Internal Standard Calibration Research Articles

Determination of Patulin in Apple Juice and Apple-Derived Products Using a Robotic Sample Preparation System and LC-APCI-MS/MS

Patulin, a toxic mycotoxin, can contaminate apple-derived products. The FDA has established an action level of 50 ppb (ng/g) for patulin in apple juice and apple juice products. To effectively monitor this mycotoxin, there is a need for adequate analytical methods that can reliably and efficiently determine patulin levels. In this work, we developed an automated sample preparation workflow followed by liquid chromatography–atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) detection to identify and quantify patulin in a single method, further expanding testing capabilities for monitoring patulin in foods compared to traditional optical methods. Using a robotic sample preparation system, apple juice, apple cider, apple puree, apple-based baby food, applesauce, fruit rolls, and fruit jam were fortified with 13C-patulin and extracted using dichloromethane (DCM) without human intervention, followed by an LC-APCI-MS/MS analysis in negative ionization mode. The method achieved a limit of quantification of 4.0 ng/g and linearity ranging from 2 to 1000 ng/mL (r2 > 0.99). Quantitation was performed with isotope dilution using 13C-patulin as an internal standard and solvent calibration standards. Average recoveries (relative standard deviations, RSD%) in seven spike matrices were 95% (9%) at 10 ng/g, 110% (5%) at 50 ng/g, 101% (7%) at 200 ng/g, and 104% (4%) at 1000 ng/g (n = 28). The ranges of within-matrix and between-matrix variability (RSD) were 3–8% and 4–9%, respectively. In incurred samples, the identity of patulin was further confirmed with a comparison of the information-dependent acquisition-enhanced product ion (IDA-EPI) MS/MS spectra to a reference standard. The metrological traceability of the patulin measurements in an incurred apple cider (21.1 ± 8.0 µg/g) and apple juice concentrate (56.6 ± 15.6 µg/g) was established using a certified reference material and calibration data to demonstrate data confidence intervals (k = 2, 95% confidence interval).

Biomonitoring urinary pesticide metabolites in preschool children by supported liquid extraction and ultra-high performance liquid chromatography-tandem mass spectrometry and their association with oxidative stress

Investigating pesticide exposure and oxidative stress in preschool children is essential for elucidating the determinants of environmental health in early life, with human biomonitoring of urinary pesticide metabolites serving as a critical strategy for achieving this objective. This study demonstrated biomonitoring of 2 phenoxyacetic acid herbicides, 2 organophosphorus pesticide metabolites, and 4 pyrethroid pesticide metabolites in 159 preschool children and evaluated their association with oxidative stress biomarker 8-hydroxydeoxyguanosine. An enzymatic deconjugation process was used to release urinary pesticide metabolites, which were then extracted and enriched by supported liquid extraction, and quantified by ultra-high performance liquid chromatography-tandem mass spectrometry with internal standard calibration. Dichloromethane: methyl tert‑butyl ether (1:1, v/v) was optimized as the solvent for supported liquid extraction, and we validated the method for linear range, recovery, matrix effect and method detection limit. Method detection limit of the pesticide metabolites ranged from 0.01 μg/L to 0.04 μg/L, with satisfactory recoveries ranging from 70.5 % to 95.5 %. 2,4,5-Trichlorophenoxyacetic acid was not detected, whereas the other seven pesticide metabolites were detected with frequencies ranging from 10.1 % to 100 %. The concentration of urinary pesticide metabolites did not significantly differ between boys and girls, with the median concentrations being 9.39 μg/L for boys and 4.90 μg/L for girls, respectively. Spearman correlation analysis indicated that significant positive correlations among urinary metabolites. Bayesian kernel machine regression revealed a significant positive association between urinary pesticide metabolites and 8-hydroxydeoxyguanosine. Para-nitrophenol was the pesticide metabolite that contributed significantly to the elevated level of oxidative stress.

