Screening of volatiles from explosive initiators and plastic-bonded explosives (PBX) using headspace solid-phase microextraction coupled with gas chromatography - mass spectrometry (SPME/GC-MS).

The detection of explosives and explosive devices based on the volatile compounds they emit is a long-standing tool for law enforcement and physical security. Toward that end, solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) has become a crucial analytical tool for the identification of volatiles emitted by explosives. Previous SPME studies have identified many volatile compounds emitted by common explosive formulations that serve as the main charge in explosive devices. However, limited research has been conducted on initiators like fuses, detonating cords, and boosters. In this study, a variety of SPME fiber coatings (i.e., polydimethylsiloxane (PDMS), polydimethylsiloxane/divinylbenzene (PDMS/DVB), divinylbenzene/carboxin/polydimethylsiloxane (DVB/CAR/PDMS), and carboxin/polydimethylsiloxane (CAR/PDMS)) were employed for the extraction and analysis of volatiles from Composition C-4 (cyclohexanone, 2-ethyl-1-hexanol, and 2,3-dimethyl-2,3-dinitrobutane (DMNB)) and Red Dot double-base smokeless powder (nitroglycerine, phenylamine). The results revealed that a PDMS/DVB fiber was optimal. Then, an assortment of explosive items (i.e., detonation cord, safety fuse, slip-on booster, and shape charge) were analyzed with a PDMS/DVB fiber. A variety of volatile compounds were identified, including plasticizers (tributyl acetyl citrate, N-butylbenzenesulfonamide), taggants (DMNB), and degradation products (2-ethyl-1-hexanol).

Quantitative analysis of smokeless powder particles in post-blast debris via gas chromatography/vacuum ultraviolet spectroscopy (GC/VUV).

Forensic analysis of smokeless powder particles recovered from the debris of an improvised explosive device can provide information about the type of smokeless powder used and can aid investigation efforts. In this study, quantitative methods were used to yield information about the difference in the chemical composition of the particles pre- and post-blast. The technique, gas chromatography/vacuum ultraviolet spectroscopy (GC/VUV), was able to quantify nitroglycerin, 2,4-dinitrotoluene, diphenylamine, ethyl centralite, and di-n-butyl phthalate in pre- and post-blast smokeless powder particles using heptadecane as an internal standard. Post-blast debris was obtained via controlled explosions with assistance from the Indiana State Police Bomb Squad. Two galvanized steel and two polyvinyl chloride pipe bombs were assembled. Two devices contained single-base smokeless powder and two contained double-base smokeless powder. 2,4-dinitrotoluene and diphenylamine were successfully quantified in the single-base smokeless powder post-blast debris while nitroglycerin, diphenylamine, and ethyl centralite were successfully quantified in the double-base smokeless powder post-blast debris. Compounds were detected at concentrations as low as 9 µg of 2,4-dinitrotoluene per mg, <3 µg of diphenylamine per mg, 131 µg of nitroglycerin per mg, and <3 µg of ethyl centralite per mg. Concentration changes between pre- and post-blast smokeless powder particles were determined as well as microscopic differences between pre- and post-blast debris for both smokeless powders in all devices. To our knowledge, this is the first use of GC/VUV for the quantification of explosives.

Gas Chromatography-Mass Spectrometry Paired with Total Vaporization Solid-Phase Microextraction as a Forensic Tool.

Gas Chromatography - Mass Spectrometry (GC-MS) is a frequently used technique for the analysis of numerous analytes of forensic interest, including controlled substances, ignitable liquids, and explosives. GC-MS can be coupled with Solid-Phase Microextraction (SPME), in which a fiber with a sorptive coating is placed into the headspace above a sample or immersed in a liquid sample. Analytes are sorbed onto the fiber which is then placed inside the heated GC inlet for desorption. Total Vaporization Solid-Phase Microextraction (TV-SPME) utilizes the same technique as immersion SPME but immerses the fiber into a completely vaporized sample extract. This complete vaporization results in a partition between only the vapor phase and the SPME fiber without interference from a liquid phase or any insoluble materials. Depending upon the boiling point of the solvent used, TV-SPME allows for large sample volumes (e.g., up to hundreds of microliters). On-fiber derivatization may also be performed using TV-SPME. TV-SPME has been used to analyze drugs and their metabolites in hair, urine, and saliva. This simple technique has also been applied to street drugs, lipids, fuel samples, post-blast explosive residues, and pollutants in water. This paper highlights the use of TV-SPME to identify illegal adulterants in very small samples (microliter quantities) of alcoholic beverages. Both gamma-hydroxybutyrate (GHB) and gamma-butyrolactone (GBL) were identified at levels that would be found in spiked drinks. Derivatization by a trimethylsilyl agent allowed for conversion of the aqueous matrix and GHB into their TMS derivatives. Overall, TV-SPME is quick, easy, and requires no sample preparation aside from placing the sample into a headspace vial.

