厄贝沙坦片在大鼠体内血药浓度的测定及药动学研究

目的:建立厄贝沙坦片给药后大鼠血药浓度的高效液相色谱测定法并进行药动学研究。方法:色谱柱为Consmosil-C18,流动相为磷酸-甲醇-乙腈(7∶80∶13,V/V/V),流速为1.0mL·min-1,检测波长为272nm;大鼠灌胃给予厄贝沙坦水溶液30mg·kg-1后分别在在给药前及给药后0、5、10、20、30、45、60、120、180、240、360、480min测定血浆中厄贝沙坦的浓度,计算药动学参数。结果:厄贝沙坦的检测浓度线性范围为0.5～50μg.mL-(1r=0.999 1);回收率为93.89%,RSD小于4.14%;t1/2、tmax、Cmax、AUC0～t分别为(48.71±4.037)h、(0.33±0.12)h、(1.33±0.18)μg·mL-1、(23.10±1.56)mg·L·h-1。结论:所建立的血药浓度测定方法简单、灵敏、专一,能够满足血药浓度和药动学研究的需要。     

高效液相色谱法用于褪黑素香口胶的含量测定

目的建立褪黑素香口胶质量控制的高效液相色谱方法。方法PolarisC18色谱柱,乙腈-0.05mol/L醋酸盐缓冲液(1∶3,pH4.6)为流动相,流速1.0ml/min,检测波长278nm。测定制剂的含量及其均匀度、稳定性和溶出度。结果在4.86~97.20μg/L范围内,褪黑素浓度与色谱峰高比呈现良好线性相关,在高、中、低浓度时平均总加样回收率分别为(103.35±3.98)%,(101.88±3.91)%和(100.46±4.42)%(n=6);平均日内和日间RSD分别为2.68%和3.86%(n=5);最低检出浓度为0.1μg/L。结论此方法适用于香口胶中腿黑素测定。     

HPLC法测定大鼠血浆中奥拉西坦的浓度

目的:建立大鼠血浆中奥拉西坦浓度的检测方法。方法:取大鼠8只,一次性灌胃给予奥拉西坦200 mg.kg-1,给药2 h后取血,采用高效液相色谱法检测其血药浓度。色谱柱:Symmetry shiedTMRP18;流动相:乙腈-水(3.2∶96.8,V/V),流速:0.8 mL.min-1;检测波长:210 nm;柱温:40℃;进样量:20μL。结果:奥拉西坦的保留时间为3.65 min;其检测浓度的线性范围为2～100 mg.L-1(r=0.999 8);低、中、高浓度血浆样品的绝对回收率分别为(91.1±5.17)%、(97.9±3.22)%、(98.4±1.99)%,方法回收率分别为(96.35±3.01)%、(101.40±3.41)%、(102.23±1.37)%;日内RSD分别为2.86%、4.38%、3.54%,日间RSD分别为3.25%、3.17%、3.14%;奥拉西坦血浆样品-40℃保存30 d及反复冻融3次后性质稳定,血浆样品处理后放置4 h对测定无影响。8只大鼠血浆样品中奥拉西坦的平均浓度为(35.58±10.83)mg.L-1。结论:本方法简便、准确、灵敏度高、重现性好,可用于大鼠血浆中奥拉西坦浓度的测定。     

高效液相色谱法测定金水宝胶囊中腺苷及腺嘌呤的含量

目的建立测定金水宝胶囊中腺苷及腺嘌呤含量的高压液相色谱方法。方法采用0.5%磷酸超声提取制备金水宝胶囊样品溶液,色谱柱为Venusil-XBP-C18(250 mm×4.6 mm,5μm);流动相采用[0.01 mol.L-1 NaH2PO4∶0.01 mol.L-1 Na2HPO4(1∶2.2,V/V)]∶甲醇(17∶3,V/V),流速0.6 mL.min-1,进样量10μL,柱温25℃,检测波长260 nm,运行时间18 min。运行时用流动相∶甲醇(85∶15,V/V)梯度洗脱。结果腺苷及腺嘌呤分别在0.62～157.07 mg.L-1和0.23～58.29 mg.L-1内线性关系良好;两者的平均回收率分别为(97.40±4.28)%和(99.10±5.03)%;3种批号的金水宝胶囊样品中腺苷的含量分别为(0.324±0.017)%、(0.318±0.025)%和(0.331±0.019)%;腺嘌呤的含量分别为(0.089±0.009)%、(0.086±0.012)%和(0.093±0.007)%。结论高效液相色谱法测定金水宝胶囊中腺苷及腺嘌呤的含量操作简便、结果准确可靠、灵敏度高、重复性好,能作为有效控制金水宝胶囊质量的测定方法之一。     

人血浆中卡马西平血药浓度的测定

目的:建立人血浆中卡马西平血药浓度的高效液相色谱-二级管阵列检测器(HPLC-DAD)测定法。方法:采用色谱柱为Symmetry C_(18)(3.9 mm×150 mm,5μm);以乙腈-含0.067%甲酸的0.077%醋酸铵溶液(0.3∶0.6)为流动相;流速为0.9 m L/min;检测波长为245 nm;柱温为40℃;以阿普唑仑为内标,血浆经无水乙醚提取后检测卡马西平血药浓度。结果:血浆中卡马西平浓度在0.6~28.8μg/m L范围内呈良好的线性关系(r=0.999 8);低、中、高3个浓度(1.2,9.6,19.2μg/m L)的日内RSD分别为3.34%,2.59%和1.99%;日间RSD分别为4.51%,2.55%和2.96%;绝对回收率分别为96.42%、95.45%和93.59%,相对回收率分别为100.60%、101.14%和102.15%。结论:该方法简便、快速、灵敏、准确,适用于人血浆中卡马西平的浓度测定。     

