Isolation and identification of degradation products of salvianolic acid A by NMR and LC-MS

Salvianolic acid A was subjected to thermal degradation condition in distilled water at 90 °C. Four degradation products, including two novel compounds (1, 2) and two known ones (5, 6), were isolated by reverse-phase semi-preparative liquid chromatography and identified on the basis of NMR and MS data. Other two degradation products (3, 4) were determined by LC-MS analysis.     

杀虫安对小鼠脑组织内神经递质的影响

研究了杀虫安经灌胃给药后对小鼠脑组织内多巴胺及氨基酸含量的影响。将60只ICR (美国癌症研究所)小鼠随机分为空白对照、杀虫安低剂量(200 mg/kg bw)及高剂量(300 mg/kg bw) 3个处理组,于灌胃后出现惊厥时断头取脑,采用高效液相色谱-电化学(HPLC-ECD)检测法测定了鼠脑组织内多巴胺(DA)及其代谢产物二羟苯乙酸(DOPAC)和高香草酸(HVA)的含量,采用液相色谱-质谱联用(LC-MS/MS)法测定了氨基酸的含量。结果发现,两个剂量处理组鼠脑组织内DA及氨基酸含量与对照相比均有显著性差异。其中:DA比对照分别升高25.25%和31.93%(PPPPγ-氨基丁酸(GABA)含量比对照降低32.55% (PP<0.05)。表明杀虫安导致惊厥时伴有鼠脑内多巴胺和氨基酸含量的变化。     

液质联用法同时测定猪粪便中16种（氟）喹诺酮类药物残留

为检测猪粪便中氟喹诺酮类药物残留,建立了高效液相色谱-串联质谱方法,可同时测定猪粪便中环丙沙星、诺氟沙星等16种氟喹诺酮类药物的残留量。样品经50%硝酸镁-4%氨水混合溶液提取,过HLB固相萃取小柱净化,HPLC-MS/MS多反应监测模式下进行定性及定量分析,16种氟喹诺酮类药物在40～200ng·mL^-1浓度范围内呈现良好线性关系,西诺沙星和沙拉沙星检出限为0.005μg·g-1,其他FQs药物检出限均为0.002μg·g-1。添加浓度为0.05、0.5、1.0μg·g-1时,方法平均回收率为51.4%～109.3%,RSD小于20%。该方法前处理简单、快速、灵敏、准确,仅使用少量有机试剂,检测成本低,对环境危害小。     

猪组织中丙硫咪唑亚砜和砜含量的液质联用测定方法的研究

采用液质联用方法检测了动物组织(肌肉、肝脏、肾脏、肌肉)中丙硫咪唑亚砜和丙硫咪唑砜含量,采用液液分配的提取方法,用乙酸乙酯、正己烷和甲醇:水(80:20,V/V)对样品进行处理和净化,welchrom-C<,18>(4.6 mm×250 mm,5μm)色谱柱进行液相色谱一质谱法分析.结果显示以S/N=3为检测限,该方法...     

Comparative analysis between endometrial proteomes of pregnant and non-pregnant ewes during the peri-implantation period

Background

Early pregnancy failure has a profound impact on both human reproductive health and animal production. 2/3 pregnancy failures occur during the peri-implantation period; however, the underlying mechanism(s) remains unclear. Well-organized modification of the endometrium to a receptive state is critical to establish pregnancy. Aberrant endometrial modification during implantation is thought to be largely responsible for early pregnancy loss.

Result

In this study, using well-managed recipient ewes that received embryo transfer as model, we compared the endometrial proteome between pregnant and non-pregnant ewes during implantation period. After embryo transfer, recipients were assigned as pregnant or non-pregnant ewes according to the presence or absence of an elongated conceptus at Day 17 of pregnancy. By comparing the endometrial proteomic profiles between pregnant and non-pregnant ewes, we identified 94 and 257 differentially expressed proteins (DEPs) in the endometrial caruncular and intercaruncular areas, respectively. Functional analysis showed that the DEPs were mainly associated with immune response, nutrient transport and utilization, as well as proteasome-mediated proteolysis.