Tracking the Progression of Triple Negative Mammary Tumors over Time by Chemometric Analysis of Urinary Volatile Organic Compounds.

Previous studies have shown that volatile organic compounds (VOCs) are potential biomarkers of breast cancer. An unanswered question is how urinary VOCs change over time as tumors progress. To explore this, BALB/c mice were injected with 4T1.2 triple negative murine tumor cells in the tibia. This typically causes tumor progression and osteolysis in 1-2 weeks. Samples were collected prior to tumor injection and from days 2-19. Samples were analyzed by headspace solid phase microextraction coupled to gas chromatography-mass spectrometry. Univariate analysis identified VOCs that were biomarkers for breast cancer; some of these varied significantly over time and others did not. Principal component analysis was used to distinguish Cancer (all Weeks) from Control and Cancer Week 1 from Cancer Week 3 with over 90% accuracy. Forward feature selection and linear discriminant analysis identified a unique panel that could identify tumor presence with 94% accuracy and distinguish progression (Cancer Week 1 from Cancer Week 3) with 97% accuracy. Principal component regression analysis also demonstrated that a VOC panel could predict number of days since tumor injection (R2 = 0.71 and adjusted R2 = 0.63). VOC biomarkers identified by these analyses were associated with metabolic pathways relevant to breast cancer.

A critical comparison of vacuum UV (VUV) spectrometer and electron ionization single quadrupole mass spectrometer detectors for the analysis of alkylbenzenes in gasoline by gas chromatography: Experimental and statistical aspects.

Recent advances in benchtop vacuum ultraviolet (VUV) spectrometers have yielded effective universal detectors for gas chromatography (GC). The ability of these detectors to acquire absorbance spectra from 125 nm to 430 nm poses an alternative to the gold standard of mass spectrometry (MS) as a sensitive and selective GC detector. The applications of GC/VUV extend into many areas. Featured here is the potential application of GC/VUV to the analysis of ignitable liquids, which may be found on debris from suspected arson fires. A particular compound class of interest is the alkylbenzenes, as they are a significant component in fuels such as gasoline, petroleum distillates, and aromatic solvents such as degreasers and cleaning solvents. To measure the sensitivity, selectivity and specificity of GC-VUV and GC-MS for alkylbenzenes we employed both library search methods and chemometric analysis using discriminant analysis. The GC-VUV detector was found to have superior specificity to the GC-MS detector in full scan mode. The GC-VUV detector was able to identify all alkylbenzenes correctly, including the correct identification of all structural isomers. LODs for both GC-VUV and GC-MS were found to be picograms on column.

Detection of ɣ-hydroxybutyric acid (GHB) and ɣ-butyrolactone (GBL) in alcoholic beverages via total vaporization solid-phase microextraction (TV-SPME) and gas chromatography-mass spectrometry.

Total Vaporization Solid-Phase Microextraction (TV-SPME) relies on the same technique as standard SPME but completely vaporizes a sample extract, and analytes are sorbed directly from the vapor phase. On-fiber derivatization may also be performed using TV-SPME, where the fiber is first exposed to the headspace of a vial containing the derivatization agent, then exposed to a new vial containing the sample. É£-Hydroxybutyric acid (GHB) and É£-butyrolactone (GBL) are drugs of concern in that they may be used in drug facilitated sexual assault by surreptitiously spiking them into a victim's beverage. These drugs cause sedation, memory loss, and are difficult to detect in biological samples. One challenge in their analysis is that they can interconvert in aqueous samples, which was demonstrated in samples allowed to stand at room temperature for long periods. A volume study of GBL in water was performed with volumes ranging from 1 to 10,000 µl to compare the efficacy of TV-SPME, headspace SPME, and immersion SPME. Lastly, water, beer, wine, liquor, and mixed drinks were spiked with either GHB or GBL with realistic concentrations (mg/ml) and microliter quantities were analyzed using a TV-SPME Gas Chromatography-Mass Spectrometry method. The GBL volume study demonstrated an increased sensitivity in GBL detection when TV-SPME was utilized. Additionally, GHB and GBL were identified in various beverages at realistic concentrations. Overall, TV-SPME is beneficial because it requires no sample preparation and uses smaller sample volumes than immersion and headspace SPME.