Simultaneous Determination of Ultra-Trace Pt, Pd, Rh, and Ir in Gold Mining Residue by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) with an Antimony-Copper Fire Assay and Microwave Digestion

The evaluation of the flow direction of platinum group elements (PGEs) during gold-copper ore mining is challenging due to the uneven distribution and low content. Therefore, it is vital to accurately and simultaneously determine ultra-trace PGEs Pt, Pd, Rh, and Ir in gold mining residue samples. In this study, a method to simultaneously determine ultra-trace Pt, Pd, Rh, and Ir in gold mining residue is reported using Sb–Cu fire assay, microwave digestion, and inductively coupled plasma-mass spectrometry (ICP-MS). The Sb–Cu fire assay was used to enrich Pt, Pd, Rh, and Ir, obtaining Sb–Cu alloy particles that concentrate these elements. An optimal microwave method was employed to digest the Sb–Cu alloy particles. In ICP-MS, the collision reaction interface was used to eliminate the interferences of other elements and improve the accuracy. Moreover, the matrix-matched multi-external standards combined with internal standard calibration were utilized to reduce the error. High concentrations of Au, Ag, and Pb showed no interferences with Pt, Pd, Rh, or Ir determination. The limits of detection for Pt, Pd, Rh, and Ir were 0.018, 0.052, 0.064, 0.035 ng·g−1, respectively, and the relative standard deviation was 5.18, 4.66, 1.55, and 3.17%. Additionally, the determination of Pt, Pd, Rh, and Ir in standard reference materials showed good agreement with the certified values. In summary, a reliable and efficient method is provided to determine various PGEs which is of great significance for gold-copper ore mining.

Removal of isobaric interference using pseudo-multiple reaction monitoring and energy-resolved mass spectrometry for the isotope dilution quantification of a tryptic peptide.

Energy-resolved mass spectrometry (ERMS) and an isotopically labelled internal standard were successfully combined to accurately quantify a tryptic peptide despite the presence of an isobaric interference. For this purpose, electrospray ionisation tandem mass spectrometry (ESI-MS/MS) experiments were conducted into an ion trap instrument using an unconventional 8m/z broadband isolation window, which encompassed both the tryptic peptide and its internal standard. Interference removal was assessed by determining an excitation voltage that was high enough to maintain a constant value for the analyte/internal standard peaks intensity ratio, thus ensuring accurate quantification even in the presence of isobaric contamination. Pseudo-multiple reaction monitoring (MRM) was employed above this excitation voltage to quantify the trypic peptide. The internal standard calibration model showed no lack of fit and exhibited a linear dynamic range from 0.5μM up to 2.5μM. The detection limit was 0.08 μM. The accuracy of the method was evaluated by quantifying the tryptic peptide of three reference samples intentionally contaminated with the isobaric interference. All the reference samples were accurately quantified with ∼1% deviation despite the isobaric contamination. Furthermore, we have demonstrated that this methodology can also be applied to quantify the isobaric peptide by standard additions down to 0.2μM. Finally, liquid chromatography ERMS (LC ERMS) experiments yielded similar results, suggesting the potential of the proposed methodology for analysing complex samples.

Development and Validation of an Analytical Method for the Determination of Fipronil and its Degradation Products in 28 Organic and Regular Honey Samples by GC-ECD

Recently, there has been a worldwide problem of increased bee mortality (Colony Collapse Disorder) and the intensive use of pesticides is suspected as one of the causes. Honey samples are one of many indicators to assess bee exposure to pesticides. In this work, a method for the simultaneous analysis of the pesticide Fipronil and its degradation products in honey samples by gas chromatography with electron capture detector (GC-ECD) is presented and validated. The extraction procedure was investigated using C18-SPE with different solvents and methanol showed the best performance. The analytical quantification was performed by internal standard matrix-matched calibration, which resulted in analytical curves presenting correlation coefficients higher than 0.99. The proposed method was validated with good results, such as recoveries around 70 - 99%, limits of detection and quantification bellow 0.014 and 0.072 μg mL-1, respectively, and relative standard deviations below 7%. The method is simple, effective and was successfully applied to 28 commercial honey samples, regular and organic, from different floral sources. The results showed the presence of fipronil desulfinyl, the main degradation product of fipronil, in some samples, even among the organic ones.

