液相色谱-串联质谱法测定肉肠中莱克多巴胺残留量的研究

建立了肉肠中莱克多巴胺残留量固相萃取-液相色谱-串联质谱分析方法。肉肠样品经甲醇提取,提取物经葡萄糖醛酸酶酶解后,固相萃取小柱净化。选用多反应监测模式进行测定。通过基质效应的考察,确定了定量校正的方法。结果显示,莱克多巴胺添加浓度在0.5～4μg/kg范围内时,其峰面积和浓度呈良好的线性范围,相关系数R2大于0.99。本方法最低检测限为0.15μg/kg。     

Tropane and ergot alkaloids in grain-based products for infants and young children in the Netherlands in 2011–2014

An LC-MS/MS multi-method was developed to simultaneously quantify ergot alkaloids (EAs) and tropane alkaloids (TAs) in 113 cereal-based food for infants and young children. To assess yearly variation, samples were collected in 2011, 2012 and 2014. EAs were detected in 54% and TAs in 22% of the samples. Mean EA levels in the three sampling years were 10.6, 6.2 and 8.6 µg kg^?1, respectively (maximum: 115.4 µg kg^?1), indicating that exposure to EAs would not have exceeded the health-based guidance values set by EFSA in 2012. Mean TA levels were 3.9, 2.4 and 0.4 µg kg^?1, respectively (maximum: 80.8 µg kg^?1). The acute reference dose for TAs, derived by EFSA in 2013, would have been exceeded by young children when consuming some of the products sampled in 2011–2012. TA levels had decreased drastically in 2014, possibly due to measures taken by producers as response to the EFSA Opinion.     

液相色谱质谱联用测定多种兽药残留方法中标准品稳定性的挑战（网络首发、推荐阅读）

采用等时测量法对可用于食品和饲料链中兽药检测的标准品混合溶液和中间混合液进行了短期和长期稳定性研究。其中短期稳定性实验考察了混合标准曲线在–20（作为基线）、4和23 ℃（避光和不避光保存）条件下存储1、2、4和7 d的稳定性,而长期稳定性实验考察了中间混合液在–20、4、23 ℃（避光和不避光保存）和–80 ℃储存条件下2、4、8和12周的稳定性。结果表明,使用不含酸溶剂配制的混合标准曲线在4 ℃下建议储存1~2 d,中性储存条件即使在室温也可短期储存1~2 d。对于含有ß-内酰胺和头孢菌素类标准品的中间混合液在4 ℃和室温下储存超过1个月会损失近90%。对于含有青霉素V和G的混和标准品中间储备液,在–20 ℃不含酸的条件下可储存2周。其他类别的兽药标准品对长期储存条件要求较少。整体而言,兽药标准品中间混合液长期储存的最佳储存条件为–20 ℃。     

一株拮抗海鱼干优势耐盐真菌芽孢杆菌HY-5筛选及其抗菌成分鉴定

目的:开发新的海鱼干耐高盐优势真菌抑制剂。方法:取用湛江市特呈岛红树林海域的海水、海泥以及红树林的根、茎和叶的样本,利用琼脂扩散法和平板点种法筛选出一株强抗菌活性菌株HY-5,通过传统细胞形态观察并结合生理生化实验及16S rDNA序列进行分析鉴定;利用杯碟法测定上清液对17株霉变海鱼干优势真菌抗菌谱,发酵液酸沉后经LC-MS检测其抗菌成分。结果:HY-5被鉴定为解淀粉芽孢杆菌,其上清液对17株霉变海鱼干优势真菌均有明显抗菌作用;LC-MS检测到HY-5发酵液中含2组抗菌脂肽surfactin(994.6、1008.6、1022.5和1036.6)和iturin(1016.5、1030.5、1044.4和1058.3)组分。结论:海洋源解淀粉芽孢杆菌HY-5的粗提液中含有抗菌脂肽,其对霉变海鱼干优势真菌有较强的拮抗作用。     

Proteome Analysis of Subsarcolemmal Cardiomyocyte Mitochondria: A Comparison of Different Analytical Platforms

Mitochondria are complex organelles that play critical roles in diverse aspects of cellular function. Heart disease and a number of other pathologies are associated with perturbations in the molecular machinery of the mitochondria. Therefore, comprehensive, unbiased examination of the mitochondrial proteome represents a powerful approach toward system-level insights into disease mechanisms. A crucial aspect in proteomics studies is design of bioanalytical strategies that maximize coverage of the complex repertoire of mitochondrial proteins. In this study, we evaluated the performance of gel-based and gel-free multidimensional platforms for profiling of the proteome in subsarcolemmal mitochondria harvested from rat heart. We compared three different multidimensional proteome fractionation platforms: polymeric reversed-phase liquid chromatography at high pH (PLRP), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and isoelectric focusing (IEF) separations combined with liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS), and bioinformatics for protein identification. Across all three platforms, a total of 1043 proteins were identified. Among the three bioanalytical strategies, SDS-PAGE followed by LC-MS/MS provided the best coverage of the mitochondrial proteome. With this platform, 890 proteins with diverse physicochemical characteristics were identified; the mitochondrial protein panel encompassed proteins with various functional roles including bioenergetics, protein import, and mitochondrial fusion. Taken together, results of this study provide a large-scale view of the proteome in subsarcolemmal mitochondria from the rat heart, and aid in the selection of optimal bioanalytical platforms for differential protein expression profiling of mitochondria in health and disease.     

