Preparing the key metabolite of Z‐ligustilide in vivo by a specific electrochemical reaction

The key in vivo metabolites of a drug play an important role in its efficacy and toxicity. However, due to the low content and instability of these metabolites, they are hard to obtain through in vivo methods. Electrochemical reactions can be an efficient alternative to biotransformation in vivo for the preparation of metabolites. Accordingly, in this study, the metabolism of Z‐ligustilide was investigated in vitro by electrochemistry coupled online to mass spectrometry. This work showed that five oxidation products of the electrochemical reaction were detected and that two of the oxidation products (senkyunolide I and senkyunolide H) were identified from liver microsomal incubation as well. Furthermore, after intragastric administration of Z‐ligustilide in rats, senkyunolide I and senkyunolide H were detected in the rat plasma and liver, while 6,7‐epoxyligustilide, a key intermediate metabolite of Z‐ligustilide, was difficult to detect in vivo. By contrast, 6,7‐epoxyligustilide was obtained from the electrochemical reaction. In addition, for the first time, 6 mg of 6,7‐epoxyligustilide was prepared from 120 mg of Z‐ligustilide. Therefore, electrochemical reactions represent an efficient laboratory method for preparing key drug metabolites.     

Quantitative screening for steroids in animal feeding water using reversed phase LC with gradient elution

Several isocratic separations for the determination of 20 steroids (STER) in animal feeding water samples (AFWS) from drinking-trough by LC using a mobile phase ACN/H(2)O (35:65 v/v) and different RP columns (Hypersil C18, Gemini C18 (GM), Purospher Star C18, Synergi Max C12, and Synergi Fusion) and UV detection were obtained. The elution order was the same: a first group of corticoids (CC) was early eluted, a second group of CC and anabolics (AAS) exhibited intermediate retention, and a third group constituted by AAS was strongly retained. To improve the separation performances of the isocratic separations an ACN gradient elution optimization was carried out for each column. The most satisfactory results were obtained using a Purospher column which allowed the separation of 19 STER in an analysis time close to 26 min. After sample preparation using SPE, method validation was performed in an AFWS spiked with STER according to the EC decision criteria established for quantitative screening methods. For this purpose calibration graphs, extraction efficiencies, decision limits, detection capabilities, precision (repeatability and within-laboratory reproducibility), accuracy, selectivity, and robustness were evaluated. The proposed method was applied to other AFWS with satisfactory results.     

Determination of selenocysteine and selenomethionine in edible animal tissues by 2D size-exclusion reversed-phase HPLC-ICP MS following carbamidomethylation and proteolytic extraction

A method was developed for the simultaneous determination of selenomethionine (SeMet) and selenocysteine (SeCys) in meat (chicken and lamb muscles) and different offal tissues (heart, liver, kidney). The analytical procedure was based on the protein extraction with urea under reducing conditions (dithiothreitol), derivatization of SeCys and SeMet by carbamidomethylation with iodoacetamide (IAM) followed by quantitative proteolysis. The mixture of the derivatized Se-amino acids was purified by size-exclusion liquid chromatography (LC) and analysed by ion-paring reversed-phase HPLC–inductively coupled plasma mass spectroscopy (ICP MS). The quantification of SeCys and SeMet was carried out by the method of standard additions. ⁷⁷SeMet was used to control the SeMet derivatization efficiency and recovery. The method was validated by the determination of the Se mass balance. The Se-amino acids accounted for 91 ± 8% of the total selenium (mean of 95 samples of seven tissues analysed over a period of 18 months). The method was applied to the discrimination of the contribution of selenoproteins (containing SeCys) and other Se-containing proteins (containing SeMet) in tissues of animals during supplementation studies (dose–effect and tolerance).     

动物肝脏中九种多溴联苯醚残留量的GC—NCI／MS分析

建立了动物肝脏中9种PBDEs残留量的气相色谱-负化学离子源/质谱(GC-NCI/MS)的分析方法。动物肝脏样品经V(正己烷)∶V(丙酮)=1∶1超声辅助提取,中性与酸性硅胶层析柱净化和V(正已烷)∶V(CH2Cl2)=1∶1洗脱和浓缩后,以PCB-103为内标物,采用GC-NCI/MS的选择离子监测方式(SIM)对其中的9种PBDEs残留量进行了定性与定量分析。当动物肝脏空白样品的加标质量浓度为5.0、20.0μg/kg(PBDE-183为6.0、24.0μg/kg)时,9种PBDEs的平均加标回收率为75.1%~88.2%,相对标准偏差为3.3%~7.9%,方法检出限均小于0.07μg/kg;线性范围除PBDE-183为0.12~600.0μg/kg外,其余8种PBDEs为0.1~500.0μg/kg,相关系数都大于0.9993。所建立的分析方法已用于5种动物肝脏的8个样品中9种PBDEs残留量的分析。     

高效液相色谱法定量分析人胎肝细胞上清液及细胞裂解液中7-酮基胆固醇

用高效液相色谱法定量分析了７－酮基胆固醇在胎肝细胞上清液及细胞裂解液中的含量。色谱柱为μ－ＰｏｒａｓｉｌＳｉＯ２柱,流动相为正己烷∶异丙醇（９１∶９,Ｖ／Ｖ）。方法回收率高、色谱重复性好、分析速度快。     