Identification method of 2-methylisoborneol and geosmin in water by purge and trap-gas chromatography-mass spectrometry

To establishe an analysis and identification method for 2-methylisoborneol(2-MIB) and geosmin(GSM) in water using purge and trap-gas chromatography-mass spectrometry. The samples were enriched and analyzed using a purge and trap system, followed by the separation on a DB-624(30 m×0.25 mm, 1.4 μm) chromatographic column. Quantification was performed using gas chromatography-mass spectrometry with the selected ion monitoring and internal standard calibration. The calibration curves for 2-MIB and GSM showed an excellent linearity in the range of 1 to 100 ng/L with R~2 values greater than 0.999. The detection limit and quantification limit for both 2-MIB and GSM were 0.33 ng/L and 1.0 ng/L, respectively. Spike recovery experiments were further carried on the source water and drinking water at three concentration levels. It showed that the average recoveries were from 82.0% to 111.0% for 2-MIB while 84.0% to 110% for GSM. Additionally, the test precision of 2-MIB and GSM ranged from 1.9% to 7.3% and 1.9% to 5.0%(n=6), respectively. The analysis of multiple samples including the local source water, treated water and distribution network water confirmed the existence of 2-MIB and GSM. Compared to the national standard(GB/T 5750.8-2023), the proposed method enables fully automated sample introduction and analysis without the extra pre-treatment. It provides the advantages of simplicity, good repeatability and high accuracy.

Phantomless Computed Tomography-Based Quantitative Bone Mineral Density Assessment: A Literature Review

The global surge in aging populations has intensified osteoporosis challenges, necessitating improved diagnostic methods. While dual X-ray absorptiometry (DXA) is the conventional standard in assessing bone mineral density (BMD), the exploration of quantitative computed tomography (QCT) has been proposed, particularly phantomless techniques. Challenges in single-energy CT, emphasizing internal calibration standards like subcutaneous fat, are discussed. Advances in PL-QCT, notably with automatic region of interest (ROI) selection, show improved accuracy. Dual-energy CT (DECT) introduces new dimensions for musculoskeletal analysis. Despite advancements, challenges persist, including interindividual variability and patient-specific factors. Evolving single-energy CT and DECT techniques show promise in refining BMD assessment and osteoporosis diagnosis, enhancing patient care. Continued research and integration into clinical practice are vital for realizing these advancements’ full benefits. In this review, we evaluate and summarize current evidence on the feasibility and different approaches to achieve analysis of BMD with phantomless QCT.

Reliable quantitative detection of uric acid in urine by surface-enhanced Raman spectroscopy with endogenous internal standard

Abnormal levels of uric acid (UA) in urine serve as warning signs for gout and metabolic cardiovascular diseases, necessitating the monitoring of UA levels for early prevention. However, the current analytical methods employed suffer from limitations in terms of inadequate suitability for home-based applications and the requirement of non-invasive procedures. In this approach, creatinine, a metabolite with a constant excretion rate, was incorporated as an endogenous internal standard (e-IS) for calibration, presenting a rapid, pretreatment-free, and accurate strategy for quantitative determination of UA concentrations. By utilizing urine creatinine as an internal reference value to calibrate the signal fluctuation of surface-enhanced Raman spectroscopy (SERS) of UA, the quantitative accuracy can be significantly improved without the need for an external internal standard. Due to the influence of the medium, UA, which carries a negative charge, is selectively adsorbed by Au@Ag nanoparticles functionalized with hexadecyltrimethylammonium chloride (CTAC) in this system. Furthermore, a highly convenient detection method was developed, which eliminates the need for pre-processing and minimizes matrix interference by simple dilution. The method was applied to the urine detection of different volunteers, and the results were highly consistent with those obtained using the UA colorimetric kit (UACK). The detection time of SERS was only 30 s, which is 50 times faster than UACK. This quantitative strategy of using urinary creatinine as an internal standard to correct the SERS intensity of uric acid is also expected to be extended to the quantitative detection needs of other biomarkers in urine.

Cell membrane-targeted surface enhanced Raman scattering nanoprobes for the monitoring of hydrogen sulfide secreted from living cells

