中药制剂中地西泮的LC/MS/MS分析方法研究

采用液相色谱串联质谱联用仪分析中药制剂中的地西泮.用乙腈-0.01 mol/L乙酸铵水溶液(6+4)超声提取试样中的地西泮,离心分离,取上清液过滤膜后用LC/MS/MS进行检测.通过对提取方式和液相色谱分离条件的优化,建立了中药制剂中地西泮的LC/MS/MS检测方法.样品加标的回收率为87.6%～96.8%,相对标准偏差为3.1%～9.6%,多反应监测(选择离子对286.0/194.2及286.0/223.1)检出限为0.5 μg/L.     

共沉淀法制备球形镍锰复合草酸盐的最佳pH值

由共沉淀法合成了锂离子电池正极材料LiNi1/2Mn1/2O2用球形镍锰复合草酸盐前驱体.通过对M2+-(Ni2+,Mn2+)-NH3-C2O42--H2O体系的热力学分析,得到了M2+-NH3-C2O42--H2O体系的lg[M2+]-pH关系图(其中M为过渡金属元素),从而得出了以NH4OH为络合剂、H2C2O4为沉淀剂,采用共沉淀法制备球形镍锰复合草酸盐的最佳共沉淀pH值为8.2.在此pH值条件下合成的镍锰复合草酸盐粉料呈球形,且分散较好.     

SiC颗粒表面金属化工艺及显微组织分析

为改善碳化硅颗粒与金属的润湿性,本文研究了在碳化硅颗粒表面金属化即沉积Ni-P层的工艺方法.首先采用表面清洁处理,继而采用粗化和活化敏化工艺对平均尺寸为5μm的Si C颗粒进行预处理,实现了在化学镀镍前在碳化硅表面产生微小的缺陷,并吸附Pd作为催化活性物质,为后序化学镀过程Ni-P的吸附、形核、长大提供了有利条件.利用正交试验方法,结合增重百分率和镍元素的相对含量等指标,研究了化学镀液的成分对镀层组织及形貌的影响.研究发现,当化学镀液中[Ni~(2+)]浓度为0.25 mol/L、[NH_4~+]浓度为0.6 mol/L,[Ni~(2+)]/[H_2PO_2~-]浓度之比为0.4、柠檬酸浓度为0.1 mol/L及pH为10,温度为45℃时,SiC颗粒表面完全包覆Ni-P.通过比较实验可知,预处理中活化敏化过程对后续Ni-P合金的包覆有重要促进作用.同时,化学镀液各成分中对施镀效果的影响顺序为:溶液中[Ni~(2+)]浓度pH柠檬酸三钠的含量温度[Ni~(2+)]/[H_2PO_2~-]浓度之比[NH_4~+]浓度.     

纳米羟基磷灰石对Ni^2＋的吸附性能及机理研究

以硝酸钙和磷酸氢二铵为原料用化学共沉淀法制备纳米羟基磷灰石(n-HA),用XRD、TEM 和BET表征样品的相组成、结晶形貌和比表面积.结果表明:制备的针状羟基磷灰石长轴约为31.9nm,短轴约为21.3nm,粒径均匀且表面活性高,具有较低结晶度,比表面积高达135m2/g.选用Ni2+作为吸附目标离子,通过Zeta电位、XRD和XPS分析n-HA吸附前后的表面电位、晶体物相和表面原子结合能的变化可知:Langmuir等温吸附模型仅适用于Ni2+初始浓度低于0.1mol/L的范围;当Ni2+初始浓度高于0.1mol/L时,Ni2+取代Ca2+与表面的O成键,吸附过程包括离子交换、静电吸附和溶解沉淀作用.     

纳米金溶胶细胞毒性的研究

文章主要研究纳米金溶胶的细胞毒性。通过[3H]-TdR掺入法,研究粒径在15—20nm的纳米金溶胶,溶胶中含有的柠檬酸钠溶液以及溶胶中含有的柠檬酸钠与PVP混合溶液对正常细胞（人体表皮细胞、人体皮肤成纤维细胞）和癌细胞（HeLa细胞、K562细胞）活性的影响,实验结果表明,纳米金溶胶中含有的柠檬酸钠与PVP对这四种细胞的活性基本没有影响,可以在一定浓度范围内促进人体正常皮肤细胞的增殖,并具有剂量（以50μmol／L浓度为临界点）和时间依赖性;对于癌细胞则需高于一定浓度（50μmol／L）才有显著的抑制作用。     

