Selected Ion Monitoring Gas Chromatography-mass Research Articles

Multi-Residual Determination of Multi-Class Pesticides in Groundwater by Direct Immersion Solid-Phase Microextraction with Gas Chromatography-Selected Ion Monitoring Mass Spectrometry (GC–MS/SIM) Detection

Multi-Residual Determination of Multi-Class Pesticides in Groundwater by Direct Immersion Solid-Phase Microextraction with Gas Chromatography-Selected Ion Monitoring Mass Spectrometry (GC–MS/SIM) Detection

A gas chromatography–mass spectrometry method for the detection of chlorpyrifos contamination in palm‐based fatty acids

AbstractChlorpyrifos is a Malaysian Pesticide Board‐approved organophosphate insecticide, which may become concentrated during fractionation. The objective of this project was to develop and validate a method to detect and quantify chlorpyrifos in food‐grade fatty acids ingredients, e.g. caprylic‐capric acid mixture and oleic acid (OLA) used to synthesize triacylglyceride based food additives and in the cosmetic industry. A selective ion monitoring gas chromatography–mass spectrometry method with a matrix‐matched calibration curve calibration was selected. The method involved the direct injection of chlorpyrifos spiked into the fatty acids matrix. The percentage recoveries at spiking levels of 0.5, 0.75, 2.5, and 4.0 μg g−1 of chlorpyrifos in OLA and caprylic‐capric acid ranged from 85.7%–101.1% to 97.2%–112%, respectively, with a relative standard deviation of within 11%. The intra‐day and inter‐day precision were deemed acceptable as indicated by an relative SD value of within 10%. A good linear relationship with a coefficient of correlation >0.99 for the matrix‐matched calibration was achieved between 0.5 and 5 μg g−1. The limit of detection and limit of quantification corresponded to 0.5 μg g−1 for OLA and 0.55 μg g−1 for caprylic‐capric acid.

Determination of 24 Pesticides Residues in Leather Products by Solid-Phase Microextraction Coupled with Gas Chromatography–Mass Spectrometry

Considering the high content of oil and complex residual additives in leather samples, a new analytical method based on the solid-phase extraction technique and gas chromatography-selected ion monitoring mass spectrometry (GC-SIM-MS) was developed to determine 24 organic compounds involving the organochlorine pesticides (OCPs), organophosphorous pesticides (OPPs) and pyrethroids pesticides residues in leather. The extraction conditions (such as the extraction solution, purification procedure and solid-phase extraction column) were optimized using the positive leather samples based on the recovery rates of the pesticides. The best extraction solution, solid-phase extraction column and chromatography column were n-hexane and ethyl acetate (1+1, volume) mixed solution, Carb-PSA (1.0 g, 6mL) and DB-1701 (length: 30 m, inside diameter: 0.25 mm, film thickness: 0.25 μm). The optimized extraction time and temperature were 20 min and 25°C, respectively. The detection limits of 24 pesticide residues range from 0.05 to 0.10 mg/kg, and the recoveries range from 74% to 116%. The relative standard deviations (RSD, n=6) range from 5.42% to 12.00%. The developed method presented a simple, rapid, sensitive, and inexpensive method to detect 24 pesticides in skin and leather and was successfully applied to the detect them in leather products (cowhide, sheep leather and pig leather).

Solid-phase extraction and gas chromatography-mass spectrometry analysis of 11-nor-9-carboxy-δ9-tetrahydrocannabinol in urine for monitoring marijuana abuse

A specimen pretreatment procedure, combining solid-phase extraction and methylation, was developed for the analysis of 11-nor-9-carboxy-Δ9- tetrahydrocannabinol (9-THC-COOH) in urine for monitoring marijuana exposure. d3-11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (d3-9-THC-COOH) was used as an internal standard, while the now universal selected ion monitoring gas chromatography-mass spectrometry approach was used for detection (m/z 313, 357, 372 and 316, 360, 375 for 9-THC-COOH and d3-9-THC-COOH, respectively) and quantitation (m/z 372 and 375 for 9-THC-COOH and d3-9-THC-COOH, respectively). The effectiveness of the overall procedure was evaluated. One of the extraction columns evaluated achieved > 90% recovery of the analyte in 5-mL drug-free urine fortified with 15 ng/mL 9-THC-COOH. Using the one-point calibration approach, the overall protocol achieved the following analytical parameters: detection limit: 2.09 ng/mL; limit of quantitation: 4.18 ng/mL; upper end of linear range: 376 ng/mL (or better); interday precision: 1.49- 8.03% CV in the 4.18-376 ng/mL concentration range studied.