Hydrogen sulfide (H2S), an important gas signal molecule, participates in intercellular signal transmission and plays a considerable role in physiology and pathology. However, in-situ monitoring of H2S level during the processes of material transport between cells remains considerably challenging. Herein, a cell membrane-targeted surface-enhanced Raman scattering (SERS) nanoprobe was designed to quantitatively detect H2S secreted from living cells. The nanoprobes were fabricated by assembling cholesterol-functionalized DNA strands and dithiobis(phenylazide) (DTBPA) molecules on core-shell gold nanostars embedded with 4-mercaptoacetonitrile (4-MBN) (AuNPs@4-MBN@Au). Thus, three functions including cell-membrane targeted via cholesterol, internal standard calibration, and responsiveness to H2S through reduction of azide group in DTBPA molecules were integrated into the nanoprobes. In addition, the nanoprobes can quickly respond to H2S within 90 s and sensitively, selectively, and reliably detect H2S with a limit of detection as low as 37 nM due to internal standard-assisted calibration and reaction specificity. Moreover, the nanoprobes can effectively target on cell membrane and realize SERS visualization of dynamic H2S released from HeLa cells. By employing the proposed approach, an intriguing phenomenon was observed: the other two major endogenous gas transmitters, carbon monoxide (CO) and nitric oxide (NO), exhibited opposite effect on H2S production in living cells stimulated by related gas release molecules. In particular, the introduction of CO inhibited the generation of H2S in HeLa cells, while NO promoted its output. Thus, the nanoprobes can provide a robust method for investigating H2S-related extracellular metabolism and intercellular signaling transmission.

Inner-Wall Coated Nanopipette Microextraction for Quantitative Analysis of Per- and Polyfluoroalkyl Substances in Single Cells Using Mass Spectrometry.

Per- and polyfluoroalkyl substances (PFASs) are a series of organic pollutants with potential cytotoxicity and biotoxicity. Accurate and sensitive detection of trace PFASs in single cells can provide insights into investigating their cytotoxicity, carcinogenicity, and mutagenicity. Here we report the development of an inner-wall coated nanopipette microextraction coupled with induced nanoelectrospray ionization mass spectrometry (InESI-MS) method and its application for rapid, sensitive, and accurate analysis of trace PFASs in single cells. A specially designed inner-wall coated nanopipette was prepared for sampling of the cytoplasm from a single cell, and the trace PFASs in the cytoplasm were selectively enriched into the coating via reversed-phase adsorption, ion bonding adsorption, and π-π interaction mechanisms. After the extraction, the cytoplasm was removed, and the enriched PFASs were then desorbed into some organic solvent, applying an alternating current (AC) voltage to the inner-wall coated nanopipette for InESI-MS analysis. The inner-wall coated nanopipette showed an exhaustive extraction to the trace PFASs in one single cell, and thus, the mass of each target analyte in the cytoplasm can be calculated via an internal standard calibration curve method, avoiding the measurement of ultrasmall volume cytoplasm for one single cell. By using the inner-wall coated nanopipette microextraction coupled with InESI-MS method, trace PFASs accumulated in the LO2 cells with pollutant exposure were successfully detected, and the accumulative behaviors and heterogeneities of PFASs in single cells were explored.

Determination of tetrodotoxin in urine by liquid chromatography-tandem mass spectrometry with internal standard calibration

An analytical method was developed for tetrodotoxin(TTX) in urine by liquid chromatography-tandem mass spectrometry(LC-MS/MS) with internal standard calibration. TTX in the sample was extracted with the mixture of acetic acid/methanol/acetonitrile(0.005 mL/0.8 mL/1.8 mL), cleaned by solid phase extraction(SPE) with cation exchange cartridge, eluted with 50% acetonitrile/water containing 0.3% hydrochloric acid, and neutralized with ammonia. The extract was separated by a Waters XBridge~(TM) BEH Amide column(150 mm×3.0mm, 1.7 μm) and measured by MS/MS. By optimizing sample extraction and SPE cleanup conditions, the problems of low recovery and strong suppression effects of MS signal for TTX in urine were resolved when cleaned with cation exchange cartridge. Quantitatively calibrated by the internal standard of Kasugamycin, good linear relationship was found for TTX in urine at the range of 0.2-200 μg/L with the correlation coefficient(r~2) of 0.997. The limits of detection and quantitation for TTX in sample matrix were 0.1 and 0.2μg/L, respectively. The average recoveries at three spiking levels(0.2, 10.0 and 200 μg/L) were 89.3%-95.3% with relative standard deviation(n=6) less than 5.1%. The concentrations of TTX in urine from 11 poisoning patients were 0.4-138 μg/L. The detection rate was 100% in urine collected within 3 days after poisoning. The established method was simple, accurate and sensitive. It can provide reliable technical support for the rapid treatment of TTX poisoning events and the study of toxin metabolism in vivo.

