UPLC-Q TOF MS~E与镜像对比分析四种麻黄炮制过程的成分变化

利用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q TOF MS~E)技术采集麻黄和其不同炮制品的复杂化学成分数据,通过对比分析,探究蜜炙、醋制、酒制、炒炭4种炮制过程中麻黄非挥发性化学成分的变化。根据时间依赖型质谱扫描模式(MS~E)采集的超高精度样品数据快速识别麻黄及其炮制品中的主要化学成分,并结合镜像对比分析,总结不同炮制工艺对化学成分与变化趋势的影响。结果显示,与生品麻黄相比,炮制品中共有4类主要成分:生物碱、黄酮、烯烃和有机酸,21种非挥发性化学成分发生变化。其中,生物碱类成分均有所下降,以酒制和炒炭炮制品下降最多;黄酮、烯烃和有机酸类成分经蜜炙后含量上升,而在其他炮制品中含量下降。麻黄及其炮制品物质基础复杂,药效是多类成分共同作用的结果,不同炮制过程会使麻黄的化学成分发生明显变化,炮制品功效可能会随之改变,该方法可为麻黄炮制的现代化研究提供物质基础,也可为临床区别用药提供借鉴。     

超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用鉴定椴树蜂蜜中的抗氧化活性肽

应用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱联用法(UPLC-Q-Exactive)鉴定椴树蜂蜜蛋白经胰蛋白酶水解后获得的肽类物质的组成和结构,并探讨椴树蜂蜜酶解多肽与其抗氧化活性的关系。椴树蜂蜜经三氯乙酸溶液沉淀,所得蛋白用胰蛋白酶酶解,酶解肽段经C18固相萃取柱富集净化后,采用0.1%甲酸(A)-乙腈(B)作为流动相进行梯度洗脱,以高分辨质谱(Q-Exactive)的Full MS/ddMS2模式对其进行鉴定。采用MaxQuant软件分析质谱结果,所得肽段在Uniprot上进行Blast序列对比,共鉴定出52种椴树蜂蜜胰蛋白酶解多肽,其中71.15%来自蜂王浆主要蛋白(MRJPs),鉴定出的肽段归属于10种蛋白质,其中6种(60%)属于MRJP家族蛋白。对椴树蜂蜜酶解多肽质谱鉴定结果进行重复性分析,2次以上重复质谱分析可实现对椴树蜂蜜多肽(87.1%)的可靠鉴定。结合文献报道的蜂蜜多肽抗氧化活性的构效关系,筛选出11条可能具有抗氧化活性的多肽。化学合成11条多肽后,进行4种不同的抗氧化活性实验。结果表明,这11条多肽的抗氧化活性存在较大差异,其中1 g/L VIYEWK多肽显示出较强的ABTS自由基清除活性(33.8%±0.6%)、DPPH自由基清除活性(96.5%±0.0%),并表现出一定的还原力(0.010±0.001)和Fe2+螯合活性(4.0%±0.2%)。该方法能快速有效鉴定椴树蜂蜜多肽结构,可为揭示椴树蜂蜜多肽结构与抗氧化活性的关系提供依据。     

UPLC-MS/MS法测定蜂蜜中磺胺类抗生素残留

应用超高效液相色谱-串联质谱(UPLC-MS/MS)法测定蜂蜜中磺胺类抗生素。针对还原糖与磺胺类抗生素结合物难以提取的问题，用添加回收率法考察0.7 mol/L盐酸、0.3 mol/L柠檬酸和磷酸溶液（pH2）对结合态磺胺的水解情况，以阳性蜂蜜样品中测得的磺胺类抗生素含量为参考指标，对3种前处理方法的水解效率进行验证。结果表明，0.7 mol/L盐酸、0.3 mol/L柠檬酸和pH2磷酸溶液水解提取磺胺类抗生素的回收率分别为59.2%~72.2%、27.8%~71.7%和0.4%~48.9%，可见提取过程存在明显损失。采用同位素稀释质谱法（IDMS）同步校正磺胺类抗生素在提取净化过程的损失及基质效应的干扰，3种提取方法的添加回收率均提高到95.4%~104.2%。但是，采用磺胺药物阳性蜂蜜样品对提取方法进行验证时，盐酸、柠檬酸和磷酸的IDMS方法测量结果分别为10.8~22.4、5.7~16.2和0.1~5.3 μg/kg，后2种提取方法的测量结果比第1种的分别偏低23.1%~57.3%和73.8%~99.6%。表明对于结合态药物残留的测定，采用添加回收率考察方法的准确性时，需要使用阳性样品进行验证。     

气相色谱-质谱法分析金雀根中脂肪酸的组成

The chemical constituents of fatty acid in the roots of Caragana sinica were determined and identified by GC/MS. Eighteen compounds were identified. The main components are 9,12,15-octadecatrienoic acid (28.41%), 9,12-octadecatrienoic acid (34.61%), Hexadecanoic acid (19.33%) and octadecatrienoic acid (6.37%).     