高效液相色谱法检测人肝组织中黄曲霉毒素

以人肝癌组织为研究材料,丙酮、氯仿为主要抽提溶剂,建立了用高效液相色谱技术检测肝组织中黄曲霉毒素Ｂ＿１的方法。采用μ－Ｐｏｒａｓｉｌ色谱往,氯仿、环己烷、乙腈、异丙醇为流动相,荧光检测器检测,激发波长３６５ｎｍ,发射波长４３５ｎｍ。方法的平均回收率为９６．８５％,变异系数为４．５２％,最低检测限为０．０５ｎｇ。１０例肝癌组织中有７例检出５。４９～７３．７０ｎｇ／ｇ不等的黄曲霉毒素Ｂ＿１。方法快速、准确、灵敏度高。     

吸附型“人工肝”辅助材料的制备及其性能研究Ⅱ.免疫吸附剂的制备及其吸附性能

本文介绍以醋酸乙烯酯为单体,二乙烯苯为交联剂制得大孔共聚物;经皂化、活化后,偶联IgG制得免疫吸附剂。讨论了不同活化剂对偶联配体的影响,观察其对人血清中的HBsAg的吸附性能。     

In Vitro 31P MR Chemical Shifts of In Vivo-Detectable Metabolites at 3T as a Basis Set for a Pilot Evaluation of Skeletal Muscle and Liver 31P Spectra with LCModel Software

Most in vivo ³¹P MR studies are realized on 3T MR systems that provide sufficient signal intensity for prominent phosphorus metabolites. The identification of these metabolites in the in vivo spectra is performed by comparing their chemical shifts with the chemical shifts measured in vitro on high-field NMR spectrometers. To approach in vivo conditions at 3T, a set of phantoms with defined metabolite solutions were measured in a 3T whole-body MR system at 7.0 and 7.5 pH, at 37 °C. A free induction decay (FID) sequence with and without ¹H decoupling was used. Chemical shifts were obtained of phosphoenolpyruvate (PEP), phosphatidylcholine (PtdC), phosphocholine (PC), phosphoethanolamine (PE), glycerophosphocholine (GPC), glycerophosphoetanolamine (GPE), uridine diphosphoglucose (UDPG), glucose-6-phosphate (G6P), glucose-1-phosphate (G1P), 2,3-diphosphoglycerate (2,3-DPG), nicotinamide adenine dinucleotide (NADH and NAD⁺), phosphocreatine (PCr), adenosine triphosphate (ATP), adenosine diphosphate (ADP), and inorganic phosphate (Pi). The measured chemical shifts were used to construct a basis set of ³¹P MR spectra for the evaluation of ³¹P in vivo spectra of muscle and the liver using LCModel software (linear combination model). Prior knowledge was successfully employed in the analysis of previously acquired in vivo data.     

Eco-friendly and Cleaner Process Using Online Microwave-assisted Steam Extraction Coupled with Solid-phase Extraction for Trace Analysis of Sulfonamides in Animal Feed

An environmentally friendly method for extracting sulfonamides(SAs) residues from animal feed was described and applied. The method used online microwave-assisted steam extraction coupled with solid phase extraction(MASE-SPE), which was followed by the analysis using high performance liquid chromatography-mass spectrometry(HPLC-MS/MS). The SAs residues were extracted successively with water steam under microwave irradiation, and thus directly introduced into an SPE column containing cation-exchange resin. The SAs were then eluted with methanol-ammonia(90:10, volume ratio) from the SPE column and followed by HPLC-MS/MS. The limits of detection(LODs) for the analytes ranged from 0.24 ng/g to 0.49 ng/g. The limits of quantification(LOQs) ranged from 0.82 ng/g to 1.63 ng/g. Average recoveries of SAs were 76.3%-92.1%. The developed method was a reliable and environmentally friendly alternative to previous methods with respect to time, solvent and labor consumption for the analysis of SAs in animal foodstuffs.     

RESEARCH ON DYNAMIC MECHANICAL RESPONSE AND CONSTITUTIVE MODEL OF PORCINE LIVER

在交通事故中,腹部器官常因冲击载荷作用而受到伤害,严重时甚至危及生命.肝损伤是腹部损伤中最为常见的一种,致死率很高,了解肝脏的动态力学性能对于事故中肝脏的损伤评估及防护设计有着重要的意义.从新鲜的猪肝组织中取肝实质部分制作试样,利用英斯特朗材料试验机对其进行两种加载率（0.004 s^-1,0.04 s^-1）和两种加载方向（垂直肝脏表面和平行于肝脏表面）的准静态压缩试验,并压缩至破坏.利用改进的分离式霍普金森压杆（split Hopkinson pressure bar,SHPB）实验装置沿平行于肝脏表面方向进行三种高应变率（1 300 s^-1,2 400 s^-1,4 500 s^-1）的动态压缩试验.结果表明：所有应变率下的猪肝压缩应力应变曲线都呈非线性凹向上特征,初始阶段应力值很低,应变约30%后应力幅值显著增大;准静态压缩时,两种应变率（0.004 s^-1,0.04 s^-1）和两种加载方向下肝脏组织破坏应力和破坏应变等力学性能无显著不同,平均破坏应变为48%,平均破坏应力为0.45 MPa.高应变率下肝脏组织的流动应力明显高于准静态下的流动应力,表现出一定的率敏感性.采用Yeoh型超弹性本构模型描述猪肝组织准静态力学性能,基于黏超弹性模型理论,提出了一个能描述肝脏组织从低应变率到高应变率范围力学性能的率相关本构模型,该模型与实验结果有很好的一致性.