湿热证动物模型微量元素、维生素E变化及清香散的调节作用

采用多因素造模方法复制成湿热证动物模型，观察了动物模型微量元素Zn、Cu、Fe、Se和维生素E代谢水平的变化。结果显示，模型动物血清Zn下降（P＜0．05），Cu升高（P＜0．05或P＜0．01），Fe变化不大（P＞0．05），血Se水平下降（P＜0．05），血浆维生素E含量减少（P＜0．01）。经清热祛湿的经验方清香散治疗后，治疗I组动物血清Zn、血Se、血浆维生素E明显升高（P＜0．05），血清Cu下降（P＜0．05）；治疗Ⅱ组血浆血清Cu有变化（P＜0．05）外，其余变化不大。     

痤疮患者血清中微量元素含量的变化

测定了52例痤疮患者血清Ca,Mg,Zn,Cu,Fe五种元素的含量。结果表明，痤疮患者血清Zn水平显著低于正常人（P＜0．001），血清Ca,Mg,Cu,Fe水平则与正常人无差别（P＞0．05）。部分病人口服补充锌剂后痤疮症状有所好转。提示痤疮患者应重视体内微量元素含量的变化，并做适当的补充。     

差热分析法测定化工品、药品纯度

研究了差热分析（DSC）法测定化工品、药品纯度的条件，测定的最佳条件为升温速度0．7℃/min，样品量2－3mg，保护气流速20－40mL/min。测定结果的RSD为0．02％，F检验及t检验的结果表明，该方法与容量法无显著差异。该方法简便、快速、准确，适合化工品、药品的定量测定。     

复方锗制剂治疗肿瘤患者及气阴两虚症病人124例疗效分析

复方锗制剂是广西肿瘤防治研究所研制的新中药制剂，具有益气养阴、补血活血之功效，适用于气阴不足引起的神疲乏力、气短自汗、口干咽燥等症的治疗和肿瘤病人放、化疗的辅助治疗，以减轻放、化疗的毒副作用，增强机体的免疫功能。共治疗124例（其中一般气阴两虚症60例，接受放、化疗的肿瘤病人64例）同时用生脉饮治疗60例一般人，用消-TU治疗32例肿瘤病人作对照。结果：一般病人和肿瘤病人的有效率分别为93．3％和95．5％；生脉饮对照组有效率为87.0％；消-TU对照组为75．0％。对一般病例的疗效复方锗制剂与生脉饮无统计学差异（P＞0．05）；对肿瘤病人的疗效明显优于消-TU对照组（P＜0．05）。对肿瘤病人因放、化疗而引起的机体免疫功能抑制有一定的辅助治疗作用，治疗后体液中IgG、IgA、IgM、WBC、NKC及ERFT均有提高作用，尤其以NKC的提高有显著意义（P＜0．05％）。     

肾炎及肾衰患者血中金属元素含量的变化

用原子吸收法或极谱法测定了对例慢性肾炎病人（A组）和24例慢性肾衰病人(B组)及40名健康人（对照组，C组）全血中的Fe、Zn、Cu、Mn、Se、Pb和Ga含量。结果表明，A组与C组比较．血中Fe的含量A组明显低于C组（P＜0.05），Zn、Mn和Se的含量A组均非常明显低于C组（P＜0.01），Pb含量A组非常明显高于C组（P＜0．01），Cu和Ca含量两组间均无明显差异（P＞0.05）；B组与C组比较，Fe含量B组明显低于C组（P＜0.06），Se含量则B组明显高于C组（P＜0.05）；Zn、Mn、Cu、Ga含量B组均非常明显低于C组（P＜0.01），而Pb含量则B组非常明显高于C组（P＜0．01）。     