A Selective Gas Chromatography-Selected Ion Monitoring Mass Spectrometry (GC‑MS/SIM) Method to Determine Polychlorinated Biphenyls in Fuller's Earth

Polychlorinated biphenyls (PCBs) are carcinogenic to humans and can be found in Fuller’s earth used for the regeneration of used transformer oil. This work describes a highly selective gas chromatographic-selected ion monitoring mass spectrometric method (GC-MS/SIM) for the simultaneous determination of three Aroclor PCB congener mixtures (1242, 1254, and 1260). The separation of the PCBs could be systematically varied by changing the temperature conditions of the chromatographic column. Methods were first selected from the literature, and the resolution of the PCBs was then optimized by adjusting k* (effective retention), a (selectivity), and N (column efficiency), enabling the detection of 83 PCBs in 35 min. The extract of PCBs from Fuller’s earth was obtained by ultrasonication (employing hexane and acetone as solvents) and cleanup with Florisil and was concentrated to about 500 µL. The ions analyzed in SIM mode were divided into groups according to the similarity (m/z) of the PCB fragmentation profiles. The limits of detection (LOD) and quantification (LOQ) were 0.19 and 0.64 µg kg-1, respectively. The method developed was used to determine the PCBs present in Fuller’s earth provided by an electric power company. The mean total PCBs concentration was found 1.9 µg kg-1 in Fuller’s earth.

Identification of potential indicators of time-dependent tequila maturation and their determination by selected ion monitoring gas chromatography–mass spectrometry, using salting-out liquid–liquid extraction

In this work, gas chromatography–mass spectrometry (GC–MS) profiles obtained for a series of rested and aged tequilas were statistically compared to identify volatile compounds indicative of time-dependent tequila maturation. For the determination of nine selected compounds, 2-acetylthiazole was added as internal standard, ethyl acetate extraction was carried out in the presence of sodium chloride (salting-out) and sodium sulfate was used to eliminate water traces. Baseline separation was achieved within 18 min and quantification was carried out by selected ion monitoring. The method detection limits were as follows: 2.10 µg L− 1 guaiacol (m/z 109), 1.78 µg L− 1 4-methylguaiacol (m/z 123), 1.97 µg L− 1 4-ethylguaiacol (m/z 137), 3.78 µg L− 1trans-whiskey lactone (m/z 99), 4.42 µg L− 1cis-whiskey lactone (m/z 99), 10.3 µg L− 1eugenol (m/z 103), 8.27 µg L− 1 vanillin (m/z 151), 14.2 µg L− 1 apocynin (m/z 151), 12.1 µg L− 1 syringaldehyde (m/z 182). For accuracy testing, 100 µg L− 1 of each analyte was added to silver tequila, yielding the recoveries in the range 92.1–116%. Data obtained in the analysis of 28 tequilas confirmed that the concentrations of individual compounds were changing among liquors at different maturation stage; however, due to many variables affecting the profile of volatile/semi-volatile compounds during aging in wooden barrels, discrimination of tequilas was not feasible. Otherwise, the proposed procedure allows for reliable determination of nine odor-active compounds in tequila at trace levels with simple sample pretreatment and short analytical run.

