Chromatography-triple Quadrupole Linear Ion Trap Research Articles

Improved analysis of folpet and captan in foods using liquid chromatography-triple quadrupole linear ion trap mass spectrometry: applying mass filtering, collision, and trapping conditions

Accurate and highly sensitive analysis of folpet and captan was accomplished using liquid chromatography-triple quadrupole linear ion trap mass spectrometry (LC-QqQIT) with selective ion mode; mass filtering, collision, and trapping condition. Dimensional mass spectrometry (MS3) parameters were optimized for the residue detection of folpet and captan in six food commodities (apples, tomatoes, sweet pepper, wheat flour, sesame seeds, and fennel seeds). The sample preparation method was based on the known QuEChERS protocol, except a mixture of acetonitrile/acetone was used for the sample extraction from the sesame seeds. The robustness and reliability of the developed MS3 method were demonstrated by performing a full validation, according to SANTE/11312/2021, at 0.01-0.25mg/kg. Recovery ranged from 83 to 118% with a relative standard deviation below 19% in all the tested commodities, and limits of quantifications (LOQs) were 0.01mg/kg in apples and tomatoes; 0.03mg/kg in sweet pepper; and 0.05mg/kg in wheat flour, sesame seeds, and fennel seeds. Monitoring results showed that about 90% of apples contained captan residue, and in sweet pepper, concentrations of captan and folpet as high as 1.57 and 0.97mg/kg were found, respectively. The novel developed MS3 method enabled more reliable identification of these commonly problematic fungicides at lower LOQs than previously reported methods.

Systematic Qualitative and Quantitative Analyses of Wenxin Granule via Ultra-High Performance Liquid Chromatography Coupled with Ion Mobility Quadrupole Time-of-Flight Mass Spectrometry and Triple Quadrupole-Linear Ion Trap Mass Spectrometry.

Wenxin granule (WXG) is a popular traditional Chinese medicine (TCM) preparation for the treatment of arrhythmia disease. Potent analytical technologies are needed to elucidate its chemical composition and assess the quality differences among multibatch samples. In this work, both a multicomponent characterization and quantitative assay of WXG were conducted using two liquid chromatography–mass spectrometry (LC-MS) approaches. An ultra-high performance liquid chromatography–ion mobility quadrupole time-of-flight mass spectrometry (UHPLC/IM-QTOF-MS) approach combined with intelligent peak annotation workflows was developed to characterize the multicomponents of WXG. A hybrid scan approach enabling alternative data-independent and data-dependent acquisitions was established. We characterized 205 components, including 92 ginsenosides, 53 steroidal saponins, 14 alkaloids, and 46 others. Moreover, an optimized scheduled multiple reaction monitoring (sMRM) method was elaborated, targeting 24 compounds of WXG via ultra-high performance liquid chromatography–triple quadrupole linear ion trap mass spectrometry (UHPLC/QTrap-MS), which was validated based on its selectivity, precision, stability, repeatability, linearity, sensitivity, recovery, and matrix effect. By applying this method to 27 batches of WXG samples, the content variations of multiple markers from Notoginseng Radix et Rhizoma (21) and Codonopsis Radix (3) were depicted. Conclusively, we achieved the comprehensive multicomponent characterization and holistic quality assessment of WXG by targeting the non-volatile components.

Rapid and selective determination of 9 nitrosamines in biological samples using ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry.

