高效液相色谱法手性拆分奥美拉唑对映异构体

目的：手性拆分奥美拉唑对映异构体,建立检查埃索美拉唑钠原料药中R-异构体的HPLC方法。方法：采用Chiral pak IC手性柱（4.6 mm×250 mm,5 μm）,以正己烷-异丙醇-甲醇-三乙胺（40∶40∶20∶0.1,v/v/v/v）为流动相,检测波长为302 nm,柱温为30 ℃,流速为1.0 mL·min^-1。结果：埃索美拉唑及其R-异构体的分离度（R）大于7。R-异构体的检测限和定量限分别为32 ng·mL^-1和80 ng·mL^-1,线性范围为0.5~30 μg·mL^-1。自制三批埃索美拉唑钠盐中R-异构体的含量均为0.02%,R-异构体的限度符合要求。结论：该方法简便,快速,可用来检查埃索美拉唑钠原料药中R-异构体的限度。     

HPLC测定盐酸达泊西汀片中的异构体

目的 建立盐酸达泊西汀片剂中异构体的HPLC检测方法。方法 采用DAICEL CHIRALPAK OZ-H色谱柱（250 mm×4.6 mm,5 μm）,柱温：40℃,检测波长：293 nm,流速：1.0 mL·min^-1,流动相：正己烷-异丙醇-二乙胺（80：20：0.05）。结果 盐酸达泊西汀及其异构体在0.01~0.50 mg·mL^-1内具有良好的线性关系（r=0.999 9）,盐酸达泊西汀定量限为0.002 mg·mL^-1（相当于0.02%）,最低检测限为0.001 mg·mL^-1（相当于0.01%）;异构体定量限浓度为0.001 mg·mL^-1（相当于0.01%）,最低检测限为0.000 5 mg·mL^-1（相当于0.005%）。结论 该方法操作简便,结果准确,可用于盐酸达泊西汀片剂中异构体的含量测定。     

HPLC同时测定黄石感冒片中阿魏酸、芦丁、大黄酸、大黄素和大黄酚的含量

目的 建立HPLC同时测定黄石感冒片中阿魏酸、芦丁、大黄酸、大黄素和大黄酚的含量。方法 采用HPLC,色谱柱为Welch Topsil-C₁₈柱(4.6 mm×250 mm,4 μm);以甲醇-0.1%磷酸溶液(78∶22)为流动相;流速：1.0 mL·min^-1;柱温：30 ℃;检测波长：280 nm。结果 阿魏酸、芦丁、大黄酸、大黄素和大黄酚峰线性范围分别为0.020 82~0.416 3 μg·mL^-1(r=0.999 7),0.011 32~0.278 3 μg·mL^-1(r=0.999 3),0.017 22~0.344 3 μg·mL^-1(r=0.999 5),0.015 79~0.315 8 μg·mL^-1(r=0.999 6)和0.051 34~1.027 μg·mL^-1(r=0.999 9),平均加样回收率分别为97.4%(RSD=1.1%),95.0%(RSD=0.88%),97.5%(RSD=1.3%),97.4%(RSD=1.4%)和96.3%(RSD=0.87%)。结论 新建方法简便、准确,可用于黄石感冒片的定量分析方法。     

HPLC测定格列美脲有关物质的改进

目的 改进格列美脲有关物质检测的高效液相色谱法。方法 采用Agilent Zorbax SB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),以磷酸二氢钠溶液(取0.5 g磷酸二氢钠,加水500 mL溶解,用磷酸调pH至2.5)-乙腈(50∶50)为流动相,流速1.0 mL· min^-1,检测波长为228 nm。结果 溶剂对杂质检测无影响,格列美脲与已知杂质分离度良好,杂质1与杂质2的定量限均为10 ng·mL^-1,杂质1在0.304 2~2.028 μg·mL^-1,杂质2在0.316 2~2.108 μg·mL^-1内呈良好线性关系(r分别为0.999 5和0.999 7),平均回收率分别为 100%和99%,RSD为1.4%和1.3%。结论 改进的方法消除了溶剂对杂质检测的干扰,可用于格列美脲中有关物质的检测。     

HPLC法测定甲磺酸沙芬酰胺中对映异构体的含量

摘 要 目的： 建立测定甲磺酸沙芬酰胺中对映异构体含量的方法。方法: 采用HPLC法,色谱柱： Chiralpak AS-H柱（250 mm×4.6 mm,5 μm）;流动相：正己烷-乙醇-二乙胺（75∶25∶0.1）;流速：1.0 ml·min^-1;检测波长：225 nm;柱温：35℃。结果: 甲磺酸沙芬酰胺与其对映异构体之间的分离度大于2.0;两者分别在1.007~2517.500 μg·mL^-1和0.909~2273.200 μg·mL^-1范围内呈良好的线性关系,r均为0.999 0;对映异构体的平均加样回收率为104.9%,RSD为2.3%（n=9）。结论:该方法准确、快速,可用于甲磺酸沙芬酰胺产品中对映异构体的含量测定。     