Application of Selected Ion Monitoring-Gas Chromatography Mass Spectrometry (SIM-GCMS) for Quantification of Poly Aromatic Hydro Carbons in Surface Water from Sarawak, Mambong River (Malaysia)

Background: Increasing activities have led to serious environmental problems due to Pullution caused by toxic materials such as poly aromatic carbons whose levels are rising in the environment.
 Objective: The study presented here carried out the ultra-trace Quantification of Poly Aromatic Hydro carbons in surface water using Selected Ion Monitoring with optimization Gas Chromatography Mass Spectrometry parameter.
 Methods: A Shimadzu QP 2010 Plus GCMS equipped with an auto-injector AOC-20i, 30 m x 0.25 mm x 0.25 um of BP-X5 capillary column (SGE, USA) was used to obtain the result of poly aromatic hydrocarbons to separate and quantify the PAH compounds as well as to identify the Polyromantic Hydrocarbon using USEPA.
 Results: The amount of polycyclic aromatic hydrocarbons (PAHs) at ultra-trace level in the surface water samples was estimated. Sixteen Poly Aromatic Hydro carbons were identified among which Naphthalene Pyrene (45.7 ng/L), Chrysene (59.38 ng/L), Benzo (g, h, i) perylene (40.35 ng/L) as the major pollutants in the water surface of Mambong River water. Total Poly Aromatic Hydro carbons (PAHs) concentrations ranged from 2.0 to 40 ng L-1 was detected in the river water.
 Conclusion: The Selected Ion Monitoring -Gas Chromatography Mass Spectrometry (SIM-GCMS) can be considered as sensitive and robust method to analyse PAHs contamination in environmental samples. Thus, help the researchers to uncover the critical areas of environmental pollution at ultra-trace level that many researchers were not able to explore.

Exploring metabolic syndrome serum free fatty acid profiles based on GC–SIM–MS combined with random forests and canonical correlation analysis

Metabolic syndrome (MetS) is a cluster of metabolic abnormalities associated with an increased risk of developing cardiovascular diseases or type II diabetes. Till now, the etiology of MetS is complex and still unknown. Metabolic profiling is a powerful tool for exploring metabolic perturbations and potential biomarkers, thus may shed light on the pathophysiological mechanism of diseases. In this study, fatty acid profiling was employed to exploit the metabolic disturbances and discover potential biomarkers of MetS. Fatty acid profiles of serum samples from metabolic syndrome patients and healthy controls were first analyzed by gas chromatography–selected ion monitoring–mass spectrometry (GC–SIM–MS), a robust method for quantitation of fatty acids. Then, the supervised multivariate statistical method of random forests (RF) was used to establish a classification and prediction model for MetS, which could assist the diagnosis of MetS. Furthermore, canonical correlation analysis (CCA) was employed to investigate the relationships between free fatty acids (FFAs) and clinical parameters. As a result, several FFAs, including C16:1n-9c, C20:1n-9c and C22:4n-6c, were identified as potential biomarkers of MetS. The results also indicated that high density lipoprotein-cholesterol (HDL-C), triglycerides (TG) and fasting blood glucose (FBG) were the most important parameters which were closely correlated with FFAs disturbances of MetS, thus they should be paid more attention in clinical practice for monitoring FFAs disturbances of MetS than waist circumference (WC) and systolic blood pressure/diastolic blood pressure (SBP/DBP). The results have demonstrated that metabolic profiling by GC–SIM–MS combined with RF and CCA may be a useful tool for discovering the perturbations of serum FFAs and possible biomarkers for MetS.

Authentication of pineapple (Ananas comosus [L.] Merr.) fruit maturity stages by quantitative analysis of γ- and δ-lactones using headspace solid-phase microextraction and chirospecific gas chromatography–selected ion monitoring mass spectrometry (HS-SPME–GC–SIM-MS)

Headspace solid phase microextraction and chirospecific gas chromatography–mass spectrometry in selected ion monitoring mode (HS-SPME–GC–SIM-MS) allowed quantitative determination of δ-lactones (δ-C8, δ-C10) and γ-lactones (γ-C6, γ-C8, γ-C10). A stable isotope dilution assay (SIDA) with d7-γ-decalactone as internal standard was used for quantitative analysis of pineapple lactones that was performed at three progressing post-harvest stages of fully ripe air-freighted and green-ripe sea-freighted fruits, covering the relevant shelf-life of the fruits. Fresh pineapples harvested at full maturity were characterised by γ-C6 of high enantiomeric purity remaining stable during the whole post-harvest period. In contrast, the enantiomeric purity of γ-C6 significantly decreased during post-harvest storage of sea-freighted pineapples. The biogenetical background and the potential of chirospecific analysis of lactones for authentication and quality evaluation of fresh pineapple fruits are discussed.

