HPLC双波长法同时测定美辛唑酮红古豆醇酯栓中的2种有效成分

摘 要 目的：建立同时测定美辛唑酮红古豆醇酯栓中呋喃唑酮和吲哚美辛含量的高效液相色谱双波长分析方法。方法: 采用ZORBAX Extend C18 色谱柱（250 mm×4.6 mm,5 μm）,以乙腈和0.035 mol·L^-1的磷酸二氢钾水溶液（冰醋酸调节pH至3.0）为流动相进行梯度洗脱,流速1.0 ml·min-1,检测波长分别为364 nm和318 nm,柱温30℃,进样量20 μl。结果: 在选定的色谱条件下,呋喃唑酮和吲哚美辛在0.005～0.05 mg·mL^-1范围内线性关系良好,r=0.999 9,检测限分别为20 ng·mL^-1和26 ng·mL^-1,定量限分别为70 ng·mL^-1和90 ng·mL^-1,平均回收率分别为99.4%（RSD=0.6%,n=9）和99.4%（RSD=0.3%,n=9）。结论: 所建立的方法简便快速,专属性强,结果准确可靠,可用于美辛唑酮红古豆醇酯栓中呋喃唑酮及吲哚美辛的检测分析。     

HPLC-MS/MS法测定人血浆中拉西地平药物浓度的方法学研究

摘 要 目的：建立测定人血浆中拉西地平浓度的方法。方法: 200 μl血浆样本采用500 μl叔丁基甲醚萃取浓缩,采用HPLC MS/MS测定,以¹3C8 拉西地平为内标,色谱柱为Agilent ZORBAX Eclipse XDB C18 （150 mm×2.1 mm, 5 μm）,流动相为水（含5 mmol·L^-1甲酸铵） 乙腈（15〖KG*9〗∶〖KG-*2〗85,v/v）,流速0.3 ml·min^-1,柱温40℃,进样量10 μl,采用电喷雾离子源,以多反应监测方式进行正离子扫描,以[M+NH4]+作为分子离子峰,用于拉西地平定量的三对反应离子对为m/z 473.5→m/z 410.3、m/z 473.5→m/z 400.1和m/z 473.5→m/z 354.3,用于内标定量的反应离子对为m/z 481.4→m/z 362.3。结果: 拉西地平在0.1 10 ng·mL^-1范围内线性关系良好（r＞0.996 9）,定量下限为0.1 ng·mL^-1;日内、日间精密度为3.15% ～7.04%,相对偏差（RE）为-8.58 ～12.17%,提取回收率为107% ～118%,RSD＜15%。血浆样品在经历3次冻融（-20 ℃到室温）循环和冷冻放置40 d等条件下均稳定,处理后样品在室温和自动进样器中（4℃）各放置24 h均稳定（RSD＜15%）。结论: 该方法快速、准确、灵敏度高、专属性强,可应用于拉西地平相关的生物等效性一致性评价和药动学研究。     

HPLC法同时测定云南红大戟中三种蒽醌类化合物的含量

摘 要 目的：建立HPLC法同时测定红大戟药材中芦西定、5 羟基巴戟醌与红大戟素的含量。方法: 采用 Waters Xbridge C18色谱柱（250 mm×4.6 mm,5 μm）,以0.05%磷酸为流动相A,以乙腈为流动相B,梯度洗脱;流速为1.0 ml·min^-1,检测波长为280 nm,柱温为30℃。 结果: 芦西定、5 羟基巴戟醌、红大戟素的线性范围分别为0.147～29.400 μg·mL^-1(r=0.999 6)、0.126～25.200 μg·mL^-1 (r=0.999 9)、0.135～27.000μg·mL^-1 (r=0.999 5),平均加样回收率分别为98.50%（RSD=1.20%）、98.72%（RSD=0.73%）、101.10%（RSD=1.12%）(n=6)。结论:本文建立的方法经方法学验证可用于评价红大戟药材质量。     