A sensitive, selective and convenient method for the simultaneous determination of 9 nitrosamines (NAs) in biological samples was developed using isotope dilution ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UPLC-QTRAP-MS). Multiple reaction monitoring-information dependent acquisition-enhanced product ion (MRM-IDA-EPI) scan mode was performed to eliminate false positive results, and the whole detection procedure was characterized by less time consuming and simple sample preparation. 9 NAs were separated through a T3 column with the gradient elution of acetonitrile and water, and detected by UPLC-QTRAP-MS with an atmospheric pressure chemical ionization (APCI) source in the positive mode. The quantitative analysis was carried out via the isotope internal standard method with a matrix calibration curve. Under the optimized conditions, good linearity for the 9 NAs was achieved in the range of 0.2-20 μg L-1 with correlation coefficients (r) higher than ≥0.9991, and the limits of detection and limits of quantitation were 0.02-0.1 μg L-1 (S/N = 3) and 0.06-0.3 μg L-1 (S/N = 10), respectively. Satisfactory recoveries ranging from 79.4% to 108.0% were obtained, and the precision of the proposed method, indicated by the relative standard deviations (RSDs), was 2.3-12.9%. The matrix effect study showed that NDMA, NMOR and NMEA presented a matrix suppression effect, NDPHA displayed a matrix enhancement effect, and the matrix effects of the other 5 analytes could be ignored. Real application of the developed method in 13 urine and 24 plasma samples demonstrated that NDBA, NPIP and NPYR occurred in both urine and plasma samples with the concentration of 0.038-0.60 μg L-1, while other NAs were not detected. Such a method was sensitive and selective, and could be applied to the rapid qualitative and quantitative analysis of the 9 NAs in biological samples.

Chemical profiling and multicomponents quantitative analysis of Panzerina lanata by ultra-fast liquid chromatography with tandem mass spectrometry.

Panzerina lanata is a Chinese medicine with the bioactivity of detumescence and detoxification. In this study, novel qualitative and quantitative methods were established by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry and ultra-high-performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry, respectively. As a result, 20 compounds were identified or tentatively characterized including flavonoids, organic acids, alkaloids, and lignans, five of which were identified for the first time based on the reference standards. The quantitative approach exhibited good linearity (R2 >0.995), precision (RSDs<4.97%), stability (RSDs<4.77%), and recovery (96.04-104.14%). Afterward, this method was implemented to determine 11 flavonoids in four batches of P. lanata. Among them, seven compounds were quantified for the first time. Narcissin was abundant in each batch of P. lanata (average of 10.890-14.230mg/g) with the highest quantities. The results provide valuable information for quality evaluation.

A high efficient adsorbent for plant growth regulators based on ionic liquid and β-cyclodextrin functionalized magnetic graphene oxide

Four types of ionic liquids which composed of cation with imidazole ring and anion with benzene, naphthalene, anthraquinone and naphthoquinone ring were prepared and named VOIm+BenSO3-, VOIm+NapSO3-, VOIm+AQSO3- and VOIm+NQSO3-, respectively. Then, ionic liquids and β-cyclodextrin functionalized magnetic graphene oxide materials were firstly synthesized for the extraction of 23 plant growth regulators in vegetable samples. The synthesized materials anticipated the unique properties of ionic liquids, β-cyclodextrin, graphene oxide and Fe3O4@SiO2, including high surface area, high supramolecular recognition capability, superparamagnetism and large delocalized π-electron system, which were successfully characterized. Among four kinds of ionic liquids functionalized adsorbents, the VOIm+AQSO3- modified material exhibited high adsorption capability to 7 plant growth regulators compared with Fe3O4@SiO2/GO/β-CD, Fe3O4@SiO2/GO and other three types of ionic liquids based adsorbents. Parameters that could affect the recoveries of 7 plant growth regulators were investigated, such as mass ratio of Fe3O4@SiO2/GO/β-CD to ionic liquids, salt concentration, pH, amount of adsorbent, extraction time and desorption solvent. Under the optimal condition, the magnetic solid phase extraction method using VOIm+AQSO3- and β-cyclodextrin functionalized magnetic graphene oxide material (Fe3O4@SiO2/GO/β-CD/IL) as adsorbent was developed and coupled with ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry for the determination of 7 plant growth regulators in vegetable samples. Validation results showed that the limit of detection and limit of quantitation were in the range of 0.01–0.18 μg/kg and 0.03–0.58 μg/kg, respectively. The satisfactory linearities were acquired in the concentration of 2–50 μg/kg with the correlation coefficients higher than 0.9982. Taken together, the synthesized Fe3O4@SiO2/GO/β-CD/IL was a highly effective magnetic solid phase extraction adsorbent for the enrichment of trace level of plant growth regulators in vegetable samples.

