RP-HPLC法检测奈韦拉平原料药中有关物质

目的 建立RP-HPLC测定奈韦拉平原料药中已知杂质A、B、C和未知杂质的方法.方法 RP-HPLC法,以Gemini C18色谱柱(4.6mm×250mm,5μm)(Phenomenex),乙腈-0.025mol·L-1磷酸二氢钾溶液(pH5.0)(20∶80)为流动相,流速1.0mL·min-1,检测波长为220nm,柱温35℃,进样量50μL.结果 奈韦拉平杂质线性范围A为0.1203 μg·mL-1 ～0.6016μg·mL-1,r =0.9998;B为0.1198μg·mL-1～0.5992μg·mL-1,r =0.9999;C为0.1195μg·mL-1～0.5976μg·mL-1,r =0.9999.平均回收率A为98.87％,RSD=1.03％;B为100.43％,RSD=0.62％;C为99.19％,RSD =1.69％.结论 本方法准确、简便、快捷,可用于奈韦拉平有关物质的测定.     

HPLC测定缩宫素原料药的含量

目的 采用HPLC测定缩宫素原料药的含量.方法 色谱柱为Agilent Zorbax SB-C18(250 mm×4.6 mm,5μn),流动相A为15.6 g·L-1磷酸二氢钠,流动相B为50％乙腈,流速1.0 mL· min-1,检测波长220 nm,进样量25 μL,外标法定量.结果 缩宫素的线性范围为150～350 μg· mL-1(r=0.9992);定量限为0.025μg· mL-1.结论 所用方法简便、快速、准确,专属性好,可用于缩宫素原料药的含量测定.     

HPLC测定注射用头孢曲松钠他唑巴坦钠的含量及其有关物质

目的 采用HPLC法测定注射用头孢曲松钠他唑巴坦钠的含量及其有关物质.方法 采用Kromasil C18色谱柱(250mm ×4.6 mm,5μm),流动相为磷酸缓冲液-乙腈(90:10),流速1.0 mL· min-1,柱温30℃,检测波长230 nm.结果 他唑巴坦、头孢曲松含量的线性范围分别为0.05 ～ 0.40、0.15～ 1.20 mg· mL-1(r=0.9999),重复性的RSD分别为1.5％、0.5％,平均回收率分别为101.0％、100.9％,定量限分别为0.12、0.04μg;他唑巴坦、头孢曲松有关物质的线性范围分别为2.5 ～25.3、7.5 ～75.0 μg·mL-1,r分别为0.9999、0.9998,精密度的RSD分别为2.0％、0.5％,检测限分别为0.04、0.01 μg,各杂质与头孢曲松及他唑巴坦两色谱峰均能完全分离.结论 所用方法可用于注射用头孢曲松钠他唑巴坦钠的质量控制.     

HPLC测定甲钴胺的含量及其有关物质

目的 采用HPLC法测定甲钴胺的含量和有关物质.方法 色谱柱为Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为0.03 mol∶ L-1磷酸二氢钾溶液(用0.2 mol·L-1氢氧化钠或磷酸调pH4.5)-乙腈(84∶16),含量测定和有关物质的检测波长分别为266、342 nm.流速为1.0 mL·min-1.结果 甲钴胺峰的分离度良好,最低检测限为0.18 ng,甲钴胺的线性范围为3.94 ～ 5.92 μg∶ mL-1(r2=0.9999,n=5)和4.93 ～ 78.90 μg· mL-1(r2 =0.9994,n=5).结论 所用方法简便、准确、重复性好,可用于甲钴胺的含量和有关物质的测定.     

HPLC同时测定左归丸中的5种成分

目的 采用HPLC法同时测定左归丸中尿囊素、薯蓣皂苷元、梓醇、麦角甾苷和吉奥诺苷B1的含量.方法 采用依利特C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.2％磷酸溶液,梯度洗脱,流速1.3 mL· min-1,柱温为30℃,检测波长为224(尿囊素)、210(薯蓣皂苷元和梓醇)和330(麦角甾苷和吉奥诺苷B1)nm.结果 尿囊素、薯蓣皂苷元、梓醇、麦角甾苷和吉奥诺苷B1的线性范围分别为7.270～145.4μg·mL-1(r=0.9999)、5.150～103.0 μg·mL-1(r=0.9992)、5.730～114.6 μg·mL-1(r =0.9998)、6.740 ～134.8 μg·mL-1(r =0.9995)和5.100～102.0 μg·mL-1(r =0.9997);其平均加样回收率分别为98.37％、96.92％、99.11％、96.65％、98.31％,RSD分别为0.89％、1.28％、1.10％、0.57％、1.34％(n=6).结论 所用方法简便快捷、回收率好、重复性好,可用于左归丸中尿囊素、薯蓣皂苷元、梓醇、麦角甾苷和吉奥诺苷B1的含量测定.     

