UHPLC-MS/MS测定人血浆中依匹哌唑浓度及其生物等效性研究

目的 建立简便灵敏的超高效液相色谱-串联质谱法(ultra performance liquid chromatography tandem mass spectrometry,UHPLC-MS/MS)测定人血浆中依匹哌唑浓度,并应用于2种片剂的生物等效性研究。方法 采用Waters Acquity UPLC BEH C₁₈色谱柱(2.1 mm×50 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈-甲醇(50∶50,含0.1%甲酸)(B)梯度洗脱,流速0.4 mL·min^–1,进样量为2μL,柱温40℃,以正离子MRM模式测定依匹哌唑(m/z 434.2→273.2)的浓度,依匹哌唑-d₈(m/z 442.4→281.3)作为内标,离子源为ESI源。血浆样本加入内标,加入甲醇后进行蛋白沉淀,取上清液稀释后进样检测。结果 依匹哌唑在0.2～50 ng·mL^–1呈线性关系,定量下限为0.2 ng·mL^–1,质控样品批内、批间精密度CV≤5.0%,准确度相对偏差在标示值–1.7...     

UPLC-MS/MS测定酒石酸伐尼克兰中基因毒性杂质N-亚硝基伐尼克兰

目的 建立酒石酸伐尼克兰原料药和片剂中基因毒性杂质N-亚硝基伐尼克兰超高效液相色谱-串联三重四级杆质谱(UPLC-MS/MS)的检测方法。方法 采用ACQUITY UPLC^? CSH^TM Phenyl-Hexyl(150 mm×3.0 mm,1.7μm)色谱柱;0.1%甲酸水溶液为流动相A,0.1%甲酸的甲醇溶液为流动相B,梯度洗脱;流速为0.45 mL·min^–1,柱温为50℃;采用ESI离子源正离子扫描,多反应监测(MRM)模式下,对基因毒性杂质进行定量检测。结果 杂质在0.10～10.04ng·mL^–1具有良好的线性关系;原料药的低、中、高3个浓度的加样回收率(n=3)分别为103.58%(RSD=3.30%),98.65%(RSD=2.73%),92.00%(RSD=1.98%);片剂的低、中、高3个浓度的加样回收率(n=3)为91.53%(RSD=0.78%),96.76%(RSD=3.12%),93.01%(RSD=2.21%);检测限与定量限分别为0.014 ng·mL^–1<...     

杏仁腈在常用溶剂中的稳定性考察及其在健儿清解液中的含量测定

目的 采用HPLC考察杏仁腈在常用溶剂中的稳定性并测定健儿清解液中杏仁腈的含量。方法 采用Agilent TC-C₁₈(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%磷酸溶液(23∶77)为流动相,流速为1.0 mL·min^–1,柱温为30℃,检测波长为207 nm。以峰面积的下降率为指标,考察以甲醇、95%乙醇、乙腈、水、pH值2.0～6.0的磷酸溶液及1.0%冰醋酸乙腈为溶剂制备的杏仁腈溶液的稳定性。结果 杏仁腈在甲醇、95%乙醇及水中不稳定,而在乙腈中相对稳定;使用pH值为2.0～3.5的磷酸溶液及1.0%冰醋酸乙腈溶液配制的杏仁腈对照品溶液,可在12 h内保持稳定。杏仁腈质量浓度在1.033～294.987μg·mL^–1内与峰面积线性关系良好(r=0.999 9),平均加样回收率为97.4%,RSD为0.6%(n=9)。5家企业生产的16批次样品中杏仁腈的含量为3.854～154.578μg·mL^–1。结论 所建立的方法简单、准确,可以用于健儿清解液的质量控制;同时应关注溶剂对...     