Cork as a new (green) coating for solid-phase microextraction: Determination of polycyclic aromatic hydrocarbons in water samples by gas chromatography–mass spectrometry

A new fiber for solid-phase microextraction (SPME) was prepared employing cork as a coating. The morphology and composition of the cork fiber was evaluated by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR), respectively. The proposed fiber was used for the determination of polycyclic aromatic hydrocarbons (PAHs) in river water samples by gas chromatography–selected ion monitoring–mass spectrometry (GC–SIM–MS). A central composite design was used for optimization of the variables involved in the extraction of PAHs from water samples. The optimal extraction conditions were extraction time and temperature of 60min and 80°C, respectively. The detection and quantification limits were 0.03 and 0.1μgL−1, respectively. The recovery values were between 70.2 and 103.2% and the RSD was ≤15.7 (n=3). The linear range was 0.1–10μgL−1 with r≥0.96 and the fiber-to-fiber reproducibility showed RSD≤18.6% (n=5). The efficiency of the cork fiber was compared with commercially available fibers and good results were achieved, demonstrating the applicability and great potential of cork as a coating for SPME.

Metabolite target analysis of isoprenoid pathway in Saccharomyces cerevisiae in response to genetic modification by GC-SIM-MS coupled with chemometrics

A highly selective, sensitive and nonradioactive analytical method for identification and quantification of intracellular metabolites involved in isoprenoid pathway has been developed by means of gas chromatography-selected ion-monitoring mass spectrometry (GC-SIM-MS). These metabolites are classified into two groups: sterols (squalene, ergosterol, lanosterol) and phosphorylated compounds (geranyl diphosphate, farnesyl pyrophosphate, geranylgeranyl pyrophosphate) based on their physicochemical properties. To quantify both groups in a single analytical run, GPP, FPP and GGPP were cleaved to the parent alcohols, geraniol, farnesol, geranylgeraniol by pyrophosphatase followed by alkaline phosphatase before extraction, separation and detection. This study evaluated several extraction procedures and determined the effects of the type of extraction solvent, times used for extraction. Under optimized GC/EI-MS conditions, six compounds were separated with high efficiency in the selected-ion monitoring (SIM) mode. Linearity of the method was good with correlation coefficients (r 2) in the range of 0.9953–0.9999 and detection limits were 1.53–151.88 ng/ml. The intra-day and inter-day precision of the method, as RSD, were less than 5.31 and 6.04%, respectively. The accuracy of six compounds varied between 87.7 and 110.8%. This assay was successfully applied to the determination of six major metabolites in the pathway for isoprenoid biosynthesis in S. cerevisiae and is sensitive to detect changes following genetic modification. By isolating statistically significant differences among metabolite levels from four biological conditions, we observed discriminatory metabolic features that hinted that the role of erg9 and coq1gene was involved in isoprenoid pathway. Integrating this analytical approach with statistical strategies, we can determine the influence of erg9 and coq1gene on isoprenoid levels of S. cerevisiae, thus leading to improved understanding of the pathway in a multitude of biological systems.

