UPLC-MS/MS法同时测定同仁牛黄清心丸中15个胆汁酸的含量

目的 建立同时测定同仁牛黄清心丸中15个胆汁酸成分含量的方法,并采用该法测定15批样品的含量。方法 以脱氢胆酸为内标,采用超高效液相色谱-串联质谱（UPLC-MS/MS）技术进行测定。色谱柱为Hypersil GOLD C18,以甲醇-0.1%甲酸溶液为流动相进行梯度洗脱,流速为0.2 mL/min,柱温为40℃,进样量为2μL;采用加热型电喷雾离子源,在负离子模式下进行平行反应监测模式扫描。采用SPSS 24.0软件对含量测定结果进行化学模式识别分析。结果 15个胆汁酸成分在各自检测质量浓度范围内与峰面积线性关系良好（R2均不小于0.998 9）,精密度、重复性、稳定性均良好（RSD均不大于5.49%）,平均加样回收率为93.8%～105.7%(RSD为0.5%～5.8%)。牛磺胆酸、7-酮-3α,12α-羟基胆烷酸、12-脱氢胆酸、甘氨胆酸、3-oxo-7α,12α-hydroxy-5β-cholanoic acid、牛磺鹅脱氧胆酸、3α-羟基-7-氧代-5β-胆烷酸、猪胆酸、牛磺脱氧胆酸钠、猪脱氧胆酸、胆酸、甘氨鹅脱氧胆酸、甘氨脱氧胆酸、鹅脱氧胆酸、脱氧胆酸的平均含量分别为670...     

HPLC-ELSD法同时测定生物转化样品中牛磺熊去氧胆酸等5种成分的含量

目的:建立同时测定生物转化样品中牛磺熊去氧胆酸、牛磺鹅去氧胆酸、熊去氧胆酸、鹅去氧胆酸、牛磺7-酮石胆酸5种成分含量的方法。方法:采用高效液相色谱法,检测器为蒸发光散射检测器。色谱柱为Agilent XBD-C18,流动相为0.05%三氟乙酸-乙腈,梯度洗脱,流速为1.0 ml/min;漂移管温度为80℃,N2流量为3.0 ml/min,柱温为40℃。结果:牛磺熊去氧胆酸、牛磺鹅去氧胆酸、熊去氧胆酸、鹅去氧胆酸、牛磺7-酮石胆酸检测质量浓度线性范围分别为10.00²00.08、10.22200.08、10.22²04.32、10.00204.32、10.00¹99.92、9.85199.92、9.85¹97.08、10.25197.08、10.25²04.92μg/ml(r=0.999 5204.92μg/ml(r=0.999 5⁰.999 8);精密度、稳定性、重复性试验的RSD均不大于2.0%。5种成分的平均回收率分别为97.9%、98.4%、97.9%、98.1%、98.3%,RSD分别为0.49%、0.63%、0.81%、0.72%、0.78%(n=3)。结论:该方法快速、准确、重复性好,可用于生物转化样品中牛磺熊去氧胆酸等5种成分的定量测定。     

RP-HPLC法同时测定猪胆粉药材中3种牛磺结合型胆酸的含量

目的:建立同时测定猪胆粉药材中牛磺猪胆酸、牛磺猪去氧胆酸和牛磺鹅去氧胆酸含量的RP-HPLC方法。方法:采用Welchrom C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-磷酸二氢钠溶液(0.03 mol·L-1)(70∶30),用磷酸调节pH为4.4,流速为1.0 mL·min-1,检测波长200 nm,柱温25℃。结果:牛磺猪胆酸、牛磺猪去氧胆酸和牛磺鹅去氧胆酸进样量分别在1.65～16.5μg(r=0.9998),2.03～20.3μg(r=0.9998),1.09～10.9μg(r=0.9996)范围内线性关系良好;检测限分别为26.4,7.6,38.2 ng;平均加样回收率(n=9)分别为101.2%(RSD=1.9%),99.1%(RSD=1.7%),100.6%(RSD=2.1%)。结论:该方法简便、准确,重复性好,在同一色谱条件下实现多指标成分的同时测定,为猪胆粉的全面质量评价提供参考。     

