RP-HPLC法测定硫唑嘌呤代谢产物6-巯基嘌呤血浆药物浓度

目的：建立RP-HPLC法检测人血浆中硫唑嘌呤(AZA)代谢产物6-巯基嘌呤(6-MP)的浓度,为临床进行血药浓度监测提供基础。方法：采用Hypersil ODS2色谱柱(250 nm * 4.6 nm,5μm),流动相为乙腈-0.3%冰乙酸化(3:97),流速为0.8 mL.min-1,检测波长323 nm,血浆样品经15%高氯酸去蛋白后加氢氧钠(NaOH)调节PH后20μL进样分析。结果：6-MP血浆样品在0.02-2μg.mL-1范围内线性关系良好,最低检测下限(LLOQ)为0.02μg.mL-1；LLOQ、高浓度(1.5μg.mL-1)、中浓度(1μg.mL-1)和低浓度(0.05μg.mL-1)提取回收率分别为85.71%、92.93%、103.53%和96.02%,质控四个浓度的日内与日间精密度均<15.0%,符合中国药典生物样本分析相关规定。结论：本实验成功建立一种简单、快速、高灵敏度的6-MP血浆浓度测定方法,符合AZA血药浓度监测和药代动力学研究的相关规定。     

HPLC法测定劳拉西泮片有关物质的含量

目的 建立劳拉西泮片的含量及有关物质的HPLC方法。方法 以十八烷基硅烷键合硅胶作为填充剂的ZORBAX SB-C₁₈柱(250 mm×4.6 mm,5 μm)为色谱柱,流动相:0.05 mol·L-1的磷酸二氢铵(含0.5%的三乙胺,用磷酸调pH至2.5)∶甲醇∶乙腈=35∶35∶30;流速:1.0 mL·min-1;检测波长:235 nm;进样量:10 μL;柱温:30℃。结果 劳拉西泮峰及各杂质峰均能良好分离。杂质A浓度与峰面积在1.5 μg·L-1 ～8.048 mg·L-1内线性关系良好。杂质B浓度与峰面积在15.0 μg·L-1～8.224 mg·L-1内线性关系良好。杂质C浓度与峰面积在1.7 μg·L-1～8.144 mg·L-1内线性关系良好;杂质D浓度与峰面积在2.5 μg·L-1～8.032 mg·L-1内线性关系良好;杂质E浓度与峰面积在3.0 μg·L-1～8.032 mg·L-1内线性关系良好。测定样品含量平均值为97.9%,有关物质均符合要求。结论 该方法快速、简单,稳定,重现性好,可作为劳拉西泮片的含量及有关物质的检测方法。     

HPLC-MS/MS法测定人血浆中左乙拉西坦浓度的不确定度分析

目的 对高效液相色谱串联质谱(HPLC-MS/MS)法测定人血浆中左乙拉西坦浓度的不确定度进行评定。方法 用HPLC-MS/MS法测定人血浆中左乙拉西坦浓度,对测试过程进行分析,对各个影响因素,包括测定精密度、标准曲线拟合、称量、标准溶液的配制、含药血浆的配制、提取回收率、基质效应、仪器等进行分析评定,计算合成不确定度和扩展不确定度。结果 血浆中低(0.100 2 mg·L-1)、中(2.004 mg·L-1)、高(30.06 mg·L-1)左乙拉西坦浓度测定的合成不确定度为0.010 50、0.123 3、1.958 0 mg·L-1,扩展不确定度分别为0.021 00、0.246 6、3.916 0 mg·L-1。结论 HPLC-MS/MS法测定人血浆中左乙拉西坦浓度的不确定度主要由标准曲线拟合(低浓度) 、仪器误差、标准溶液的配制和含药血浆的配制引入。     