A capillary gas chromatography-selected ion monitoring mass spectrometry method for the analysis of atractylenolide I in rat plasma and tissues, and application in a pharmacokinetic study

The aim of this paper is to investigate the characteristics of atractylenolide I (AO-I) in the body by a GC–MS method. All bio-samples were cleared up with a liquid–liquid extraction procedure. The calibration curves were linear within a range of 5–1000 ng/mL for plasma samples, 0.06–16.00 μg/g for cerebellum samples, and 0.03–8.00 μg/g for other tissue samples. The limit of quantification (LOQ) for AO-I was 1.0 ng/mL or 1.0 ng/g (S/N ≥ 10) in the bio-samples. In the applications, the main pharmacokinetic parameters were firstly obtained as follows: T max = 0.37 ± 0.19 h, C max = 0.26 ± 0.05 μg/mL, AUC = 1.95 ± 0.30 μg h/mL and k a = 10.08 ± 5.60 h −1. The tissue distribution of AO-I in rats after the oral administration of 50.0 mg/kg was from 0.225 to 0.031 μg/g with a decreasing tendency in different tissues like liver > kidney > spleen > cerebellum > heart > cerebrum > lung. The protein binding in rat plasma, human plasma and bovine serum albumin was 80.8 ± 3.9, 90.6 ± 3.1 and 60.9 ± 5.1%, respectively.

Sensitive profiling of chemically diverse bioactive lipids

Here, we present an improved method for sensitive profiling of lipids in a single high-performance liquid chromatography-electrospray ionization-quadrupole time of flight mass spectrometry experiment. The approach consists of i) sensitive isocratic elution, which takes advantage of C18 column material that is resistant to increased pH values induced by piperidine, ii) chemometric alignment of mass spectra followed by differential analysis of ion intensities, and iii) semiquantitative analysis of extracted ion chromatograms of interest. A key advantage of this method is its wide applicability to extracts that harbor lipids of considerable chemical complexity. The method allows qualitative and semiquantitative analysis of fatty acyls, glycerophospholipids (such as glycerophosphatidylinositols, glycerophosphatidylserines, and glycerophosphatidylcholines in brain extracts), phosphatidylinositol mannosides, acylated glycerophospholipids, sphingolipids (including ceramides and gangliosides in brain extracts), and, for the first time with ESI, prenols and mycolic acids (MAs). MAs are targets in antimycobacterial therapy, and they play an important immunomodulatory role during host-pathogen interactions. We compared high-resolution mass spectra of MAs derived from Mycobacterium bovis Bacille Camette-Guérin during entry into nonreplicative conditions induced by oxygen deprivation (hypoxic dormancy). Although the overall composition is not drastically altered, there are pronounced differences in individual MAs. alpha-MAs accumulate during entry into dormancy, whereas a subpopulation of keto-MAs is almost entirely eliminated. This effect is reversed upon resuscitation of dormant mycobacteria. These results provide detailed chemical information with relevance to drug development and immunobiology of mycobacteria.

A robust and selective method for the quantification of glycosylphosphatidylinositols in biological samples.

We have developed an assay for the quantification of glycosylphosphatidylinositol (GPI)-anchored glycoconjugates. The method is based on nitrous acid deamination and sodium borodeuteride reduction of the glucosamine residue, common to all GPI structures, to yield [1-2H]-2,5-anhydromannitol. Following acid methanolysis and trimethylsilyl derivatization, detection is by selected ion monitoring gas chromatography-mass spectrometry. The unnatural inositol isomer scyllo-inositol is used as an internal standard and the [1-2H]-2,5-anhydromannitol trimethylsilyl derivative is detected by following a characteristic electron-impact fragment ion at m/z 273. This method is more selective for GPIs than assays based on measuring myo-inositol content, which are often confounded by contaminating inositol-phospholipids. We show that the method can be applied to measure total GPI content in crude total lipid extracts and even in whole trypanosome ghosts. The method was applied to whole cell lysates of wild-type, GPI-deficient, and glycosaminoglycan-deficient CHO cells. The data revealed that proteoglycans did not interfere with total glucosamine estimates but that there are background levels of non-GPI and nonproteoglycan glucosamine-containing material in CHO cells.