Conclusion

These analysis imply that dysfunction of these biological processes or pathways of DEP in the endometrium is highly associated with early pregnancy loss. In addition, many proteins that are essential for the establishment of pregnancy showed dysregulation in the endometrium of non-pregnant ewes. These proteins, as potential candidates, may contribute to early pregnancy loss.

Electronic supplementary material

The online version of this article (doi:10.1186/s40104-015-0017-0) contains supplementary material, which is available to authorized users.     

液相色谱-串联质谱法测定猪肉中四环素类药物残留量的不确定度评定

评定液相色谱-串联质谱法（LC-MS/MS）测定猪肉中四环素类药物残留量的不确定度。依据GB/T 21317-2007《动物源性食品中四环素类兽药残留量检测方法 液相色谱-质谱/质谱法与高效液相色谱法》,建立相关数学模型,对实验过程中的不确定度来源进行分析与评定。当采用LC-MS/MS法测定猪肉中土霉素、四环素、金霉素、强力霉素的残留量分别为50.05、42.81、42.45、41.60μg/kg时,扩展不确定度分别为 5.44、6.72、2.86、4.80μg/kg (k=2)。检测结果的不确定度主要受回收率、标准溶液配制、工作曲线拟合的影响。     

Complex Toxin Profile of French Mediterranean Ostreopsis cf. ovata Strains,Seafood Accumulation and Ovatoxins Prepurification

Ostreopsis cf. ovata produces palytoxin analogues including ovatoxins (OVTXs) and a putative palytoxin (p-PLTX), which can accumulate in marine organisms and may possibly lead to food intoxication. However, purified ovatoxins are not widely available and their toxicities are still unknown. The aim of this study was to improve understanding of the ecophysiology of Ostreopsis cf. ovata and its toxin production as well as to optimize the purification process for ovatoxin. During Ostreopsis blooms in 2011 and 2012 in Villefranche-sur-Mer (France, NW Mediterranean Sea), microalgae epiphytic cells and marine organisms were collected and analyzed both by LC-MS/MS and hemolysis assay. Results obtained with these two methods were comparable, suggesting ovatoxins have hemolytic properties. An average of 223 μg·kg⁻¹ of palytoxin equivalent of whole flesh was found, thus exceeding the threshold of 30 μg·kg⁻¹ in shellfish recommended by the European Food Safety Authority (EFSA). Ostreopsis cells showed the same toxin profile both in situ and in laboratory culture, with ovatoxin-a (OVTX-a) being the most abundant analogue (~50%), followed by OVTX-b (~15%), p-PLTX (12%), OVTX-d (8%), OVTX-c (5%) and OVTX-e (4%). Ostreopsis cf. ovata produced up to 2 g of biomass per L of culture, with a maximum concentration of 300 pg PLTX equivalent cell⁻¹. Thus, an approximate amount of 10 mg of PLTX-group toxins may be produced with 10 L of this strain. Toxin extracts obtained from collected biomass were purified using different techniques such as liquid-liquid partition or size exclusion. Among these methods, open-column chromatography with Sephadex LH20 phase yielded the best results with a cleanup efficiency of 93% and recovery of about 85%, representing an increase of toxin percentage by 13 fold. Hence, this purification step should be incorporated into future isolation exercises.     

Largazole Pharmacokinetics in Rats by LC-MS/MS

A highly sensitive and specific LC-MS/MS method for the quantitation of largazole thiol, the active species of the marine-derived preclinical histone deacetylase inhibitor, largazole (prodrug), was developed and validated. Largazole thiol was extracted with ethyl acetate from human or rat plasma along with the internal standard, harmine. Samples were separated on an Onyx Monolithic C18 column by a stepwise gradient elution with 0.1% formic acid in methanol and 0.1% aqueous formic acid employing multiple reaction monitoring (MRM) detection. Linear calibration curves were obtained in the range of 12.5–400 ng/mL with 200 µL of human plasma. The overall intra-day precision was from 3.87% to 12.6%, and the inter-day precision was from 7.12% to 9.8%. The accuracy at low, medium and high concentrations ranged from 101.55% to 105.84%. Plasma protein bindings of largazole thiol in human and rat plasma as determined by an ultrafiltration method were 90.13% and 77.14%, respectively. Plasma drug concentrations were measured by this LC-MS/MS method. The pharmacokinetics of largazole thiol in rats was studied following i.v. administration at 10 mg/kg and found to follow a two-compartment model. Largazole thiol was rapidly eliminated from systemic circulation within 2 h. The established LC-MS/MS method is suitable for the analysis of largazole thiol in human plasma, as well.     

