HPLC法测定阿巴卡韦双夫定片中的含量

摘 要 目的： 建立HPLC法测定阿巴卡韦双夫定片中阿巴卡韦、拉米夫定、齐多夫定的含量。方法: 采用Hypersil C₁₈(200mm×4.6mm,5μm)色谱柱,以甲醇 庚烷磺酸钠溶液（取庚烷磺酸钠2.02 g,加三乙胺5 ml,加水至700 ml,用磷酸调pH至3.9）（30∶70）为流动相,流速为1.0 ml·min^-1,检测波长270 nm。结果: 拉米夫定、齐多夫定、阿巴卡韦分别在0.08～1.60 μg (r=0.999 4),0.24～4.80 μg (r=0.999 9),0.24～4.80 μg (r=0.999 9)范围内线性关系良好, 平均回收率分别为99.6%（RSD=0.9%）,99.7%(RSD=0.9%),99.9%（RSD=1.0%）。结论: 本方法简便、可靠、准确,可用于同时测定三组分含量。     

HPLC测定人工关节假体感染患者关节液中万古霉素浓度

目的 建立HPLC测定人工关节假体周围感染患者关节液中万古霉素浓度的方法。方法 以替硝唑为内标,采用Hypersil C₁₈柱（250 mm×4.6 mm,5 μm）,流动相为乙腈-0.012 5mol·L^-1磷酸二氢钾缓冲液（pH=3.2）（10：90）,流速1 mL·min^-1,柱温35℃,检测波长236 nm,进样量10 μL。结果 关节液中万古霉素浓度在0.906 8~90.681 8 μg·mL^-1线性良好。批内、批间精密度RSD均<15%,准确度85.89%~103.51%。结论 本法易于操作,精密度和准确度良好,适用于人工关节假体周围感染患者关节液中的万古霉素浓度监测。     

HPLC测定三苯甲基氯沙坦的有关物质

目的 建立高效液相色谱法测定三苯甲基氯沙坦有关物质。方法 采用Lichrospher RP-select B 60A(250 mm×4.0 mm,5 μm)色谱柱;流动相：采用0.01 mol·L^-1的磷酸二氢钠溶液(20%的磷酸调节pH 2.5)和乙腈为流动相进行梯度洗脱;流速：1.5 mL·min^-1;柱温：35 ℃;检测波长：230 nm。结果 样品中的杂质均能很好检出,各杂质与主峰之间的分离度良好。结论 该方法简单快速,专属性强,结果准确,可用作三苯甲基氯沙坦有关物质检查。     

HPLC法测定辛伐他汀滴丸的含量和有关物质

目的: 建立辛伐他汀滴丸含量测定及有关物质的检查方法。方法: 采用反相高效液相色谱法,色谱柱为SUPELCO C₁₈柱(4.6 mm×33 mm,3μm),以乙腈-0.1%磷酸(50:50)为流动相A,0.1%磷酸乙腈溶液为流动相B,梯度洗脱,流量3.0 mL·min^-1,检测波长238 nm。结果: 辛伐他汀与辛伐他汀酸、洛伐他汀及强制破坏产生的降解产物均分离良好,辛伐他汀浓度在20.94～188.5μg·mL^-1范围内,与峰面积呈良好的线性关系,回归方程A=9 414.7C 7 327.2,r=1.000(n=7);平均回收率为99.8%,RSD为0.77%(n=9)。结论: 该法专属性强,准确、灵敏,可用于辛伐他汀滴丸的含量测定和有关物质检查。     

少腹逐瘀丸中松香酸的HPLC-MS检测方法研究

目的：建立少腹逐瘀丸中添加松香酸的HPLC和HPLC-MS/MS检测法。方法：采用HPLC、HPLC-PDA、HPLC-MS等。色谱柱为C18柱（250 mm×4.6 mm,5μm）,体积流量1.0 mL·min^-1,流动相为乙腈-0.1%甲酸水（82：18）,检测波长为241 nm。液质联用验证采用C18柱,流动相为乙腈-0.1%甲酸水,体积流量0.2 mL·min^-1,梯度洗脱。结果：松香酸进样量在0.1582~1.582μg·mL^-1与峰面积呈良好的线性关系;平均加样回收率为99.8%,RSD为0.23%。结论：该方法简便、快速、准确,专属性稳定性及重复性均良好。本文所建立的检测方法,可满足定性定量检测少腹逐瘀丸中非法添加松香酸的要求。     

HPLC-MS/MS测定替加环素脑脊液浓度及临床应用

摘 要 目的： 建立测定人脑脊液中替加环素(TGC)浓度的液质联用方法,验证后应用于临床样本的TGC浓度测定。方法: 使用Kromasil C₁₈ (50 mm×2.1 mm,5 μm)色谱柱,使用含0.05%三氟乙酸的水和乙腈为流动相梯度洗脱,流速为0.6 ml · min^-1,柱温40℃。TGC和内标d9 替加环素(IS)在正离子电喷雾电离模式下的定量离子对分别为m/z 586.3>513.2和m/z 595.3>514.3。结果: TGC浓度在250 ~ 100 000 ng · ml^-1范围内线性良好。TGC在低、中、高浓度的回收率在72.82%~ 82.86%之间,内标归一化的基质效应在101.13%~103.58%之间,所有质控样品在考察的条件下均稳定。结论:本研究建立的使用液质联用测定TGC脑脊液浓度的方法准确并且稳定,可用于TGC的药动学研究。     