Validated semiquantitative/quantitative screening of 51 drugs in whole blood as silylated derivatives by gas chromatography-selected ion monitoring mass spectrometry and gas chromatography electron capture detection

A comprehensively validated procedure is presented for simultaneous semiquantitative/quantitative screening of 51 drugs of abuse or drugs potentially hazardous for traffic safety in serum, plasma or whole blood. Benzodiazepines (12), cannabinoids (3), opioids (8), cocaine, antidepressants (13), antipsychotics (5) and antiepileptics (2) as well as zolpidem, zaleplon, zopiclone, meprobamate, carisoprodol, tizanidine and orphenadrine and internal standard flurazepam, were isolated by high-yield liquid-liquid extraction (LLE). The dried extracts were derivatized by two-step silylation and analyzed by the combination of two different gas chromatographic (GC) separations with both electron capture detection (ECD) and mass spectrometry (MS) operating in a selected ion-monitoring (SIM) mode. Quantitative or semiquantitative results were obtained for each substance based on four-point calibration. In the validation tests, accuracy, reproducibility, linearity, limit of detection (LOD) and limit of quantitation (LOQ), selectivity, as well as extraction efficiency and stability of standard stock solutions were tested, and derivatization was optimized in detail. Intra- and inter-day precisions were within 2.5–21.8 and 6.0–22.5%, and square of correlation coefficients of linearity ranged from 0.9896 to 0.9999. The limit of quantitation (LOQ) varied from 2 to 2000 ng/ml due to a variety of the relevant concentrations of the analyzed substances in blood. The method is feasible for highly sensitive, reliable and possibly routinely performed clinical and forensic toxicological analyses.

Clinical spectrum of succinic semialdehyde dehydrogenase deficiency.

Succinic semialdehyde dehydrogenase (SSADH) deficiency is a rare autosomal recessive disorder affecting CNS gamma-aminobutyric acid (GABA) degradation. SSADH, in conjunction with GABA transaminase, converts GABA to succinate. In the absence of SSADH, GABA is converted to 4-OH-butyrate. The presence of 4-OH-butyrate, a highly volatile compound, may be undetected on routine organic acid analysis. Urine organic acid testing was modified at the authors' institution in 1999 to screen for the excretion of 4-OH-butyrate by selective ion monitoring gas chromatography-mass spectrometry in addition to total ion chromatography. Since then, five patients with 4-hydroxybutyric aciduria have been identified. The authors add the clinical, neuroimaging, and EEG findings from a new cohort of patients to 51 patients reported in the literature with clinical details. Ages ranged from 1 to 21 years at diagnosis. Clinical findings include mild-moderate mental retardation, disproportionate language dysfunction, hypotonia, hyporeflexia, autistic behaviors, seizures, and hallucinations. Brain MRI performed in five patients at the authors' institution revealed symmetric increased T2 signal in the globus pallidi. SSADH deficiency is an under-recognized, potentially manageable neurometabolic disorder. Urine organic acid analysis should include a sensitive method for the detection of 4-hydroxybutyrate and should be obtained from patients with mental retardation or neuropsychiatric disturbance of unknown etiology.

Effects of Culture Conditions on the Mycolic Acid Composition of Isolates of Rhodococcus spp. from Activated SludgeFoams

The influence of two different carbon sources and three incubation temperatures on the mycolic acid compositions of three Rhodococcus isolates from activated sludge was examined using Selective Ion Monitoring (SIM) gas chromatography-mass spectrometry (GC-MS). Considerable qualitative and quantitative differences were detected in the mycolic acid compositions of the three very closely related isolates grown under the same conditions. Culture age also affected both the chain lengths and proportions of saturated mycolic acids detected in cell extracts, but not in the same manner for each isolate. Mycolic acids generally were of shorter chain lengths in cells grown with Tween 80 compared to glucose grown cells in strain 11R but the opposite situation occurred with strains A7 and D5. In all three, the proportion of unsaturated mycolic acids decreased with increasing growth temperatures. The taxonomic relevance of these observations and possible explanations for the observed changes in mycolic acid composition under various culture conditions are discussed.

