双内标毛细管GC-MS定量分析尼古丁

建立了以樟脑和间甲酚为双内标物毛细管GC-MS定量分析尼古丁的方法.色谱分析数据表明:色谱条件不变时,双内标得到的尼古丁定量曲线的线性相关系数明显优于单内标法.这可能是由于双内标可使样品分压在进样室快速均匀分布所致.文中还讨论了内标物浓度以及内标物相对分析物的出峰位置对方法线性的影响.结果表明,采用双内标法进行毛细管定...     

MALDI-TOF监控高分子合成反应

基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)自发展以来,虽多用于多肽、蛋白质等生物分子的分析测定.但实践证明,MALDI-TOF还是高分子化合物的有效表征和结构鉴定手段.本文将MALDI-TOF用于监控甲基丙烯酸丁酯末端酯基的水解反应,实验结果表明:MALDI-TOF为高分子反应的监测与控制,提供了快速、灵敏、有效的手段.通过不同条件下反应物与产物比例的监测,考察不同反应条件、不同反应时间对反应产率的影响,利用MALDI-TOF测定数据优化了高分子反应的实验条件.     

叠加内标法色谱定量分析

 叠加内标法是指在色谱定量分析中将内标法与叠加法结合的一种新的定量方法。叙述了叠加内标法定量的理论依据 ,规定了其操作步骤 ,并详细说明了这种方法的适用条件和优缺点。     

Quantitative Determination of Uric Acid in Serum by Reversed-Phase Liquid Chromatography Using an Internal Standard

Abstract

A rapid and sensitive high-performance liquid chromatography procedure is reported for the analysis of uric acid in serum. The method employs a single treatment of serum with cyclohexane and an excess of ammonium sulfate to eliminate endogenous serum components which easily contaminate the chromatographic system. Reversed-phase chromatography is performed at room temperature using a binary-solvent gradient, monitoring the effluent at 290 nm. Quantitation is based on peak-height ratio of uric acid to internal standard (β-hydroxyethyltheophylline). A linear response is obtained to 160 mg/1. Within-day and between-day precisions for the mean of this range varied from 0.59 to 1.1% and from 0.71 to 1.2%, respectively. Numerous potential interfering substances tested do not interfere.     

液相色谱-串联质谱外标法定量分析中基质效应的评估

以苯巯基尿酸为目标化合物,考察了色谱分离条件(即强洗脱、弱洗脱及梯度洗脱)对采用外标法定量的液相色谱-串联质谱(LC-MS/MS)分析中基质效应的影响。试验表明:①对无基质标准溶液的分析,以强洗脱条件下的灵敏度最高,且可实现高通量分析;②对同一基质不同浓度的试液,弱洗脱条件的基质效应明显改善,而梯度洗脱的结果最好;③对不同来源的基质和不同浓度的试验,在3种不同的色谱分离条件中以梯度洗脱效果最好,提高了分离度,减少了共流出,有效地降低了基质效应对目标物分析的影响。因此,在LC-MS/MS分析中,色谱分离条件的改善对降低基质效应对测定的影响十分重要。     

C60衍生物的MALDI-TOFMS分析

C_(60)衍生物在超导、非线性光学、催化、材料和生物活性等方面有巨大的潜在应用价值。C_(60)衍生物大多为固体,蒸汽压较低,采用需要加热才能够使样品气化电离的质谱或"硬"电离质谱方法进行测定,易造成C_(60)衍生物分解并释放出配体。近年来国内外应用基体辅助激光解吸软电离质谱法成功分析了许多不同类型的C_(60)衍生物如卤化C_(60)酰胺化C_(60)芳基化C_(60)、C_(60)部花菁、煤基C_(60)烟灰萃取产物、金属C_(60)衍生物以及C_(60)乙二胺膜等。本文报道采用MALDI-TOFMS法分析C_(60)酯衍生物和C_(60)吡咯烷衍生物的结果。     

