HPLC法测定益心酮片中五种成分的含量

目的:建立HPLC同时测定益心酮片中牡荆素-4″-O-葡萄糖苷、牡荆素鼠李糖苷、牡荆素、芦丁、金丝桃苷的含量。方法:采用HPLC法,色谱柱为Sepax HP C18柱(4.6 mm×250 mm,5μm);流动相:A为乙腈-四氢呋喃(20∶1),B为0.5%甲酸溶液,采用梯度洗脱;0~10 min,A为14%~17%,B为86%~83%;10~26 min,A为17%~18%,B为83%~82%;26~28 min,A为18%~14%,B为82%~86%;检测波长360 nm,流速1.0 mL.min-1;柱温32℃。结果:该方法加样回收率牡荆素-4″-O-葡萄糖苷为99.61%(RSD=0.64%),牡荆素鼠李糖苷为99.30%(RSD=0.49%),牡荆素为98.97%(RSD=1.03%),芦丁为99.45%(RSD=0.85%),金丝桃苷为99.27%(RSD=0.96%)。结论:该方法准确可靠,重现性好,可用于益心酮片的质量控制。     

HPLC法同时测定山楂叶提取物中的7种主要成分

目的同时测定山楂叶提取物中牡荆素葡萄糖苷(Ⅰ)、牡荆素鼠李糖苷(Ⅱ)、芦丁(Ⅲ)、牡荆素(Ⅳ)、金丝桃苷(Ⅴ)、熊果酸(Ⅵ)和槲皮素(Ⅶ)的含量。方法采用HPLC法,色谱柱为Agilent ZORBAX SB-C18柱(5μm,4.6×250mm,柱号:880975-902)流动相A为1%冰醋酸水溶液,B为乙腈。梯度洗脱。0~5min,流动相A由95%~90%,B由5%~10%;5~15min,A由90%~80%,B由10%~20%;15~35min,A由80%~0%,B由20%~100%;35~45min,A 0%,B 100%,流速1.0 mL.min-1;检测波长0~20min,检测波长为360nm;20~30min,检测波长为220nm;30~45min,检测波长为360nm,参比波长均为385nm,柱温40℃。结果Ⅰ~Ⅶ的平均加样回收率分别为99.87%、99.86%、101.23%、100.65%、101.58%、101.45%和98.37%,其RSD分别为1.26%、1.28%、1.35%、1.49%、1.32%、1.51%和1.44%(n=6)。结论该方法简便、快速、准确、可靠,可用于山楂叶提取物的质量控制。     

HPLC法同时测定山楂叶提取物中4种成分的含量

目的建立HPLC法同时测定山楂叶提取物中牡荆素-2″-O-鼠李糖苷、金丝桃苷、槲皮素、山奈酚的含量。方法色谱柱为DIAMONSIL C18柱(250 mm×4.6 mm,5μm);流动相为甲醇-体积分数为0.5%乙酸水溶液,梯度洗脱[0～15 min,φ(甲醇)=40%～65%;15～20 min,φ(甲醇)=65%～100%;20～40 min,φ(甲醇)=100%];流速为1.0 mL.min-1;波长为365 nm。结果牡荆素-2″-O-鼠李糖苷、金丝桃苷、槲皮素、山奈酚的线性关系良好,其质量浓度分别在14.0～112 mg.L-1、3.50～28.0 mg.L-1、6.25～50.0 mg.L-1、0.550～4.40 mg.L-1内与峰面积呈良好的线性关系。平均回收率分别为101.3%、98.6%、107.0%、98.1%,RSD分别为1.1%、2.2%、2.6%、2.1%(n=5)。结论该法简便、可行,重现性好,结果准确,可作为山楂叶提取物的含量测定方法,为其质量控制提供参考依据。     