基于一测多评法鉴别蓝芩制剂中的黄柏与关黄柏

目的 建立蓝芩制剂中盐酸黄柏碱、盐酸巴马汀及盐酸小檗碱的HPLC一测多评法,并基于含量测定结果建立蓝芩制剂中黄柏与关黄柏的鉴别方法。方法 采用Agilent 1260色谱仪,Waters Atlantis T3(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.2%磷酸溶液(每100 mL加十二烷基磺酸钠0.2 g)梯度洗脱,流速为1 mL·min^–1,检测波长为280 nm,柱温30℃,以盐酸小檗碱为参照物,计算盐酸黄柏碱及盐酸巴马汀的相对校正因子,所得结果与外标法结果进行比较,判断该方法的可靠性。利用聚类分析含量测定结果,讨论黄柏与关黄柏对3种生物碱含量的影响,建立鉴别方法,对不同厂家的蓝芩制剂进行测定。结果 盐酸黄柏碱、盐酸巴马汀及盐酸小檗碱分别在1.087～54.35μg·mL^–1,0.100 9～20.18μg·mL^–1,0.536 5～26.82μg·mL^–1内线性关系良好;平均加样回收率分别为91.57%(RSD=3.0%),97.96%(RSD=2.1%),100.6%(RSD...     

二维高效液相色谱法检测达托霉素血药浓度的可行性考察

目的：建立二维高效液相色谱结合中间柱拦截技术测定达托霉素血药浓度的方法。方法：样品通过一维色谱柱Aston SRC(4.6 mm×50 mm,5μm)初步分离后,被中间柱ASTON-PLI(20 mm×4.6 mm,3μm)截取到二维色谱柱Aston SCB(4.6 mm×250 mm,5μm)进行二次分离;一维流动相为LC1移动相,流速：0.6 mL·min^–1。二维流动相为LC2移动相,流速：1.2 mL·min^–1,检测波长260 nm,温度40°C,进样量200μL。结果：达托霉素在1.00～40.06μg·mL^–1范围内线性关系良好,r=0.999 9,最低检测限为0.03μg·mL^–1,绝对回收率为99.2%～101.9%,日间、日内RSD均小于5.94%。结论：该方法新颖,自动化程度高,样本检测简单快速、准确稳定,适用于临床达托霉素血药浓度的测定。     

液质联用技术用于醋酸阿托西班注射液中杂质结构鉴定

目的:采用高效液相色谱(HPLC)、超高效液相色谱-四极杆串联静电场轨道阱高分辨质谱(UPLC-Q Orbitrap MS)技术对醋酸阿托西班注射液中杂质结构进行鉴定。方法:HPLC法采用Inertsil ODS-2 C₁₈色谱柱(250 mm×4.6 mm, 5μm),流动相A为乙腈-甲醇-三氟乙酸溶液(pH 3.2)(15∶10∶75),流动相B为乙腈-甲醇(60∶40),梯度洗脱，流速1.2 mL·min^-1,检测波长220 nm。UPLC-Q Orbitrap MS法采用BEH300 C₁₈色谱柱(150 mm×2.1 mm, 1.7μm),流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B),梯度洗脱，流速0.2 mL·min^-1;采用电喷雾离子源，选择正离子模式进行Full MS/dd-MS²扫描。结果:对5家企业各1批醋酸阿托西班注射液进行了有关物质测定，以面积归一化计算，含量>0.1%的杂质峰个数分别为9、10、13、9和6,总杂含量分别为0.35...     

太白茶HPLC指纹图谱的建立及UHPLC-QExactive Focus MS/MS联用分析

目的:建立太白茶药材高效液相色谱(HPLC)指纹图谱分析方法,并运用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q Exactive Focus MS/MS)法对太白茶中的特征峰进行定性分析。方法:采用Agilent TC-C₁₈(250 mm×4.6 mm, 5μm)色谱柱,以甲醇(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min^-1,柱温30℃,检测波长254 nm,进样量10μL。采用“中药色谱指纹图谱相似度评价系统2012年版”,建立指纹图谱并进行相似度评价。化学成分分析选用UHPLC-Q Exactive Focus MS/MS技术,采用Waters Acquity UPLC BEH C₁₈(50 mm×2.1 mm, 1.7μm)色谱柱,以乙腈(A)-0.1%甲酸水(B)为流动相,梯度洗脱,流速0.3 mL·min^-1,柱温30℃;质谱条件为电喷雾离子源(ESI),负离子模式扫描,扫描范围m/z 80～1 200。结果:建立的太白茶特征指纹图谱中...     