Determination of psilocybin and psilocin content in multiple Psilocybe cubensis mushroom strains using liquid chromatography – tandem mass spectrometry

A method for clinical potency determination of psilocybin and psilocin in hallucinogenic mushroom species Psilocybe cubensis was developed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Five strains of dried, intact mushrooms were obtained and analyzed: Blue Meanie, Creeper, B-Plus, Texas Yellow, and Thai Cubensis. An extraction protocol was developed; this included an evaluation of sample milling technique, extraction solvents, and recovery/stability. Reversed phase chromatography on fused-core particle phases was developed for the determination of the two analytes using internal standard calibration with deuterated isotopologues of each analyte. The separation takes less than 5 min. Matrix effects were investigated by comparing signal response of calibration samples in neat solution and several mushroom matrices; no significant matrix effects were observed. The limit of detection for psilocybin was 1.5 ng/mL (1.5 pg on-column; 300 ng/g mushroom) and for psilocin was 0.15 ng/mL (0.15 pg on-column; 30 ng/g mushroom) using a Shimadzu LCMS-8050 triple quadrupole mass spectrometer. Assessment of the accuracy and precision of the method indicated percent error and RSD were <6 % at all concentration levels. Three whole, intact mushrooms from each strain were analyzed individually to obtain average content differences both between strains and between mushrooms of the same strain. From most to least potent, the study found that the average total psilocybin and psilocin concentrations for the Creeper, Blue Meanie, B+, Texas Yellow, and Thai Cubensis strains were 1.36, 1.221, 1.134, 1.103, and 0.879 % (w/w), respectively. A subset of these mushrooms was also tested in a separate non-affiliated laboratory, and the results were comparable between the two laboratories. Results from the secondary laboratory showed improved precision when multiple mushrooms were homogenized together, prior to extraction.

Variability of internal standard method calibration factors estimated with a multifactorial Taguchi experiment in the analysis of alcoholic products by gas chromatography with flame ionization detection.

This study compares the variability of relative response factors (RRFs) using Taguchi's multifactorial analysis for two internal standard (IS) methods in gas chromatography (GC) for quality control of alcoholic products. Methods where either ethanol or pentan-1-ol was used as an IS were compared. For ten volatile substances prescribed by legislation, the RRFs of both methods were compared under 27 different experimental conditions. The influence of parameters (control factors) such as ethanol content in the matrix, analyte concentration, injected volume, injector temperature, split ratio, and flame ionization detector temperature was evaluated. The selected control factors had values at one of the three levels to cover the commonly used ranges of their settings in the measuring system and to characterize the majority of alcoholic products commonly analyzed in practice. The obtained results showed that the biggest differences in the variability of the results between the two methods were found for the most strictly controlled substances in alcoholic products, acetaldehyde, and methanol, where the application of ethanol as an IS provides clearly better results. For both methods, the way control factors affect the repeatability of GC measurements expressed in the form of relative deviation was also evaluated.

Separation of bile acid isomer plays a pivotal role in bioequivalence evaluation of ursodeoxycholic acid

Based on our experiences in bile acid profiling, this work developed and validated a liquid chromatography electrospray ionization tandem mass spectrometry method to separate endogenous bile acid isomers and quantitatively determine ursodeoxycholic acid (UDCA), glycoursodeoxycholic acid (GUDCA) and tauroursodeoxycholic acid (TUDCA) in human plasma. The separation was performed on a CORTECS C18 column with the mobile phase consisting of 1.0 mM ammonium acetate and acetonitrile-methanol (80:20, v/v). UDCA, GUDCA and TUDCA were detected in the negative mode on a triple-quadrupole mass spectrometer at the ion transitions of m/z 391 > 391, m/z 448 > 74, m/z 498 > 80, respectively. Phosphate buffer was employed as the surrogate matrix to establish the isotope internal standard corrected calibration curves of analytes. The background-method with a linearity range of 10–200 ng/mL was partially validated to determine the endogenous levels of analytes in blank human plasma, which was incorporated into the validation of bioequivalence-method with a linearity range of 50–10000 ng/mL. The bioequivalence (BE)-method was fully validated with special focus on matrix effects, which have been critically evaluated using the precision and accuracy of quality control samples prepared from the blank human plasma of 12 individuals. It is disclosed for the first time that the BE results of UDCA formulation may yield false results when the method is insufficient to separate UDCA from isoursodeoxycholic acid, a microbial metabolite of both endogenous and exogenous UDCA. The present method has established a milestone for the evaluation of UDCA formulations and is expected to provide a valuable reference for the bioanalytical development of endogenous medicinal products.