Toxicity assessment of nano‐TiO2 in Apis mellifera L., 1758: histological and immunohistochemical assays

The aim of this study was to evaluate the toxicity of titanium dioxide nanoparticles (TiO₂NPs) by short‐term toxicity tests in Apis mellifera, considered an excellent bioindicator organism mainly due to its sensitivity. Bees have been exposed to several concentrations of TiO₂NPs (1 × 10^?3, 1 × 10^?4, 1 × 10^?5, 1 × 10^?6 mg/10 ml) for 10 days. Morphostructural and histological assays were done on gut and honey sac. The research of exposure biomarkers like metallothioneins 1 (MT1) and Heat Shock Protein 70 (HSP70) was performed to verify if a detoxification mechanism has been activated in the exposed animals. No histological alteration on the epithelium of the gut and honey sac were observed in exposed samples. A significant positivity for anti‐MT1 antibody was observed only in the honey sac cells. A weak positivity for HSP70 was observed in both structures analyzed. In several studies have shown the non‐toxicity of TiO₂NPs on other model organisms, in our study, titanium dioxide nanoparticles was proven to be highly toxic at the highest concentration tested (100% of lethality to 1 × 10^?3 mg/10 ml) and moderately toxic at lower concentrations. Honey bees proved to be excellent models for study of NPs toxicity and for monitoring environment.     

HPLC-ESI-MS/MS分析黄芪蜜炙前后活性成分差异

建立了HPLC-ESI-MS/MS联用技术分析生黄芪与炙黄芪的成分，并比较两者成分差异。采用Agilent ZORBAX SB-C18色谱柱（4.6 mm×250 mm×5 μm），以0.1%甲酸乙腈溶液-0.1%甲酸水溶液为流动相进行梯度洗脱，流速1 mL/min，检测波长203 nm，柱温25 ℃；使用ESI离子源，分别在正、负离子模式下进行全扫描和二级扫描，质量扫描范围m/z 100~2000。采用该方法鉴定了生黄芪与炙黄芪中的62种成分，并对黄芪蜜炙前后的成分进行了比较，实验结果可为黄芪炮制方法的选择提供参考依据。     

Morphostructural and immunohistochemical study on the role of metallothionein in the detoxification of heavy metals in Apis mellifera L., 1758

Honey is a valuable food produced by bees from sugary substances that they gather in nature. The transformation the nectar into honey, by bees, is long and complex. Except for honey, where heavy metals are absent or are found only in traces, the bees and their products have always been considered excellent biomarkers of such contaminants. We have assumed that the absence of heavy metals in honey is due to the presence of a detoxification system in the digestive system of bees, which involves metallothioneins, proteins that have a role in the homeostatic control of essential and non‐essential metals. We have placed the beehives in three different zones: industrial, urban and rural. Investigations were carried out with ICP‐MS method for the detection of heavy metals in the guts of honey bees and honey. The metallothioneins have been identified by Immunohistochemical and Western‐blotting analisys. The investigations have shown the presence of heavy metals only in bees guts but not in honey, while the presence of metallothionein has been highlighted only in epithelium of the honey sac, demonstrating the existence of an efficient system of detoxification of heavy metals.     

Distribution of dopamine receptors and dopamine receptor homologs in the brain of the honey bee, Apis mellifera L

In the brain of the honey bee, Apis mellifera, the radioligands [3H]-SCH23390 and [3H]-spiperone recognise D1- and D2-like receptors, respectively. In addition to being pharmacologically distinct and exhibiting significantly different expression profiles during the lifetime of the bee, [3H]-SCH23390- and [3H]-spiperone-binding sites differ markedly in their distribution within the brain. Estimates of [3H]-SCH23390-binding site density are highest in the somatal rind, whereas [3H]-spiperone-binding sites are most concentrated in the beta lobe neuropil of the mushroom bodies. Molecular cloning techniques have been used to identify two honey bee genes encoding dopamine receptor homologs. The first is the honey bee counterpart of a Drosophila D1-like dopamine receptor and is expressed in the mushroom bodies of both workers and drones. The second is related to D2-like dopamine receptors from vertebrates and is expressed in the brain of the bee, but the precise distribution of expression is not yet known.     

基于ARM处理器的蜂蜜蔗糖快速检测仪的研制与应用

研制了一种基于ARM处理器的新型蜂蜜蔗糖快速检测仪，阐述了其工作原理以及软硬件设计。对实际样品的检测结果表明，该仪器检测速度快，检测时间为10～30分钟，检测结果与国家标准方法相比无显著性差异。     

麻黄中主要化合物的串联质谱研究

The major chemical constituents of Ephedra（ephedrine, pseudoephedrine, methylephedrine, norephedrine）were analyzed by electrospray ionization tandem-mass spectrometry. Major chemical components, including Ephedrine, Pseudoephedrine, Methylephedrine, Norephedrine were studied. Using tandem mass spectrometry, useful fragments of Ephedrine, Pseudoephedrine, Methylephedrine, Norephedrine were obtained. The experimental results demonstrate that Ephedrine and Pseudoephedrine show the similar fragmentation in ESI-MS.