Thermal and spectroscopic analysis of nitrated compounds and their break-down products using gas chromatography/vacuum UV spectroscopy (GC/VUV).

Gas chromatography/vacuum UV spectroscopy (GC/VUV) was utilized to study various explosives and pharmaceuticals in the nitrate ester and nitramine structural classes. In addition to generating specific VUV spectra for each compound, VUV was used to indicate the onset of thermal decomposition based upon the appearance of break-down products such as nitric oxide, carbon monoxide, formaldehyde, water, and molecular oxygen. The effect of temperature on decomposition could be fit to a logistical function where the fraction of intact compound remaining decreased as the transfer line/flow cell temperature was increased from 200 °C to 300 °C. Utilizing this relationship, the decomposition temperatures for the nitrate ester and nitramine compounds were determined to range between 244 °C and 277 °C. It was also discovered that the decomposition temperature was dependent on the GC carrier gas flow rate and, therefore, the residence time of the compounds in the transfer line/flow cell. For example, the measured decomposition temperature of nitroglycerine ranged from 222 °C to 253 °C across four flow rates. Tracking the appearance/disappearance of decomposition products across this temperature range indicated that NO, CO, and H2CO are final decomposition products while O2 and H2O are intermediate products. The decomposition temperatures for all explosives were highly correlated to similar decomposition measurements taken by differential scanning calorimetry (DSC) (r = 0.91) and thermal gravimetric analysis (TGA) (r = 0.90-0.98). In addition, the decomposition temperatures for all explosives were negatively correlated to the heat of explosion at constant volume (r = -0.68) and strongly positively correlated to the oxygen balance (r = 0.92).

Optimization of the qualitative and quantitative analysis of cocaine and other drugs of abuse via gas chromatography - Vacuum ultraviolet spectrophotometry (GC - VUV).

Gas Chromatography-Vacuum UV Spectroscopy (GC-VUV) has seen increased attention in many areas, however, a statistical optimization of VUV method parameters has not been published. This article presents the first statistical optimization of parameters influencing analytes such as cocaine in the VUV flow-cell. Flow-cell temperature, make-up gas pressure, and carrier gas flow rate from the GC were examined and optimized for the detection of controlled substances. The accuracy, precision, linearity, and optimized detection limits for drugs such as cocaine (98.5%, 1.2%, 0.9998, 1.5 ng), heroin (99.3%, 0.94%, 0.9998, 2.0 ng), and fentanyl (98.5%, 1.7%, 0.9752, 9.7 ng) are reported. In general, the limits of detection for cocaine, heroin, fentanyl, and methamphetamine after optimization were comparable to gas chromatography-mass spectrometry (GC-MS) in "scan mode", which had detection limits of 1.1-38 ng on column. The VUV absorption spectra of cocaine, PCP, lorazepam, and HU-210 are also reported. And three samples of "real world" cocaine are analyzed to demonstrate applicability to forensic drug analysis.

Identifying Thermal Decomposition Products of Nitrate Ester Explosives Using Gas Chromatography-Vacuum Ultraviolet Spectroscopy: An Experimental and Computational Study.

Analysis of nitrate ester explosives (e.g., nitroglycerine) using gas chromatography-vacuum ultraviolet spectroscopy (GC-VUV) results in their thermal decomposition into nitric oxide, water, carbon monoxide, oxygen, and formaldehyde. These decomposition products exhibit highly structured spectra in the VUV that is not seen in larger molecules. Computational analysis using time-dependent density functional theory (TDDFT) was utilized to investigate the excited states and vibronic transitions of these decomposition products. The experimental and computational results are compared with those in previous literature using synchrotron spectroscopy, electron energy loss spectroscopy (EELS), photoabsorption spectroscopy, and other computational excited state methods. It was determined that a benchtop GC-VUV detector gives comparable results to those previously reported, and TDDFT could predict vibronic spacing and model molecular orbital diagrams.