不同厂家蛇胆川贝胶囊中牛磺胆酸钠的含量比较

［摘要］目的比较不同厂家蛇胆川贝胶囊中牛磺胆酸钠的含量。方法采用高效液相色谱法,以蛇胆川贝胶囊中牛磺胆酸钠的含量作为评价指标,比较5个不同厂家蛇胆川贝胶囊的质量差异。结果5个厂家生产的蛇胆川贝胶囊中牛磺胆酸钠的含量差异显著。结论建立蛇胆川贝胶囊中有效成分的含量测定方法以评价药物的质量必不可少。     

一测多评法测定熊胆粉中多种结合型胆酸的含量

目的：建立熊胆粉中3种胆酸类成分牛磺熊去氧胆酸（TUDCA）、牛磺鹅去氧胆酸（TCDCA）和牛磺胆酸（TCA）的一测多评法,验证该方法在熊胆粉质量评价中的准确性和可行性。方法以牛磺熊去氧胆酸为内参物,确定其他两种成分相对于牛磺熊去氧胆酸的相对校正因子（Relative correction factor,RCF）,计算熊胆粉中牛磺鹅去氧胆酸和牛磺胆酸的量,实现一测多评;同时采用外标法测定熊胆粉中3种结合型胆酸的含量,比较计算值与实测值的差异。结果各相对校正因子重复性良好,一测多评法与外标法测定结果无显著差异。结论本文建立的一测多评方法操作简单,测定结果准确,可用于熊胆粉中结合型胆酸成分的定量分析。     

HPLC-ELSD法测定龙泽熊胆胶囊中牛磺熊去氧胆酸和牛磺鹅去氧胆酸的含量

以牛磺熊去氧胆酸和牛磺鹅去氧胆酸为指标,建立龙泽熊胆胶囊中熊胆粉的含量测定方法。方法：采用高效液相色谱串联蒸发光检测器,色谱柱为ChromCore AQ C18（4.6 mm×250 mm,5 μm）,乙腈（A）-5 mmol·L-1醋酸铵溶液（B）为流动相,梯度洗脱（0~40 min,25%A;40~50 min,25%A→29%A;50~80 min,29%A;80~100 min,29%A→40%A）,流速1.0 mL·min-1,柱温30 ℃,ELSD漂移管温度110 ℃,氮流量2.5L·min-1。结果：牛磺熊去氧胆酸进样量在1.069~9.57μg内、牛磺鹅去氧胆酸进样量在0.740 46~7.404 64 μg内,进样量的对数与峰面积的对数呈良好的线性关系;仪器精密度、重复性、稳定性试验的RSD＜2.0%;经低、中、高3个浓度的准确度试验考察,牛磺熊去氧胆酸的回收率为95.2%~97.7%,牛磺鹅去氧胆酸的回收率为91.9%~95.9%。测定样品42批次,牛磺熊去氧胆酸和牛磺鹅去氧胆酸的含量分别为0.18~0.43、0.10~0.44 mg·粒-1。结论：本法适用于龙泽熊胆胶囊中熊胆粉的质量控制,可为完善龙泽熊胆胶囊的质量标准提供科学的依据。     