Determination of corticosterone in rat and mouse plasma by gas chromatography-selected ion monitoring mass spectrometry

A simple, highly sensitive and specific method based on gas-chromatography-selected ion monitoring (SIM) mass spectrometry has been developed for the quantitation of corticosterone in rat and mouse plasma. After extraction of the plasma with ethyl acetate, the residue was trimethy-silylated with pentafluorobenzyl hydroxylamine-trimethylsilyl (PFBO-TMS). Detection of the derivatives was accomplished by a quadruple mass spectrometer in the selected ion monitoring mode ( m/ z of 316, 648, 663 and 678). The detection limit of the assay was 0.1 pg on column. The results show that in the plasma of non-stressed animals, only minor amounts of corticosterone were found; whereas in the plasma of stressed animals, it was dramatically increased. The method developed here can be used to examine corticosterone levels as a marker of stress in rats and mice and may also be used for estimation of the effect of stress-release medications.

Sequences of polypeptide antibiotics stilboflavins, natural peptaibol libraries of the mold Stilbella flavipes.

From the culture broths of the mold Stilbella flavipes CBS 146.81, a mixture of polypeptides could be isolated by adsorption on XAD polystyrene resin and purified by Sephadex LH-20 chromatography. Using preparative thin-layer chromatography (TLC) three groups of peptides, named stilboflavins (SF) A, B, and C could be separated. Each of the groups showed microheterogeneity when investigated by high-performance liquid chromatography (HPLC). Employing on-line HPLC-electrospray ionization tandem mass spectrometry in the positive and negative ionization mode, together with gas chromatography-selected ion monitoring mass spectrometry, enantioselective GC and quantitative amino acid analysis, the sequences of stilboflavins A and B could be determined. Exchange of Glu in stilboflavins A peptides (acidic) against Gln in stilboflavins B peptides (neutral) is the rational for different polarity of the peptide groups and their separatability by TLC. Since SF A and B are bioactive N-acetylated 20-residue peptides with a high proportion of alpha-aminoisobutyric acid and C-terminal bonded amino alcohols (either leucinol, isoleucinol or valinol) the peptides belong to the group of peptaibol antibiotics.

Isotopic analogues as internal standards for quantitative analyses of drugs and metabolites by GC-MS--nonlinear calibration approaches.

In order to achieve accurate quantitation of drugs and metabolites (analytes) in complex matrices, 2H- (and less commonly 13C-) labeled analogues of the analytes are now routinely adapted as the internal standards (IS) using linear calibration models to fit data generated by selected ion monitoring gas chromatography-mass spectrometry (GC-MS) protocols. In this study, the effects of cross-contribution (contribution of the IS to the intensity of the ion designated for the analyte and vice versa) on the linearity of the calibration data are examined. Nonlinear approaches that may address this problem are also studied. Two ion pairs (one with least and one with significant cross-contribution) from each of the following analyte/IS pairs are used as the exemplar systems for this study: butalbital/13C4-butalbital, butalbital/2H5-butalbital, secobarbital/13C4-secobarbital, and secobarbital/2H5-secobarbital. Analyte/IS ion intensity ratios of a series of standard solutions are correlated with the analyte/IS concentration ratios using one-point, multiple-point (unweighted and weighted) linear, and hyperbolic functions. The one-point calibration approach produces excellent calibration results in treating data derived from ion pairs with no significant cross contribution. In cases where significant cross-contribution exists, results derived from the one-point approach show, as expected, significant deviations at both ends of the concentration range. With the cross-contribution phenomenon accounted for, the hyperbolic calibration model is clearly more effective in fitting calibration data at both the lower and higher analyte concentration ends, thus significantly lowering the detection limit and extending the calibration range to a higher level. However, the calibration range cannot be extended indefinitely. At the low concentration end, noise-to-signal ratio and the cross-contribution of the IS to the intensity of the ion designated for the analyte, however insignificant, will incrementally reduce the quality of the observed ion intensity and intensity ratio data. At the high concentration end, detection saturation and the cross-contribution of the analyte to the intensity of the ion designated for the IS, however insignificant, will incrementally decrease the "slope" of the calibration curve. Thus, acceptable sensitivity (increase in analyte/IS ion-pair intensity ratio per unit increase in analyte concentration) of the calibration curve will become the limiting factor.