X-射线衍射内标法定量滑石粉中的温石棉

对比α-氧化铝、氧化镁、硅粉三者的结晶属性,选择晶相良好且与温石棉特征峰无重叠的单质硅为内标物,建立温石棉衍射吸收标准曲线,该曲线在温石棉含量0%～9.3%具有较好的回归性。使用曲线分析已知温石棉含量的分析样品,测得温石棉含量为1.70%,与理论值(温石棉添加量:1.68%)的相对误差RE为0.99%。分析该方法检出限与定量限分别为0.10%和0.30%。与以金属基底建立曲线的石棉定量方法相比,内标法有较好灵敏度和准确度,是一种准确可靠的石棉定量方法。     

液相色谱-串联质谱法检测水产品中磺胺类和喹诺酮类药物残留

建立了高效液相色谱-串联质谱(HPLC-MS/MS)法检测水产品中磺胺类、喹诺酮类等合成抗菌剂残留的方法。样品以乙腈为提取剂,提取物经脱脂、净化、浓缩后,用流动相溶解。用氘代试剂内标法定量,高效液相色谱-串联质谱法测定。本方法通过梯度洗脱将12种磺胺类、喹诺酮类进行良好分离。标准曲线线性范围0.05~0.8 mg/L,线性相关系数r=0.987 5~0.999 1,回收率为61%~104%,相对标准偏差(RSD)为4.75%~6.12%(n=5),检出限为10~50μg/kg。     

Analysis of Bacterial Lipooligosaccharides by MALDI-TOF MS with Traveling Wave Ion Mobility

Lipooligosaccharides (LOS) are major microbial virulence factors displayed on the outer membrane of rough-type Gram-negative bacteria. These amphipathic glycolipids are comprised of two domains, a core oligosaccharide linked to a lipid A moiety. Isolated LOS samples are generally heterogeneous mixtures of glycoforms, with structural variability in both domains. Traditionally, the oligosaccharide and lipid A components of LOS have been analyzed separately following mild acid hydrolysis, although important acid-labile moieties can be cleaved. Recently, an improved method was introduced for analysis of intact LOS by MALDI-TOF MS using a thin layer matrix composed of 2,4,6-trihydroxyacetophenone (THAP) and nitrocellulose. In addition to molecular ions, the spectra show in-source “prompt” fragments arising from regiospecific cleavage between the lipid A and oligosaccharide domains. Here, we demonstrate the use of traveling wave ion mobility spectrometry (TWIMS) for IMS-MS and IMS-MS/MS analyses of intact LOS from Neisseria spp. ionized by MALDI. Using IMS, the singly charged prompt fragments for the oligosaccharide and lipid A domains of LOS were readily separated into resolved ion plumes, permitting the extraction of specific subspectra, which led to increased confidence in assigning compositions and improved detection of less abundant ions. Moreover, IMS separation of precursor ions prior to collision-induced dissociation (CID) generated time-aligned, clean MS/MS spectra devoid of fragments from interfering species. Incorporating IMS into the profiling of intact LOS by MALDI-TOF MS exploits the unique domain structure of the molecule and offers a new means of extracting more detailed information from the analysis.

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Flash Pyrolysis-Gas Chromatography Using an Internal Standard For Quantitative Determination Of Resin Content in A Model Thermosetting Composite System

Abstract

It has been demonstrated, by operation on a series of thermosetting phenolic-asbestos composites, that pyrolysis gas chromatography may be applied as a general method for estimating polymer content in cured composite materials. A platinum filament is employed for flash pyrolysis and quantitative estimation is based on the internal standard method.     

Synthesis and MALDI-TOF MS Analysis of Protected Oligosaccharide Components of N-Glycoproteins

Essential components of N-glycoproteins were synthesized in octyl glycoside form starting from 3-O-allyl-D-glucose derivative. The β-mannosidic linkage was formed by the oxidation reduction method. MALDI-TOF mass spectrometry and its post-source decay (PSD) mode were used for identification and structural elucidation of protected synthetic oligosaccharides related to N-glycoproteins. Most fragments, identified in the PSD spectra, were products of the cleavage of glycosidic bonds.     

Internal Standard Quantitative Method Of Tetrodotoxin By Capillary Electrophoresis

ABSTRACT

An internal standard quantitative method of tetrodotoxin by capillary electrophoresis was developed, ephedrine hydrochloride being chosen as the internal standard of tetrodotoxin. The calibration curve was built with the correlation coefficient of 0.9997, and the linear range was from 0.01 mg.ml^?1 to 10.0 mg-ml^?1 with the detection limit of 1.0 x 10^?3 mg.m1^?1. RSD of the migration time of ephedrine hydrochloride and tetrodotoxin were all less than 1.0 %. The method was applied to determine tetrodotoxin in the poisonous frilled fish samples.     