高效液相梯度洗脱法测定灵杞黄斑颗粒中毛蕊花糖苷和丹酚酸B含量

目的建立同时测定灵杞黄斑颗粒中毛蕊花糖苷和丹酚酸B含量的反相高效液相梯度洗脱法。方法采用Agilent Eclipse XDB-C18(250mm×4.6mm,5μm)色谱柱,流动相:乙腈(A)-0.1%磷酸水溶液(B)进行梯度洗脱,梯度洗脱程序为进样0～8min,调节流动相A∶B(20∶80,V/V);8～20min,流动相A∶B(30∶70,V/V);20～23min,流动相A∶B(20∶80,V/V);流速:1.0mL·min-1,检测波长:334nm,柱温:30℃,进样量:10μL。结果毛蕊花糖苷和丹酚酸B分别在1.50～14.97mg·L-1和18.92～189.23mg·L-1范围内与峰面积呈良好的线性关系,相关系数分别为0.9998和0.9999,平均加样回收率毛蕊花糖苷为(99.7±s1.1)%,RSD=1.12%,丹酚酸B为(99.5±1.0)%,RSD=1.02%,3批样品中毛蕊花糖苷平均含量均不低于36.9μg·g-1,RSD均不大于1.54%,丹酚酸B平均含量均不低于344μg·g-1,RSD均不大于1.94%。结论该方法简便、快速、准确、灵敏度高、专属性强,可用于灵杞黄斑颗粒剂的质量控制。     

柱前衍生化法测定血浆中醋酸精氨酸的浓度

目的:建立柱前衍生化高效液相色谱法测定血浆样品中醋酸精氨酸浓度的方法。方法:衍生剂为2,4—二硝基氟苯,色谱柱为氨基酸分析专用ODS色谱柱,流动相为50%乙腈-0.1mol/L醋酸钠溶液(29∶71),流速为1.2ml/min,检测波长为360nm,柱温为30℃。结果:醋酸精氨酸检测浓度线性范围为10~160μg/ml(r=0.9 998),高、中、低3种浓度血浆样品的回收率分别为(96.64±9.69)%、(90.94±4.70)%、(92.72±4.48)%,精密度<3.91%。含药血浆在冰冻条件(—24℃)下至少可以稳定15d。结论:本法简便可行,适用于醋酸精氨酸的血药浓度监测及其药动学研究。     

HPLC法测定狼疮肾病患者口服吗替麦考酚酯后的血药浓度

目的:建立测定狼疮肾病患者口服吗替麦考酚酯后血药浓度的方法。方法:狼疮肾病患者血样处理后采用高效液相色谱法测定吗替麦考酚酯体内代谢物霉酚酸。色谱柱为SGE Protecol C18,流动相为甲醇-乙腈-0.01%磷酸(23∶37∶40,V/V/V),检测波长为216nm,流速为1.0ml/min,进样量为20μl,柱温为30℃。结果:霉酚酸血药浓度在0.125～10mg/L范围内线性关系良好(r=0.9999);方法回收率为98.67%～104.31%;萃取回收率为83.43%～97.54%,内标(尼美舒利)萃取回收率为79.35%;日内、日间RSD分别为2.6%～7.7%、2.3%～8.5%。结论:本法简单、准确,可用于狼疮肾病患者服用吗替麦考酚酯后霉酚酸的血药浓度检测。     

高效液相色谱法测定青霉素V钾片中主药的含量

目的:建立以高效液相色谱法测定青霉素V钾片中主药含量的方法。方法:色谱柱为DiamonsilC18,流动相为乙腈-0.02mol/L醋酸钠(70∶30),流速为1.0ml/min,检测波长为268nm,灵敏度为0.01AUFS,柱温为(26±1)℃,进样量为20μl。结果:青霉素V钾检测浓度在10.0～120.0μg/ml范围内与峰面积呈良好的线性关系(r=0.9999,n=5);平均加样回收率为100.2%(RSD=0.42%)。结论:本方法快速、简便、准确,可用于青霉素V钾片的质量控制。     

高效液相色谱法测定复方醋酸地塞米松搽剂中2组分的含量

目的建立高效液相色谱法测定复方醋酸地塞米松搽剂中地塞米松、樟脑的含量。方法 CAPCELL PAK C18色谱柱(4.6 mm×150 mm,5μm),流动相:乙腈-水(65∶35,V/V),柱温:30℃,流速:1.0 mL.min-1,检测波长:298 nm。结果地塞米松、樟脑进样量分别在0.375～6.00μg(r=0.999 9)、9.69～155.04μg(r=0.999 9)范围内有良好线性关系,平均回收率分别为(100.75±0.83)%、(99.61±0.71)%,RSD分别为0.81%、0.69%(n=9)。结论本方法简便、准确、重复性好,可用于复方醋酸地塞米松搽剂的质量控制。     

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.

---

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.