HPLC波长切换联合梯度洗脱法同时测定消疲灵颗粒中的7种成分

目的 建立同时测定消疲灵颗粒中7种成分含量的HPLC波长切换联合梯度洗脱方法。方法 采用Venusil MP-C₁₈色谱柱,流动相为乙腈-1%冰醋酸溶液,流速为0.9 mL·min^-1,梯度洗脱,柱温为30 ℃,进样量为10 μL。结果 牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、金丝桃苷、芒柄花苷、毛蕊异黄酮和芒柄花素检测浓度分别在2.56~51.20 μg·mL^-1,14.87~297.40 μg·mL^-1,2.14~42.80 μg·mL^-1,3.16~63.20 μg·mL^-1,3.80~76.00 μg·mL^-1,2.14~42.80 μg·mL^-1,4.81~ 96.20 μg·mL^-1内与峰面积呈良好的线性关系（r≥0.999 1）,平均回收率97.0%~100.0%,RSD 0.55%~1.67%,精密度和重复性良好,供试品溶液在室温条件下12 h内稳定。结论 该方法操作简便,精密度、稳定性、重复性好,可用于消疲灵颗粒中7种有效成分含量的同时测定。     

HPLC法测定脾约丸中厚朴酚与和厚朴酚的含量

目的建立脾约丸中厚朴酚与和厚朴酚的含量测定方法。方法采用HPLC法测定脾约丸中厚朴(厚朴酚与和厚朴酚)的含量,色谱柱为Thermo BDS HYPERSIL C18,5μm,4.6 mm×250 mm,流动相为甲醇-水(75∶25),检测波长为294 nm,柱温25℃。结果和厚朴酚在6.056~90.84μg/m L的范围内、厚朴酚在6.704~100.56μg/m L的范围内呈良好线性关系,脾约丸中和厚朴酚平均回收率为99.58%(RSD为1.4%,n=9),厚朴酚平均回收率为99.62%(RSD为1.3%,n=9)。结论所建立的HPLC方法专属性强,重复性好,可用于脾约丸的质量控制。     

山楂叶不同采收期多种有效成分含量的比较

目的探索山楂叶最佳采收期。方法 HPLC条件:金丝桃苷检测采用Shim-pack C18(150mm×4.6mm,5μm);乙腈-甲醇-四氢呋喃-5 mL·L~(-1)醋酸溶液(1∶1∶19.4∶78.6)为流动相,检测波长为363nm。牡荆素鼠李糖苷检测采用Diamonsil C18柱(250mm×4.6mm,5μm);四氢呋喃-甲醇-乙腈-乙酸-水(38∶3∶3∶4∶152)为流动相;检测波长为330nm。总黄酮检测利用可见分光光度法,以芦丁对照品为对照,NaNO2-Al(NO3)3-NaOH体系作显色剂,在500nm波长处测定总黄酮含量。结果金丝桃苷进样量在0.080 5~0.402 4μg范围内,质量浓度与峰面积线性关系良好(r=0.999 3);牡荆素鼠李糖苷进样量在0.418~2.090μg范围内,质量浓度与峰面积线性关系良好(r=0.999 1)。结论 5月采收的山楂叶中牡荆素鼠李糖苷含量最高;8~11月采收的山楂叶中金丝桃苷及总黄酮的含量较高。     

HPLC法同时测定定喘止咳糖浆中5种成分的含量

目的建立高效液相色谱法同时测定定喘止咳糖浆中苦杏仁苷、和厚朴酚、厚朴酚、橙皮苷和川陈皮素含量的方法。方法采用依利特C_(18)柱,流动相为乙腈(A)-0.2%冰醋酸溶液(B),梯度洗脱;流速:0.8 mL/min;柱温:30℃;检测波长:λ_1=210 nm(苦杏仁苷),λ_2=254 nm(和厚朴酚、厚朴酚),λ_3=283 nm(橙皮苷、川陈皮素)。结果苦杏仁苷、和厚朴酚、厚朴酚、橙皮苷、川陈皮素的线性范围分别为12.10~242.00μg/mL(r=0.999 8)、7.65~153.00μg/mL(r=0.999 3)、6.37~127.40μg/mL(r=0.999 7)、4.29~85.80μg/mL(r=0.999 9)、3.90~78.00μg/mL(r=0.999 2);5种成分的平均加样回收率(n=6)分别为98.62%、96.94%、99.17%、96.85%、97.84%,RSD分别为0.88%、1.12%、1.10%、1.04%、1.55%。结论定喘止咳糖浆中5种成分的含量测定方法准确、可靠。     