Instrumental and chemometric analysis of opiates via gas chromatography-vacuum ultraviolet spectrophotometry (GC-VUV).

Since its introduction, gas chromatography (GC) coupled to vacuum ultraviolet spectrophotometry (VUV) has been shown to complement mass spectrometry (MS) for materials such as petrochemicals, explosives, pesticides, and drugs. In forensic chemistry, opioids are commonly encountered but rarely are the samples pure. This work focuses on GC-VUV analysis applied to naturally occurring (e.g., morphine), semi-synthetic (e.g., heroin), and synthetic (fentanyl) opioids as well as common adulterants and diluents (e.g., lidocaine and quinine). The specificity of the VUV spectra were examined visually as well as via descriptive statistical methods (e.g., correlation coefficients and sums of square residuals). Multivariate pattern recognition techniques (principal component analysis and discriminant analysis (DA)) were used to prove the opioid spectra can be reliably differentiated. The accuracy of the DA model was 100% for a test set of VUV spectra. Finally, three "street" heroin samples were analyzed to show "real-world" performance for forensic analyses. These samples contained adulterants such as caffeine, as well as by-products of heroin manufacture.

Preparation and characterization of micro-bore wall-coated open-tubular capillaries with low phase ratios for fast-gas chromatography-mass spectrometry: Application to ignitable liquids and fire debris.

Fast Gas Chromatography (GC) allows for analysis times that are a fraction of those seen in traditional capillary GC. Key modifications in fast GC include using narrow, highly efficient columns that can resolve mixtures using a shorter column length. Hence, a typical fast GC column has an inner diameter of 100-180 µm. However, to maintain phase ratios that are consistent with typical GC columns, the film thickness of fast GC stationary phases are also low (e.g., 0.1-0.18 µm). Unfortunately, decreased film thickness leads to columns with very low sample capacity and asymmetric peaks for analytes that are not sufficiently dilute. This paper describes micro-bore (50 µm i.d.) capillary columns with thick films (1.25 µm), and low phase ratios (10). These columns have greater sample capacity yet also achieve minimum plate heights as low as 110 µm. Hence, separation efficiency is much higher than would be obtained using standard GC columns. The capillary columns were prepared in-house using a simple static-coating procedure and their plate counts were determined under isothermal conditions. The columns were then evaluated using temperature programming for fast GC-MS analysis of ignitable liquids and their residues on fire debris exemplars. Temperature ramps of up to 75 °C min-1 could be used and separations of ignitable liquids such as gasoline, E85 fuel, and lighter fluid (a medium petroleum distillate) were complete within 3 min. Lastly, simulated fire debris consisting of ignitable liquids burned on carpeting were extracted using passive headspace absorption-elution and the residues successfully classified.

Generating highly specific spectra and identifying thermal decomposition products via Gas Chromatography / Vacuum Ultraviolet Spectroscopy (GC/VUV): Application to nitrate ester explosives.

Gas chromatography/mass spectrometry (GC/MS) is a "workhorse" instrument for chemical analysis, but it can be limited in its ability to differentiate structurally similar compounds. The coupling of GC to vacuum ultraviolet (VUV) spectroscopy is a recently developed technique with the potential for increased detection specificity. To date, GC/VUV has been demonstrated in the analysis of volatile organic compounds, petroleum products, aroma compounds, pharmaceuticals, illegal drugs, and lipids. This paper is the first to report on the utility of GC/VUV for explosives analysis in general, and the first to report on thermal degradation within the VUV cell and its analytical utility. The general figures of merit and performance of GC/VUV were evaluated with authentic standards of nitrate ester explosives (e.g., nitroglycerine (NG), ethylene glycol dinitrate (EGDN), pentaerythritol tetranitrate (PETN), and erythritol tetranitrate (ETN)). In addition, the explosive analytes were thermally degraded in the VUV cell, yielding reproducible, complex and characteristic mixtures of gas phase products (e.g., nitric oxide, carbon monoxide, and formaldehyde). The relative amounts of the degradation products were estimated via spectral subtraction of library spectra. Lastly, GC/VUV was used to analyze milligram quantities of intact and burned samples of double-base smokeless powders containing nitroglycerine, diphenylamine, ethyl centralite, and dibutylphthalate.

