HPLC-FLD法测定清火片中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚

目的建立高效液相色谱–荧光检测(HPLC-FLD)法测定清火片中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚。方法采用Agilent C18色谱柱(250 mm34.6 mm,5μm);流动相:甲醇–0.1%磷酸溶液(78∶22);体积流量为1 mL/min;柱温30℃;进样量为20μL。对比HPLC-FLD法与HPLC-UV法的测定结果。结果芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚分别在100.6～1 609.6、86.8～1 388.8、359.4～5 750.4、81.2～1 299.2、89.4～1 430.4 ng线性关系良好;游离蒽醌中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的平均回收率分别为93.9%、90.8%、96.2%、92.9%、89.2%,RSD值分别为2.2%、2.4%、1.0%、2.4%、0.6%;总蒽醌中各成分平均回收率分别为100.2%、85.7%、99.2%、104.5%、104.2%,RSD值分别为2.5%、2.2%、3.7%、0.5%、0.7%。结论本方法具有重复性好、灵敏度高、专属性强、内源物干扰小等优点,与HPLC-UV法测定结果一致性较高,可用于中成药清火片的质量控制。     

HPLC法测定通腑胶囊中大黄素、大黄酸、大黄酚、大黄素甲醚及芦荟大黄素的含量

目的建立高效液相色谱法测定通腑胶囊中大黄素、大黄酸、大黄酚、大黄素甲醚及芦荟大黄素含量的方法。方法采用HPLC法,色谱柱:Waters C18(250×4.6mm,5μm),以0.1%磷酸:甲醇(15∶85)为流动相,流速1.0μL/min,检测波长:254nm,柱温:35℃。结果大黄素、大黄酸、大黄酚、大黄素甲醚及芦荟大黄素进样量分别在0.0162~0.323μg、0.016~0.318μg、0.0164~0.328μg、0.008~0.16μg、0.020~0.406μg范围内进样量与峰面积积分值呈良好的线性关系,相关系数均为0.9999,平均回收率分别为:99.0%、99.0%、99.2%、98.3%、99.4%(n=6)。结论该方法专属性强,重现性好,可用于通腑胶囊中蒽醌类成分的含量测定。     

用HPLC法测定牛黄上清片中各大黄素成分的含量

目的:建立HPLC同时测定牛黄上清片中芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚含量的方法.方法:色谱柱为Shimadzu VP-ODS柱(4.6 mm×250 mm,5 mm),流动相为乙腈-0.1%磷酸梯度洗脱,流速为1.0 ml/min,检测波长为254 nm,柱温30℃.结果:芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚分别在0.042～0.840μg(r=0.9996)、0.168～3.352μg(r=0.9998)、0.084～1.680 μg(r=0.9997)、0.093～1.852μg(r=0.9999)和0.174～3.488μg(r=0.9994)内呈良好线性关系,平均回收率分别为99.8%(RSD=1.34%)、99.8%(RSD=1.72%)、100.0%(RSD=0.89%)、99.5%(RSD=0.97%)和100.2%(RSD=1.04%).结论:本方法简便、快速、准确,可用于牛黄上清片的质量控制.     

HPLC法同时测定痔痛安搽剂中三种蒽醌类成分的含量

目的建立同时测定痔痛安搽剂中三种蒽醌类成分大黄素、大黄酚和大黄素甲醚含量的方法,提高制剂的质量控制标准。方法采用HPLC法。色谱柱为Diamonsil C18柱,流动相为甲醇-质量分数为0.1%的磷酸水溶液(体积比75∶25),流速1.0 mL.min-1,检测波长254 nm,柱温35℃。结果大黄素、大黄酚、大黄素甲醚质量浓度分别在0.64~20.48 mg.L-1(r=0.9995)、0.31~9.84 mg.L-1(r=0.9997)、0.50~16.08 mg.L-1(r=0.999 8)内与峰面积呈良好的线性关系;大黄素平均加样回收率为98.8%,大黄酚平均加样回收率为99.6%,大黄素甲醚平均加样回收率为98.5%。结论采用HPLC法可同时测定痔痛安搽剂中三种蒽醌类成分大黄素、大黄酚和大黄素甲醚的含量,且操作方法简便,分离效果好,运行成本低。该法可用于痔痛安搽剂的质量评价。     

大败毒胶囊中总蒽醌的含量测定

目的建立测定大败毒胶囊中总蒽醌含量的高效液相色谱法。方法以ZORBAX C18（250 mm×4.6 mm，5 μm）为色谱柱，甲醇 0.1%磷酸溶液（85：15）为流动相，流速1.0 mL•min－1，柱温25 ℃，检测波长254 nm。 结果色谱峰分离度良好，大黄酸在5.724×10－2～0.572 μg、大黄素在4.584×10－2～0.458 μg、大黄酚在6.368×10－2～0.637 μg、大黄素甲醚在6.548×10－2～0.655 μg范围内呈良好线性关系。大黄酸、大黄素、大黄酚、大黄素甲醚平均加样回收率分别为97.8%，100.7%，98.6%，97.0%。结论该方法简单、快速、重复性好，可用于大败毒胶囊中总蒽醌的质量控制。     