Simultaneous determination of amantadine and rimantadine in feed by liquid chromatography-Qtrap mass spectrometry with information-dependent acquisition.

A sensitive method for simultaneous determination of amantadine and rimantadine in feed was developed using an ultra-high-performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UHPLC-Qtrap-MS) in the multiple reaction monitoring information-dependent acquisition-enhanced product ion (MRM-IDA-EPI) mode, and employing the mixed cation exchange (MCX) solid-phase extraction column as sample cleanup and amantadine-d15 and rimantadine-d4 as internal standards, respectively. Compared to traditional MRM mode, for the targeted drugs in feed simultaneously both the secondary mass spectra and MRM information can be obtainedusing UHPLC-Qtrap-MS with MRM-IDA-EPI mode, and thus more accurate qualitative confirmation results achievedeven at lower concentration of 0.2μg/L in acceptable purity fit values. After optimization of sample preparation, good linearities (R > 0.9994) were obtained over the concentration range from 1 to 200μg/L for amantadine and rimantadine. The precision was validated by intra-day and inter-day, and the relative standard deviations were all within 9.61%. Mean recoveries ranged from 76.1 to 112% at spiked concentrations of 0.5-100 μg/kg in three types of feed samples, including formula feed and complex concentrated feed for pigs and premix feed for chicken. The limits of detection (LODs) and quantification (LOQs) were 0.2 and 0.5μg/kg for both drugs, respectively. The application in real feed samples further proved the accuracy and reliability of the developed method. This method provides an important tool to detect illegal uses of amantadine and rimantadine in feed. Graphical abstract Simultaneous quantitation and qualitative confirmation of amantadine and rimantadine in feed by MRM-IDA-EPI.

Fabrication of a high selectivity magnetic solid phase extraction adsorbent based on β-cyclodextrin and application for recognition of plant growth regulators

An adsorbent, consisting of silica-coated Fe3O4 grafted graphene oxide and β-cyclodextrin (Fe3O4@SiO2/GO/β-CD), which possessed the merits of antioxidation, superparamagnetism, high surface area, high supramolecular recognition and environment friendly, was successfully fabricated. Considering the synergy between β-CD and graphene oxide in adsorption mechanism, the synthesized adsorbent could grasp compounds especially with aromatic structures through π-π interaction, hydrophobic interaction and host-guest inclusion complex forming. Based on the advantages, a magnetic solid phase extraction (MSPE) method for 9 PGRs using Fe3O4@SiO2/GO/β-CD as adsorbents was developed in this study. The characterizations of Fe3O4@SiO2/GO/β-CD were performed on Fourier transform infrared spectrometry (FT-IR), X-ray photoelectron spectroscopy (XPS), CHNS/O elemental analyzer, scanning electron microscopy (SEM) and vibrating sample magnetometry (VSM). Under the optimal MSPE condition, the Fe3O4@SiO2/GO/β-CD exhibited selectivity capability toward 9 PGRs when compared with Fe3O4@SiO2/GO. Meanwhile, the selectivity capability of Fe3O4@SiO2/GO/β-CD was higher than that of Fe3O4@SiO2/GO/α-CD except for 4-FPA. When the developed MSPE procedure was coupled with ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UHPLC-QTrap-MS/MS) to quantitative analysis of 9 PGRs, linearities ranging from 2 to 50 μg/kg were achieved for 9 PGRs with the correlation coefficients (r2) in the range of 0.9975-0.9999. The limits of detection (LODs) for 9 analytes varied from 0.04 to 0.29 μg/kg. Finally, the proposed technique was applied to analyze PGRs residues in mutiple vegetable samples.