钠基膨润土对废水中Ni2+,Cu2+的吸附效果

为了提高膨润土吸附废水中Ni2+,Cu2+的效果,对其改性制备了钠基膨润土,考察了吸附时间、废水pH值以及Ni2+和Cu2+初始浓度对自制钠基膨润土吸附Ni2+,Cu2+效果的影响,分析了吸附等温线,并探讨了吸附机理。结果表明:钠基膨润土对Ni2+,Cu2+都有较好的吸附性能;初始阶段(前10 min)吸附很快,之后随时间延长吸附量提高甚微;pH值为6.2时,处理后的水质即可达国家排放标准;随着Ni2+,Cu2+初始浓度增加,去除率先增加后降低,而吸附量迅速增加,Ni2+和Cu2+初始浓度分别为25 mg/L和40 mg/L时,钠基膨润土对Ni2+,Cu2+的去除率均可达98.6%;钠基膨润土对Ni2+,Cu2+的吸附等温线符合Freundlich吸附等温式。     

流化床化学气相沉积法制备CNT/Fe-Ni/TiO2及其光催化性能研究

以Fe-Ni/TiO2为催化剂,采用流化床化学气相沉积法(FBCVD)在TiO2表面原位生长碳纳米管(CNT),得到CNT/Fe-Ni/TiO2复合光催化剂.通过SEM、XRD、UV-Vis等方法表征其结构和性能,以亚甲基蓝溶液降解为模型考察其光催化性能.结果表明:Fe-Ni/TiO2催化剂在FBCVD过程中,镍主要起到了CNT生长催化活性位的作用;在生长CNT后的复合光催化剂中,比例较低的Fe3+主要作为电子俘获剂,抑制TiO2光生电子空穴的复合;Ni和CNT共同起到将电子迅速地从TiO2中导出,从而降低光生电子空穴复合几率的作用.三者的协同作用显著改善了TiO2的光催化性能.其中Fe和Ni掺杂量分别为0.25mol%和4.75mol%样品的光催化活性较高,生长CNT后得到的复合光催化剂对亚甲基蓝的降解效率较纯TiO2提高约70%.     

TiO2纳滤膜对重金属离子的截留性能

采用孔径为1.5 nm的TiO2纳滤膜,在压力0.4～0.8 MPa,pH值3～7的条件下,考察了膜对浓度范围为50～500 mg/L的Cu(NO3)2、Ni(NO3)2、ZnC12和CdC12四种单组份重金属溶液的截留性能.结果表明:除Cd2+外,膜对重金属离子的截留率随溶液浓度的增大先增大,当浓度达到200 mg/L后趋于稳定;升高压力膜的离子截留率会略有增加;当pH=6时,膜对Ni2+和Cd2+的截留率最低,而对Cu2+和Zn2+的截留率在pH=5～6时达到最高.TiO2膜对Cu2+、Ni2+、Zn2+和Cd2+的最高截留率分别可以达到96.9％、95.9％、92.5％和83.2％.     

碳纳米管化学镀镍及镍层形貌控制研究

利用胶体钯溶液对碳纳米管(CNTs)进行活化处理,再采用化学镀的方法在碳纳米管表面沉积金属镍镀层,得到镀镍碳纳米管(Ni/CNTs)。通过场发射扫描电子显微镜(FESEM)、能量色散谱(EDS)、X射线衍射(XRD)和分光光度法对Ni/CNTs和镀液进行表征,分析了镍镀层形成的热力学条件与过程。结果表明:CNTs化学镀镍层为非晶态；CNTs浓度及反应温度对Ni²⁺沉积速率无明显影响；NaOH和NaH₂PO₂浓度与Ni²⁺沉积速率呈线性关系；镍镀层生长符合球形融合模型,通过改变CNTs浓度,可对镀层形貌进行有效调控。     

