LC-MS/MS测定人血浆中的辛伐他汀

目的建立测定人血浆中辛伐他汀的方法。方法采用LC-MS/MS法,血浆样品中加入内标洛伐他汀,经乙腈沉淀蛋白后,以正离子多反应监测(MRM)方式测试,用于定量的离子为m/z441.2→325.2(辛伐他汀)和m/z427.2→325.2(洛伐他汀)。色谱柱为Restek Allure C18(50 mm×2.1 mm,5μm),流动相为乙腈-0.5%甲酸水溶液(70:30)。结果血浆中辛伐他汀的线性范围为0.1-50.0 ng.ml-1,定量限为0.1 ng.ml-1,方法回收率为98%~102%,日内RSD<3.6%,日间RSD<5.6%。结论所建方法专属性好、灵敏度高,符合血浆样品测定的要求,适用于辛伐他汀的临床药物动力学研究。     

HPLC法测定比格犬血浆中的辛伐他汀及辛伐他汀羟基酸

目的:建立血浆中辛伐他汀及其主要代谢活性成分辛伐他汀羟基酸的含量测定方法。方法:以洛伐他汀为内标物质,流动相采用甲醇-水-冰醋酸(74:26:0.5),检测波长为238nm,流速为1.0mL·min~(-1)。血浆样品采用乙腈沉淀蛋白处理。并测定了10只比格犬单剂量口服辛伐他汀自乳化胶囊和市售片后的血药浓度。结果:血浆中内源性物质对辛伐他汀及辛伐他汀羟基酸的测定无干扰;最低检出限辛伐他汀为0.5ng·mL~(-1),辛伐他汀羟基酸为0.75ng·mL~(-1);辛伐他汀羟基酸在2～100ng·mL~(-1)浓度范围内线性关系良好,r=0.9989,辛伐他汀在1～50ng·mL~(-1)浓度范围内线性关系良好,r=0.9993;绝对回收率为79.68%～95.3%,方法回收率为89.1%～95.3%;日内精密度 RSD≤2.9%、日间精密度 RSD≤4.6%。结论:本方法处理简单,无干扰,灵敏度高,适合测定生物样本中的辛伐他汀及辛伐他汀羟基酸。     

LC-MS/MS法测定人血浆中的氨氯地平

目的 采用LC-MS/MS测定人血浆中的氨氯地平.方法 血浆样品经氢氧化钠碱化,乙酸乙酯萃取后进行分析.使用Gmini C18 110A(50 mm×3 mm,5 μm)色谱柱,流动相为甲醇-水-甲酸(55:45:0.025),流速0.2 ml·min-1,通过电喷雾离子化四极串联质谱,以多反应离子方式检测.氨氯地平m/z 409.4→237.9和莫沙比利m/z 422.1→198.0.结果 线性范围为0.0313～20 ng·ml-1,最低定量限为0.0313 ng·ml-1,萃取回收率为74.4%～83.1%,方法回收率为96.1%～108.7%,日内和日间RSD均<8%.结论 所用方法快速、简便、灵敏度高,适用于人血浆中药物浓度的测定及药物动力学和生物利用度的研究.     

LC-MS/MS法同时测定人血浆中的辛伐他汀和辛伐他汀酸

目的 采用LC-MS/MS法同时测定人血浆中的辛伐他汀和辛伐他汀酸.方法 采用BEH C18色谱柱(50 mm ×2.1mm,1.7 μm),流动相为乙腈-0.01 mol·L-1醋酸铵(72∶28),流速0.15 mL·min-1,柱温40℃,进样量8μL.辛伐他汀、辛伐他汀酸及内标洛伐他汀的检测离子对分别为:m/z 419.43→199.12、437.38→303.26、405.45→199.14.结果 辛伐他汀、辛伐他汀酸的线性范围分别为0.241 ～61.76 ng·mL-1(r =0.999)、0.344 ～ 88.16 ng·mL-1(r =0.997).在人血浆基质中,高、中、低浓度(0.482、3.86、30.88 ng· mL-1)的日内、日间RSD均小于15％,方法回收率分别为95％～104％、97％ ～108％.样品预处理方法对血浆中的辛伐他汀和辛伐他汀酸测定无干扰.结论 所用方法处理简单、灵敏、特异性高,定量准确,可为辛伐他汀制剂的药动学研究提供方法.     

液相色谱-电喷雾串联质谱法测定人血浆中辛伐他汀浓度

目的:建立液相色谱-电喷雾串联质谱联用测定人血浆中辛伐他汀浓度的方法。方法:色谱条件为 Discovery ODS C_(18)(2.1 mm×100 mm,3μm)色谱柱;流动相为乙腈-0.1%甲酸溶液(65:35,v/v);柱温30℃;流速0.2 mL·min~(-1);进样量5μL。质谱条件为电喷雾电离源(ESI),选择性检测定量离子为 m/z 441.3/325.0(辛伐他汀)和 m/z 427.2/325.0(洛伐他汀,内标)带正电荷的离子峰。样品用乙酸乙酯提取。结果:血浆中辛伐他汀浓度在0.20～20.0 ng·mL~(-1)内呈良好线性关系,r=0.9996。提取回收率为90.0%～92.4%(RSD 6.0%～14.9%),方法回收率在85%～115%之内,日内和日间精密度试验的RSD 均<15%,最低检测浓度为0.2 ng·mL~(-1)。结论:该测定方法经全面考察符合血浆样品的测定要求,可以应用于辛伐他汀的人体药动学研究和生物等效性评价。     