Stereoselective effects of ibuprofen on adult zebrafish brain tissues based on sphingolipidomics

The aim of this study was to investigate the stereoselective effects of a popular pharmaceutical and personal care product component ibuprofen (IBU) on adult zebrafish using sphingolipidomics. Sphingolipidomics is an effective method developed to determine sphingolipids present in biological samples using ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry. Using this method, 46 sphingolipids in adult zebrafish brain samples were quantified after rac-/R-(-)-/S-(+)-IBU exposure, and the biomarkers were selected using chemometric approaches, including principal component analysis and orthogonal partial least squares discrimination analysis. Besides, combined with the results of the Duncan post hoc test and Dunnett post hoc comparison, sphingolipidomic perturbations were found to be induced after exposure to ibuprofen at environmental concentrations, and enantiomers of ibuprofen had stereoselective effects on zebrafish. This study may provide a new insight on the influence of ibuprofen on aquatic organisms.

Determination of 17 Plant Growth Regulator Residues by Ultra-High Performance Liquid Chromatography-Triple Quadrupole Linear Ion Trap Mass Spectrometry Based on Modified QuEChERS Method

A new method combining modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) with ultra-high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry (UHPLC-QTrap-MS/MS) was developed for simultaneous determination of 17 plant growth regulator (PGR) residues in vegetables. Quantitative data acquisition was accomplished in multiple reaction monitoring (MRM) mode, and qualitative analysis and identification of target compounds was achieved by using linear ion trap scan functions. Qualitative and quantitative analysis can be simultaneously achieved by a pair of ions in UHPLC-QTrap-MS/MS. More importantly, QTrap system can minimize the risk of false positive and negative results. To achieve the optimum performance towards the target analytes, several parameters including the content of formic acid in mobile phase and scanning mode were investigated. In the optimized conditions, 17 PGRs showed good linearity over the range of 2~50 μg/L with correlation coefficients of 0.9902~0.9999. The limits of detections (LODs) were in the range of 0.01~1.32 μg/kg. The recoveries of 17 PGRs ranged from 78.5 to 119.2% with the spiked levels of 1, 2, and 5 μg/kg into homogenized radish, and the relative standard deviations (RSDs) ranged from 0.8 to 14.8%. The method illustrated is suitable for routine quantitative and qualitative analyses of PGRs in vegetable samples.

Cleaning Up Vegetable Samples Using a Modified “QuEChERS” Procedure for the Determination of 17 Plant Growth Regulator Residues by Ultra High Performance Liquid Chromatography–Triple Quadrupole Linear Ion Trap Mass Spectrometry

A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) sample pretreatment method coupled with ultra high performance liquid chromatography–triple quadrupole linear ion trap mass spectrometry (UHPLC-QTrap-MS/MS) was developed for simultaneous determination of 17 plant growth regulator (PGR) residues in vegetables. Generik ChlorF (GCF) was used as one of the adsorbents in modified QuEChERS procedure, which can selectively remove the chlorophyll of acetonitrile extract without causing the losses of PGRs that have planar structure. To achieve the optimum performance toward the target analytes, extraction solvent, the type of adsorbent, and the amount of the adsorbent were investigated. Under the optimized conditions, good linearities with correlation coefficients (γ) higher than 0.9954 were obtained. The new method provided sufficient sensitivity as reflected by the values of limit of detection (LOD) and limit of quantification (LOQ). The LOD varied between 0.01 and 0.70 μg/kg and the LOQ varied between 0.04 and 2.32 μg/kg. Acceptable recoveries ranging from 78.5 to 116.8 % with the relative standard deviations (RSDs) of 0.9–11.9 % were achieved. The evaluated method provided reliable screening, quantification, and identification of 17 PGRs in vegetables.

Unequivocal Enantiomeric Identification and Analysis of 10 Chiral Pesticides in Fruit and Vegetables by QuEChERS Method Combined With Liquid Chromatography-Quadruple/Linear Ion Trap Mass Spectrometry Determination.

