixtures, such as toxaphene, PCAs and PCTs, for which about 10 times as many peaks can easily be identied. For less complex mixtures, such as PCBs and PBDEs, the gain is self-evidently more modest, but still some 50% more congeners were resolved than by 1D-GC. For the class of PCNs, with only 75 congeners, the improvement was limited, irrespective of the stationary phase used. However, one should always consider that the separation of the target analytes from matrix constituents is another key issue and, here, GC GC contributes to a satisfactory overall improvement also in the case of limited numbers of congeners. Between-class separations of the various groups of organohalogens come in more or less the same category, and improvement is pronounced because of the larger differences in physicochemical properties between classes than amongst congeners of the same class. A striking example is the
393
Trends
essentially complete separation of the technical mixtures of toxaphene, short-chain PCAs and PCTs in one GC GC run. The selection of a suitable column combination is essential to achieve the desired separation. Certainly for the fairly hydrophobic and essentially non-polar organohalogens, the best option is to use a non-polar stationary phase in the rst dimension, because then a truly orthogonal system can be set up. Pure dimethylpolysiloxane phases containing 08% diphenyl groups or those stabilized with polycarborane are preferred and are easily available from various producers. The separation mechanism in the second dimension should differ as much as possible from that in the rst dimension. Today, commercial columns with geometry suitable for use in the second dimension are coated with one of the following stationary phases polyethylene glycol or dimethylpolysiloxane with up to 70% diphenyl groups, up to 100% cyanopropyl groups, up to 50% triuoropropyl groups or up to 50% liquid crystals. Detailed experimental work has shown that all of these, except the triuoropropyl phase, effect a signicantly improved separation of the organohalogens. One problem common to all second-dimension stationary phases, except the phenyl phase, is their low temperature stability. Further attention needs to be paid urgently to the production of low-bleeding polar/shapeselective phases that are stable up to 300C. Another challenge for the manufacturers is to produce columns containing two different stationary phases, with a discrete in-between border. Coupling columns by means of press-ts, which is a rather laborious and delicate job, will then become superuous. A notable exception to the (non-polar) (polar/shapeselective) set-up is encountered in the analysis of enantiomers such as the 19 pairs of atropisomeric CBs. The shape-selective (i.e. chiral) column now has to be used in the rst dimension, because the rather marginal separation usually obtained on such phases, makes it absolutely necessary to use a long column of, typically, 2060 m: the 12 m length of a second-dimension column would have little effect. Analyte detectability can also be affected by the column combination selected. Low-bleeding columns should be used in the rst dimension to keep noise as low as possible. This is another reason why non-polar stationary phases are preferred in the rst dimension. In addition, the second-dimension separation is performed under isothermal conditions. Consequently, if a study mainly aims to improve LODs, phases that hardly, or do not at all, retain the target analytes are preferred in the second dimension, in order to generate very narrow and, thus, very high peaks. In this way, an impressive iLOD of 0.2 pg was obtained for 2,3,7,8-TCDD with TOF-MS detection. However, we would agree that the key aspect of the analytical approach is now lost it is essentially a 394
1D-GC operation and there is no or very limited improvement of the separation. As regards the detection of organohalogens, today the lECD is the most sensitive detector available for these compounds with iLODs typically down to 10 fg. The data-acquisition rate of 50 Hz is satisfactory for nearly all applications, but there is considerable peak broadening due to the relatively large volume of the detector cell (150 ll). In other words, the development of a lECD with a 3050 ll cell volume would be a distinct bonus. For the rest, whatever the merits of lECD detection (and these are signicant), for analyte identication/ conrmation, one has to use a mass spectrometer. Today, the work-horse is the Leco TOF-MS, which can acquire up to 500 spectra per second over the entire mass range and is described in the literature for a wide variety of analytes and sample types. With this instrument, excellent deconvolution of mass spectra can be obtained. As can be seen from the present review, this is also true for most organohalogen analyses. However, it is also clear that, for ultra-trace studies, satisfactory results cannot always be obtained. In such instances, the use of ECNI instead of EI (see below) offers a solution. However, the Leco TOF-MS does not have this option, and the recently marketed ThermoElectron TOFMS is, in our experience, rather unstable and will require further upgrading to allow use in routine analysis. A medium-resolution TOF-MS (resolution up to 5000) with a data-acquisition rate up to 25 Hz and with ECNI, was recently marketed by Jeol. Evaluation of this instrument for GC GC is eagerly awaited, and proper comparison should be made with the other MS instruments available. In this context, the introduction of a new generation of rapid-scanning quadrupole MS instruments is of marked interest. They are much faster than the state-ofthe-art conventional qMSs and can be coupled successfully to GC GC: a mass range of 300 Da can be acquired with a data-acquisition rate of 25 Hz and, in the SIM mode, two ions can be acquired at 45 Hz. With ECNI in the SIM mode, analyte detectability for higher halogenated compounds is similar to that of an ECD (i.e. down to 10 fg); however, for compounds with four or fewer halogens, the detector rapidly becomes less sensitive. Although the rapid-scanning qMSs enable quantication, the 79 data points per peak do not always sufce for a proper functioning of the deconvolution software, mainly due to spectral skewing. Neither can they be used over very wide mass ranges (e.g., as required when the search for unknowns is an aspect of key interest); a TOFMS is then indispensable. One more aspect should be mentioned here. Next to detection, as commonly understood, there is in GC GC the phenomenon of ordered structures, bands or clusters. The rapid recognition, upon visual inspection
Trends