ASE-HPLC检测某型号球形发射药中的叠氮硝胺、硝化甘油、Ⅱ号中定剂含量的研究

采用快速溶剂萃取（ASE）技术和高效液相色谱法测定某球形药中叠氮硝胺（DIANP）、硝化甘油（NG）和Ⅱ号中定剂（C2）的含量．ASE提取条件：二氯甲烷做萃取溶剂,萃取温度100℃,静态萃取10min,萃取2次．HPLC测定条件：YWGC18柱（150×4．6mm,10μm）,以甲醇和水作为流动相,梯度洗脱,流速1 mL／min,检测波长210nm．测定结果表明DIANP、NG、C2平均回收率分别为99．6％、100．3％、99．4％,RSD分别为0．7％、0．8％、0．9％（n=5）,检出限分别为2.1、1.5和0．2mg／L,线性范围分别为0．02～0．98g／L,0．03～1．38g／L,0．002～0．124g／L．用此方法共检测某批球形发射药样品5份,检测结果与滴析-HPLC法检测结果相当．     

Cy-5标记脱氧核糖核酸荧光毛细分析法的研究

在荧光毛细分析法（FCA）的基础上开发了一种用于DNA快速检测的DNA-FCA法。在毛细管内表面将DNA探针固定化，制成荧光毛细生物反应器（DNA-CBR）。测定时，用DNA-CBR吸入含Cy-5标记的靶DNA样品液进行杂交反应，然后在646nm激发波长、664nm发射波长下进行荧光测定；Cy-5标记的靶DNA浓度在0．1～1．0μmol／L之间线性良好（y=139．73x＋39．613，r=0．9985）；RSD〈5．5％；检出限为0．17pmol，样品用量10μL；DNA-CBR能够重复使用6次。本方法可用于靶DNA的定性和定最检测。     

高效液相色谱法测定复方板蓝根颗粒剂中靛玉红含量

为通过测定复方板蓝根颗粒中靛玉红的含量控制制剂内在质量，采用高效液相色谱法，以V（甲醇）＋V（水）=75＋25为流动相，流速0．5mL／min，检测波长为290nm。结果表明，靛玉红含量与峰面积在0．0172～0．1370μg范围内，线性关系良好（r=0．9999），平均回收率为99．4％（RSD=2．60％，n=5），方法重现性好（RSD=2．90％），可避免制剂中其他化学成分的干扰，专属性好，且操作简便。     

高硒情况下维生素E对老龄大鼠血清中抗氧化作用的影响

研究了高Se或高维生素E下，老龄大鼠体内抗氧化活性的变化以及高Se下维生素E对其体内抗氧化水平的影响。将十二月龄Wistar大鼠32只随机分成4组：（1）正常饲料组；（2）高Se饲料组；（3）高维生素E饲料组；（4）高Se高维生素E饲料组。饲至十周末，处死大鼠，按试剂盒要求取其血液制成血清测GSH－Px,SOD活性，MDA含量。结果表明，高Se或高维生素E下饲养下的大鼠与对照组比较，血清中GSH－Px活性升高（P＜0．05）。SOD活性未见改变，MDA含量分别减少50％和56％，有显著差异（P＜0．01）；维生素E与高Se合用与高Se组比较能进一步同GSH－Px,SOD活性（P＜0．01）。结论：高Se会升高老龄大鼠的抗氧化活性，而与维生素E合用更加升高老龄大鼠血液中抗氧化活性。     

微量元素锌、血清胱抑素 C在糖尿病肾病早期诊断的意义

目的：探讨早期糖尿病肾病患者（ DN）的微量元素锌、血清胱抑素C（ CysC）、血清肌酐（ SCr）和血清尿素（ BUN）水平的变化程度及意义。方法将糖尿病肾病患者分为3组：正常蛋白尿组,微量蛋白尿组,临床蛋白尿组。所有患者和对照组均空腹抽取静脉血检测微量元素锌、血清胱抑素C、血清尿素、血清肌酐。结果正常蛋白尿组、微量蛋白尿组、临床蛋白尿组微量元素锌均明显低于正常对照组,差异有统计学意义（ P＜0.05）。正常蛋白尿组、微量蛋白尿组、临床蛋白尿组血清胱抑素C水平均明显高于正常对照组,差异有统计学意义（ P＜0.05）。微量蛋白尿组、临床蛋白尿组血清肌酐检测值和正常对照组有明显差异,差异有统计学意义（ P＜0.05）。结论检测微量元素锌、血清胱抑素C对糖尿病肾病的早期肾损害有较高的临床价值,可作为早期糖尿病肾病诊断的参考指标。     

Quantification of trimethylsilyl derivatives of amino acid disease biomarkers in neonatal blood samples by gas chromatography-mass spectrometry

