HPLC法测定参乌益肾片中8种成分及其化学计量学综合评价

目的 建立HPLC法同时测定参乌益肾片中β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷8种成分,联合化学计量学方法对参乌益肾片的产品质量进行综合评价。方法 采用Sepax Bio-C₁₈色谱柱（250 mm×4.6 mm,5 μm）;以乙腈–0.1%甲酸进行梯度洗脱;检测波长分别为250 nm（0～20 min测定β-蜕皮甾酮、25R-牛膝甾酮和25S-牛膝甾酮）、330 nm（20～60 min测定大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷）;柱温30℃;体积流量1.0 mL/min;进样量10 μL。采用聚类分析、主成分分析和偏最小二乘法–判别分析法对8种指标成分检测结果相关性进行分析评价。结果 β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷分别在1.47～36.75、0.66～16.50、0.74～18.50、5.80～145.00、15.77～394.25、4.58～114.50、1.29～32.25、8.78～219.50 μg/mL有良好的线性关系;平均回收率分别为97.78%、96.90%、97.96%、99.67%、100.01%、99.54%、98.03%、100.11%,RSD值分别为1.17%、0.82%、1.48%、0.93%、0.77%、0.69%、1.13%、0.65%。成分8（2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷VIP＝1.503）、成分4（大车前苷AVIP＝1.404）、成分5（毛蕊花糖苷VIP＝1.274）、成分1（β-蜕皮甾酮VIP＝1.030）是影响参乌益肾片样品质量的关键成分。结论 建立的方法简便快捷、准确可靠,可用于参乌益肾片中多指标成分的质量综合评价模式。     

HPLC法同时测定肉苁蓉饮片中8种成分的含量

摘要：目的:采用HPLC法同时测定肉苁蓉饮片中京尼平苷酸、松果菊苷、肉苁蓉苷A、管花苷A、毛蕊花糖苷、异毛蕊花糖苷、2’-乙酰毛蕊花糖苷和管花苷B的含量。方法:采用HPLC法,色谱柱为Agilent ZORBAX C18柱（250 mm×4.6 mm,5μm）,流动相为乙腈-0.5%乙酸,梯度洗脱。检测波长为237 nm,流速为1.0 ml·min-1,柱温为30℃。采用SPSS 19.0软件对24批次肉苁蓉饮片中8种成分含量进行相关性分析。结果:京尼平苷酸、松果菊苷、肉苁蓉苷A、管花苷A、毛蕊花糖苷、异毛蕊花糖苷、2’-乙酰毛蕊花糖苷和管花苷B的进样质量分别在0.098～9.800μg、0.105～10.520μg、0.104～10.440μg、0.095～9.530μg、0.099～9.860μg、0.108～10.780μg、0.118～11.810μg、0.099～9.860μg范围内与峰面积呈良好的线性关系;平均加样回收率分别为95.4%（RSD=1.86%）,99.2%（RSD=1.50%）,96.5%（RSD=0.99%）,94.8%（RSD=1.75%）,102.5%（RSD=1.13%）,97.2%（RSD=1.46%）,105.1%（RSD=1.39%）,96.8%（RSD=1.97%）（n=6）。松果菊苷和毛蕊花糖苷含量呈显著正相关（P<0.01）。管花苷A、异毛蕊花糖苷含量与松果菊苷、毛蕊花糖苷呈显著正相关（P<0.01）。2’-乙酰毛蕊花糖苷与毛蕊花糖苷的含量呈正相关（P<0.05）。结论:该方法操作简便快捷,准确有效,专属性强,有助于全面评价肉苁蓉饮片质量。     

一测多评法同时测定结石通片中7种成分含量

目的 建立同时测定结石通片中7种成分含量的一测多评（QAMS）法。方法 采用高效液相色谱法，色谱柱为Waters SunFire C18柱（250 mm×4.6 mm,5μm），流动相为乙腈-0.15%甲酸水溶液（梯度洗脱），流速为1.0 mL/min，检测波长分别为330 nm（夏佛塔苷、异夏佛塔苷、异牡荆苷、绿原酸、芒果苷和大车前苷）和210 nm（茯苓酸），柱温为35℃，进样量为10μL。以绿原酸为内参物，计算其他6种成分的相对校正因子，测定含量并与外标法测定结果比较。结果 夏佛塔苷、异夏佛塔苷、异牡荆苷、绿原酸、芒果苷、大车前苷和茯苓酸质量浓度分别在1.54～38.61μg/mL、0.53～13.14μg/mL、1.17～29.24μg/mL、2.05～51.28μg/mL、1.61～40.37μg/mL、1.16～29.03μg/mL、0.92～23.09μg/mL范围内与峰面积线性关系良好（r> 0.999 1,n=6）；检测限分别为0.13,0.06,0.11,0.19,0.15,0.10,0.08μg/mL，定量限分别为0.46,0.19,0.37,0.65,0....     