全血及血清中羟氯喹HPLC测定方法的建立及其应用

目的：建立高效液相色谱法测定人全血及血清中羟氯喹浓度。方法：200 μL人全血（血清）样品经400 μL的10%甲酸-乙腈（1∶9）沉淀后,取上清液进行色谱分析。采用Zorbax SB-C₁₈为色谱柱,以乙腈-0.2 mol·L^-1磷酸二氢钾（15∶85,v/v,磷酸调pH=3.0）为流动相,检测波长：254 nm,流速：0.8 mL·min^-1,进样量：30 μL。结果：对46例长期（>1年）服用硫酸羟氯喹（200 mg·d^-1）的系统性红斑狼疮患者的血药谷浓度进行检测分析。羟氯喹在10~3000 ng·mL^-1线性关系良好,批内、批间精密度RSD均小于8.6%,全血及血清样品的提取回收率分别为74.5%~78.6%和80.7%~85.4%。患者平均全血/血清谷浓度分别为（707.6±398.4） ng·mL^-1、（282.2±153.8） ng·mL^-1。结论：本法操作简便,灵敏,准确度高,适用于羟氯喹血药浓度检测及药效学研究。     

西红花酸在大鼠的药代动力学研究

目的建立高效液相色谱法测定大鼠血浆中西红花酸的浓度。方法用HPLC法,色谱柱为Hypersil C₁₈柱(5 μm,4.6 mm×200 mm),流动相为甲醇-水-冰醋酸(75∶24.5∶0.5),流速1.0 mL·min^-1,柱温30℃,检测波长423 nm。结果西红花酸浓度0.49～7.87 μg·mL^-1与峰面积呈良好的线性关系(γ=0.9996),最低检测浓度为0.14 μg·mL^-1(S/N=3)。样品的平均加样回收率为105.2%。日内、日间精密度的RSD均小于5%。不同时间、不同温度及冻融处理的稳定性考察,RSD分别为7.4%,4.9%和3.3%。结论本法稳定、简单、可靠,可用于西红花酸血药浓度分析及其药代动力学研究。     

HPLC同时测定四季三黄片中芦荟大黄素、黄芩苷、黄柏酮和西红花苷-I的含量

目的 采用HPLC同时测定四季三黄片中芦荟大黄素、黄芩苷、黄柏酮和西红花苷-I的含量。方法 采用welch Topsil-C₁₈色谱柱(4.6 mm×250 mm,4 μm);流动相：甲醇-0.2%磷酸水溶液;梯度洗脱;检测波长分别为254 nm(芦荟大黄素、黄芩苷、黄柏酮)和440 nm(西红花苷-I);流速：1.0 mL·min^-1;柱温：30 ℃。结果 芦荟大黄素、黄芩苷、黄柏酮和西红花苷-I线性范围分别为0.079 10~1.582 μg·mL^-1(r=0.999 5),0.167 5~3.350 μg·mL^-1(r=0.999 8),0.097 92~1.958 μg·mL^-1(r=0.999 7)和0.033 57~0.671 5 μg·mL^-1(r=0.999 4),平均加样回收率分别为95.9%(RSD=0.7%),97.5%(RSD=0.8),96.4%(RSD=1.0%),95.5%(RSD=1.3%)。结论 该方法简便、准确、专属性强、重复性好,可用于四季三黄片中芦荟大黄素、黄芩苷、黄柏酮和西红花苷-I的定量分析。     

固相萃取高效液相色谱法测定人血浆中依那普利浓度

建立了用固相萃取高效液相色谱法测定依那普利血药浓度的方法。色谱柱为200mm×4.6mm不锈钢柱,内填 Spherisorb C₈(5μm),流动相为乙醇—水—10% H₃PO₄—三乙胺(30∶70∶1.5∶0.1);流速1.0ml·min^-1,紫外检测波长215nm。血样用固相小柱预处理。此法线性范围25～150ng·mL^-1。最小检测浓度1.5ng·mL^-1,日内及日间误差<8.8%,平均回收率>91.6%。用此法测定了8例健康志愿者po国产依那普利片后的血药浓度并计算了药代动力学参数。     

LC-MS/MS 法测定人血浆中倍他米松

建立测定人血浆中倍他米松的LC-MS/MS方法。采用Venusil XBP C₈ (200 mm×3.9 mm ID, 5 μm)色谱柱，流动相为甲醇-水(含甲酸铵5 mmol·L^-1)(80∶20)，流速0.4 mL·min^-1；质谱仪离子源为电喷雾离子源(ESI)，正离子模式检测，监测离子为393.3→355.2(倍他米松)和361.3→343.2(泼尼松龙,内标)。血浆样本用乙酸乙酯处理。倍他米松在0.5~80.0 ng·mL^-1线性关系良好(r=0.999 2)， 血浆低、 中、 高3种浓度(1.0， 10.0， 60.0 ng·mL^-1)平均提取回收率为88.24%，定量限为0.5 ng·mL^-1。本方法操作简便、准确、灵敏，适用于复方倍他米松注射液人体药代动力学研究。     

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.     

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.     

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.     

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.