[11] R.A. Shellie, P. Marriott, C.W. Huie, J. Sep. Sci. 26 (2003) 1185. [12] M. Kallio, T. Hyotylainen, M. Lehtonen, M. Jussila, K. Hartonen, M. Shimmo, M.-L. Riekkola, J. Chromatogr., A 1019 (2003) 251. [13] J. Dalluge, R.J.J. Vreuls, D.J. van Iperen, M. van Rijn, U.A.Th. Brinkman, J. Sep. Sci. 25 (2002) 608. [14] P. Korytar, J. Parera, P.E.G. Leonards, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 1067 (2005) 255. [15] M. Adahchour, M. Brandt, H.-U. Baier, R.J.J. Vreuls, A.M. Batenburg, U.A.Th. Brinkman, J. Chromatogr., A 1067 (2005) 245. [16] J. Grainger, Organohal. Compd. 27 (1996) 354. [17] J. Grainger, J.M.D. Dimandja, V.E. Green, Z. Liu, D.G. Patterson Jr., Organohal. Compd. 35 (1998) 28A. [18] D.G. Patterson, S.M. Welch, W.E. Turner, J.-F. Focant, Organohal. Compd. 67 (2005) CD ROM ID 1924. [19] P. Korytar, C. Danielsson, P.E.G. Leonards, P. Haglund, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 1038 (2004) 189. [20] P. Korytar, P.E.G. Leonards, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 958 (2002) 203. [21] M. Harju, C. Danielsson, P. Haglund, J. Chromatogr., A 1019 (2003) 111. [22] J.-F. Focant, A. Sjodin, D.G. Patterson Jr., J. Chromatogr., A 1040 (2004) 227. [23] P. Korytar, P.E.G. Leonards, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 1086 (2005) 29. [24] P. Korytar, J. Parera, P.E.G. Leonards, F.J. Santos, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 1086 (2005) 71. [25] M. Harju, P. Haglund, J. Microcol. Sep. 13 (2001) 300. [26] M. Harju, A. Bergman, M. Olsson, A. Roos, P. Haglund, J. Chromatogr., A 1019 (2003) 127. [27] L.R. Bordajandi, P. Korytar, J. de Boer, M.J. Gonzalez, J. Sep. Sci. 28 (2005) 163. [28] L.R. Bordajandi, L. Ramos, M.J. Gonzalez, J. Chromatogr., A 1078 (2005) 128. [29] J.W. Cochran, G.M. Frame, J. Chromatogr., A 843 (1999) 323. [30] D.G. Patterson Jr., J.R. Barr, E.S. Di Pietro, J. Grainger, V.E. Green, C.R. Lapeza, V.L. Maggio, P.C. McClure, S. Sirimanne, W.E. Turner, Organohal. Compd. 27 (1996) 309. [31] H.-J. de Geus, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 767 (1997) 137. [32] J.B. Phillips, J. Xu, Organohal. Compd. 31 (1997) 199. [33] H.-J. de Geus, J. de Boer, U.A.Th. Brinkman, Chromatographia 55 (2002) 339. [34] T. Hyotylainen, M. Kallio, K. Hartonen, M. Jussila, S. Palonen, M.-L. Riekkola, Anal. Chem. 74 (2002) 4441. [35] J.-F. Focant, A. Sjodin, D.G. Patterson Jr., J. Chromatogr., A 1019 (2003) 143. [36] J.-F. Focant, E.J. Reiner, K. MacPherson, T. Kolic, A. Sjodin, D.G. Patterson Jr., S.L. Reese, F.L. Dorman, J.W. Cochran, Talanta 63 (2004) 1231. [37] J.-F. Focant, A. Sjodin, W.E. Turner, D.G. Patterson Jr., Anal. Chem. 76 (2004) 6313. [38] J.-F. Focant, G. Eppe, M.L. Scippo, A.C. Massart, C. Pirard, G. Maghuin-Rogister, E. de Pauw, J. Chromatogr., A 1086 (2005) 45. [39] G.M. Frame, Fresenius J. Anal. Chem. 357 (1997) 701. [40] C. Rappe, Environ. Sci. Technol. 18 (1984) 78A. [41] C. Danielsson, K. Wiberg, P. Korytar, J. de Boer, P. Haglund, Organohal. Compd. 60 (2003) 395. [42] W. Vetter, M. Oehme, in: J. Paasivirta (Editor), The Handbook of Environmental Chemistry, Anthropogenic Compounds, Part K: New Types of Persistent Halogenated Compounds, SpringerVerlag, Berlin, Germany, 2000, p. 237. [43] R.L. Holmstead, S. Khalifa, J.E. Casida, J. Agric. Food Chem. 22 (1974) 939. [44] B. Jansson, Separation av Toxafenkomponenter (in Swedish), Statens naturvaardsverk, Rapport 82-04, Specialanalytiska Laboratoriet, Wallenberglaboratoriet, Stockholm, Sweden, 1982.
of a 2D chromatogram, of clearly ordered groups and sub-groups, is a valuable tool in ngerprinting studies and, especially with the organohalogens, is a powerful aid when tracing specic halogen-substitution patterns (e.g., the partial unraveling of the composition of toxaphene and the PCAs). To sum up, from the instrumental point of view, GC GC can be called an established technique even though most analysts will add that designing more robust and/or user-friendly modulators and column couplings should receive further attention, while their bosses will clamor for less expensive TOF-MS instruments to be available on the market. A wide variety of applications, both in the eld of organohalogen analysis and in many other areas, convincingly demonstrates the practicability of the technique, for both qualitative and quantitative analysis. In most instances, analytical performance data are fully satisfactory down to, but not yet below, the low-ng/g level. Nevertheless, there remain challenges to be met before GC GC can be recommended for routine analysis: the wealth of information that is generated per GC GC run, and especially when using MS detection, distinctly shows that the development of user-friendly software packages for (semi-) automated data handling and interpretation has a high probably, top priority; and, proper training programmes should be set up, and protocols made available to educate the 1D-GC experts and make them recognize that switching to the 2D alternative creates not only many more, but also more detailed and reliable, results and that, to give one example, HS-SPMEGC GCEI-TOF-MS may seem intimidating, but is a fascinating acronym.
References
[1] S. Jensen, New Scientist 32 (1966) 612. [2] J. Dalluge, J. Beens, U.A.Th. Brinkman, J. Chromatogr., A 1000 (2003) 69. [3] M. Adahchour, J. Beens, U.A.Th. Brinkman, Trends Anal. Chem. to be submitted (2006).. [4] E.M. Kristenson, P. Korytar, C. Danielsson, M. Kallio, M. Brandt, J. Makela, R.J.J. Vreuls, J. Beens, U.A.Th. Brinkman, J. Chromatogr., A 1019 (2003) 65. [5] P. Haglund, M. Harju, C. Danielsson, P. Marriott, J. Chromatogr., A 962 (2002) 127. [6] E. Jover, M. Adahchour, J.M. Bayona, R.J.J. Vreuls, U.A.Th. Brinkman, J. Chromatogr., A 1086 (2005) 2. [7] P. Haglund, M. Harju, R. Ong, P. Marriott, J. Microcol. Sep. 13 (2001) 306. [8] H.-J. de Geus, A. Schelvis, J. de Boer, U.A.Th. Brinkman, J. High Resolut. Chromatogr. 23 (2000) 189. [9] C. Danielsson, K. Wiberg, P. Korytar, S. Bergek, U.A.Th. Brinkman, P. Haglund, J. Chromatogr., A 1086 (2005) 61. [10] G.S. Frysinger, R.B. Gaines, J. High Resolut. Chromatogr. 22 (1999) 251.
395
Trends
[45] J. Zhu, M.J. Mulvihill, R.J. Norstrom, J. Chromatogr., A 669 (1994) 103. [46] J. de Boer, H.-J. de Geus, U.A.Th. Brinkman, Environ. Sci. Technol. 31 (1997) 873. [47] J. Burhenne, D. Hainzl, L. Xu, B. Vieth, L. Alder, H. Parlar, Fresenius J. Anal. Chem. 346 (1993) 779. [48] D. Hainzl, J. Burhenne, H. Barlas, H. Parlar, Fresenius J. Anal. Chem. 351 (1995) 271. [49] V.A. Nikiforov, V.G. Tribulovich, V.S. Karavan, Organohal. Compd. 26 (1995) 393. [50] P.G. Wester, H.-J. de Geus, J. de Boer, U.A.Th. Brinkman, Chemosphere 35 (1997) 1187. [51] P.G. Wester, H.-J. de Geus, J. de Boer, U.A.Th. Brinkman, Chemosphere 35 (1997) 2857. [52] P. Andrews, W. Vetter, Chemosphere 31 (1995) 3879. [53] M. Oehme, R. Kallenborn, Chemosphere 30 (1995) 1739.