不同胆汁制胆南星中胆酸类成分及其解热作用比较

目的比较猪、牛、羊胆汁制胆南星中胆酸类成分及其对发热小鼠体温的影响,为规范胆南星发酵工艺提供依据。方法分别采用猪、牛、羊胆汁与天南星生品发酵制备胆南星。采用HPLC-CAD法检测3种胆南星中胆酸类成分胆酸、去氧胆酸、猪去氧胆酸、鹅去氧胆酸。以干酵母制备小鼠发热模型,观察猪、牛、羊胆汁制胆南星对发热小鼠体温的影响。结果不同胆汁制胆南星中胆酸类成分存在差异,牛胆汁制胆南星中胆酸类成分质量分数最高,胆酸、猪去氧胆酸、鹅去氧胆酸、去氧胆酸的质量分数分别为0.035 9%、0.843 4%、2.260 2%、0.037 2%。猪胆汁制胆南星中鹅去氧胆酸和去氧胆酸升高,胆酸和猪去氧胆酸降低;羊胆汁制胆南星中胆酸类成分除去氧胆酸外均降低。不同胆汁制胆南星对发热小鼠均有一定的清热作用,猪胆汁制胆南星组小鼠体温先升高后降低,且与模型组比较差异有显著性,牛胆汁制胆南星亦表现出清热作用,但与模型组比较无显著性差异。羊胆汁制胆南星清热作用较弱。结论综合比较猪、牛、羊胆汁制胆南星中胆酸类成分质量分数和清热作用,作为制备胆南星的辅料,牛胆汁、猪胆汁优于羊胆汁。     

人工引流熊胆粉中结合胆汁酸成分的研究

目的研究人工引流熊胆粉中的结合型胆汁酸成分。方法采用反相树脂柱层析法对人工引流熊胆粉进行分离,然后通过IR1、H-NMR和13C-NMR等波谱数据进行结构鉴定。结果从人工引流熊胆粉中分离得到3个化合物,鉴定为:牛磺熊去氧胆酸(Ⅰ)、牛磺鹅去氧胆酸(Ⅱ)和牛磺熊去氧酮胆酸(Ⅲ)。结论直接得到结合型胆汁酸Ⅰ和Ⅱ,化合物Ⅲ为首次从熊胆中分离。     

HPLC法测定熊去氧胆酸片有关物质的含量

[摘要]目的：建立高效液相色谱法测定熊去氧胆酸片有关物质含量的方法。方法：用十八烷基硅烷键合硅胶为填充剂，以磷酸盐缓冲液（pH3.0）-甲醇-乙腈（35:37:28）为流动相，示差折光检测器测定。结果：在选定的色谱条件下，各成分可达到很好的分离，7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸分别在0.05~2.00，0.05~2.00，0.05~2.00，0.10~2.00mg·mL-1的范围内线性关系良好(r>0.9989)；精密度、稳定性试验的RSD均不大于0.5%；回收率分别为100.1%、99.8%、100.7%和101.9%（n=9,RSD<1.4%）。7-酮基胆石酸、胆酸、鹅去氧胆酸、胆石酸定量限分别为3.0、2.5、7.0、20μg·mL-1，检测限分别为0.8、0.7、2.0、3.5μg·mL-1。测定供试品两批，供试品一7-酮基胆石酸检出量为0.03%，胆酸和胆石酸均未检出，鹅去氧胆酸检出量为0.66%；供试品二7-酮基胆石酸、鹅去氧胆酸检出量均为0.10%，胆酸和胆石酸均未检出。结论：该方法灵敏度高，专属性强，适于测定熊去氧胆酸片中有关物质的含量。     

HPLC-ELSD法测定蛇胆川贝液中牛磺胆酸钠的含量

目的：建立HPLC—ELSD法测定蛇胆川贝液中牛磺胆酸钠的含量。方法：采用Inertsil ODS-3 C18柱（250mm×4．6mm,5μm）,以乙腈（A）-0．003％三乙胺溶液（B）为流动相进行梯度洗脱,流速1．0ml·min^-1,柱温35℃,漂移管温度110℃,气体流速3．5L·min^-1。结果：牛磺胆酸钠在0．102～1．024μg的范围内线性关系良好,r=0．9994,平均回收率为103．69％,RSD为1．04％（n=9）。结论：该方法操作简便,准确性、重复性好,可用于蛇胆川贝液中的牛磺胆酸钠的含量测定。     

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.