HPLC法同时测定不同产地荔枝草中齐墩果酸和熊果酸含量

摘 要 目的：建立HPLC法同时测定不同产地荔枝草中齐墩果酸和熊果酸的含量。方法: 采用Agilent C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇-水-冰醋酸 三乙胺（90∶10∶0.03∶0.06）,检测波长：210 nm,流速：1.0 ml·min^-1,柱温：25 ℃。结果: 齐墩果酸在7.50～75.00 μg· mL^-1、熊果酸在5.00～50.00 μg · mL^-1 浓度范围内线性关系良好,r均为0.999 7。齐墩果酸和熊果酸的平均加样回收率分别为100.15%（RSD=1.14%,n=6）、99.40%（RSD=1.67%,n=6）。结论:该方法简便、准确,适用于荔枝草中齐墩果酸和熊果酸的含量测定。     

UPLC-MS/MS同时检测消癌平注射液中通关藤苷A、I、H的含量

目的 建立UPLC-MS/MS同时测定消癌平注射液中通关藤苷A、通关藤苷I、通关藤苷H的含量。方法 采用Phenomenex Kinetex XB-C₁₈色谱柱（2.1 mm×50 mm,2.6 μm）,以0.1%甲酸水-乙腈为流动相,梯度洗脱分离,流速为0.2 mL·min^-1,通过电喷雾离子源,多反应监测（MRM）,正离子模式。结果 通关藤苷A、I、H分别在0.05~10 ng·mL^-1,0.025~10 ng·mL^-1,0.025~10 ng·mL^-1浓度内呈良好的线性关系,r>0.998,检测限为0.012 5~0.025 ng·mL^-1,平均回收率分别为97.9%,95.7%,96.1%,RSD<3.4%。结论 方法简单快速,准确灵敏,可用于消癌平注射液中通关藤苷A、I、H的定量测定。     

HPLC测定达比加群酯中的有关物质

目的 建立达比加群酯原料药中有关物质的测定方法。方法 采用Agilent SB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈(A)-0.2%的醋酸铵(B,用冰醋酸调节pH值至4.4)为流动相进行梯度洗脱：0~18 min,90%→40%B;18~30 min,40%B。流速为1.0 mL·min^-1,检测波长为340 nm。结果 各杂质与主峰之间的分离度良好。5个已知杂质：杂质A浓度在0.117 0~1.872 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 7;杂质B浓度在0.126 5~2.024 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 5;杂质C浓度在0.113 0~1.808 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 5;杂质D浓度在0.120 5~1.928 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 9;杂质E浓度在0.123 0~1.968 μg·mL^-1内与峰面积呈良好的线性关系,r为0.999 6;杂质A、杂质B、杂质C、杂质D和杂质E加样回收率的平均值分别为98.75%,98.91%,98.39%,99.0%和99.73%;RSD分别为0.91%,1.09%,1.22%,1.35%和1.18%。结论 本方法简便、准确可靠,适用于达比加群酯中有关物质的控制。     

LC-MS/MS及HPLC法检测利奈唑胺血药浓度及临床应用

摘 要 目的：建立LC-MS/MS与HPLC法分析人血浆中利奈唑胺浓度,并将其应用于患者的治疗药物监测(TDM)与药动学研究。方法: 100 SymbolmA@l血浆样本加入蛋白沉淀剂(含内标呋喃唑酮乙腈),振荡离心后取上清液进样：① 乙腈-水(80∶20)为流动相经Eclipse XDB C18(100mm×2.1mm,3.5μm)色谱柱分离,流速0.3ml·min^-1,质谱的定量离子对分别为利奈唑胺：m/z338.1→296.2、呋喃唑酮：m/z226.1→122.0。②以乙腈 水(含0.1% 甲酸) (20〖KG*9〗∶〖KG-*2〗80)为流动相,经Eclipse XDB C18 (250 mm×4.6 mm, 5 μm)分离,流速：1 ml·min^-1,检测波长：254 nm。采用建立的方法分析重症患者用药后血浆标本。结果: 利奈唑胺在0.05~30 g·ml^-1 (LC-MS/MS)及0.25~30 SymbolmA@g·ml^-1(HPLC)范围内线性良好(r²>0.999),方法提取回收率、基质效应分别为82.1%～91.3%与74.0%～82.3%;相对回收率分别为91.2%～106.4%和100.1%～111.6%。日内、日间精密度RSD均小于20%,两种方法相关性良好。12例患者谷浓度为（1.77SymbolqB@1.23）μg·ml^-1,5例患者给药后2h浓度为（13.36SymbolqB@2.63）μg·ml^-1。结论: LC-MS/MS法简便、快速,专属性强且灵敏准确,适用于利奈唑胺的TDM与药动学研究。     

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.     

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.