[54] P. Korytar, L.L.P. van Stee, P.E.G. Leonards, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 994 (2003) 179. [55] G.T. Tomy, A.T. Fisk, J.B. Westmore, D.C.G. Muir, Rev. Environ. Contam. Toxicol. 158 (1998) 53. [56] M. Reth, M. Oehme, Anal. Bioanal. Chem. 378 (2004) 1741. [57] G.T. Tomy, J.B. Westmore, G.A. Stern, D.C.G. Muir, A.T. Fisk, Anal. Chem. 71 (1999) 446. [58] M. Coelhan, Anal. Chem. 71 (1999) 4498. [59] A. Covaci, S. Voorspoels, J. de Boer, Environ. Int. 29 (2003) 735. [60] P. Korytar, A. Covaci, J. de Boer, A. Gelbin, U.A.Th. Brinkman, J. Chromatogr., A 1065 (2005) 239. [61] P. Korytar, A. Covaci, P.E.G. Leonards, J. de Boer, U.A.Th. Brinkman, J. Chromatogr., A 1100 (2005) 200. [62] C. Debonneville, A. Chaintreau, J. Chromatogr., A 1027 (2004) 109. [63] J.-F. Focant, C. Pirard, G. Eppe, E. de Pauw, J. Chromatogr., A 1067 (2005) 265.
396

