液相色谱-三重串联四极杆质谱测定粮油中的黄曲霉毒素

建立了超声提取-液相色谱-电喷雾三重串联四极杆质谱测定玉米、大米、大豆等粮油固体样品中黄曲霉毒素B1、B2、G1和G2(AFB1、AFB2、AFG1和AFG2)的方法。分析前对样品进行超声提取,优化得到最佳超声提取条件: 溶剂为甲醇-水(含40 g/L NaCl) (80:20, v/v)溶液、料液比为1:3(g:mL)、温度为50 ℃、时间为3 min。然后对提取的样品进行免疫亲和特异性净化。最后与液相色谱-电喷雾三重串联四极杆质谱联用,使用C18反相色谱柱,流动相为甲醇-10 mmol/L乙酸铵水溶液梯度洗脱,以黄曲霉毒素M1(AFM1)作为内标进行定量测定。结果表明,AFB1、AFB2、AFG1和AFG2的检出限分别为0.002、0.004、0.004和0.012 μg/kg。方法的加标回收率为87%～111%,日内相对标准偏差(RSD)和日间RSD分别不大于6.7%和5.6%。实验结果表明该方法可以有效地降低基质效应的影响,相比于外标法能极大地提高方法的准确度。     

Effect of hapten structures on specific and sensitive enzyme-linked immunosorbent assays for N-methylcarbamate insecticide metolcarb

Five different haptens of the N-methylcarbamate insecticide metolcarb were designed and synthesized. All of the haptens were conjugated with ovalbumin (OVA) for the coating antigen, and one hapten containing all of the structure of metolcarb was conjugated with bovine serum albumin (BSA) for the immunogen. Two polyclonal antisera were raised against the BSA conjugate, and ten antibody/coating conjugate combinations were selected for studies of assay sensitivity and specificity for metolcarb. A class-specific combination was found, with the I₅₀ of the assay ranged from 0.64 to 20.98 μg mL⁻¹ for seven tested N-methylcarbamate insecticides except for pirimicarb. Considering titer, I₅₀ and cross-reactivity of all combinations of antibody/coating conjugate, a competitive indirect enzyme-linked immunosorbent assay (ELISA) in a homologous system, whose limit of detection (LoD) reached 1.4 ng mL⁻¹, was presented. The results of competitive ELISAs indicated that coating hapten structure can significantly affect not only assay sensitivity but also its specificity.     

Identification of triacylglycerol using automated annotation of high resolution multistage mass spectral trees

High complexity of identification for non-target triacylglycerols (TAGs) is a major challenge in lipidomics analysis. To identify non-target TAGs, a powerful tool named accurate MSⁿ spectrometry generating so-called ion trees is used. In this paper, we presented a technique for efficient structural elucidation of TAGs on MSⁿ spectral trees produced by LTQ Orbitrap MSⁿ, which was implemented as an open source software package, or TIT. The TIT software was used to support automatic annotation of non-target TAGs on MSⁿ ion trees from a self-built fragment ion database. This database includes 19108 simulate TAG molecules from a random combination of fatty acids and corresponding 500582 self-built multistage fragment ions (MS ≤ 3). Our software can identify TAGs using a “stage-by-stage elimination” strategy. By utilizing the MS¹ accurate mass and referenced RKMD, the TIT software can discriminate unique elemental composition candidates. The regiospecific isomers of fatty acyl chains will be distinguished using MS² and MS³ fragment spectra. We applied the algorithm to the selection of 45 TAG standards and demonstrated that the molecular ions could be 100% correctly assigned. Therefore, the TIT software could be applied to TAG identification in complex biological samples such as mouse plasma extracts.     

Optimization of ultrasonic-assisted extraction of 3-monochloropropane-1,2-diol (MCPD) and analysis of its esters from edible oils by gas chromatography-mass spectrometry

In this paper, ultrasonic-assisted extraction of 3-chloropropane-1,2-diol and its esters from edible oils was studied with isotope dilution GC-MS. Effects of several experimental parameters, such as types and concentrations of extracting solvent, ratios of liquid to material, extraction temperature, time of ultrasonic treatment on the extraction efficiency of 3-chloropropane-1,2-diol and its esters from edible oils and sample preparation for calibration were compared and optimized. The optimal extraction conditions were suggested as 66 mg oil sample in mixture of 0.5 mL MTBE/ethyl acetate (20% v/v) and 0.5 mL of sulfuric acid/n-propanol (0.3% v/v), being extracted for 30 min at 45°C under ultrasonic irradiation. Good linearity was gained in the range of 0.020-5.000 μg/g with the limit of detection (LOD) of 0.006 μg/g (S/N = 3) and the limit of quantification (LOQ) of 0.020 μg/g (S/N = 10). The recoveries at five spiked concentrations were ranged from 91.9 to 109.3% with RSD less than 9.4%. The method was successfully applied to the determination of 3-chloropropane-1,2-diol and its esters amounts in rapeseed, sesame, peanut, camellia, and soybean oils.     