复方地锦胶囊中8种成分的HPLC法测定及其化学计量学综合评价

目的 建立复方地锦胶囊中槲皮素、山柰酚、木犀草素、芹菜素、大车前苷、毛蕊花糖苷、异毛蕊花糖苷和木通苯乙醇苷B的高效液相色谱测定方法,并结合化学计量学综合评价对其进行综合质量评价.方法 采用Acclaim 120 C18色谱柱(250 mm×4.6 mm,5μm);乙腈–0.1％甲酸为流动相,梯度洗脱;体积流量1.0 m...     

基于HPLC-QAMS法多指标成分定量控制和化学计量学的穿龙骨刺片综合质量评价

摘要：目的:建立高效液相一测多评法同步检测穿龙骨刺片中焦地黄苯乙醇苷A1、毛蕊花糖苷、焦地黄苯乙醇苷B1、杯苋甾酮、淫羊藿苷、宝藿苷Ⅰ、原薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、薯蓣皂苷含量的方法。方法:以Agilent SB-C18（250 mm×4.6 mm, 5μm）为色谱柱,流动相选择乙腈-0.1%磷酸,采用梯度洗脱。检测波长分别为330 nm（检测焦地黄苯乙醇苷A1、毛蕊花糖苷和焦地黄苯乙醇苷B1）、243 nm（检测杯苋甾酮）、270 nm（检测淫羊藿苷和宝藿苷Ⅰ）和203 nm（检测原薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷、薯蓣皂苷）。以淫羊藿苷为内参物,建立其它9种成分的相对校正因子,并计算各成分含量。采用SPSS 26.0统计软件对穿龙骨刺片中10种成分含量进行聚类分析和主成分分析。结果:焦地黄苯乙醇苷A1、毛蕊花糖苷、焦地黄苯乙醇苷B1、杯苋甾酮、淫羊藿苷、宝藿苷Ⅰ、原薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷和薯蓣皂苷线性范围良好,相关系数（r=0.999 1～0.999 4）,加样回收率为96.94%～100.12%（RSD≤1.60%,n=9）。一测多评法所测结果与外标法差异无统计学意义。结论:该方法可用于同时检测穿龙骨刺片中10种成分含量。     

HPLC同时测定抗宫炎片中的6种成分

目的 采用HPLC法同时检测抗宫炎片中去甲异波尔定、盐酸益母草碱、连翘酯苷B、毛蕊花糖苷、金石蚕苷、异毛蕊花糖苷的含量。方法 采用资生堂反向C₁₈柱,以乙腈-0.5%磷酸溶液为流动相,梯度洗脱,检测波长依次为280 nm(去甲异波尔定)、218 nm(盐酸益母草碱)、332 nm(连翘酯苷B、毛蕊花糖苷、金石蚕苷、异毛蕊花糖苷),流速1.0 mL·min^-1。结果 去甲异波尔定、盐酸益母草碱、连翘酯苷B、毛蕊花糖苷、金石蚕苷、异毛蕊花糖苷的线性范围分别为4.69～93.79、1.06～21.21、16.04～320.9、4.79～95.79、18.78～375.6、2.05～40.97μg·mL^-1(r均≥0.9995),平均加样回收率为98.76%～102.00%,RSD为1.32%～1.73%。结论 所用方法操作简便、结果准确,可用于抗宫炎片中6种成分的含量测定。     

HPLC同时测定裸花紫珠片中3种苯乙醇苷类成分含量

目的 建立同时测定裸花紫珠片中连翘酯苷B、毛蕊花糖苷与异毛蕊花糖苷3种苯乙醇苷类成分的高效液相色谱法。方法 用50%乙醇超声提取样品,乙腈-0.4%磷酸水溶液(18.2∶81.8)为流动相,色谱柱为Agilent ZORBAX SB-C₁₈(4.6 mm×250 mm,5 μm),柱温为30 ℃,流速为0.8 mL·min^-1,进样量为10 μL,检测波长为327 nm,检测时间为25 min。结果 连翘酯苷B、毛蕊花糖苷与异毛蕊花糖苷分别在0.936~4.680,1.852~9.260,2.140~10.700 μg内线性关系良好(r分别为0.999 1,0.999 9,0.999 9),加样回收率分别为100.07%,99.98%,97.78%。结论 该方法简便、快速、准确,适用于裸花紫珠片等裸花紫珠各类产品中苯乙醇苷类成分的含量检测。     

HPLC法测定软脉灵口服液中毛蕊花糖苷、焦地黄苯乙醇苷B1、迷迭香酸、紫草酸、丹酚酸B、2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷和虎杖苷