RP-HPLC法测定替勃龙原料药有关物质及含量

目的:建立RP-HPLC法测定替勃龙原料药的有关物质及含量。方法:采用Promosil-C18柱(250 mm×4.6 mm,5μm),以甲醇-乙腈-水(50:30:20)为流动相,流速1.0 mL·min-1,检测波长204 nm,柱温为室温,进样量10μL。结果:替勃龙浓度在0.5～250.0μg·mL-1范围内呈良好的线性关系(r=0.9999);以主成分自身对照法检测供试品中的相关物质,线性范围是0.2～20.0μg·mL-1(r=0.9994);检出限为0.05μg·mL-1。替勃龙低、中、高浓度平均加样回收率(n=3)分别为99.7%,99.9%,99.9%;RSD分别为0.2%,0.2%,0.1%。结论:本方法简便、灵敏,重复性好,结果准确可靠,可用于替勃龙原料药有关物质及含量的检测与分析以及质量控制。     

HPLC-PAD法同时测定精制银翘解毒胶囊中对乙酰氨基酚、绿原酸、连翘苷和牛蒡苷的含量

目的:采用HPLC-PAD法同时测定精制银翘解毒胶囊中对乙酰氨基酚、绿原酸、连翘苷和牛蒡苷的含量.方法:色谱柱:Capcell Pak C18(150 mm×4.6 mm,5μm);流动相:乙腈-水(含0.25％的冰醋酸)梯度洗脱;检测波长:300 nm和228 nm;流速:1mL· min-1;柱温:30C.结果:对乙酰氨基酚测定的线性范围28.15～84.45 μg (r=0.9999),平均含量为235.06 mg·g-1,RSD为0.17％;绿原酸测定的线性范围0.2048～0.6144 μg (r=0.9998),平均含量为1.91 mg·g-1,RSD为0.21％;连翘苷测定的线性范围0.1054～0.3162μg (r=0.9994),平均含量为1.00 mg·g-1,RSD为0.32％;牛蒡苷测定的线性范围1.044～3.132 μg (r=0.9998),平均含量为7.04 mg·g-1,RSD为0.16％.结论:该法简便、灵敏、准确,适用于精制银翘解毒胶囊的质量控制.     

高效液相色谱法测定药用辅料日落黄的含量

目的 建立高效液相色谱法测定药用辅料日落黄的含量.方法 采用安捷伦C18柱(4.6 mm×250 mm,5 μm),以甲醇-0.02 mol·L-1乙酸铵溶液(30∶70)为流动相,流速为1.0 mL· min-1,检测波长为236 nm,柱温为35℃.结果 日落黄线性范围为5.55～111.03 μg·mL-1,r=0.999 9,平均回收率为100.0％,RSD=0.23％.结论 改进后的方法简单、准确,能控制本品的质量.     

HPLC法测定奥拉西坦原料及其注射剂中奥拉西坦的含量和有关物质

目的：建立用高效液相色谱法测定奥拉西坦原料及其注射剂中有关物质和含量的方法.方法：色谱柱为Agilent Eclipse XDB C18（250 mm×4.6 mm,5μm）,有关物质测定采用0.01 mol·L-1磷酸二氢钾溶液（磷酸调节pH至2.5）为流动相A,流动相A-乙腈（50∶ 50）为流动相B的梯度洗脱法,含量测定采用流动相A的等度洗脱法,检测波长为214 nm,流速为0.5ml·min-1,柱温为30℃,进样量为20 μl.结果：主峰和相邻杂质峰能完全分离,杂质A浓度在0.734 0 ～9.787 1 μg·ml-1范围内与其峰面积呈良好的线性关系,r =0.999 0,平均回收率为95.6％（RSD =1.5％,n=9）;奥拉西坦在60.71 ～121.41 μg·ml-范围内与峰面积呈良好线性关系,r =0.999 9,平均回收率为100.0％（RSD =0.6％,n=9）.结论：本方法专属性强,能检出更多的杂质,准确度高,能有效质控奥拉西坦原料及注射剂中的有关物质和含量.     

离子对色谱法测定乙酰谷酰胺铝的含量与有关物质

目的:建立离子对色谱法测定乙酰谷酰胺铝(化学名:N-乙酰-L-谷氨酰胺铝复合物)的含量与有关物质.方法:采用Diamonsil C18柱(150 mm×4.6 mm,5 μm),流动相为甲醇-水(含4 mmol·L-1四丁基氢氧化铵与30mmol·L-1磷酸二氢钾,磷酸调pH 5.0)(1:100),柱温:室温,流速:1 mL·min-1,检测波长210 nm,进样量20 μL.结果:乙酰谷酰胺铝的线性范围为20～70 μg·mL-1(r=0.999 9),平均回收率为99.89%,RSD为1.1%.结论:本法操作简便,专属性好,适合于测定乙酰谷酰胺铝的含量与有关物质.     

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.

---

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.     

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.