基于HPLC多指标成分定量控制联合化学计量学的山菊降压胶囊质量评价研究

目的建立山菊降压胶囊中多指标成分高效液相色谱测定方法,并联合化学计量学方法对其进行综合质量评价。方法以Eclipse Plus C18柱(250 mm×4.6 mm,5μm)为色谱柱,乙腈–0.1%磷酸为流动相进行梯度洗脱;检测波长分别为340 nm(0～22 min检测牡荆素葡萄糖苷、牡荆素鼠李糖苷和牡荆素)、284 nm(22～41 min检测橙黄决明素、黄决明素和美决明子素)和208 nm(41～75 min检测泽泻醇A、24-乙酰泽泻醇A和23-乙酰泽泻醇B);柱温30℃;体积流量0.9 mL/min;进样量10μL。采用SPSS 26.0统计软件对10批山菊降压胶囊含量测定结果进行主成分分析和聚类分析。结果 9种成分牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、橙黄决明素、黄决明素、美决明子素、泽泻醇A、24-乙酰泽泻醇A和23-乙酰泽泻醇B线性范围良好,平均加样回收率96.90%～100.02%,RSD值均小于2.0%;聚类分析和主成分分析结果一致,10批山菊降压胶囊聚为2类,主成分1～3是影响山菊降压胶囊质量评价的主要因子。结论所建立的HPLC法可用于山菊降压胶囊中9种指标成分牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、橙黄决明素、黄决明素、美决明子素、泽泻醇A、24-乙酰泽泻醇A和23-乙酰泽泻醇B的定量控制和综合质量评价。     

2种半夏炮制品-厚朴药对HPLC特征图谱与多成分含量测定研究

目的:建立2种半夏炮制品-厚朴药对的HPLC特征图谱分析方法及鸟苷、尿苷、肌苷、紫丁香苷、绿原酸、金丝桃苷、和厚朴酚、厚朴酚的含量测定的方法。方法:采用Agilent Zorbax SB-C₁₈色谱柱(4.6 mm×250 nm, 5μm),以0.5%磷酸水溶液(A)-乙腈(B)为流动相,梯度洗脱,流速为1.0 mL·min^-1,检测波长为210 nm,柱温为30℃。采用“中药色谱指纹图谱相似度评价系统”(2012版)软件建立20批样品的HPLC特征图谱并进行相似度评价,使用SPSS 24.0、SIMCA 14.1软件进行主成分分析(PCA)和偏最小二乘聚类分析(PLS-DA)。结果:从供试品溶液中未检出肌苷,鸟苷、尿苷、紫丁香苷、绿原酸、金丝桃苷、和厚朴酚、厚朴酚在一定浓度范围内呈良好的线性关系(r=0.999 4～0.999 8),平均回收率(n=6)分别为97.1%、96.2%、97.1%、98.2%、96.1%、98.3%及96.9%(RSD<2.0%)。方法的精密度RSD均小于3.0%,重复性RSD小于3.0%,供试品溶液放置...     

高效液相色谱波长切换法同时测定百效丸中6种主成分的含量

目的 建立高效液相色谱波长切换法同时测定百效丸中6种主成分(哈巴苷、安格洛苷C、哈巴俄苷、巴豆苷、和厚朴酚、厚朴酚)含量的方法.方法 采用Venusil MP C18(4.6 mm×250 mm,5.0μm)色谱柱;流动相A:乙腈-甲醇(1:4),流动相B:0.2%磷酸溶液,梯度洗脱;检测波长210 nm(哈巴苷)、280 nm(安格洛苷C、哈巴俄苷)、294 nm(巴豆苷、和厚朴酚、厚朴酚).结果 哈巴苷、安格洛苷C、哈巴俄苷、巴豆苷、和厚朴酚、厚朴酚分别在3.98~79.60 mg·L-1、2.65~53.00 mg·L-1、2.45~49.00 mg·L-1、6.91~138.20 mg·L-1、5.29~105.80 mg·L-1、9.37~187.40 mg·L-1范围内线性关系良好;平均回收率96.90%~100.05%,RSD值0.89%~1.59%;精密度和重复性良好;供试品溶液在室温条件下10 h内稳定.结论 该方法操作简便,可用于百效丸中6种主成分的同时测定.     