基于代谢组学的‘永春佛手’乌龙茶化学成分解析

【目的】不同茶树品种制作的乌龙茶风味品质和内含成分具有明显差异。本研究结合非靶向代谢组学、化合物定量分析、多元统计学等方法比较‘永春佛手’‘铁观音’和‘水仙’乌龙茶的化学成分差异,分析‘永春佛手’乌龙茶的特征性化学成分,并探讨茶树品种对乌龙茶内含成分和感官品质的影响。【方法】以‘永春佛手’品种为主要研究对象,‘铁观音’和‘水仙’品种为对照,三者均加工制成清香型、浓香型和陈香型乌龙茶,采用超高效液相色谱-四级杆-飞行时间质谱（UHPLC-Q-TOF/MS）进行非靶向代谢组学分析,对不同品种乌龙茶的差异化合物进行鉴定和筛选,并采用超高效液相色谱仪、氨基酸分析仪和超高效液相色谱-四极杆-静电场轨道阱质谱（UHPLC-Q-Orbitrap-MS）对茶叶中主要化学成分和不同品种乌龙茶间差异化合物进行定量分析。【结果】相比于同一香型的对照品种乌龙茶,‘永春佛手’具有相对较高的氨基酸总量和茶氨酸含量,儿茶素类化合物总量和咖啡碱含量在清香型和浓香型的不同品种间无显著差异,在陈香型中均以‘永春佛手’乌龙茶含量最高,没食子酸含量在3种香型中均呈现‘水仙’>‘永春佛手’>‘铁观音’。代谢组学结合多变量统计表明,不同品种制成的乌龙茶化合物表型具有明显差异。清香型、浓香型和陈香型乌龙茶中,‘永春佛手’与对照品种乌龙茶间的差异化合物分别为50、59、47个,其中共同差异化合物有23个,包括14个黄酮（醇）糖苷、5个儿茶素类化合物及其衍生物、1个脂类、1个生物碱、1个有机酸和1个氨基酸类化合物。进一步对茶叶中常见的20个黄酮（醇）糖苷组分进行定量分析,结果显示‘永春佛手’乌龙茶中14个黄酮（醇）糖苷组分的含量显著高于对照品种乌龙茶,其含量是相同香型‘铁观音’品种乌龙茶的1.4—14.6倍,是‘水仙’品种乌龙茶的1.3—18.0倍。在定量的4类黄酮（醇）糖苷中,槲皮素糖苷均为主要的黄酮（醇）糖苷组分,含量高于山柰酚糖苷、杨梅素糖苷和芹菜素糖苷。感官审评分析表明较高含量的黄酮（醇）糖苷并未显著增强茶汤的涩味。【结论】代谢组学方法可以有效地对不同品种制作的乌龙茶进行化合物表征。‘永春佛手’‘铁观音’和‘水仙’按照相同工艺制成的乌龙茶具有较为明显的化学成分差异,其中黄酮（醇）糖苷类化合物差异最为显著,具有较高含量的黄酮（醇）糖苷类成分是‘永春佛手’乌龙茶在化学成分方面的一个重要特征。黄酮（醇）糖苷类成分有望作为判别指标用于乌龙茶制作品种的判别。     

替米考星在黄羽肉鸡组织中残留消除规律研究

在常规饲养条件下,以200 mg/kg替米考星剂量水平添加到饲料中饲喂4种黄羽肉鸡,连续饲喂5d,停药后6h,1、3、5、7、10、14 d每1品种肉鸡分别宰杀5只,采集每只鸡的肌肉和肝脏组织,用高效液相色谱-串联质谱法进行残留量测定.结果表明,替米考星主要残留于肉鸡的肝脏组织中;肌肉、肝脏中替米考星残留消除速率相似.停药14 d后,肌肉、肝脏中替米考星残留量均低于最高残留限量.建议替米考星在黄羽肉鸡临床休药期以20 d为宜.