In this research, 10 chiral pesticides in fruits and vegetables were simultaneously determined using chiral liquid chromatography triple quadrupole-linear ion trap hybrid mass spectrometry (LC-QqLIT). The QuEChERS method was applied for sample preparation, and an enhanced product ion (EPI) scan was used to acquire tandem mass spectrometry (MS/MS) spectra for the library search. Parameters including limit of detection (LOD), limit of quantification (LOQ), linearity, relative standard deviation (RSD), and matrix effects were evaluated in five representative matrices (strawberry, leek, cowpea, tomato, and eggplant). Good linearity with coefficient of determination (r(2) ) ≥0.997 was obtained for all 20 enantiomers in these five matrices over the range from 1.0 to 250 µg L(-1) . All the recoveries at 5 and 50 µg kg(-1) (n = 5) ranged between 70% and 120% with RSD below 20%, indicating satisfactory precision. The LOQ for the enantiomers ranged between 0.05 and 1 µg kg(-1) . Based on the proposed method, 135 commonly consumed fruits and vegetables taken from markets in Guizhou province, China, were analyzed. Enantioselective degradation for the selected chiral pesticides was observed in most of the positive samples.

Dynamic variation of bioactive compounds and aflatoxins in contaminated Radix Astragali during extraction process.

Although increasing attention has been paid to the health threat caused by mycotoxins in commodities such as food or medicines, mycotoxin transfer processes from crude material to products have raised little concern so far. Radix Astragali is a commonly used edible and medicinal herbal plant that is susceptible to contamination with aflatoxins from Aspergillus flavus. There have been no studies on mycotoxin transfer into pharmaceutical preparations or derivative products. To facilitate the aflatoxin reduction and bioactivity retention, the dynamic variations of aflatoxins as well as herbal compounds, namely calycosin-7-glucoside, astragaloside and formononetin, in Radix Astragali contaminated by A. flavus during water decoction and ethanol refluxing treatments were evaluated simultaneously by an ultra-fast liquid chromatography-triple quadrupole linear ion trap mass spectrometry method. After the extraction processes, although the amount of alfatoxins was reduced remarkably, aflatoxin residuals in preparation still exceed recommended limits, manifesting the great need to establish a limit for aflatoxins in herbal extractions or derivative products. Meanwhile, due to the hydrolysis of glucoside, water decoction period should be no longer than 4 h. This investigation would benefit from the determination of the dynamic variation of aflatoxins in infected herbs in preparation treatments, in order to further develop aflatoxin limits in herbal preparations.

Rapid identification and quantification of five major mogrosides in Siraitia grosvenorii (Luo-Han-Guo) by high performance liquid chromatography–triple quadrupole linear ion trap tandem mass spectrometry combined with microwave-assisted extraction

An effective and rapid method involving high performance liquid chromatography–triple quadrupole linear ion trap tandem mass spectrometry (HPLC–QTrap–MS/MS) was developed for identification and quantification of five major mogrosides (mogroside III, mogroside IV, siamenoside I, mogroside V and 11-oxomogroside V) contained in real samples of Siraitia grosvenorii (Luo-Han-Guo) for the first time. High extraction efficiency was obtained for the five mogrosides by using microwave-assisted extraction compared with other common extraction methods during sample pretreatment prior to determination. Mass measurement for molecular ions and characteristic fragment ions well accorded with corresponding identification criteria. Quantitative data acquisition was accomplished in multiple reaction monitoring (MRM) mode, and further identification of target compounds was achieved by using ultra performance liquid chromatography–quadrupole-time-of-flightmass spectrometry. The proposed method could be used as a rapid and reliable approach for effective assessment of Luo-Han-Guo, and also provided use for reference in terms of real analysis of other traditional Chinese medicines in the future.

Degradation study of enniatins by liquid chromatography–triple quadrupole linear ion trap mass spectrometry

Enniatins A, A1, B and B1 (ENs) are mycotoxins produced by Fusarium spp. and are normal contaminants of cereals and derivate products. In this study, the stability of ENs was evaluated during food processing by simulation of pasta cooking. Thermal treatments at different incubation times (5, 10 and 15min) and different pH (4, 7 and 10) were applied in an aqueous system and pasta resembling system (PRS). The concentrations of the targeted mycotoxins were determined using liquid chromatography coupled to tandem mass spectrometry. High percentages of ENs reduction (81–100%) were evidenced in the PRS after the treatments at 5, 10 and 15min of incubation. In contrast to the PRS, an important reduction of the ENs was obtained in the aqueous system after 15min of incubation (82–100%). In general, no significant differences were observed between acid, neutral and basic solutions. Finally, several ENs degradation products were identified using the technique of liquid chromatography–triple quadrupole linear ion trap mass spectrometry.