Simultaneous determination of vitexin-4″-O-glucoside and vitexin-2″-O-rhamnoside from hawthorn leaves flavonoids in rat plasma by HPLC method and its application to pharmacokinetic studies

The present study was to investigate the pharmacokinetics of the two similar flavonoid glycosides, vitexin-4″-O-glucoside (VGL) and vitexin-2″-O-rhamnoside (VRH) in rats after intravenous administration of hawthorn leaves flavonoids (HLF). Blood samples were collected via tail vein at time intervals after drug administration and the plasma concentrations of the studied ingredients were analyzed by HPLC after the plasma protein was precipitated directly with methanol. VGL and VRH were successfully separated using a C₁₈ column with a UV detection at 330 nm and a mobile phase of methanol–acetonitrile–tetrahydrofuran–0.5% acetic acid (1:1:19.4:78.6, v/v/v/v). The assay linearities of VGL and VRH were confirmed over the range 0.23–138.42 and 0.36–218.49 μg/ml, respectively. The accuracy and precision of the two analytes at high, medium and low concentration were within the range of −3.13% to 3.51% and below 4%, the mean assay recoveries of them (n = 5) ranged from 96.87% to 101.75% and 96.88% to 103.51% for intra- and inter-day assays and the mean extraction recoveries of them (n = 5) varied from 92.68% to 95.74% for VGL and 93.45% to 99.26% for VRH, respectively. After intravenous administration of HLF to rats over the doses range of 10–40 mg/kg, the plasma concentration–time curves of VGL and VRH were both conformed to the three-compartment open pharmacokinetic model and linear pharmacokinetic characteristics.     

山玫胶囊的HPLC特征图谱研究

目的 建立山玫胶囊的HPLC特征图谱.方法 采用Zorbax SB-C18色谱柱(250 mm × 4.6 mm,5μm),柱温30℃,检测波长采用切换法(0～15 min时检测波长为260 nm、15～ 65 min时检测波长为370 nm),流动相为甲醇-乙腈-四氢呋喃-0.5％甲酸水溶液,梯度洗脱,流速1.0 mL· min-1.结果 HPLC特征图谱中的主要成分得到了有效分离,共得到16个共有峰,确定了8个特征峰的峰归属.结论 建立了山玫胶囊的特征图谱,所用方法简单、准确、重复性好,可为山玫胶囊的质量控制提供有效的测定方法.     

HPLC determination of malondialdehyde in ECV304 cell culture medium for measuring the antioxidant effect of vitexin-4″-<Emphasis Type="Italic">O</Emphasis>-glucoside

To investigate the antioxidant effect of vitexin-4'-O-glucoside, a flavone glycoside, isolated from the leaves of Crataegus pinnatifida Bge. var. major, we developed a simple and sensitive high-performance liquid chromatography (HPLC) method to determine levels of malondialdehyde (MDA) in ECV304 cell culture medium after induction by tert-butyl-hydroperoxide (TBHP). The preparation of analyzed samples involved a one-step derivatization with thiobarbituric acid (TBA). HPLC analysis was performed on a Synergi Hydro-RP, a polar end-capped C18 column (250 x 4.6 mm, 4 mum), using an acetonitrile-ammonium acetate aqueous solution (10 mM, pH 6.8) as the mobile phase under linear gradient conditions with UV detection at 532 nm. The calibration curve was linear over 0.0125-1.25 microM MDA (r = 0.9951). Relative standard deviations (RSDs) of intra-day and inter-day precision were less than 6.1% and 5.0%, respectively. The mean recovery was 96.9 +/- 1.6%. The lower limit of quantification (LLOQ) of MDA was 0.0125 microM. This chromatographic method was successfully applied to investigating the in vitro antioxidant effect of vitexin-4'-O-glucoside. Vitexin-4'-O-glucoside (120 M) protected ECV304 cells from peroxidation induced by TBHP.     

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.     

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.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.