心脏移植患者磺胺甲噁唑血药浓度快速检测分析方法的构建及临床应用

目的 探索建立心脏移植患者磺胺甲噁唑血药浓度分析方法,以指导临床合理用药。方法 采用柱切换技术的新型二维液相色谱仪(2D-LC-UV),使用一维色谱柱Aston SC2(3.5 mm×25 mm,5μm)对血浆中磺胺甲噁唑进行在线固相萃取,然后经中间柱Aston SBR(3.5 mm×10 mm,5μm)截取保留,随后转移至第二维色谱柱Aston SNX4(4.6 mm×130 mm,5μm)对目标分析物进行完全分离检测。色谱条件：一维流动相为乙腈-甲醇-水(10∶10∶70),流速0.8 mL·min^–1;二维流动相比例为BPI-1碱性移动相-API-3酸性移动相-甲醇(20∶40∶40),流速为1.2 mL·min^–1;紫外检测波长为240 nm。结果磺胺甲噁唑浓度在9.96～200.04μg·mL^–1内与峰面积呈良好的线性关系(相关系数R2=0.999 6);低、中、高3个浓度的日内和日间精密度RSD均<15%、相对回收率为85%～115%。对在院56例患者血药浓度进行测定,仅有30例(53.57%)患者血样...     

UPLC-MS/MS同时测定乳核内消液中的贝母素甲、贝母素乙、西贝母碱和湖贝甲素

目的 采用超高效液相色谱-串联质谱法同时测定乳核内消液中贝母素甲、贝母素乙、西贝母碱、湖贝甲素的含量。方法 经超声离心后,采用YMC-Triart C₁₈色谱柱(150 mm×2.1mm, 3μm),流动相为乙腈-0.1%甲酸溶液,梯度洗脱,流速0.3 mL·min^-1,柱温35℃;质谱采用ESI⁺离子源,多反应监测模式检测。结果 贝母素甲、贝母素乙、西贝母碱、湖贝甲素等的线性范围分别为0.119～23.72μg·mL^-1(r=0.9958)、0.120～23.94μg·mL^-1(r=0.9951)、0.105～20.93μg·mL^-1(r=0.9942)、0.109～21.72μg·mL^-1(r=0.9900),平均回收率(n=6)分别为99.4%(RSD=2.00%)、98.3%(RSD=2.20%)、100.9%(RSD=1.90%)、98.7%(RSD=2.50%)。结论 所用方法同时测定了乳核内消液中4种生物碱成分的含量,方...     

HPLC法结合LC-MS法测定硫酸鱼精蛋白含量及肽段鉴定

目的:建立了一种同时测定硫酸鱼精蛋白中主成分纯度和含量的HPLC方法，并采用LC-MS法对肽段氨基酸序列进行鉴定。方法:HPLC方法采用的色谱柱为ThermoHypersil GOLD色谱柱(250 mm×4.6 mm, 5μm),流动相A为0.1 mol·L^-1磷酸二氢钠溶液(用磷酸调节pH至1.8),流动相B为乙腈-0.1 mol·L^-1磷酸二氢钠溶液(用磷酸调节pH至1.8)(6.5∶93.5),梯度洗脱，流速1.0 mL·min^-1,柱温55℃,紫外检测波长为214 nm,进样量100μL。LC-MS法先采用馏分收集方式对各肽段进行提取富集，然后液质联用采用Waters ACQUITY UPLC Peptide BEH C₁₈色谱柱(15 cm×0.21 cm, 1.7μm),流速为0.2 mL·min^-1,流动相A为0.1%甲酸溶液，流动相B为5%乙腈-0.1%甲酸溶液，梯度洗脱；Thermo Q-Exactive Plus, ESI离子源，阳离子检测模式，雾化电压为...     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.