Untargeted fatty acid profiles based on the selected ion monitoring mode

Fatty acids are potential biomarkers of some diseases and also key markers and quality parameters of different dietary fats and related products. Thus, untargeted fatty acid profiles are important in the study of dietary fat quality and fat-related diseases, as well as in other fields such as bioenergy. In addition, accurate identification of unknown components is a technological breakthrough for the selected ion monitoring (SIM) mode for untargeted profiles. In this study, we developed untargeted fatty acid profiles based on SIM. We also investigated mass spectral characteristics and equivalent chain lengths (ECL) to eliminate the influence of non-FAMEs for identifying fatty acids in samples. As an application example, fatty acid profiles were used to classify three edible vegetable oils. The results indicated that SIM-based untargeted fatty acid profiles could yield accurate qualitative and quantitative results for more fatty acids and benefit related studies of metabolite profiles.     

Current development of microfluidic immunosensing approaches for mycotoxin detection via capillary electromigration and lateral flow technology

Mycotoxin contamination in the food chain has caused serious health issues in humans and animals. Thus, a rapid on-site and lab-independent detection method for mycotoxins, such as aflatoxins (AFTs), is desirable. Microfluidic chip based immunosensor technology is one of the most promising methods for fast mycotoxin assays. In this review, we cover the major microfluidic immunosensors used for mycotoxin analysis, via flow-through (capillary electromigration) and lateral flow technology. Sample preparation from different matrices of agricultural products and foodstuffs is summarized. The choice of materials, fabrication strategies, and detection methods for microfluidic immunosensors are further discussed in detail. The sensors application in mycotoxin determination is also outlined. Finally, future challenges and opportunities are discussed.     

3D nanoporous gold scaffold supported on graphene paper: Freestanding and flexible electrode with high loading of ultrafine PtCo alloy nanoparticles for electrochemical glucose sensing

Recent advances in on-body wearable medical apparatus and implantable devices drive the development of light-weight and bendable electrochemical sensors, which require the design of high-performance flexible electrode system. In this work, we reported a new type of freestanding and flexible electrode based on graphene paper (GP) supported 3D monolithic nanoporous gold (NPG) scaffold (NPG/GP), which was further modified by a layer of highly dense, well dispersed and ultrafine binary PtCo alloy nanoparticles via a facile and effective ultrasonic electrodeposition method. Our results demonstrated that benefited from the synergistic effect of the electrocatalytically active PtCo alloy nanoparticles, the large-active-area and highly conductive 3D NPG scaffold, and the mechanically strong and stable GP electrode substrate, the resultant PtCo alloy nanoparticles modified NPG/GP (PtCo/NPG/GP) exhibited high mechanical strength and good electrochemical sensing performances toward nonenzymatic detection of glucose, including a wide linear range from 35 μM– to 30 mM, a low detection limit of 5 μM (S/N = 3) and a high sensitivity of 7.84 μA cm⁻² mM⁻¹ as well as good selectivity, long-term stability and reproducibility. The practical application of the proposed PtCo/NPG/GP has also been demonstrated in in vitro detection of blood glucose in real clinic samples.     

Pharmacokinetic study of dendrobine in rat plasma by ultra‐performance liquid chromatography tandem mass spectrometry

Dendrobine, considered as the major active alkaloid compound, has been used for the quality control and discrimination of Dendrobium which is documented in the Chinese Pharmacopoeia. In this work, a sensitive and simple ultra‐performance liquid chromatography tandem mass spectrometry (UPLC‐MS/MS) method for determination of dendrobine in rat plasma is developed. After addition of caulophyline as an internal standard (IS), protein precipitation by acetonitrile–methanol (9:1, v/v) was used to prepare samples. Chromatographic separation was achieved on a UPLC BEH C₁₈ (2.1 ×100 mm, 1.7 µm) column with acetonitrile and 0.1% formic acid as the mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring mode was used for quantification using target fragment ions m/z 264.2 → 70.0 for dendrobine and m/z 205.1 → 58.0 for IS. Calibration plots were linear throughout the range 2–1000 ng/mL for dendrobine in rat plasma. The RSDs of intra‐day and inter‐day precision were both <13%. The accuracy of the method was between 95.4 and 103.9%. The method was successfully applied to pharmacokinetic study of dendrobine after intravenous administration. Copyright © 2015 John Wiley & Sons, Ltd.