HPLC双波长法同时测定炎热清片中6种有效成分的含量

目的 建立HPLC双波长法同时测定炎热清片中龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素和汉黄芩素6种成分的含量。方法 采用Agilent TC-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为甲醇∶0.2%磷酸水溶液（梯度洗脱）,柱温为35℃,流速为1.0 mL·min^-1,检测波长0~20 min为254 nm,同时检测龙胆苦苷、栀子苷、芒果苷,20~50 min为280 nm,同时检测黄芩苷、黄芩素、汉黄芩素。结果 龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素、汉黄芩素分别在18.92~189.2 ng（r=0.999 9）,107.44~1 074.4 ng（r=0.999 9）,9.82~98.2 ng（r=0.999 9）,767.68~7 676.8 ng（r=1.000 0）,100.94~1 009.4 ng（r=0.999 9）,49.84~498.4 ng（r=0.999 9）线性良好,平均回收率（n=9）分别为98.31%,98.35%,98.40%,98.03%,98.46%,98.24%,RSD分别为0.4%,0.3%,0.3%,0.4%,0.4%,0.3%。结论 该方法灵敏、简便、准确,可用于炎热清片的质量控制。     

HPLC法同时测定复方利胆解毒颗粒中绿原酸和连翘酯苷A

目的 建立复方利胆解毒颗粒中绿原酸和连翘酯苷A的定量测定方法。方法 采用HPLC法,Diamonsil-C₁₈柱（250 mm×4.6 mm,5 μm）,以甲醇-0.05%磷酸水（33：67）为流动相,在330 nm检测波长下同时对复方利胆解毒颗粒中有效成分绿原酸和连翘酯苷A进行定量测定。结果 绿原酸在0.044 72～0.894 4 μg线性关系良好,r=1.000 0,平均回收率为98.92%,RSD为1.56%;连翘酯苷A在0.052 16～1.043 2 μg线性关系良好,r=0.999 9;平均回收率为98.36%,RSD为1.44%。结论 本方法专属性强、简便、准确,可有效控制该制剂的质量。     

UPLC同时测定清热止咳颗粒中7种黄酮类成分含量

目的 建立同时测定清热止咳颗粒中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷含量的方法。方法 采用超高效液相色谱法。采用ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm,1.7 μm）,流动相乙腈-0.1%甲酸水溶液(梯度洗脱) ,流速0.3 mL·min^-1,检测波长280 nm,柱温30 ℃。结果 芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷分别在0.324～6.480（r=0.999 9）,1.337～26.75（r=0.999 7）,0.687～13.80（r=0.999 6）,1.347～26.95（r=0.999 8）,2.770～55.40（r=0.999 2）,0.330～6.600（r=0.999 8）,0.242～4.840（r=0.999 6）μg·mL^-1范围内与峰面积呈良好的线性关系;并且芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷平均加样回收率分别为99.41%、101.80%、99.42%、98.47%、101.71%,102.27%和97.64%,RSD范围分别是2.32%、1.69%、2.38%、2.45%、1.24%、1.77%和1.91%。结论 建立的超高效液相色谱(UPLC)方法操作简便、稳定、准确且可行,可用于清热止咳颗粒的质量控制。     

HPLC同时测定坤泰胶囊中毛蕊花糖苷、芍药苷和黄芩苷含量

目的 建立HPLC同时测定坤泰胶囊中毛蕊花糖苷、芍药苷和黄芩苷含量的方法，为完善现有质量标准提供参考。方法 采用Phenomsil C₁₈（4.6 mm×250 mm，5 μm）色谱柱；以乙腈为流动相A，以0.1%乙酸为流动相B，采用梯度洗脱；流速为1.0 mL·min^-1；柱温为35℃；毛蕊花糖苷的检测波长为334 nm，芍药苷检测波长为230 nm，黄芩苷的检测波长为280 nm。结果 毛蕊花糖苷在3.27~32.68 μg·mL^-1内线性关系良好，平均回收率为98.4%，RSD为0.68%（n=6）；芍药苷在17.65~176.51μg·mL^-1内线性关系良好，平均回收率为99.2%，RSD为1.08%（n=6）；黄芩苷在48.78~487.77 μg·mL^-1内线性关系良好，平均回收率为97.7%，RSD为0.87%（n=6）。结论 该方法简便、快捷、结果准确、重复性好，可应用于坤泰胶囊的质量控制。     