山波苓搞癌作用分析

目的:探讨山波苓体外抗肿瘤作用的活性部位。方法:MTT染色法对山波苓五种溶剂提取物体外抗肿瘤活性进行初步的研究。结果:氯仿提取物对乳腺癌细胞(Bre-04)、神经癌细胞(N-04)、肺癌细胞(Lu-04)有较好的抑制生长作用,IC50分别为0.2699mg/ml、0.2634mg/ml、0.4961mg/ml,对肝癌细胞HepG2抑制作用差,IC50为0.9379mg/ml;山波苓石油醚提取部位对乳腺癌细胞(Bre-04)、神经癌细胞(N-04)、肺癌细胞(Lu-04)、肝癌细胞HepG2的IC50分别为0.5902mg/ml、0.5725mg/ml、0.7938mg/ml、0.6374mg/ml;乙酸乙酯提取物对肺癌细胞(Lu-04)有一定的抑制作用,IC50为0.7343mg/ml;水溶性提取物以及正丁醇提取物无明显抑制肿瘤作用。结论:山波苓氯仿提取部位、石油醚提取部位体外抗肿瘤作用较好,值得对其提取部位进一步分离纯化并研究其抗肿瘤作用机制。     

Liquid chromatography/mass spectrometry methods for distinguishing N-oxides from hydroxylated compounds

This study describes the application of liquid chromatography/mass spectrometry (LC/MS) methods for distinguishing between aliphatic and aromatic hydroxylations and between hydroxylations and N-oxidations. Hydroxylations and N-oxidations are common biotransformation reactions of drugs. Electrospray (ESI) and atmospheric pressure chemical ionization (APCI) were used to generate ions from liquid chromatographic effluents. ESI-MS, ESI-MS/MS, APCI-MS, and APCI-MS/MS experiments were performed on several metabolites and derivatives of loratadine (a long-acting and nonsedating tricyclic antihistamine) using an ion trap mass spectrometer (LCQ) and a triple-quadrupole mass spectrometer (TSQ). The observations are as follows: (1) LC/ESI-MS produced predominantly [M + H]+ ions with minor fragmentation. (2) LC/ESI-MS/MS data, however, showed a predominant loss of water from metabolites with aliphatic hydroxylation while the loss of water was not favored when hydroxylation was phenolic. N-Oxides (aromatic and aliphatic) showed only a small amount of water loss in the MS/MS spectra. (3) Under LC/APCI-MS conditions, aliphatic hydroxylation could be readily distinguished from aromatic hydroxylation based on the extent of water loss. In addition, N-oxides produced distinct [M + H - O]+ ions. These [M + H - O]+ ions were not produced in the APCI-MS spectra of hydroxylated metabolites. (4) Similar to the ESI-MS/MS spectra, the APCI-MS/MS spectra from the (M + H)+ ions of N-oxides yielded a small amount of water loss but no [M + H - O]+ ions. These results indicate that LC/APCI-MS can be used to distinguish between hydroxylated metabolites and N-oxides.     

Co2+与(Co2++Ni2+)摩尔浓度比对Ni-Co-P合金化学镀层组织与性能的影响

通过改变化学镀液中金属离子Co2+与(Co2++Ni2+)的摩尔浓度比获得了不同的Ni-Co-P合金化学镀层。研究了钴的引入对Ni-Co-P合金镀层的沉积速率及组织性能的影响。结果表明:随着Co2+与(Co2++Ni2+)摩尔浓度比的增加,镀层的沉积速率不断下降,镀层中P含量逐渐减小,镀层表面的胞状组织由大到小;镀层硬度随Co2+与(Co2++Ni2+)摩尔浓度比的增加先升高后降低,Co2+与(Co2++Ni2+)摩尔浓度比为0.4时,硬度最大达554.12HV,镀层具有优良的耐硫酸腐蚀性能,Co2+与(Co2++Ni2+)摩尔浓度比为0.6时,镀层耐蚀性最强。     

Development and evaluation of a candidate reference method for the determination of total cortisol in human serum using isotope dilution liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry

