HPLC-QAMS同时测定益肾养元颗粒中9种成分的含量

建立一测多评法（QAMS）同时测定益肾养元颗粒中2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、虎杖苷、大黄素、大黄素甲醚、薯蓣皂苷元、齐墩果酸、乌索酸、蔷薇酸和坡模酸的含量,验证此方法在制剂中应用的准确性和可行性。采用高效液相色谱法,以Spursil C18（4.6 mm×250 mm,5 μm）为色谱柱,柱温为30 ℃;甲醇-0.1%甲酸溶液为流动相,梯度洗脱,流速为1.0 mL·min-1;检测波长分别为280 nm（检测2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、虎杖苷、大黄素和大黄素甲醚）和210 nm（检测薯蓣皂苷元、齐墩果酸、乌索酸、蔷薇酸和坡模酸）。以益肾养元颗粒为研究对象,选定薯蓣皂苷元为内参物,建立其与其他组分的相对校正因子,计算各成分含量,同时将外标法实测值与一测多评法计算值进行比较,验证一测多评方法的准确性和可靠性。2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、虎杖苷、大黄素、大黄素甲醚、薯蓣皂苷元、齐墩果酸、乌索酸、蔷薇酸和坡模酸的线性范围分别为23.89～477.80、8.67～173.40、4.26～85.20、2.99～59.80、9.86～197.20、0.69～13.80、2.17～43.40、1.28～25.60、1.66～33.20 μg·ml-1,r为0.999 4～0.999 9;平均回收率分别为99.45%、98.49%、97.94%、98.91%、100.03%、96.90%、99.02%、97.86%和98.35%,RSD分别为1.01%、0.87%、1.37%、1.19%、0.70%、0.98%、1.03%、0.82%和1.53%;2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、虎杖苷、大黄素、大黄素甲醚、薯蓣皂苷元、齐墩果酸、乌索酸、蔷薇酸和坡模酸含量的计算值和实测值之间无明显差异。所建立的一测多评法操作便捷、结果准确,可同时测定益肾养元颗粒中9种成分含量。     

益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量测定

目的 建立益肾乌发口服液中2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、大黄素、大黄素甲醚含量的测定方法,从指标成分角度来探讨益肾乌发口服液毒性与何首乌的关系,并为益肾乌发口服液的质量控制提供方法.方法 采用HPLC法,C18柱(4.6mm×250mm,5μm);测定二苯乙烯苷的流动相为乙腈-水(25:75),检测波长为320hm,流速为1.0 mL·min-1,柱温:25℃;测定大黄素大黄素甲醚的流动相为甲醇-0.1%磷酸溶液(80:20),检测波长为254nm,流速为1.0 mL·min-1,柱温:25℃.结果 二苯乙烯苷在0.1μg～10μg、大黄素在0.0168μg～1.68μg、大黄素甲醚在0.0208～1.04μg范围内呈良好的线性关系,r分别为0.9989(n=8)、0.9984(n=8)、0.9966(n=8);平均回收率:二苯乙烯苷为99.33%,大黄素为99.998%,大黄素甲醚为99.576%;RSD:二苯乙烯苷为0.68%,大黄素为1.11%,大黄素为1.29%.结论 高效液相色谱法测定益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量方法操作简单、灵敏度高、干扰少、重现性好、回收率高,可用于益肾乌发口服液的含量测定.初步证实益肾乌发口服液的毒性可能与君药制何首乌中二苯乙烯苷、大黄素、大黄素甲醚的含量有关,应控制其剂量范围,避免不良反应的发生.     

维血宁颗粒的质量标准研究

目的:提高维血宁颗粒的质量标准.方法:采用薄层色谱法 (TLC法 )对处方中虎杖、白芍、太子参进行定性鉴别;采用反相高效液相色谱法 (RP- HPLC法 )测定颗粒中大黄素的含量.结果: TLC鉴别方法专属性强、大黄素在 0.014 98～ 0. 074 90 μ g范围内进样量与峰面积积分值呈良好的线性关系, r=0.999 3,平均回收率为 99.89% (n=6),RSD=0.97%.结论:本质量标准可有效控制维血宁颗粒的质量.     

薄层扫描法测定龟鹿补肾丸中大黄素的含量

目的:研究龟鹿补肾丸中大黄素的含量测定方法.方法:采用薄层扫描法测定龟鹿补肾丸中大黄素的含量.结果:以对照品点样量为横坐标,峰面积积分值为纵坐标,作回归分析,得回归方程Y=715.05 X 30.07,r=0.999 4,大黄素点样量在0.21-1.05 μg范围内与峰面积积分值呈良好的线性关系;稳定性试验.RSD为1.98%;重现性考查RSD为2.33%;精密度测定RSD为2.37%;平均回收率为97.96%,RSD为1.98%;测定龟鹿补肾丸大黄素的平均含量为0.02696 mg/g.结论:该测定方法结果稳定、重现性好,可作为该制剂的质量控制方法.     