Endothelial disruptive proinflammatory effects of nicotine and e-cigarette vapor exposures.

The increased use of inhaled nicotine via e-cigarettes has unknown risks to lung health. Having previously shown that cigarette smoke (CS) extract disrupts the lung microvasculature barrier function by endothelial cell activation and cytoskeletal rearrangement, we investigated the contribution of nicotine in CS or e-cigarettes (e-Cig) to lung endothelial injury. Primary lung microvascular endothelial cells were exposed to nicotine, e-Cig solution, or condensed e-Cig vapor (1-20 mM nicotine) or to nicotine-free CS extract or e-Cig solutions. Compared with nicotine-containing extract, nicotine free-CS extract (10-20%) caused significantly less endothelial permeability as measured with electric cell-substrate impedance sensing. Nicotine exposures triggered dose-dependent loss of endothelial barrier in cultured cell monolayers and rapidly increased lung inflammation and oxidative stress in mice. The endothelial barrier disruptive effects were associated with increased intracellular ceramides, p38 MAPK activation, and myosin light chain (MLC) phosphorylation, and was critically mediated by Rho-activated kinase via inhibition of MLC-phosphatase unit MYPT1. Although nicotine at sufficient concentrations to cause endothelial barrier loss did not trigger cell necrosis, it markedly inhibited cell proliferation. Augmentation of sphingosine-1-phosphate (S1P) signaling via S1P1 improved both endothelial cell proliferation and barrier function during nicotine exposures. Nicotine-independent effects of e-Cig solutions were noted, which may be attributable to acrolein, detected along with propylene glycol, glycerol, and nicotine by NMR, mass spectrometry, and gas chromatography, in both e-Cig solutions and vapor. These results suggest that soluble components of e-Cig, including nicotine, cause dose-dependent loss of lung endothelial barrier function, which is associated with oxidative stress and brisk inflammation.

Design and optimization of a total vaporization technique coupled to solid-phase microextraction.

Solid-phase microextraction (SPME) is a popular sampling technique in which chemical compounds are collected with a sorbent-coated fiber and then desorbed into an analytical instrument such as a liquid or gas chromatograph. Typically, this technique is used to sample the headspace above a solid or liquid sample (headspace SPME), or to directly sample a liquid (immersion SPME). However, this work demonstrates an alternative approach where the sample is totally vaporized (total vaporization SPME or TV-SPME) so that analytes partition directly between the vapor phase and the SPME fiber. The implementation of this technique is demonstrated with polydimethylsiloxane-divinylbenzene (PDMS-DVB) and polyacrylate (PA) coated SPME fibers for the collection of nicotine and its metabolite cotinine in chloroform extracts. The most important method parameters were optimized using a central composite design, and this resulted in an optimal extraction temperature (96 °C), extraction time (60 min), and sample volume (120 µL). In this application, large sample volumes up to 210 µL were analyzed using a volatile solvent such as chloroform at elevated temperatures. The sensitivity of TV-SPME is nearly twice that of liquid injection for cotinine and nearly 6 times higher for nicotine. In addition, increased sampling selectivity of TV-SPME permits detection of both nicotine and cotinine in hair as biomarkers of tobacco use where in the past the detection of cotinine has not been achieved by conventional SPME.

The effect of microbial degradation on the chromatographic profiles of tiki torch fuel, lamp oil, and turpentine.

Biodegradation can result in selective removal of many of the compounds required for the identification of an ignitable liquid. In this study, the effects of microbial degradation on tiki torch fuel, lamp oil, and turpentine are reported. Samples of soil spiked with 20 µL of the liquids were stored at room temperature for up to 7 days. The ignitable liquids were then recovered using passive headspace concentration onto charcoal strips followed by solvent elution using pentane. Microbial degradation of tiki torch fuel resulted in the loss of the n-alkanes relative to the branched alkanes. Changes in the profile of the lamp oil were minor due to the highly branched nature of its alkanes. Microbial degradation of turpentine resulted in the selective loss of limonene and o-cymene. Overall, significant degradation by microbial action could result in the inability to identify the presence of an ignitable liquid or misclassify the ignitable liquid found.

Chemical characterization of dissolvable tobacco products promoted to reduce harm.