高效液相色谱法测定稳心律合剂中4种丹参类成分含量

目的 建立高效液相色谱法(HPLC)测定稳心律合剂中4种丹参类成分的含量。方法 采用Agilent Zorbax SB C₁₈色谱柱(250 mm×4.6 mm,5 μm),柱温35 ℃,流动相A为乙腈,流动相B为0.03 mol·L-1乙酸铵(甲酸调pH至2.4),进行梯度洗脱,流速1.0 mL·min-1,检测波长280 nm。结果 4种丹参类成分(丹参素钠在15.41~154.1 mg·L-1;原儿茶醛在4.79~47.9 mg·L-1;丹酚酸B在7.37~73.7 mg·L-1;丹参酮Ⅱa在4.22~42.2 mg·L-1浓度范围内)均呈良好线性关系(r=0.999 9);平均回收率为95.79%~97.48%;RSD均小于3.0%。结论 该方法灵敏度高、准确性好、具有较好的重复性和稳定性,可用于稳心律合剂的质量控制。     

高效液相色谱梯度洗脱法同时测定冠心苏合滴丸中5个成分的含量

目的 建立高效液相色谱(HPLC)梯度洗脱法同时测定冠心苏合滴丸中的马兜铃酸Ⅰ、荭草苷、异荭草苷、牡荆苷和异牡荆苷含量。方法 采用Diamonsil C₁₈色谱柱(250 mm×4.6 mm,5 μm),以甲醇(A)-0.5％冰醋酸溶液(B)为流动相,进行梯度洗脱,流速0.9 mL·min-1,柱温30 ℃,波长切换法检测(0～21 min,390 nm,马兜铃酸Ⅰ;21～60 min,338 nm,荭草苷、异荭草苷、牡荆苷和异牡荆苷);进样量为10 μL。结果 马兜铃酸Ⅰ、荭草苷、异荭草苷、牡荆苷和异牡荆苷5个成分的线性范围分别为4.16～83.20 mg·L-1(r=0.999 6)、6.35～127.00 mg·L-1(r=0.999 8)、4.98～99.60 mg·L-1(r=0.999 2)、14.33～286.60 mg·L-1(r=0.999 7)、16.37～327.40 mg·L-1(r=0.999 4);平均加样回收率及相应的RSD分别为98.26%(1.40%),99.23%(1.35%),97.90%(1.58%),96.97%(1.14%),99.11%(1.58%)。结论 该文建立的方法符合方法学验证要求,可用于冠心苏合滴丸中的5个指标性成分的同时定量测定。     

HPLC法测定人血浆中曲唑酮的浓度

目的 :建立测定人血浆中曲唑酮 (TZD)浓度的RP HPLC法。方法 :以美国Dikma公司DiamonsilTMC18反相柱 (2 5 0mm×4 6mm ,5 μm)为色谱柱 ,流动相为甲醇 超纯水 (85∶15 ,V/V) ,流速为 0 8mL·min-1,检测波长 2 5 6nm ,以乙酸乙酯为提取剂。结果 :TZD高 (83 33mg·L-1)、中 (16 6 7mg·L-1)、低 (1 6 7mg·L-1) 3个浓度的平均回收率分别为 10 0 19% ,97 4 5 % ,95 5 1% ;日内、日间RSD均 <5 % (n =5 ) ;分析方法的最低检测浓度为 0 0 8mg·L-1。线性范围为 0 83～ 16 6 6 7mg·L-1,回归方程为Y =0 0 2 16X - 0 0 0 333,r=0 9999(n =10 )。结论 :该方法可用于临床TZD血药浓度监测和药动学研究     

高效液相色谱法测定“天竺痫康宁胶囊”中胆红素的含量

目的 建立天竺痫康宁胶囊胆红素含量的高效液相色谱(HPLC)测定方法。方法 采用Intersil-C₁₈色谱柱,流动相为甲醇-三氯甲烷-1%磷酸(80∶14∶6),流速1.0 mL·min-1,检测波长为450 nm,柱温为室温,进样量为10 μL。结果 在该条件下,浓度为2.26～45.2 mg·L-1范围内,胆红素峰面积与其浓度呈良好线性关系(r=0.999 6),平均回收率为97.4%,RSD为2.02%(n=6)。结论 该方法灵敏、简便、准确、重复性好,可作为天竺痫康宁胶囊质量标准的控制指标之一。     