Using MALDI-TOF MS coupled with a high-mass detector to directly analyze intact proteins in thyroid tissues

Protein analysis is vital for biological and clinical research, but the measurement of unseparated, intact and high-mass proteins is also a challenging task by mass spectrometry-based methods. Here, we present a protocol for rapid and high-throughput analysis of intact proteins in tissue samples using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDITOF MS) combined with a high-mass detector platform. The method involves tissue specimens that undergo a simple protein extraction before MALDI-MS analysis. Using this method, the high abundance proteins in human thyroid carcinoma and paracarcinoma tissues were successfully investigated, and the mass spectra of the tissues of the 30 illustrated thyroid cancers showed remarkable differences. The peak intensity revealed a significant increase in human albumin in thyroid carcinoma tissues(p0.05). To validate the feasibility and credibility of this method, label-free proteomics quantitative analysis and Western blotting were used to relatively quantify the proteins in these tissues. Those results demonstrated a nearly 3-fold difference in human albumin levels between thyroid carcinoma and para-carcinoma tissues, which were consistent with the results of our method. The advantages of our method are easy sample handling, remarkable reproducibility and the ability to analyze high-mass proteins without digestion, which make them have the potential to be used in biological research and in clinical practice.     

表面增强拉曼光谱内标法测定水杨酸浓度

采用电化学方法处理银电极表面,使之产生具有表面增强拉曼光谱(SERS)的活化效应,进而利用水杨酸(SA)吸附于粗糙活化的银电极表面所产生的SERS光谱,进行SA含量的测定。选择硫氰化钾(KSCN)为内标物,以SA的苯环呼吸振动峰(1 036 cm-1)与SCN-的特征峰(2 120 cm-1)强度比值作为相对强度,其相对强度与水杨酸的浓度在4×10-6~3×10-4mol/L范围内呈良好的线性关系,其线性系数为0.993 1;检出限为2.04×10-6mol/L;加标回收率为85.4%~114.6%。方法具有测定简便、内标物干扰小、绿色无害等特点,可进一步推广于其他特殊物质的测定。     

激光拉曼光谱内标法直接测定甲醇浓度

利用甲醇溶液中的CH3对称伸缩峰(2843 cm-1)的激光拉曼光谱峰,同时以本底溶液水为内标物,组成相对强度,对溶液中的甲醇含量进行了测定研究,建立了激光拉曼光谱直接法测定甲醇含量的方法,其线性范围为5%~40%,线性相关系数为0.9966,检出限为0.29%。并对模拟样品和制备液相色谱待回收废液进行了甲醇浓度的测定,其浓度分别为13.7%、9.41%和58.8%,RSD分别为0.78%、0.75%和1.34%。该方法简单、操作方便,无需添加其它任何化学试剂等优点。     

Synthesis and characterization by MALDI-TOF MS of polycyclic siloxanes derived from p-substituted novolac resins by MALDI-TOF MS

Novel polycyclic siloxane resins were prepared from phenol-formaldehyde novolac type resins by reacting them with dialkyl or diaryl dichlorosilanes under anhydrous and high dilution conditions. The formation of polycyclic species was confirmed by the detection of absolute masses by MALDI-TOF mass spectrometry. ¹H- and ²⁹Si-NMR confirmed the substitutions of phenolic hydroxy groups by siloxane bonds. Curing studies were conducted on the polycyclic siloxane resins as well as on the polycyclic siloxane resins incorporated into two types of polysiloxane gums. A trace amount of potassium hydroxide was used as a catalyst for the crosslinking of these systems. The blend of polysiloxane with 30 wt % polycyclic siloxane was found to be stable at the curing temperature. Differential scanning calorimetry and thermogravimetric analysis techniques were used to study the thermal profiles of these systems. © 1998 John Wiley & Sons, Inc. J. Polym. Sci. A Polym. Chem. 36: 2429–2437, 1998     