Cortisol is an important diagnostic marker for the production of steroid hormones, and accurate measurements of serum cortisol are necessary for proper diagnosis of adrenal function. A candidate reference method involving isotope dilution coupled with liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed and critically evaluated. An isotopically labeled internal standard, cortisol-d(3), was added to serum, followed by equilibration and solid-phase and ethyl acetate extractions to prepare samples for liquid chromatography/mass spectrometry electrospray ionization (LC/MS-ESI) and liquid chromatography/tandem mass spectrometry electrospray ionization (LC/MS/MS-ESI) analyses. (M + H)(+) ions at m/z 363 and 366 for cortisol and its labeled internal standard were monitored for LC/MS. The transitions of (M + H)(+) --> [(M + H)(+) - 2H(2)O] at m/z 363 --> 327 and 366 --> 330 were monitored for LC/MS/MS. The accuracy of the measurement was evaluated by a comparison of results of this candidate reference method on lyophilized human serum reference materials for cortisol [Certified Reference Materials 192 and 193] with the certified values determined by gas chromatography/mass spectrometry reference methods and by a recovery study for the added cortisol. The results of this method for total cortisol agreed with the certified values within 1.1%. The recovery of the added cortisol ranged from 99.8% to 101.0%. This method was applied to the determination of cortisol in samples of frozen serum pools. Excellent precision was obtained with within-set CVs of 0.3%-1.5% and between-set CVs of 0.04%-0.4% for both LC/MS and LC/MS/MS analyses. The correlation coefficients of all linear regression lines ranged from 0.998 to 1.000. The detection limits (at a signal-to-noise ratio of approximately 3-5) were 10 and 15 pg for LC/MS and LC/MS/MS, respectively. This method, which demonstrates good accuracy and precision, and is free from interferences from structural analogues, qualifies as a candidate reference method and can be used as an alternative reference method to provide an accuracy base to which the routine methods can be compared.     

A screening method for antigen-specific IgE using mast cells based on intracellular calcium signaling

A simple screening method is presented for the measurement of antigen-specific IgEs in sera in which mast cells are used. This method is based on the intracellular calcium signal in mast cells induced by cross-linking the surface high-affinity Fc receptors (FcepsilonRIs) with IgEs and multivalent antigens. When a serum containing various antigen-specific IgEs is added to the mast cell suspension, various antigen-specific IgEs are captured by FcepsilonRIs on the cell surface. However, the required antigen-specific IgE can be specifically detected after the addition of the corresponding antigen. The resulting increase in intracellular calcium concentration ([Ca2+]i), monitored by Ca2+-fluorometry, was found to be an analytical measure for the screening of IgEs. Two kinds of rodent mast cells, cell-lined RBL2H3 cells and primary cultured BMMCs, were used as a representative model system of mast cells. A DNP hapten (DNP35-HSA) and ovalbumin (OVA) were chosen for illustrative antigens, and these antigen-specific IgEs (DNP-specific IgE, OVA-specific IgE) in the corresponding rodent sera were target antibodies. It was found that [Ca2+]i increased linearly with IgE concentrations ranging from 25 to 5000 ng/mL for DNP-specific IgE and from 5 to 50 ng/mL for OVA-specific IgE. For these dynamic ranges, optimum concentrations of antigens were found to be 10 ng/mL and 1 microg/mL for DNP35-HSA and OVA, respectively. It was concluded that by monitoring the increase of [Ca2+]i in mast cells, we could determine the antigen-specific IgEs. The present immunological assay based on the Ca2+ signal transduction in mast cells offers new possibilities for efficient screening of antigen-specific IgEs and the immunogenicity of IgE in sera.     

The importance of chromatographic separation in LC/MS/MS quantitation of drugs in biological fluids: detection,characterization, and synthesis of a previously unknown low-level nitrone metabolite of a substance P antagonist

The observation of an interference peak in plasma samples from dogs dosed with compound I led to the discovery of an unidentified metabolite. The unknown metabolite had the same molecular weight as the parent drug, and their fragmentation profiles were also quite similar. LC/MS/ MS analysis of the plasma extracts of dogs and rats dosed with I and its deuterium-labeled analogue suggested a nitrone structure for the unknown metabolite. Synthesized nitrone matched the unknown metabolite with identical retention time and nearly identical fragmentation profile. The nitrone slowly decomposed in acidic aqueous solution at ambient temperature and also underwent in-source, thermal-induced hydrolysis during electrospray ionization mass spectrometric analysis. The reaction of the nitrone with diethyl acetylenedicarboxylate readily generated a [2 + 3] cycloaddition product. The example shown here clearly demonstrates that precautions must be taken when LC/MS/MS quantitation is conducted in the selected reaction monitoring mode.     