In 2009, the R. J. Reynolds Tobacco Co. released a line of dissolvable tobacco products that are marketed as an alternative to smoking in places where smoking is prohibited. These products are currently available in Indianapolis, IN, Columbus, OH, and Portland, OR. This paper describes the chemical characterization of four such products by gas chromatography-mass spectrometry (GC-MS). The dissolvable tobacco products were extracted and prepared by ultrasonic extraction using acetone, trimethylsilyl derivatization, and headspace solid phase microextraction (SPME). The following compounds were identified in the dissolvables using either ultrasonic extractions or trimethylsilyl derivatization: nicotine, ethyl citrate, palmitic acid, stearic acid, sorbitol, glycerol, and xylitol. The following compounds were identified in the dissolvables using headspace SPME: nicotine, ethyl citrate, cinnamaldehyde, coumarin, vanillin, and carvone. With the exception of nicotine, the compounds identified thus far in the dissolvables are either flavoring compounds or binders. The concentration of free nicotine in the dissolvables was determined from the Henderson-Hasselbalch equation and by measuring the pH and nicotine concentration by GC-MS. The results presented here are the first to reveal the complexity of dissolvable tobacco products and may be used to assess potential oral health effects.

Discriminating Hodgdon Pyrodex(®) and Triple Seven(®) using gas chromatography-mass spectrometry.

Pyrodex(®) and Triple Seven(®) are black powder substitutes that often find use as fillers in improvised explosive devices, such as pipe bombs. These propellants have essentially the same overall appearance and oxidizers, but different fuels. For example, Pyrodex(®) contains sulfur, sodium benzoate, and dicyandiamide (DCDA), whereas Triple Seven(®) lacks sulfur but also contains 3-nitrobenzoic acid. In this method, intact particles and postblast solid residues were reacted with bis(trimethylsilyl)trifluoroacetamide + 1% trimethylchlorosilane in acetonitrile for 30 min at 60°C. The resultant trimethylsilyl derivatives of the organic fuels were then analyzed by gas chromatography-mass spectrometry. Each derivative was clearly resolved from other components, and high-quality mass spectra were obtained. In addition, characteristic fragments resulting from loss of a methyl radical from the molecular ion (m/z 163 for sulfur, m/z 171 for DCDA, m/z 179 for benzoic acid, and m/z 224 for nitrobenzoic acid) were able to be monitored.

Characterization of surface organic components of human hair by on-line supercritical fluid extraction-gas chromatography/mass spectrometry: a feasibility study and comparison with human identification using mitochondrial DNA sequences.

This paper discusses results of a supercritical fluid extraction-gas chromatography/mass spectrometry (SFE-GC/MS) study of small samples ( 100 microg to 1 mg) of human scalp hair. The method offers a number of benefits including greater sensitivity than liquid extraction methods because the entire extractable mass is transferred to the analytical system, compared with only a few percent from a conventional liquid extraction/injection. The project's goals were to determine if SFE-GC/MS analyses of the surface-extractable components of an individual's hair yield consistent chemical profiles and to investigate if the profiles are sufficiently different to distinguish them from those of other individuals. In addition, the mtDNA sequences from ten of the same individuals used in the SFE-GC/MS study from four family units were determined, and, while the families were distinguishable, the maternal relations yielded identical sequences. In tandem, SFE-GC/MS and mtDNA techniques may provide valuable complementary data from forensic hair samples.

Evaluation of extraction techniques for the forensic analysis of human scalp hair using gas chromatography/mass spectrometry (GC/MS).

Preliminary research using on-line supercritical fluid extraction/gas chromatography-mass spectrometry (SFE/GC-MS) has shown that the natural and artificial surface components of human scalp hair are reproducible and differentiable. Therefore, these components may be useful for individualization or determining demographic characteristics or both. However, it is not known how the efficiency and selectivity of on-line SFE/GC-MS compares to other extraction methods. In this study, ultrasound, Soxhlet, and pressurized-fluid extraction were used to extract 1 mg to 1.3 g portions of a composite hair sample taken from an Asian male between the ages of 10 and 18. Percent extractables ranged from 0.9% to 5.6%, depending on the solvent used, and tended to increase with solvent polarity. Chemical analysis using GC/MS showed that the extracts contained large proportions of free fatty acids, squalene, cholesterol, and various wax esters. Finally, comparisons to SFE/GC-MS showed that this method possesses adequate efficiency, no observable differences in selectivity, and greater potential for miniaturization.