液相色谱-串联质谱法测定进出口人参、西洋参中蚜灭磷等6种农药

建立了人参、西洋参中凝胶色谱-固相萃取-液相色谱-串联质谱法测定蚜灭磷、吡虫啉、涕灭威、甲基内吸磷、甲硫威、辛硫磷6种农药的分析方法。样品经丙酮超声提取,凝胶色谱(GPC)净化后,固相萃取(SPE)净化,在HPLC/MS/MS多反应监测(MRM)模式下进行了定性和定量分析方法。在上述检测方法条件下,所测的6种农药的最低检出限为0.01 mg/kg,回收率范围为79.2%~107.3%,线性范围为0.01~0.2 mg/kg。     

Application of Hyphenated Chromatography–Mass Spectrometry Techniques to Plant Allelopathy Research

Plant allelopathy offers hope as an additional means of weed control in modern agriculture. Its mechanisms and molecular basis are not yet well understood. Research on the chemical basis for allelopathy has often been hindered by the complexity of plant and soil matrices, making it difficult to track active compounds. Recent improvements in the cost and capabilities of bench-top chromatography–mass spectrometry instruments make these tools more powerful and more widely available to assist with molecular studies conducted in today's expanding field. Such instrumental techniques are herein recommended as economically efficient means of advancing the rigor of allelopathy research and assisting the development of a better understanding of the chemical basis for the allelopathy phenomenon.     

大米中环已烯酮类除草剂残留量的测定

建立了大米中环己烯酮类除草剂残留量同时测定的液相色谱-质谱/质谱法。试样中残留的环己烯酮类除草剂用酸性乙腈高速匀浆提取,提取液经N-丙基乙二胺（PSA）、十八烷基硅烷（ODS）和石墨化炭黑净化,用液相色谱-质谱／质谱仪检测和确证,外标法定量。环己烯酮类除草剂的浓度在0．0025～0．1000μg·mL^-1范围内时,线性关系良好,8种环己烯酮类除草剂的相关系数为0．9947～0．9992。在0．005～0．050mg·kg^-1浓度范围内,样品平均加标回收率在73．2％～107．0％之间,相对标准偏差为5．14％～10．15％。8种环己烯酮类除草剂的最低检出限均为0．005mg·kg^-1。     

An improved LC-MS/MS method for the quantitation of acrylamide in processed foods

An improved liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of acrylamide in processed foods. The homogenized samples, spiked with ¹³C₃-acrylamide as an internal standard, were extracted with water and centrifuged. D₅-3-chloropropanediol as a recovery standard was added to 1-ml aliquots, and the sample was purified with a C₁₈-cartridge column. The extract was directly analyzed using LC-MS/MS without derivatization. The ion transitions of 72–55 m/z (acrylamide), 75–58 m/z (¹³C₃-acrylamide), and 116–98 m/z (d₅-3-chloropropanediol) were found to be the most reliable for the identification and quantification of acrylamide in multiple reaction monitoring. The limit of quantification for acrylamide, defined as a signal-to-noise ratio of 10:1, was 2 μg/kg. The use of d₅-3-chloropropanediol minimized the effects of variation in the sample matrixes and increased the quality of analysis. This method could be applied to the quantification of acrylamide in processed foods.     

Antioxidant activity and polyphenol content of Turkish thyme (Thymus vulgaris) monitored by liquid chromatography and tandem mass spectrometry

Like tea, the leaves of Turkish thyme (Thymus vulgaris) can be boiled in water to produce an extract. This is widely used as syrup for the treatment of coughs and bronchitis at alternative medicine clinics in many parts of the world. In the current study, we assessed the phenolic content and antioxidant activity of thyme. The antioxidant activities of both ethanol and aqueous extracts of thyme were determined using various in vitro methods. The total phenolic and total flavonoid contents were determined to be a gallic acid equivalent and a quercetin equivalent, respectively. Finally, the quantities of the phenolic compounds were detected using high-performance liquid chromatography and tandem mass spectrometry. The total phenolic compounds in the aqueous extract and ethanol extracts of Turkish thyme were 256.0 μg gallic acid equivalent/mg dried extract and 158.0 μg gallic acid equivalent/mg dried extract, respectively. Conversely, the total flavonoid compounds in both extracts were 44.2 μg and 36.6 μg quercetin equivalent/mg dried extract, respectively. For the first time, we determined phenolic contents and investigated the antioxidant potential of thyme. The results indicate that Turkish thyme is a good dietary source with phenolic properties.