目的 建立HPLC法测定软脉灵口服液中毛蕊花糖苷、焦地黄苯乙醇苷B1、迷迭香酸、紫草酸、丹酚酸B、2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷和虎杖苷。方法 采用Venusil XBP C₁₈色谱柱（250 mm×4.6 mm,5μm）;以乙腈–0.1%甲酸水溶液为流动相,梯度洗脱;检测波长分别为330 nm（0～17 min检测毛蕊花糖苷和焦地黄苯乙醇苷B1）、280 nm（17～50 min检测迷迭香酸、紫草酸、丹酚酸B、2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷和虎杖苷）;体积流量1.0 mL/min;柱温30℃;进样量10μL。结果 毛蕊花糖苷、焦地黄苯乙醇苷B1、迷迭香酸、紫草酸、丹酚酸B、2,3,5,4’-四羟基二苯乙烯-2-O-β-D-葡萄糖苷和虎杖苷分别在1.23～30.75、0.59～14.75、0.71～17.75、1.47～36.75、9.78～244.50、6.57～164.25、1.19～29.75 μg/mL线性关系良好（r≥0.999 1）;平均回收率分别为99.14%、97.62%、96.84%、98.96%、100.11%、99.46%、98.92%,RSD值分别为1.15%、1.27%、1.02%、0.96%、0.65%、1.41%、0.87%。结论 该方法操作简便、重复性好,为软脉灵口服液中多指标质量评价和临床应用提供参考依据。     

芪明颗粒中8种成分的含量测定及聚类分析

摘要：目的:建立HPLC梯度洗脱法同时测定芪明颗粒中毛蕊花糖苷、焦地黄苯乙醇苷B1、3’-羟基葛根素、葛根素、3’-甲氧基葛根素、决明子苷B2、红镰霉素-6-O-β-龙胆二糖苷、决明子苷C的含量。方法:采用Kromasil C18色谱柱（250 mm×4.6 mm,5μm）,柱温30℃,流动相为乙腈-0.1%甲酸溶液,梯度洗脱,体积流量1.0 ml·min-1,检测波长为330 nm（检测毛蕊花糖苷和焦地黄苯乙醇苷B1）、250 nm（检测3’-羟基葛根素、葛根素和3’-甲氧基葛根素）和284 nm（检测决明子苷B2、红镰霉素-6-O-β-龙胆二糖苷、决明子苷C）。采用SPSS 26.0统计软件对含量测定结果进行聚类分析。结果:毛蕊花糖苷、焦地黄苯乙醇苷B1、3’-羟基葛根素、葛根素、3’-甲氧基葛根素、决明子苷B2、红镰霉素-6-O-β-龙胆二糖苷、决明子苷C分别在1.76～35.20μg·ml-1,0.64～12.80μg·ml-1,4.48～89.60μg·ml-1,13.59～271.80μg·ml-1,3.52～70.40μg·ml-1,2.18～43.60μg·ml-1,4.66～93.20μg·ml-1和1.29～25.80μg·ml-1范围内线性关系良好（r≥0.999 1）;平均加样回收率分别为99.24%,96.85%,99.69%,100.02%,98.89%,99.22%,98.11%和97.64%,RSD分别为0.95%,1.06%,0.81%,0.74%,1.45%,1.34%,1.17%和1.25%（n=9）。10批样品聚类分析为2类。结论:所建立的方法操作简便、重复性好,通过聚类分析提出了含量差异的影响因素,可用于芪明颗粒的质量控制。     

HPLC梯度洗脱法测定抗饥消渴片中7种成分的含量

目的建立HPLC梯度洗脱法同时测定抗饥消渴片中原儿茶醛、儿茶素、表儿茶素、毛蕊花糖苷、焦地黄苯乙醇苷B1、麦冬甲基黄烷酮A和甲基麦冬二氢高异黄酮B的含量,为抗饥消渴片质量标准提升提供数据支持。方法采用Diamonsil C18色谱柱(4.6 mm×250 mm,5μm),柱温35℃;流动相:乙腈-0.2%磷酸溶液,梯度洗脱,流速1.0 mL·min-1;检测波长分别为280 nm(0～24.0 min检测原儿茶醛、儿茶素和表儿茶素)、330 nm(24.0～36.0 min检测毛蕊花糖苷和焦地黄苯乙醇苷B1)和296 nm(36.0～50.0 min检测麦冬甲基黄烷酮A和甲基麦冬二氢高异黄酮B)。结果原儿茶醛、儿茶素、表儿茶素、毛蕊花糖苷、焦地黄苯乙醇苷B1、麦冬甲基黄烷酮A和甲基麦冬二氢高异黄酮B分别在5.96～119.20、1.18～23.60、8.76～175.20、2.87～57.40、1.59～31.80、1.26～25.20、0.88～17.60μg·mL-1内与峰面积线性关系良好(r≥0.9991);7种测定成分的平均回收率均> 96.0%,RSD均≤2.0%。结论该方法操作简便、重复性好,可用于抗饥消渴片中多种指标性成分的质量控制。     

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.

     

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.