HPLC-MS/MS法测定比格犬血浆中PA-824的浓度及其药动学研究

摘 要 目的：建立高效液相色谱 质谱连用（HPLC MS/MS）测定比格犬血浆中PA 824的浓度,以及进行PA 824在比格犬体内的药动学研究。方法: 以卡马西平为内标,生物样品预处理采用乙酸乙酯萃取。色谱柱为Eclipse Plus C18柱（100 mm×2.1 mm,3.5 μm）,流动相为甲醇 水（90∶10）,流速为0.6 ml·min^-1,柱温为30℃,进样量为5 μl,样品分析时间为5 min;电喷雾电离源（ESI）,正离子多反应监测扫描分析,PA 824离子对为m/z360.1→m/z 175.0（碰撞能量：35,解簇电压：65）,卡马西平离子对为m/z 237.2→m/z 194.0（碰撞能量：28,解簇电压：83）。比格犬口服给药之后,在不同的时间点取血测定血浆中PA 824的含量,然后再用DAS 2.0软件进行药代动力学参数的计算。结果: 比格犬口服给药之后,PA 824在50～10 000 ng·mL^-1 （r=0.999 1）范围内呈线性关系,回收率为97.7%～105.1%,日内、日间精密度RSD均<5.0%。PA 824 3个不同给药量（100,200,500 mg）的AUC0 t分别为（5 735.18±1 918.76）,（11 548.47±1 838.04）,（21 987.88±4 587.58）ng·min·ml^-1,t^1/2分别为（14.17±5.97）,（11.11±4.39）,（13.13±5.46）h,Cmax分别为（626.66±188.48）,（2 399.13±516.51）,（4 861.33±2 253.61）ng·ml^-1。结论:该方法简单、准确、快速、重复性好,适用于比格犬血浆PA 824的药代动力学研究。     

液相色谱-串联质谱法同时测定对乙酰氨基酚及其相关代谢产物

摘 要 目的：建立一种检测小鼠血浆中对乙酰氨基酚（APAP）及其5个代谢产物的液相色谱 串联质谱（LC-MS/MS）方法,将之应用于对乙酰氨基酚的代谢研究。方法: 血浆样品经甲醇沉淀蛋白后,以对氨基苯甲酸（PABA）为内标,采用LC-MS/MS法,C18色谱柱,流动相为甲醇 5 mmol·L^-1乙酸铵水溶液（含0.1%甲酸）（55∶45）,流速为0.5 ml·min^-1,柱温为25℃,采用电喷雾离子化（ESI）,正离子多反应监测（MRM）检测方式进行定量分析：APAP,[M-H]^-,m/z 152.0→110.0;APAP cys,[M-H]^-,m/z 271.2→140.1;APAP glut,[M-H]^-,m/z 457.0→328.0;APAP NAC,[M-H]^-,m/z 313.4→208.0;APAP sulf,[M-H]^-,m/z 232.4→152.1;APAP-gluc,[M-H]^-,m/z 328.2→152.1;IS,[M-H]^-,m/z 138.2→120.0。结果: 在0.2~10 μg·mL^-1（APAP）、1.0~20 μg·mL^-1（APAP-gluc）、1.0~20 μg·mL^-1（APAP sulf）、1.0~20 μg·mL^-1（APAP glut）、0.4~15 μg·mL^-1（APAP NAC）和0.2~10 μg·mL^-1（APAP cys）范围内线性良好(r≥0.990 0)。各测定物的日内精确度均在85%~115%间,精密度均小于15%,加样回收率均在85%~115%之间,RSD均低于15%。结论:该方法快速、灵敏、准确,适用于小鼠血浆中对乙酰氨基酚及其5个代谢产物的定量测定。     

雷公藤口服液中雷公藤甲素、总二萜内酯和总生物碱的含量测定

摘 要 目的：建立雷公藤口服液中雷公藤甲素、总二萜内酯以及总生物碱的含量测定方法,为质量控制提供依据。 方法: 采用高效液相色谱（HPLC）法,Eclipse XDB C₁₈色谱柱（150 mm×4.6 mm,5 μm）,以乙腈 水为流动相进行梯度洗脱,流速1.0 ml·min^-1,柱温40℃,检测波长218 nm,测定雷公藤口服液中雷公藤甲素的含量。采用紫外可见分光光度法,分别以雷公藤甲素、雷公藤次碱为对照,测定雷公藤总二萜内酯、总生物碱的含量。结果: 雷公藤甲素、总二萜内酯（以雷公藤甲素计）和总生物碱（以雷公藤次碱计）的线性关系良好（r≥0.999 8）,平均加样回收率分别为91.96%、90.73%、99.18%,RSD均小于3%。结论:本研究所建立的方法稳定可靠,重复性良好,可用于雷公藤口服液的质量控制。     