A survey of mycotoxins in random street-vended snacks from Lagos, Nigeria, using QuEChERS-HPLC-MS/MS

A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg−1. The concentrations of aflatoxin B1 (AFB1) and AFB2 reached 23 μg kg−1 and 3 μg kg−1, respectively. Moreover a sample of baked coconut contained α-zearalenol (α-ZOL), which was up to 54 μg kg−1 in coconut candy. As considers prevalence, aflatoxins and α-ZOL were not detected in lafun and groundnut-based snacks (donkwa and kulikuli), whereas each of the three mycotoxins contaminated 12.5% (1/8) of the coconut-based samples. This is the first report of α-ZOL in cassava and coconut, and their products. AFB1 and total aflatoxins (TAFs) concentrations exceeded the maximum allowable limit recommended by National Agency for Food and Drug Administration and Control Nigeria (NAFDAC) in one sample of baked coconut (AFB1 = 23 μg kg−1 and TAFs = 26 μg kg−1) and donkwa (AFB1 = 19 μg kg−1 and TAFs = 21 μg kg−1).

Rapid quantification of four major bioactive alkaloids in Corydalis decumbens (Thunb.) Pers. by pressurised liquid extraction combined with liquid chromatography-triple quadrupole linear ion trap mass spectrometry

A new method based on pressurised liquid extraction (PLE) followed by liquid chromatography–triple quadrupole linear ion trap mass spectrometry (LC–QTrap-MS) analysis has been developed for the identification and quantification of four major alkaloids in extracts of Corydalis decumbens (Thunb.) Pers. PLE extractions were performed using 90% ethanol; temperature was set at 100°C and pressure at 1500psi. HPLC analysis was performed on a Waters XBridge™ C18 column (150mm×2.1mm i.d., 3.5μm) eluted by a mobile phase of acetonitrile and 0.2% acetic acid. Data acquisition was carried out in multiple reaction monitoring transitions (MRMs) mode, monitoring two MRM transitions to ensure an accurate identification of target compounds in the samples. Additional identification and confirmation of target compounds were performed using the enhanced product ion modus (EPI) of the linear ion trap. The novel LC–QTrap-MS platform offers the best sensitivity and specificity for characterization and quantitative determination of the four alkaloids in C. decumbens (Thunb.) Pers. and fulfils the quality criteria for routine laboratory application.

Proteomics of the Autographa californica Nucleopolyhedrovirus Budded Virions

Baculoviruses produce two progeny phenotypes during their replication cycles. The occlusion-derived virus (ODV) is responsible for initiating primary infection in the larval midgut, and the budded virus (BV) phenotype is responsible for the secondary infection. The proteomics of several baculovirus ODVs have been revealed, but so far, no extensive analysis of BV-associated proteins has been conducted. In this study, the protein composition of the BV of Autographa californica nucleopolyhedrovirus (AcMNPV), the type species of baculoviruses, was analyzed by various mass spectrometry (MS) techniques, including liquid chromatography-triple quadrupole linear ion trap (LC-Qtrap), liquid chromatography-quadrupole time of flight (LC-Q-TOF), and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF). SDS-PAGE and MALDI-TOF analyses showed that the three most abundant proteins of the AcMNPV BV were GP64, VP39, and P6.9. A total of 34 viral proteins associated with the AcMNPV BV were identified by the indicated methods. Thirteen of these proteins, PP31, AC58/59, AC66, IAP-2, AC73, AC74, AC114, AC124, chitinase, polyhedron envelope protein (PEP), AC132, ODV-E18, and ODV-E56, were identified for the first time to be BV-associated proteins. Western blot analyses showed that ODV-E18 and ODV-E25, which were previously thought to be ODV-specific proteins, were also present in the envelop fraction of BV. In addition, 11 cellular proteins were found to be associated with the AcMNPV BV by both LC-Qtrap and LC-Q-TOF analyses. Interestingly, seven of these proteins were also identified in other enveloped viruses, suggesting that many enveloped viruses may commonly utilize certain conserved cellular pathways.