A novel analytical procedure was developed for the rapid determination of disease biomarkers of maple syrup urine disease (MSUD),L-valine,L-leucine,L-isoleucine, andL-phenylalanine in dried blood spots. Amino acids extracted from neonatal dried blood spots were rapidly derivatized with bis-(trimethylsilyl)trifluoroacetamide (BSTFA) and then analyzed by gas chromatography-mass spectrometry (GC-MS). Derivatization conditions and the method validation were studied: optimal derivatization conditions were acetonitrile as reaction solvent, a temperature of 100°C, and a reaction time of 30 min. The proposed method provided a detection limit lower than 2.0 μM, recovery between 92% and 106%, and relative standard deviation less than 8.0%. The method was further tested in screening for neonatal MSUD by determination ofL-valine,L-leucineL-isoleucine, andL-phenylalanine in blood samples. The experimental results show that GC-MS following BSTFA derivatization is a rapid, simple, and sensitive method for the determination of amino acid disease biomarkers in blood samples, and is a potential tool for fast screening of MSUD. Figure     

Hydroxyproline quantification for the estimation of collagen in tissue using multiple reaction monitoring mass spectrometry

Collagens are highly abundant mammalian proteins that contain a high content of hydroxylated amino acids such as hydroxyproline. We have exploited the high hydroxyproline content of collagen and developed a method for hydroxyproline quantification as a measure of collagen content in muscle samples. The novel method utilizes a highly selective and sensitive method of multiple reaction monitoring (MRM) by mass spectrometry. The analytical method is simple, rapid (5min), convenient (no derivatization), precise (<17% RSD), accurate (90-108%), sensitive (4.88nmol/L) and linear (R(2)>0.999) over three orders of magnitude (5-5000nmol/L).     

流动注射-抑制化学发光法测定岩白菜素

本文基于岩白菜素对鲁米诺-牛血清白蛋白体系化学发光信号显著的抑制作用,建立了快速灵敏测定岩白菜素的流动注射-化学发光分析新方法。实验发现,化学发光强度的降低值与岩白菜素质量浓度对数值呈良好的线性关系,方法的线性范围为3.0~5.0×10~5 pg/mL,检出限(3σ)为1.0pg/mL。当溶液流速为2.0mL/min时,完成一次分析过程仅需30s,采样频率120/h。本法用于片剂、人血清和尿液中岩白菜素的含量测定,回收率为98.1%~102.7%,相对标准偏差小于2.0%(n=7)。同时对化学发光反应机理进行了探讨。     

Microderivatization of 4-[18O]hydroxyproline and quantitation with a benchtop mass spectrometer

Accurate estimation of in vivo turnover rates of collagen is complicated by amino acid reutilization. It was previously shown that the ideal, non-recycling tracer was [18O]hydroxyproline synthesized in vivo. The analytical method for measuring turnover rates with [18O]hydroxyproline must include analyte quantitation for pool size determination and isotope ratio measurement for determining levels of label incorporation. For ease of use and widest availability, a benchtop gas chromatograph-mass spectrometer in the electron-impact ionization mode was chosen. Here we present a versatile procedure for hydroxyproline derivatization that is well suited for routine, large-scale determination of analyte concentrations and relative levels of 18O incorporation.     

Specific and Sensitive High Performance Liquid Chromatographic Determination of Alprazolam or Triazolam

Abstract

A specific and highly sensitive liquid chromatographic procedure has been developed for the rapid determination of intact alprazolam or triazolam in dog serum, using these structurally similar triazolobenzodiazepines as mutual internal standards. The procedure consists of (1) extracting one ml of alkali buffered serum with toluene, (2) evaporating an aliquot of the toluene to dryness, and (3) quantitating the redissolved residue by HPLC using ultraviolet detection (221 nm). Samples were chromatographed on a microparticulate reverse phase column using a mobile phase composed of acetonitrile: isopropanol: water (94:5:1) and a flow rate of 0.75 ml/min. Metabolites of alprazolam and triazolam did not interfere in the assay. The lower limit of detection was approximately 1 ng/ml of serum extracted. The utility of the analytical methodology for the determination of alprazolam or triazolam in pharmacokinetic studies in the dog was demonstrated.     