UPLC-MS/MS法检测大鼠血浆中阿法替尼浓度及其药动学研究

摘 要 目的：建立一种检测大鼠血浆中阿法替尼浓度的UPLC MS/MS方法。方法: 选择用乙腈沉淀的蛋白质样品进行样品处理并运用Waters XEVO TQD三重四级杆液质联用仪和CORTECS BEH C18柱(50 mm×2.1 mm, 1.6 μm)分离分析物。流动相由乙腈和水（0.1％甲酸）组成,流速为0.4 ml·min^-1,柱温为40℃,内标为来那替尼。采用正离子电喷雾多反应监测（MRM）模式对分析物进行定量,目标碎片离子为阿法替尼m/z 486.19→112.1,来那替尼（IS）m/z 557.3→112.15。结果:阿法替尼在1 200 ng·ml^-1范围内线性关系良好(r=0.998 1),定量下限为1 ng·ml^-1。日内精密度和日间精密度均≤9.51％,阿法替尼从血浆中回收率高于77.1％。大鼠灌胃给药和静脉给药阿法替尼后,t1/2分别为7.19 h和2.69 h,Cmax分别为97.78 ng·ml^-1和123.37 ng·ml^-1,AUC(0-∞)分别为1 505.4 ng·h·ml^-1和405.55 ng·h·ml^-1。结论:该方法准确可靠,操作简便,重复性好,适用于灌胃和静脉注射10和2 mg·kg^-1剂量的阿法替尼的药动学研究。     

酮康他索乳膏中酮康唑和丙酸氯倍他索含量测定方法的改进

摘 要 目的：建立同时测定酮康他索乳膏中酮康唑和丙酸氯倍他索含量的方法。方法: 采用HPLC法,色谱柱为Kromasil C18（250 mm×4.6 mm,5 μm）柱,流动相为甲醇 0.05 mol·L^-1醋酸钠溶液（用冰醋酸调pH至5.5）,采用梯度洗脱,流速为1.0 ml·min^-1,检测波长为239 nm,柱温为25℃,进样量为10 μl。结果:酮康唑和丙酸氯倍他索分别在160.30 ~1 282.40 μg· mL^-1和4.03~32.24 μg· mL^-1范围内呈良好的线性关系（r均为1.000 0）。平均回收率分别为100.9%（RSD=0.52%,n=9）和100.2%（RSD=0.56%,n=9）。结论:该方法专属性好、灵敏度高、定量准确,可用于酮康他索乳膏中酮康唑和丙酸氯倍他索的含量检测。     

HPLC法测定两性霉素B中乙二胺四乙酸二钠含量

摘 要 目的： 建立两性霉素B注射液中乙二胺四乙酸二钠（EDTA-2Na）的HPLC检测方法。方法: 采用Waters C18柱（50 mm×4.6 mm,5 μm）,流动相A为醋酸溶液（取冰醋酸1.5 ml,加水1 000 ml,加四丁基氢氧化铵41 ml）,B为乙腈,梯度洗脱流速0.8 ml·min^-1,柱温30℃,检测波长为260 nm,进样体积25 μl。结果: EDTA 2Na峰与其他色谱峰分离度良好,在0.92~7.37 μg·ml^-1浓度范围内线性关系良好（r=0.999 9）,最低检测限（LOD）为1.93 ng·ml^-1,最低定量限（LOQ）为6.45 ng·ml^-1,平均回收率分别为102.5%,RSD为2.8%（n=9）。结论:本方法操作简便,精密度好,结果准确可靠,可用于两性霉素B原料药中EDTA 2Na的质量控制。     

HPLC法同时测定祛喘胶囊中木兰脂素、欧前胡素、异欧前胡素的含量

摘 要 目的： 建立祛喘胶囊中对木兰脂素、欧前胡素和异欧前胡素的HPLC测定方法。方法: 采用日本资生堂CAPCELL PAK C₁₈ 色谱柱（150 mm×4.6 mm,5 μm）,以乙腈-水为流动相进行梯度洗脱,柱温为30℃,流速为1.0 ml·min^-1,检测波长在0～25 min为278 nm,25～70 min为248 nm,进样量为10 μl。结果: 木兰脂素在6.126～55.134 μg·mL^-1范围内线性关系良好（r=0.999 7）,平均回收率为101.5%,RSD为1.4%;欧前胡素在9.862～88.758 μg·mL^-1范围内线性关系良好（r=0.999 8）,平均回收率为99.6%,RSD为1.2%;异欧前胡素在4.830～43.470μg·mL^-1范围内线性关系良好（r=0.999 8）,平均回收率为100.2%,RSD为1.7%。结论:本法操作简便、结果准确,可用于祛喘胶囊的质量控制。     

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.