LC-MS accurate quantification of a tryptic peptide co-eluted with an isobaric interference by using in-source collisional purification.

Interferences from isobaric and isomeric compounds represent a common problem in liquid chromatography coupled to mass spectrometry (LC-MS). In this paper, in-source purification and chromatographic separation were combined with the aim of identifying isobaric contamination and quantifying accurately a compound despite the presence of an isobaric co-eluted interference. This is achieved by totally fragmenting in-source the precursor ions of the isobaric interference providing then LC-pseudo-MS2 capability, which allows an accurate quantification without the need for optimizing the chromatographic conditions to separate the co-eluted interference. To illustrate this concept, mixtures of tryptic and non-tryptic peptides were used. The ratio of peak areas of the tryptic peptide and its isotopically labelled internal standard was used not only for quantification with an internal standard calibration curve but also to know (1) if an isobaric interference co-eluted with the tryptic peptide; and (2) what is the minimum cone voltage necessary to ensure the complete removal of isobaric interference. This strategy was applied to quantify the tryptic peptide of two standards with known concentrations and, intentionally contaminated with the isobaric interference. The confidence intervals of the concentrations calculated with the internal standard calibration curve were 8.0 ± 0.5 μM (prepared at 8.0 μM) and 15.7 ± 0.5 μM (prepared at 16.1 μM) that confirm the tryptic peptide can be correctly quantified by in-source purification without the need for improving the chromatographic separation from its isobaric interference.

Determination of tetrodotoxin in bivalve mollusks by hydrophilic interaction liquid chromatography-triple quadrupole mass spectrometry with internal standard calibration and its contamination in Zhejiang province, China

A method was established for determining tetrodotoxin (TTX) in bivalve mollusks by hydrophilic interaction liquid chromatography-triple quadrupole mass spectrometry (HILIC-MS/MS) using kasugamycin as the internal standard for quantification. Samples were subjected to ultrasonic extraction with methanol–water (1:1, v/v) containing 0.5% acetic acid, protein precipitation with acetonitrile, clean-up using a cation exchange solid phase extraction cartridge, elution with acetonitrile:water (1:1, v/v) containing 0.3% hydrochloric acid, neutralization with ammonia before HILIC-MS/MS analysis. The average recovery of the samples spiked at 3 levels ranged in 84.6%-98.1% with the relative standard deviation less than 7.2%. Using this method, the contamination of TTX in 429 bivalve mollusk samples collected in the local markets during 2018 and 2020 was investigated. The detection rates were 12.0–18.8%, following the order of oyster > mussel > clam > scallop. High contaminated oysters and mussels with TTX were found in July to August. Moreover, TTX analogs found in bivalve mollusks included 4-epiTTX, 5,6,11-trideoxyTTX, 4,9-anhydroTTX, and 5-deoxyTTX/11-deoxyTTX.

GC–MS determination of 7-dehydrocholesterol and cholesterol in milk to verify eco-friendly conditions in cow housing

Dairy products are consumed worldwide in large quantities and play an important role in a diet. Labelling of these products as “eco”, “bio” or “organic” has become a trend in recent years, and verification of such advertised properties directly from the dairy products themselves, without the possibility of examining the environment where they were produced, is often next to impossible. However, it is crucial for combating potential false advertising. Herein, a simple approach to distinguish between indoor and outdoor rearing based on quantitation of cholesterol and 7-dehydrocholesterol in milk samples by GC–MS was evaluated. Repeated liquid–liquid extraction into n-heptane resulted in 99% extraction yield. Quantitation by internal standard calibration method with epicoprostanol provided limits of quantitation of 0.814 µg/100 g and 5.60 µg/100 g in milk for cholesterol and 7-dehydrocholesterol, respectively. Values of parameter defined as thousandfold 7-dehydrocholesterol to cholesterol ratio ranged from 0.35 to 1.34 for outdoor reared cows in comparison with indoor category (2.14 to 2.85). A statistically significant difference (at 99% confidence level) between indoor and outdoor rearing was observed. The developed method is an efficient tool for verification of livestock living conditions and, hence, for verification of eco-friendly labelling by dairy producers.