Comprehensive Two Dimensional

Caricato da

Informazioni sul documento

Descrizione originale:

Copyright

Formati disponibili

Condividi questo documento

Condividi o incorpora il documento

Opzioni di condivisione

Hai trovato utile questo documento?

Questo contenuto è inappropriato?

Copyright:

Formati disponibili

Comprehensive Two Dimensional

Caricato da

Copyright:

Formati disponibili

Trends in Analytical Chemistry, Vol. 25, No.

Comprehensive two-dimensional gas chromatography for the analysis of organohalogenated micro-contaminants

Corresponding author. E-mail: Peter.Korytar@wur.nl

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

PCBs PCDDs PCDFs Toxaphene

OCPs PCTs PCDEs PCNs PCAs PBBs PBDEs

Enantiomers not included.

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Figure 1. Schematic of a GC GC system with different modulator types.

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

n.a. n.a. n.a.

210 390 580

qMS (EI, scan 50 Da) qMS (ECNI, SIM)

n.a., not available.

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

38/47/62a 20/21/33 66/155 84/89 90/101 107/123

31/53a 47/62/65 42/59 57/94a 86/112 106/107 175/182

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

RS in second dimension. For classication code, see Table 5.

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Trends in Analytical Chemistry, Vol. 25, No. 4, 2006

Potrebbero piacerti anche