Method development for metaproteomic analyses of marine biofilms

The large-scale identification and quantitation of proteins via nanoliquid chromatography (LC)-tandem mass spectrometry (MS/MS) offers a unique opportunity to gain unprecedented insight into the microbial composition and biomolecular activity of true environmental samples. However, in order to realize this potential for marine biofilms, new methods of protein extraction must be developed as many compounds naturally present in biofilms are known to interfere with common proteomic manipulations and LC-MS/MS techniques. In this study, we used amino acid analyses (AAA) and LC-MS/MS to compare the efficacy of three sample preparation methods [6 M guanidine hydrochloride (GuHCl) protein extraction + in-solution digestion + 2D LC; sodium dodecyl sulfate (SDS) protein extraction + 1D gel LC; phenol protein extraction + 1D gel LC] for the metaproteomic analyses of an environmental marine biofilm. The AAA demonstrated that proteins constitute 1.24% of the biofilm wet weight and that the compared methods varied in their protein extraction efficiencies (0.85-15.15%). Subsequent LC-MS/MS analyses revealed that the GuHCl method resulted in the greatest number of proteins identified by one or more peptides whereas the phenol method provided the greatest sequence coverage of identified proteins. As expected, metagenomic sequencing of the same biofilm sample enabled the creation of a searchable database that increased the number of protein identifications by 48.7% (≥1 peptide) or 54.7% (≥2 peptides) when compared to SwissProt database identifications. Taken together, our results provide methods and evidence-based recommendations to consider for qualitative or quantitative biofilm metaproteome experimental design.     

超声波化学镀Co-Ni-P工艺

Co-Ni-P化学镀层具有很多优点,在超声波辅助下化学镀层更光亮、细腻。在超声波辅助下在铝黄铜表面化学镀Co-Ni-P合金镀层,研究了镀液主盐浓度比、还原剂浓度、配位剂浓度、缓冲剂浓度、温度、pH值对沉积速率的影响,确定了超声波辅助下获得最佳沉积速率的化学镀Co-Ni-P工艺。结果表明:最佳工艺为11.0g/LNiCl2.6H2O,5.5g/LCoCl2.6H2O,17g/LNaH2PO2.H2O,50g/LNa3C6H5O7.2H2O,48g/LNH4Cl,温度75℃,pH值9.0;超声波化学镀层为非晶态结构,分布更均匀,表面更光亮。     

Molecular weight distributions of heavy aromatic petroleum fractions by Ag+ electrospray ionization mass spectrometry

The ability of electrospray ionization mass spectrometry (ESI MS) to analyze heavy aromatic petroleum fractions using silver nitrate as a reagent compound to form characteristic adduct ions has been examined. The complexation of aromatic compounds containing long alkyl substituents with the silver ion leads to the formation of abundant adduct ions such as [M + Ag]+ and [2M + Ag]+. The concentration of the [2M + Ag]+ ions can be reduced by increasing the sampling cone voltage. Molecular ions and other adduct ions may also be formed depending on the structure of the aromatic molecule. Results obtained from the analysis of representative heavy petroleum fractions and vacuum residues by the Ag+ ESI MS method and conventional ionization methods were in good agreement. The current method extends the applicability of electrospray ionization to the analysis of neutral hydrocarbons in heavy aromatic petroleum fractions. It is simple and compatible with widely available LC/MS instrumentation. The extreme complexity of the Ag     

Microfluidic selection and retention of a single cardiac myocyte, on-chip dye loading, cell contraction by chemical stimulation, and quantitative fluorescent analysis of intracellular calcium

A microfluidic method to study the contraction of a single cardiac myocyte (heart muscle cell) has been developed. This method integrates various single-cell operations as well as on-chip dye loading, and quantitative analysis of intracellular calcium concentration, [Ca2+]i. After the channel enlargement by on-chip etching to accommodate large-sized cardiac myocytes, a single cell is selected and retained at a V-shaped cell retention structure within the microchip. Owing to the fragile property of the cardiac myocytes that could easily be damaged by centrifugation, the calcium-sensitive fluorescent dye was loaded in the cell by on-chip dye loading. This on-chip method minimized the damage to the cells from the use of a centrifuge in the conventional method and provided a way of cellular analysis of fragile cells. Subsequently, quantitative analysis of [Ca2+]i of a single cardiac myocyte by fluorescence measurement was achieved for the first time in a microfluidic chip, thanks to the intracellular calcium stimulant of ionomycin. The resting [Ca2+]i of the cardiomyocyte determined was consistent with the literature value. From the spontaneous contraction study, it was found that fluorescence intensity cannot represent the [Ca2+]i variation accurately, which implied the importance of the quantitative analysis of [Ca2+]i.