基于超高效液相色谱指纹图谱和多成分含量测定的黄芪益肾颗粒质量评价

目的:建立黄芪益肾颗粒超高效液相色谱(UPLC)指纹图谱及多成分含量测定方法,并进行化学计量学分析,为其质量控制提供依据。方法:采用Waters CORTECS UPLC T3色谱柱(2.1 mm×150 mm, 1.6μm),柱温30℃;流动相为乙腈-0.1%冰醋酸,梯度洗脱,体积流量为0.25 mL·min^-1;检测波长270 nm。利用中药色谱指纹图谱相似度评价系统(2012版)建立黄芪益肾颗粒的UPLC指纹图谱,并对11批黄芪益肾颗粒进行质量评价;对淫羊藿苷、朝藿定A1、朝藿定B、朝藿定C、宝藿苷I、毛蕊异黄酮葡萄糖苷、大黄酸的含量测定方法进行方法学验证。结果:黄芪益肾颗粒UPLC指纹图谱确认了17个共有峰,指认了7个共有峰并建立其含量测定方法,11批黄芪益肾颗粒样品相似度均大于0.95;7种成分在各自的浓度范围内线性关系良好(R²≥0.999 6),平均加样回收率为98.67%～100.06%,RSD分别为1.4%～2.1%。结论:该方法简便高效,可用于黄芪益肾颗粒的质量控制。     

益肾补骨颗粒剂中大黄素的HPLC测定

建立高效液相色谱法测定益肾补骨颗粒剂中大黄素的含量.用Hypersil ODS(5μm)色谱柱,以甲醇-水(80:20)为流动相,检测波长254nm.大黄素线性范围0.2～1μg(r=0.9999),平均回收率为98.0%,RSD为2.2%.     

基于网络药理学及液质联用技术的三黄益肾颗粒医院制剂质量标志物预测研究

目的:采用网络药理学结合液质联用技术预测分析三黄益肾颗粒医院制剂的质量标志物。方法:采用网络药理学方法分析三黄益肾颗粒治疗糖尿病肾病的作用机制,并筛选出活性成分;通过超高效液相-四级杆-飞行时间串联质谱(UPLC-Q-TOF-MS/MS)技术对三黄益肾颗粒中的化学成分进行全面鉴定;通过质谱鉴定的成分与网络药理学筛选的成分对比,结合中医药组方理论,辨识出与功效相关的成分作为三黄益肾颗粒的质量标志物。结果:网络药理学模块化筛选得到君药组20个潜在质量标志物;臣药组筛选得到27个潜在Q-Marker;佐使药组筛选得到48个潜在质量标志物。与通过液质联用技术辨识的30种成分比对出16个交集成分,结合三黄益肾方中医药组方理论,初步预测黄芪甲苷、芒柄花黄素、阿魏酸、芦荟大黄素4种成分为三黄益肾颗粒的质量标志物。结论:通过网络药理学及液质联用技术并结合中医药组方理论,筛选出三黄益肾颗粒的质量标志物,可为提升三黄益肾颗粒的质量标准提供依据,为建立以中医药理论为指导,以临床疗效为基础的医院制剂整体质量评价体系提供思路。     

HPLC法测定转阴灵颗粒中大黄素的含量

目的建立HPLC法测定转阴灵颗粒中大黄素含量的方法。方法采用C18柱(250 mm×4.6 mm,5μm);流动相为甲醇-1 g.L-1磷酸溶液(80∶20);检测波长254 nm;流速为1 mL.min-1。结果大黄素进样量在0.101 2~0.506μg范围内,与峰面积积分值呈良好线性关系。大黄素平均回收率为98.5%,RSD为2.9%(n=5)。结论该方法简单、快速、准确、稳定,可作为转阴灵颗粒质量控制的方法之一。     