Micro Assay for Malondialdehyde in Human Serum by Extraction-Spectrophotometry Using an Internal Standard

Malondialdehyde (MDA) is a widely accepted biomarker of lipid peroxidation. Thus, the measurement of MDA in clinical samples is useful in the evaluation of oxidative stress. In this study, a micro-extraction-spectrophotometric assay was developed, based on the formation of MDA–thiobarbituric acid (TBA) adduct. To enhance the analytical performance, Erioglaucine A was used as an internal standard (IS), and the first derivative spectra were obtained. The volume of serum sample was 20µL and the total volume of aqueous phase 420µL (200µL of 0.6% TBA in acetic acid, pH 2.5 and 200µL of 6.25·10^–4% IS). The extraction of adduct and IS was carried out with 300µL of aliquat 336 (0.06%) in ethyl acetate. The analytical signal S was defined as the ratio between the first derivative absorbances measured at 543.1nm (adduct) and 644.4nm (IS). In the calibration range up to 10µmolL^–1 MDA, the linear regression coefficient was 0.9998. The quantification limit was 0.19µmolL^–1 and the CV values for 2µmolL^–1 and 5µmolL^–1 MDA, respectively, were 0.8% and 0.7%. The procedure was applied to the analysis of diabetic sera and the results compared with those obtained by HPLC (derivatization with 2,4-dinitrophenylhydrazine). Lower HPLC results (about 15%) indicated that interferences from other TBA reactive substances had not been completely eliminated by the extraction procedure and derivatization of spectral data. On the other hand, the micro-procedure presents important advantages: it is simple, precise and environmentally friendly (small amounts of reagents), which makes it readily adaptable to the analysis of large sample series. The feasibility of micro-assay in the evaluation of lipid peroxidation status was demonstrated in the analysis of 156 serum samples from diabetic patients divided into three groups according to the stage of development of typical complications.     

Using oxidized carbon nanotubes as matrix for analysis of small molecules by MALDI-TOF MS

Oxidized carbon nanotubes are tested as a matrix for analysis of small molecules by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Compared with nonoxidized carbon nanotubes, oxidized carbon nanotubes facilitate sample preparation because of their higher solubility in water. The matrix layer of oxidized carbon nanotubes is much more homogeneous and compact than that of nonoxidized carbon nanotubes. The efficiency of desorption/ionization for analytes and the reproducibility of peak intensities within and between sample spots are greatly enhanced on the surface of oxidized carbon nanotubes. The advantage of the oxidized carbon nanotubes in comparison with alpha-cyano-4-hydroxycinnamic acid (CCA) and carbon nanotubes is demonstrated by MALDI-TOF-MS analysis of an amino acid mixture. The matrix is successfully used for analysis of synthetic hydroxypropyl beta-cyclodextrin, suggesting a great potential for monitoring reactions and for product quality control. Reliable quantitative analysis of jatrorrhizine and palmatine with a wide linear range (1-100 ng/mL) and good reproducibility of relative peak areas (RSD less than 10%) is achieved using this matrix. Concentrations of jatrorrhizine (8.65 mg/mL) and palmatine (10.4 mg/mL) in an extract of Coptis chinensis Franch are determined simultaneously using the matrix and a standard addition method.     

表面增强拉曼光谱内标法测定亚胺硫磷农残含量

采用柠檬酸钠还原法制备了具有表面增强拉曼散射(SERS)活性的纳米银溶胶,进而利用亚胺硫磷(Phosmet)吸附于纳米银胶表面使其产生SERS光谱,并对Phosmet的含量进行了测定。选择硫氰化钾(KSCN)为内标物,通过不同浓度亚胺硫磷的拉曼特征峰(S—P伸缩振动503 cm-1)与内标物SCN-的特征峰(2 120 cm-1)峰高的相对强度比,建立线性回归方程。结果表明:Phosmet的线性范围为5.0×10-7~1.2×10-5mol·L-1,相关系数为0.997 8;按7次空白实验所对应响应值IR的3倍标准偏差,计算得检出限为2.82×10-7mol·L-1;回收率为88.7%~110.2%,相对标准偏差(RSD)不大于8.0%。方法测定简单、操作方便、内标物干扰小,可应用于其他农残的分析测定。