Improvements on the analysis methods of asphalt fumes generated in asphalt production and construction

Asphalt fumes are generated when producing and processing asphalt materials, and polycyclic aromatic hydrocarbons (PAHs) are commonly used as indicators to evaluate the hazardous potentials of asphalt fumes. However, the quantitative analysis of PAHs in asphalt fumes remains a challenge. Particularly, the influences of matrix compounds in asphalt fumes on PAH quantification have not been evaluated, even though matrix effect may lead to biased results. To address such a research gap, this study (1) examined the matrix effect in analyzing PAHs in asphalt fumes, (2) evaluated the effectiveness of clean-up methods, and (3) assessed the effectiveness of different methods for matrix effect compensation. Asphalt fumes from seven asphalt materials or conditions were collected and analyzed. Four clean-up methods were evaluated for their influences on PAH identification and matrix effect mitigation. Two approaches were experimented for matrix effect compensation. This study found an obvious matrix effect in the quantitative analysis of PAHs in asphalt fumes. None of the clean-up methods can effectively remove the matrix effect, but both the standard addition calibration (SAC) method and the internal standard calibration (ISC) method can compensate such effect. The findings are expected to assist in accurate quantification and evaluation of PAHs in asphalt fumes.

HS-SPME-GC-MS analysis of Cyprus vineyard soil for VOCs determination

In line with green chemistry principles, a headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) method was developed and optimized to study the profile of biogenic volatile organic compounds (BVOCs) in soil samples from vineyards. The 75 µm CAR/PDMS fiber was selected for the identification of BVOCs in soil. The analytical method showed very good linearity (R2 ≥ 0.9914), sensitivity, and precision. Subsequently, soil matrix effect (ME) was examined on the BVOCs emission, through combined standard addition and internal standard (IS) calibration curves. The validated and optimized method was further applied in real soil samples from three vineyard areas of Cyprus (Omodos, Polemi, and Agros). Thirty-eight BVOCs were identified in total; among them acetone (0.65–5.96 μg/kg), hexane (0–0.50 μg/kg), heptane (0–0.18 μg/kg), trichloromethane (0.13–0.21 μg/kg), 1,2-dichloropropane (0.05–1.37 μg/kg), cyclohexane (0–0.26 μg/kg), benzene (0–0.51 μg/kg), toluene (0–0.54 μg/kg), p-xylene (0.15–0.97 μg/kg), and 1,3-di-tert-butyl-benzene (7.91–29.55 μg/kg) were quantified. The results revealed that the sampling vineyard location affects the BVOCs emissions in each soil. Also, vineyard elevation, soil pH, Ca, N, Zn, and Fe showed a significant correlation with the BVOCs emissions, which explains why vineyard soil regions are distinguished from each other, concluding that sample from Agros is the most abundant in BVOCs emissions.

Analysis of 16 mycotoxins in genuine traditional Chinese medicine for five medicinal parts: Classification of analytical method based on PANI@CS extraction-UPLC-MS/MS

A novel PANI@CS solid-phase dispersive extractant combined with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed for the first time, which was used for high-throughput, multi-component, real-time online rapid pretreatment and quantitative classification of 16 mycotoxins from five different medicinal parts of 13 genuine traditional Chinese medicines (TCMs). Ultra performance liquid chromatography combined with triple quadrupole mass spectrometry was used for separation and ESI detection. An internal standard isotope matching calibration was used for quantification purposes to compensate for matrix effects. The limits of detection (LOD) of 16 mycotoxins ranged from 0.1 to 6.0 μg/kg. The linear coefficients (R2) were ≥0.996 in the linear range from 10.0 to 200 μg/L. The recoveries of the 16 mycotoxins ranged from 90.1% to 105.8%, and the relative standard deviations (RSDs) ranged from 1.3% to 4.1%. Thirteen TCMs from five representative medicinal parts were selected and tested under the best sample preparation procedure and chromatographic analysis conditions. The results showed that the method could improve the sensitivity and accuracy of the sample analysis, improve the selectivity and reproducibility of the decolorization and purification of TCMs, which is suitable for the practical application of mycotoxin in trace analysis. This method can also provide a new idea for accurate, efficient, rapid and multi-component online detection of mycotoxins for quality and safety control of TCMs.