Phenolic Compounds in Different Barley Varieties: Identification by Tandem Mass Spectrometry (QStar) and NMR; Quantification by Liquid Chromatography Triple Quadrupole-Linear Ion Trap Mass Spectrometry (Q-Trap)

Barley (Hordeum vulgare) is an important cereal that has many applications; as a human food, in malt products and as livestock feed. The content of soluble health-promoting fibers, beta-glucans, varies substantially among barley varieties. In the present study, the content of secondary metabolites with potential positive health effects in different high- and low-beta-glucan barley varieties was studied. Five different flavanols were isolated and identified: (2R,3S)-catechin-7-O-beta-D-glucopyranoside (1), prodelphinidin B3 (2), procyanidin B3 (3), (+)-catechin (4) and procyanidin B1 (5). Procyanidin B1 has never been reported in barley grains before. The compounds were identified using 1H NMR and quadrupolar time-of-flight mass spectrometry. A quantitative analytical method was developed for prodelphinidin B3, procyanidin B3 and (+)-catechin in liquid chromatography triple quadrupole-linear ion trap mass spectrometry and these compounds were quantified in all varieties, together with four phenolic acids: ferulic acid, vanillic acid, p-coumaric acid and p-hydroxybenzoic acid. Catechin was the compound that was present at the highest concentration in all varieties. The variation, between cultivars, in catechin concentration varied four fold. A Principal Component Analysis indicated no correlation between concentrations of beta-glucan and secondary metabolites. Concentrations of catechin and prodelphinidin B3 were strongly correlated, whereas the concentration of procyanidin B3 was not correlated with that of catechin or prodelphinidin B3. Either two different enzymes could be responsible for the dimerization of prodelphinidin B3 and procyanidin B3, or the affinity of the enzyme could be different whether the dimerization is between two catechin units or between units of gallocatechin and catechin.

Rapid quantification of steroid patterns in human serum by on-line solid phase extraction combined with liquid chromatography–triple quadrupole linear ion trap mass spectrometry

Background The determination of steroids is important for the diagnosis and monitoring of endocrine diseases and infertility workup. We developed a rapid and reliable mass spectrometric method for the simultaneous quantification of steroid patterns in human serum. Methods An on-line solid phase extraction (SPE)-liquid chromatography–triple quadrupole linear ion trap (LC-QTrap) method utilizing atmospheric pressure chemical ionization was developed. Following protein precipitation of 100 µL serum, on-line SPE and chromatographic separation was performed for 13 steroids in 1.8 min. Analytes were confirmed by the characteristic fragment patterns. Results The total run time of the method was 4 min. Detection limits ranged between 0.02 µg/L (testosterone) and 9 µg/L (dehydroepiandrosterone sulfate). The method was linear up to 7000 µg/L for dehydroepiandrosterone sulfate, 500 µg/L for cortisol, 125 µg/L for 11-deoxycortisol, and 25 µg/L for aldosterone, 17-hydroxyprogesterone, progesterone, testosterone, androstenedione and β-estradiol, respectively. Accuracy ranged between 80 and 114%. Between-day variance at three different concentration levels was < 15%. Excellent correlations with immunoassays were observed for testosterone, cortisol and β-estradiol with Pearson's correlation coefficient r = 0.967, 0.963, and 0.998, respectively. Conclusion The novel on-line SPE-LC-MS/QTrap platform offers a very fast, reliable, and sensitive quantification of steroid patterns and fulfils the quality criteria for routine laboratory application.