高效液相色谱-质谱联用测定胶原蛋白中的羟脯氨酸

建立了一种简单、快速地测定胶原蛋白中羟脯氨酸(HYP)的高效液相色谱-电喷雾质谱(HPLC-ESI/MS)方法。胶原蛋白经酸水解后,以乙腈-0.05%的三氟乙酸水溶液(体积比为5∶95)为流动相,以1.0 mL/min的流速在C8反相柱上进行分离。以茶氨酸为内标,利用质谱定性定量测定HYP。在电喷雾正离子模式下,对m/z 132和m/z 175离子进行选择离子监测。在11.7～117 mg/L范围内,HYP与内标物茶氨酸的峰面积比和HYP的质量浓度呈良好的线性关系,相关系数为0.9993。含量测定的相对标准偏差为1.87%,回收率为97.85%～101.76%。此方法流动相简单,分析时间短且无需衍生处理,抗干扰能力强,能准确快速地测定胶原蛋白中HYP的含量。     

Rapid determination of levofloxacin in pharmaceuticals and biological fluids using a new chemiluminescence system

A novel chemiluminescence method for the determination of levofloxacin is presented, which is based on the inhibitory effect of levofloxacin on the chemiluminescence reaction between luminol and myoglobin in a flow-injection system. The decrement of chemiluminescence intensity is linear with the logarithm of levofloxacin concentration over the range from 0.07 to 100.0 ng/mL (r ² = 0.9994), with the detection limit of 0.02 ng/mL (3σ). At a flow rate of 2.0 mL/min, a complete analytical process could be performed within 0.5 min, including sampling and washing, with a relative standard deviation of less than 3.0% (n = 5). The proposed procedure was applied successfully to the determination of levofloxacin in pharmaceutical preparations, human urine and serum without any pretreatment procedure.     

Voltammetric method for ultra-trace determination of total mercury and toxic metals in vegetables. Comparison with spectroscopy

A new procedure for the determination of mercury(II), copper(II), lead(II), cadmium(II) and zinc(II) traces in food matrices by square wave anodic stripping voltammetry and standard addition method is proposed. A rapid, inexpensive and multi-analyte analytical method suitable for food safety control is provided. Comestible vegetables were chosen as samples. A two-step, sequential determination was defined, employing two working electrodes: a gold electrode (GE) for mercury(II) and copper(II), and subsequently a hanging mercury drop electrode (HMDE) for copper(II), lead(II), cadmium(II) and zinc(II). No sample pre-treatment was needed. Spinach leaves, tomato leaves and apple leaves were employed as standard reference materials to optimize and defined the analytical procedure. The new method shows good selectivity, sensitivity, detectability and accuracy. A critical comparison with spectroscopic measurements is discussed. Spinach, lettuce and tomato samples sold on the market were analysed as real samples. Lead(II) and cadmium(II) concentration exceeded the relevant legal limits.      

Development of an electrospray ionization mass spectrometric method for the quantification of theophylline in horse serum

A rapid and selective method has been developed for the determination of theophylline in horse serum by LC-ESI/MS/MS. The analytical method includes a protein precipitation extraction for sample preparation, liquid chromatography separation technique and ionspray tandem mass spectrometry. The drug was extracted from serum using a protein precipitation with acetonitrile and the supernatants were analyzed using an LC-ESI/MS/MS instrument. The chromatography was performed using a 50 x 2.1 mm C(8) analytical column and an isocratic mobile phase composes of 60:40 acetonitrile-0.5% formic acid in water with a flow rate fixed at 350 microL/min. A linear (weighted 1/concentration) relationship was used to perform the calibration over an analytical range of 0.1-20 ppm. The intra-batch precision and accuracy at LLOQ, medium and high concentration were 11.7, 6.9 and 5.4% and 95.8, 107.8 and 95.8%, respectively, and the inter-batch precision and accuracy at LLOQ, medium and high concentration were 10.4, 7.9 and 7.3% and 97.3, 105.2 and 95.9%, respectively. This LC-ESI/MS/MS method for the determination of theophylline in horse serum has been proved to within generally accepted criteria used for bioanalytical assay and was used successfully during clinical investigation.