高效液相色谱法测定气管炎颗粒中黄芩苷的含量

目的 建立高效液相色谱法测定气管炎颗粒中黄芩苷的含量。 方法 采用HPLC法,色谱柱:ZORBAX SB-C₁₈柱(4.6 mm×150 mm,5 μm),以甲醇-0.1%磷酸(47:53)为流动相,流速为1 ml/min,检测波长为280 nm。 结果 黄芩苷在6.63~210 μg/ml范围内呈良好线性关系(r=0.999 8,n=6),平均加样回收率为98.16%,RSD为1.89%。 结论 本法简便、灵敏、准确、重复性好,结果可靠,可有效地控制气管炎颗粒的质量。     

HPLC法测定妥布霉素滴眼液中5种抑菌剂

目的 建立同时测定妥布霉素滴眼液中5种抑菌剂(羟苯乙酯、硫柳汞、羟苯丙酯、苯扎溴铵和苯扎氯铵)的HPLC方法。方法 采用Agilent Zorbax SB-C₁₈ (4.6 mm×150 mm,5 μm)色谱柱,流动相A为1%三乙胺（磷酸调节pH值至3.5±0.05）;流动相B为甲醇,梯度洗脱,流速为1.0 mL·min^-1,柱温35 ℃,检测波长为214 nm。结果 羟苯乙酯、硫柳汞、羟苯丙酯、苯扎溴铵/苯扎氯铵各峰均分离良好;羟苯乙酯在0.77～192.5 μg·mL-1范围内线性关系良好(r=0.999 5),平均回收率为100.0%(RSD=0.9%,n=9);硫柳汞在2.16～541.0 μg·mL^-1范围内线性关系良好(r=0.999 7),平均回收率为100.1%(RSD=0.6%,n=9);羟苯丙酯在0.84～209.8 μg·mL^-1范围内线性关系良好(r=1.000 0),平均回收率为100.3%(RSD=1.0%,n=9);苯扎溴铵在4.36～1090 μg·mL^-1范围内线性关系良好(r=0.9992),平均回收率为100.2%(RSD=0.5%,n=9);苯扎氯铵（n-C12H25）3.22～805.0 μg·mL^-1范围内线性关系良好(r=0.999 8),平均回收率为100.1%(RSD=1.1%,n=9);苯扎氯铵（n-C14H29）1.66～413.9 μg·mL^-1范围内线性关系良好(r=1.000 0),平均回收率为100.1%(RSD=0.6%,n=9)。结论 该方法准确、灵敏、简便,可用于妥布霉素滴眼液中羟苯乙酯、硫柳汞、羟丙丙酯、苯扎溴铵及苯扎氯铵5种抑菌剂的检查。     

HPLC法测定黄芩茎及叶中野黄芩苷的含量

目的 建立黄芩茎及叶中野黄芩苷的含量测定方法。方法 采用高效液相色谱(HPLC)法,C₁₈色谱柱(4.6 mm×250 mm,5 μm),以甲醇-0.02%甲酸（28∶72）为流动相等度洗脱,流速1.0 mL·min^-1,检测波长335 nm。结果 野黄芩苷在0.138 1～1.657 2 μg范围内与峰面积呈良好的线性关系,r=0.999 8,平均回收率为98.24%,RSD为0.96%;黄芩茎的野黄芩苷含量为0.86%,黄芩叶的野黄芩苷含量为2.83%。结论 该法简单方便,可用于测定黄芩茎及叶野黄芩苷的含量,对深入研究黄芩茎叶化学成分生物合成机理,确定黄芩茎叶药材最佳采收期以及制定黄芩茎叶药材质量标准提供理论依据。     

