HPLC测定奥美沙坦酯中的基因毒性杂质

目的 建立HPLC测定奥美沙坦酯中潜在的基因毒性杂质[杂质1 ：N-（三苯基甲基）-5-（4''-溴甲基联苯-2-基）四氮唑,杂质2 ：N-三苯甲基-5-（4'',4''-二溴甲基联苯-2-基）四氮唑]的含量和限度。方法 采用Phenomenex C₁₈柱（250 mm×4.6 mm,5 μm）;流动相：0.1%冰乙酸水溶液-0.1%冰乙酸乙腈溶液（15：85）;检测波长：254 nm;流速：1.5 mL·min^-1;柱温：25℃。结果 杂质1 和杂质2 均在0.030 97~0.247 7 μg·mL^-1内线性良好（r分别为0.999 6和0.998 7）,平均回收率分别为94.37%和94.43%,RSD分别为2.38%和2.72%（n=9）。结论 该方法专属性强,准确、灵敏,可以作为奥美沙坦酯中基因毒性杂质1 和杂质2 的液相分析方法。     

坎地沙坦酯脂质体冻干粉的制备及在大鼠体内药动学研究

目的 制备并评价泊洛沙姆407修饰的坎地沙坦酯脂质体（CC/PLip）,考察其口服给药后在大鼠体内药动学过程。方法 用薄膜分散法制备CC/PLip,经冷冻干燥制成冻干粉。动态光散射（DLS）测定其zeta电位和粒径,透射电镜（TEM）观察其形态,超滤离心法测定包封率;采用口服给药,观察肠道粘膜渗透行为,分析药物在大鼠体内的药代动力学参数。结果 泊洛沙姆407修饰的坎地沙坦酯脂质体呈现球形,粒径为（182.67±4.10）nm,Zeta电位为（-8.98±0.19）mV,包封率为98.94%±0.60%;体外释放试验结果表明其具有明显的缓释效果;肠道粘膜渗透试验表明CC/PLip在小肠易被吸收;药动学实验中,参比坎地沙坦酯片和CC/PLip的AUC_0→48h分别为（13 112.6±5 343.7）ng·h·mL^-1、（19 522.8±3 973.7）ng·h·mL^-1;C_max分别为（656.8±345.0）ng·mL^-1、（1 199.1±330.7）ng·mL^-1;T_max分别为（7.3±3.0）h、（2.7±1.4）h。结论 CC/PLip包封率较高,粒径较小,具有缓释效果,能提高在小肠的吸收,可显著提高在大鼠体内的生物利用度。     

HPLC法测定黄连上清片中连翘酯苷A的含量

目的 采用高效液相色谱法测定黄连上清片中连翘酯苷A的含量。方法 色谱柱：VP-ODSC₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相：乙腈-0.4%冰醋酸溶液（15:85）,流速1.0 mL·min^-1,检测波长330 nm。结果 连翘酯苷A在10.24 ~81.92 µg·mL^-1范围内与峰面积线性关系良好(r=0.999 8),精密度试验RSD为0.85%,重复性试验RSD为1.14%,平均回收率为97.91%,RSD为1.55%(n=6)。结论 该方法操作简单,精密度良好,结果准确可靠,适用于黄连上清片中连翘酯苷A的含量测定。     

GC-MS法测定甲磺酸达比加群酯中的致突变杂质

目的 建立一种气相色谱-质谱联用法（GC-MS）同时测定甲磺酸达比加群酯中甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的方法。方法 采用DB-624色谱柱（30 m×0.25 mm,1.4 μm）;程序升温：起始温度100 ℃,以15 ℃·min^-1的速率升温至200 ℃,再以25 ℃·min^-1的速率升温至240 ℃;进样口温度为220 ℃,载气为氦气,流速为1.0 mL·min^-1,进样量为2.0 μL。采集模式为选择离子监测（SIM）,离子源温度为230 ℃。通过内标法进行计算。结果 三种甲磺酸酯之间的分离度均大于2.0,甲磺酸甲酯、甲磺酸乙酯、甲磺酸异丙酯的线性回归方程分别为Y=0.0130X+0.0524（r=0.999 0）、Y=0.0249X+0.0633（r=0.999 3）、Y=0.0188X+0.0906（r=0.998 5）,均在5.0～120.0 ng·mL^-1浓度范围内线性关系良好。检出限为1.5 ng·mL^-1（0.15 ppm）,定量限为5 ng·mL^-1（0.5 ppm）。甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的平均回收率分别为96.28%、97.91%和95.87%,RSD分别为2.83%、2.93%和1.73%。结论 本方法简便、快速、准确、专属性强、灵敏度高,适用于甲磺酸达比加群酯中三种甲磺酸酯的检测。     

HS-GC-MS同时测定沙芬酰胺中磺酸酯类基因毒性杂质

目的 建立同时测定沙芬酰胺原料药中5种磺酸酯类基因毒性杂质（甲磺酸甲酯、甲磺酸乙酯、甲磺酸异丙酯、甲磺酸丙酯、甲磺酸丁酯）的方法。方法 采用顶空进样气相色谱-质谱联用法,在线衍生,RESTEK Rxi-624Sil毛细管柱（30 m×0.25 mm,1.4 μm）,四级杆质量检测器,离子化模式为电子轰击离子化（EI）模式,采集模式为选择离子监测。结果 5种杂质检测限为0.3~1.3 ng·mL^-1,定量限为0.9~4.3 ng·mL^-1;精密度、稳定性、重复性试验的RSD均<5%;浓度在5~300 ng·mL^-1内,5种杂质的峰面积与其相对应的浓度有良好的线性关系,相关系数（r²）均>0.995;回收率为99.4%~100.6%,RSD为0.8%~2.6%（n=9）。结论 该法操作简便,重复性好,结果准确可靠,可用于沙芬酰胺中磺酸酯类基因毒性杂质的测定。     