正交试验优选复方红藤颗粒的制备工艺及质量标准研究

目的:优选复方红藤颗粒的制备工艺,并建立其质量标准。方法:以总固体量、绿原酸含量、大黄素含量、综合评分为评价指标,加水量、提取时间、提取次数为考察因素,通过正交试验优选复方红藤颗粒的制备工艺。采用薄层色谱法鉴别绿原酸,高效液相色谱法同时测定绿原酸和大黄素的含量。结果:筛选的最优提取方案为加8倍量水,提取3次,每次提取2 h;薄层色谱鉴别特征明显,专属性强;绿原酸、大黄素的进样量分别在0.103⁴.120、0.0404.120、0.040⁰.520μg范围内与各自峰面积积分值呈良好线性关系(r均为0.999 9);精密度、稳定性、重复性平均加样回收率试验的RSD均小于2%。结论:该试验科学地筛选了复方红藤颗粒提取工艺,所建立的质控方法操作简便、结果准确,可以作为复方红藤颗粒的质量控制标准。     

RP-HPLC法测定益肾康颗粒中淫羊藿苷的含量

目的:建立以反相高效液相色谱法测定益肾康颗粒中淫羊藿苷含量的方法。方法:色谱柱为C18(150mm×4.6mm,5μm),流动相为乙腈-水(24∶76),流速为1.0mL·min-1,检测波长为270nm。结果:淫羊藿苷进样量在430.4～1506.4μg范围内与峰面积积分值呈良好线性关系(r=0.9998);平均回收率为99.56%,RSD=1.20%(n=9)。结论:本方法简便、准确、重复性好,可用于益肾康颗粒的质量控制。     

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.

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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.

     

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.     

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.     

2-(Acetoxyphenyl)-(Z)-styryl sulfides as cyclooxygenase inhibitors

2-(Acetoxyphenyl)-(Z)-styryl sulfides are described as selective cyclooxygenase-2 (COX-2) inhibitors, useful for treating inflammation and COX-2-mediated disorders including neoplasia. 2-(Acetoxyphenyl)-(Z)-styryl sulfide is claimed to be the most potent COX inhibitor in the series with a COX-2 selectivity ratio of 33. This compound is also claimed to be superior to celecoxib (Celebrex^®, Pfizer) in inhibiting cell growth of colorectal carcinoma cells. In this evaluation, the COX inhibitory activity of this compound is compared to that previously disclosed for diarylheterocycles and 2-(acetoxyphenyl)alkyl sulfides. The validity of the DLD-1 cell line in the growth inhibition studies is questioned based on recent literature reports indicating the lack of COX-2 expression in this cell line.     

Sleep-disordered breathing with chronic opioid use

Chronic opioid use for pain relief or as substitution therapy for illicit drug abuse is prevalent in our societies. In the US, retail distribution of methadone and oxycodone has increased by 824 and 660%, respectively, between 1997 and 2003. μ-Opioids depress respiration and deaths related to illicit and non illicit chronic opioid use are not uncommon. Since 2001 there has been an emerging literature that suggests that chronic opioid use is related to central sleep apnoea of both periodic and non-periodic breathing types, and occurs in ～ 30% of these subjects. The clinical significance of these sleep-related abnormalities are unknown. This review addresses the present knowledge of control of ventilation mechanisms during wakefulness and sleep, the effects of opioids on ventilatory control mechanisms, the sleep-disordered breathing found with chronic opioid use and a discussion regarding the future research directions in this area.     

Drug targets for cognitive enhancement in neuropsychiatric disorders

The investigation of novel drug targets for treating cognitive impairments associated with neurological and psychiatric disorders remains a primary focus of study in central nervous system (CNS) research. Many promising new therapies are progressing through preclinical and clinical development, and offer the potential of improved treatment options for neurodegenerative diseases such as Alzheimer's disease (AD) as well as other disorders that have not been particularly well treated to date like the cognitive impairments associated with schizophrenia (CIAS). Among targets under investigation, cholinergic receptors have received much attention with several nicotinic agonists (α7 and α4β2) actively in clinical trials for the treatment of AD, CIAS and attention deficit hyperactivity disorder (ADHD). Both glutamatergic and serotonergic (5-HT) agonists and antagonists have profound effects on neurotransmission and improve cognitive function in preclinical experiments with animals; some of these compounds are now in proof-of-concept studies in humans. Several histamine H3 receptor antagonists are in clinical development not only for cognitive enhancement, but also for the treatment of narcolepsy and cognitive deficits due to sleep deprivation because of their expression in brain sleep centers. Compounds that dampen inhibitory tone (e.g., GABA_A α5 inverse agonists) or elevate excitatory tone (e.g., glycine transporter inhibitors) offer novel approaches for treating diseases such as schizophrenia, AD and Down syndrome. In addition to cell surface receptors, intracellular drug targets such as the phosphodiesterases (PDEs) are known to impact signaling pathways that affect long-term memory formation and working memory. Overall, there is a genuine need to treat cognitive deficits associated with many neuropsychiatric conditions as well as an increasingly aging population.