HPLC法同时测定三黄片中4种成分的含量

目的同时测定三黄片中的盐酸小檗碱、大黄素、大黄酚和大黄素甲醚含量。方法高效液相色谱法,色谱柱Dik-ma Diamondsil C18色谱柱(250mm×4.6mm,5μm),柱温25℃,检测波长255nm,流动相为乙腈:0.5%甲酸水溶液(三乙胺调pH=3.5)梯度洗脱,流速1.0mL·min-1。结果保留时间分别为盐酸小檗碱19.4min、大黄素26.9min、大黄酚31.1min、大黄素甲醚33.0min。结论该方法能够准确、快速地测定三黄片中4种有效成分。     

HPLC法同时测定清瘟解毒散中芦荟大黄素、大黄素、大黄酚及大黄素甲醚的含量

目的建立清瘟解毒散中芦荟大黄素、大黄素、大黄酚、大黄素甲醚的含量测定方法。方法采用HPLC法,以Wondasil C18(250 mm×4.6 mm,5μm)柱为色谱柱,流动相为甲醇-体积分数为0.1%的磷酸溶液(体积比为85∶15),以1.0 mL·min~(-1)为流速,以254 nm为检测波长,柱温为30℃。结果芦荟大黄素、大黄素、大黄酚、大黄素甲醚分别在5~30、5~30、10~60和5~30 mg·L~(-1)内呈良好线性关系,相关系数分别为0.999 5、0.999 8、0.999 8和0.999 8,平均回收率分别为99.0%、99.2%、98.6%和98.9%。结论该方法可以作为清瘟解毒散质量的控制指标之一。     

HPLC法测定胆石通胶囊中芦荟大黄素、大黄酸、大黄素、大黄酚及大黄素甲醚的含量

目的建立同时测定胆石通胶囊中芦荟大黄素、大黄酸、大黄素、大黄酚及大黄素甲醚的含量测定方法。方法以UltimateTM XB-C18(250 mm×4.6 mm,5μm)为色谱柱,柱温:35℃;流动相:甲醇-1 g·L-1磷酸溶液,梯度洗脱;流速:1.0 mL·min-1;检测波长:430 nm;进样量10μL。结果芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚分别在60.02~1 200.4,70.32~1 406.4,62.24~1 244.8,71.04~1 420.8和30.48~609.6 ng范围内呈良好线性关系,相关系数分别为0.999 9,0.999 9,0.999 9,0.999 5和0.999 9,平均回收率分别为98.1%,99.3%,98.4%,99.0%和99.2%。结论该方法专属性好,准确度高,可用于综合评价胆石通胶囊的质量。     

HPLC法测定胃力片中大黄酸、大黄素、大黄酚和大黄素甲醚的含量

目的建立胃力片中大黄酸、大黄素、大黄酚、大黄素甲醚的含量测定方法。方法采用HPLC法,色谱柱:Diamonsil C18(4.6 mm×200 mm,5μm),以甲醇-体积分数为0.1%的磷酸水溶液(体积比为82∶18)为流动相,流速:1.0 mL.min-1,柱温:35℃,检测波长:254 nm。结果大黄酸、大黄素、大黄酚、大黄素甲醚分别在1.42~12.79、2.02~18.14、1.28~11.52、0.76~8.36 mg.L-1内呈良好的线性关系,相关系数分别为0.999 1、0.999 4、0.999 0和0.999 6,平均回收率分别为102.7%(RSD=1.0%,n=9)、101.2%(RSD=2.7%,n=9)、99.4%(RSD=1.6%,n=9)和97.9%(RSD=2.8%,n=9)。结论本方法可作为胃力片质量控制方法之一。     

HPLC法测定金花消痤丸中大黄酸、大黄素、大黄酚及大黄素甲醚的含量

目的建立金花消痤丸中大黄酸、大黄素、大黄酚、大黄素甲醚的含量测定法。方法采用HPLC法测定含量:Agilent Eclipse XDB-C18(4.6 mm×250 mm,5μm)色谱柱,0.1%磷酸—甲醇(24∶76)为流动相,检测波长254 nm,流速1.0 mL·min-1;柱温:30℃。结果大黄酸、大黄素、大黄酚、大黄素甲醚分离完全,平均加样回收率分别为96.6%、95.7%、97.4%、95.1%。结论该方法简便,快速,准确。     

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.     

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.     

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.     

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.