离子色谱法测定氟达拉滨中三氟甲磺酸酯类遗传毒性杂质

目的 采用离子色谱法对氟达拉滨中的三氟甲磺酸酯类遗传毒性杂质进行测定。方法 采用十八烷基硅烷键合硅胶为填充剂,洗脱液：[0.15 mmol·L^-1四丁基氢氧化铵-0.090 mmol·L^-1苹果酸（pH 6.5）]-乙腈（70：30）,流速为1.0 mL·min^-1,柱温为35℃,检测方式为直接电导检测。结果 有效测定了氟达拉滨中的三氟甲磺酸酯类遗传毒性杂质,检测限为2.5 μg·mL^-1,定量限为7.4 μg·mL^-1,线性范围为1.917~22.344 mg·L^-1,r=0.997 4,方法耐用性好。结论 本法可有效检测氟达拉滨中的三氟甲磺酸酯类遗传毒性杂质,方法灵敏度高,准确度高,耐用性好。     

HPLC测定达比加群酯中的有关物质

目的 建立达比加群酯原料药中有关物质的测定方法。方法 采用Agilent SB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈(A)-0.2%的醋酸铵(B,用冰醋酸调节pH值至4.4)为流动相进行梯度洗脱：0~18 min,90%→40%B;18~30 min,40%B。流速为1.0 mL·min^-1,检测波长为340 nm。结果 各杂质与主峰之间的分离度良好。5个已知杂质：杂质A浓度在0.117 0~1.872 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 7;杂质B浓度在0.126 5~2.024 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 5;杂质C浓度在0.113 0~1.808 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 5;杂质D浓度在0.120 5~1.928 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 9;杂质E浓度在0.123 0~1.968 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 6;杂质A、杂质B、杂质C、杂质D和杂质E加样回收率的平均值分别为98.75%,98.91%,98.39%,99.0%和99.73%;RSD分别为0.91%,1.09%,1.22%,1.35%和1.18%。结论 本方法简便、准确可靠,适用于达比加群酯中有关物质的控制。     

HPLC法测定化妆品原料2-羟乙基脲及杂质尿素的含量

目的 建立化妆品原料2-羟乙基脲的高效液相色谱（high performance liquid chromatography,HPLC）含量测定方法,并检测其杂质尿素的含量,以实现对2-羟乙基脲的质量控制。方法采用Capcell PAK ADME C_{18色谱柱（4.6 mm×250 mm,5 μm）;以水为流动相;柱温30℃;流速为1.0 mL·min^-1;进样量10 μL;检测波长194 nm。结果 2-羟乙基脲和尿素分别在1.168～116.8 μg·mL^-1（r=0.999 7）和1.032～206.5 μg·mL^-1（r=0.9992）范围内浓度与峰面积有良好的线性关系,平均回收率分别为98.0%和97.1%,检出限分别为0.12 μg·mL^-1和0.21 μg·mL^-1,定量限分别为0.36 μg·mL^-1和0.52 μg·mL^-1。结论 本方法准确度高,重复性好,检测灵敏度符合检测要求,可用于化妆品原料2-羟乙基脲及其杂质尿素的含量测定方法。}     

GC-MS测定碳酸司维拉姆中环氧氯丙烷的残留量

目的 建立气相色谱-质谱联用法测定碳酸司维拉姆中环氧氯丙烷残留量的方法,以满足原料药质量标准中的限度要求。方法 采用DB-17MS毛细管柱（30 m×0.25 mm,0.25 μm）;进样口温度为200℃;离子源温度为230℃;MS四极杆温度为150℃;载气为氦气;柱流量为1.0 mL·min^-1;程序升温;分流比为10：1;定量离子m/z 57。结果 在0.087~0.260 μg·mL^-1内环氧氯丙烷浓度与峰面积呈现良好的线性关系,定量限为0.045 μg·mL^-1,检测限为0.017 μg·mL^-1;平均加样回收率为98.4%。结论 该方法简单快速,专属性强,准确度好,灵敏度高,可以满足碳酸司维拉姆原料药标准中的限度要求。     

GC-MS测定硫酸氢氯吡格雷中硫酸二甲酯的残留量

目的 采用顶空气相色谱-质谱联用法(HS-QC-MS)测定硫酸氢氯吡格雷中硫酸二甲酯杂质的含量。方法 选用DB-624毛细管柱(30 m×0.32 mm，1.5 μm)，程序升温，初始温度为40 ℃，保持8 min，以30 ℃·min^–1升到220 ℃，保持2 min，以氦气为载气，流速为2.0 mL·min^–1，采用电子轰击电离(EI)源，在选择离子监测模式下选择m/z 45，56，88进行检测。结果 硫酸二甲酯浓度在0.15~3.0 μg·mL^–1内与峰面积线性关系良好(r²=0.994 2)；检测限为0.05 μg·mL^–1，定量限为0.15 μg·mL^–1；杂质硫酸二甲酯平均回收率为83.9%，RSD(n=9)为4.0%。经检测，硫酸氢氯吡格雷样品中硫酸二甲酯有少量检出。结论 本方法操作简便，结果准确，灵敏度高，可用于硫酸氢氯吡格雷样品中硫酸二甲酯的测定。     

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.     

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.     

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.     

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.     

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.