HPLC-串联四级杆质谱法测定功能食品中的利尿剂

目的 采用HPLC-串联四级杆质谱法专属测定功能食品中违禁添加的呋塞米、氢氯噻嗪、布美他尼利尿剂.方法 样品在甲醇一超声波提取、微孔滤膜过滤后,经HPLC分离、串联四级杆质谱一负离子多反应监测方式检测,通过相对分子质量、二级质谱碎片信息定性、标准曲线法定量,同时测定了功能食品中的呋塞米、氢氯噻嗪和布美他尼.结果 呋塞米和氢氯噻嗪的方法检测限(S/N=3)为0.5 ng·ml-1,布美他尼的为1.0 ng·ml-1;呋塞米的最低定量限(S/N=10)为4 ng·ml-1,氢氯噻嗪的为1 ng·ml-1,布美他尼的为4.5 ng·ml-1.方法 的RSD均小于0.31%,平均回收率为86.0% ～107.0%.在8种被检功能食品中,1种被检出掺有氢氯噻嗪.结论 所建方法选择性强、灵敏度高,可作为检测功能食品中非法掺人呋塞米、氢氯噻嗪和布美他尼利尿剂的有效方法.     

人血浆中西酞普兰的HPLC-荧光法测定及其药物动力学

建立了HPLC-荧光法测定人血浆中西酞普兰的浓度,并进行了药物动力学研究.以盐酸普萘洛尔为内标,采用Dikma柱,以乙腈-0.03mol/L醋酸铵缓冲液(33:67)为流动相,激发波长和发射波长分别为236和306nm.血浆中西酞普兰线性范围为1～75ng/ml,定量限为1ng/ml,方法回收率92.8%～98.8%.18名健康志愿者口服西酞普兰40mg后,血浆中的Cmax为(54.54±7.83)ng/ml,Tmax为(3.06±0.42)h,AUC0～132为(2351.1±389.9)ng·ml-1·h,AUC0～∞为(2636.5±444.9)ng·ml-1·h,t1/2为(40.45±8.68)h.     

UPLC-MS/MS法同时测定人血浆中辛伐他汀和辛伐他汀羟基酸的浓度

目的:建立UPLC-MS/MS法同时测定人血浆中辛伐他汀和辛伐他汀羟基酸的浓度。方法:血浆样品用甲基叔丁基醚提取,离心后取上清液氮气吹干,流动相复溶后进行UPLC-MS/MS测定。色谱柱:BEH C18(2.1 mm×50 mm,1.7μm);流动相:乙腈-0.01 mol.L-1醋酸铵(72∶28);流速:0.15 mL.min-1;柱温:40℃,进样量:8μL。电喷雾离子化(ESI),正离子模式多重反应选择离子检测(MRM),辛伐他汀、辛伐他汀羟基酸及内标洛伐他汀的检测离子对分别为:m/z 419→199,437→303,405→199。结果:血浆样品中辛伐他汀、辛伐他汀羟基酸线性范围分别为0.241～61.76 ng.mL-1(r=0.999,n=5)和0.344～88.16 ng.mL-1(r=0.997,n=5),日内、日间精密度(RSD)均小于15%,方法的平均回收率分别为100.6%和106.0%,血浆基质对血浆中的辛伐他汀和辛伐他汀羟基酸测定无干扰。结论:建立的UPLC-MS/MS法处理简单、灵敏、特异性高,定量准确,为辛伐他汀制剂的临床药代动力学研究提供了简便、准确的分析测定方法。     

高效液相色谱串联质谱法测定人血浆中辛伐他汀浓度及药动学研究

目的:建立高效液相色谱串联质谱法测定人血浆中辛伐他汀浓度,并进行辛伐他汀片的药动学研究。方法:以洛伐他汀为内标,血浆样品经液-液萃取后,经HPLC-MS/MS分离-分析。采用DAS2.1程序计算有关药动学参数。结果:辛伐他汀血浆浓度测定方法线性范围为0.1016～20.32ng·ml^-1,r=0.9981。定量下限为0.1016ng·ml^-1,方法相对回收率85%～115%,日内和日间RSD<11%;绝对回收率>75%。辛伐他汀片主要药动学参数AUC_0～24、C_max、t_max、t_1/2z、CL_z/F分别为（34.34±19.57）ng·h·ml^-1、（11.15±11.58）ng·ml^-1、（1.89±1.52）h、（5.99±4.93）h、（703.71±428.48）L·h^-1。结论:本试验所建立起来的人血浆中辛伐他汀浓度测定法灵敏、准确、可靠,适用于辛伐他汀的人体内药动学研究。     

Beagle犬血浆中辛伐他汀的LC-MS测定

建立了LC-MS法测定Beagle犬血浆中的辛伐他汀。采用C_(18)柱,流动相为乙腈-0.1%甲酸溶液(60:40),以洛伐他汀为内标,采用电喷雾离子化法(ESI)采集正离子,单离子方式检测m/z 419(辛伐他汀)和m/z 405(洛伐他汀)。辛伐他汀在0.5～160ng/ml浓度范围内线性良好,检测限为0.25ng/ml。血浆样品的日内、日间RSD均小于7.7%,方法回收率为92.2%～99.2%,提取回收率为92.8%～100.9%。     

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.     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.