HPLC法同时测定跌打丸中3种有效成分的含量

目的 探讨建立同时测定跌打丸中芍药苷、川续断皂苷Ⅵ和丹皮酚含量的方法。方法 采用Agilent ZORBAX C₁₈色谱柱（4.6 mm×250 mm,5 μm）;流速为0.8 mL·min^-1,以乙腈（A）-0.1%磷酸水溶液（B）为流动相,进行梯度洗脱;检测波长为230,212,274 nm。结果 芍药苷进样量在0.514 8~1.544 5 μg与峰面积线性关系良好（r=0.999 7）,平均回收率为97.01%（RSD=0.94%,n=6）。川续断皂苷Ⅵ在3.023 8~9.071 4 μg线性关系良好（r=0.999 2）,平均回收率为98.44%（RSD=0.26%,n=6）。丹皮酚在0.101 2~0.303 6 μg线性关系良好（r=0.999 8）,平均回收率为97.84%（RSD=0.42%,n=6）。结论 建立的方法简便可行,重复性好,灵敏度高,可有效控制跌打丸的质量。     

UHPLC测定芎菊上清丸中9种成分含量及化学模式分析

目的 建立UHPLC波长切换法同时测定芎菊上清丸中9种成分的含量方法。方法 采用Agilent Ecilipse C₁₈（2.1 mm×100 mm,1.6 μm）色谱柱,流动相：甲醇-0.05%磷酸水溶液,梯度洗脱;流速为0.3 mL·min^-1;检测波长：327,237,320,345,278,254 nm;柱温30℃;进样量2 μL;并采用SPSS 22.0统计软件对含量测定结果进行主成分分析与聚类分析。结果 绿原酸、3,5-二咖啡酰奎宁酸、栀子苷、甘草苷、阿魏酸、盐酸小檗碱、黄芩苷、升麻素苷、5-O-甲基维斯阿米醇苷线性范围分别为4.30~68.80 μg·mL^-1（r=0.999 0）、6.66~106.56 μg·mL^-1（r=0.999 2）、7.67~122.72 μg·mL^-1（r=0.999 4）、4.88~78.08 μg·mL^-1（r=0.999 1）、2.37~37.92 μg·mL^-1（r=0.999 1）、6.50~103.92 μg·mL^-1（r=0.999 2）、8.85~141.60 μg·mL^-1（r=0.999 4）、0.88~14.08 μg·mL^-1（r=0.999 7）、0.74~11.92 μg·mL^-1（r=0.999 3）;平均加样回收率（n=9）均在99.42%~103.10%,RSD均<2.0%。主成分分析与聚类分析均可将不同生产厂家的芎菊上清丸很好地分类,且分类结果一致。结论 所建立的多成分方法快捷、准确、重复性好,可用于芎菊上清丸的质量控制。     

HPLC法测定六味地黄胶囊中的毛蕊花糖苷、马钱苷、芍药苷和丹皮酚

目的 建立同时测定六味地黄胶囊中毛蕊花糖苷、马钱苷、芍药苷和丹皮酚4种成分含量的高效液相色谱（HPLC）测定方法。方法 应用HPLC-梯度洗脱法,采用Kromasil C₁₈（250 mm×4.6 mm,5 μm）色谱柱,以1%冰醋酸为流动相A,乙腈为流动相B进行梯度洗脱,体积流量为1.0 mL/min,检测波长为230 nm。结果 毛蕊花糖苷、马钱苷、丹皮酚和芍药苷测定浓度的线性范围分别为6～60、15～150、50～500 μg/mL和5～50 μg/mL,r值均大于0.999,平均加样回收率分别为98.3%、99.3%、99.0%和99.0%,相对标准偏差（RSD）值均小于0.9%（n=9）。结论 该实验方法简便、结果准确,可有效地用于六味地黄胶囊的质量控制。     

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.

     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.