高效液相色谱法测定人血浆中麦考酚酸的血药浓度

目的 :建立测定麦考酚酸 (MPA)血药浓度的方法并对肾移植病人给药后的血药浓度进行测定。方法 :采用高效液相色谱 (HPLC)法测定血药浓度 ,检测波长 30 3nm。乙腈 (含有 0 .1mol·L- 1磷酸 )作为血样的蛋白沉淀剂。色谱柱 :AgilentXDBC18(15 0mm× 4 .6mm ,5 μm) ;流动相 :乙酸钠缓冲液 (pH =3.0 ,30mmol·L- 1)∶乙腈 =5 7∶4 3(V/V) ;流速 :1mL·min- 1。结果 :MPA血药浓度线性范围为 0 .6～ 30mg·L- 1(r =0 .9995 ,n =6 ) ,其血浆最低检测浓度为 0 .2mg·L- 1;低、中、高 3种浓度MPA的方法回收率为 (98.9± 1.5 ) % ,提取回收率为 (10 6 .73± 0 .2 7) % ;相应 3种浓度MPA的精密度RSD均小于 5 %。结论 :HPLC法能快速、准确对MPA进行血药浓度测定 ,可以用于MPA的临床血药浓度监测和药动学研究。     

人血浆中阿莫西林-克拉维酸浓度的测定

目的 :建立阿莫西林 -克拉维酸血药浓度的HPLC法。方法 :固定相为C18对称反相色谱柱 ;流动相为磷酸 -四丁基氢氧化铵缓冲液∶甲醇 (4 8∶5 2 ,V/V) ;流速 0 8mL·min-1;荧光检测波长 :Ex 386nm ,Em 4 6 0nm ;血浆标本以高氯酸去蛋白 ,再以苯甲醛荧光衍生后上柱分析 ,进样量 5 0 μL。 结果 :阿莫西林和克拉维酸的色谱峰分离良好 ,最低检测质量浓度分别为 0 0 0 9和 0 0 1mg·L-1;线性范围分别为 0 12 5～ 2 5和 0 0 6 2 5～ 8mg·L-1;平均日内、日间RSD均 <9% ,提取回收率近 10 0 %。结论 :本方法适用于阿莫西林 -克拉维酸血药浓度测定     

万古霉素血药浓度监测256例次分析

目的 分析万古霉素血药浓度监测和临床应用情况,为临床合理用药提供参考。方法 回顾性调查安徽医科大学第一附属医院2016年7月至2017年6月进行万古霉素血药浓度监测的病人信息,统计分析血药浓度、感染部位、肾功能等数据。结果 纳入141例病人、256例次浓度监测,总血药浓度(19.22±11.33)μg/mL,首次浓度(18.40±11.88)μg/mL,浓度在10[KG-*3]～20 μg/mL的构成比分别为32.81%、30.05%,两组差异无统计学意义。eGFR 75[KG-*3]～110 mL·min-1·1.73 m-2组病人万古霉素浓度达标率45.20%。58.73%的肾功能严重受损病人血药浓度高于20 μg/mL(eGFR<40 mL·min-1·1.73 m-2),然而eGFR≥110 mL·min-1·1.73 m-2的病人中,45.24%血药浓度低于10 μg/mL。用药前后,5例病人肾损害可能与药物相关。结论 万古霉素首次与总血药浓度达标率均较低,尤其是肾功能不全或亢进的病人人群,监测结果未能在临床应用中充分发挥作用。临床应用加强血药浓度监测,同时综合病人临床情况,选择适宜的给药方案,实现个体化治疗,提高药物的安全、有效性。     

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.     

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.