HPLC法测定复方芦丁片中芦丁的含量

目的 建立HPLC测定复方芦丁片中芦丁的含量.方法 采用反相高效液相色谱法;色谱柱为HYPERSIL ODS-C18柱(4.6 mm ×250 mm,5μm);流动相为甲醇-0.1%冰醋酸(15:35);检测波长为359 nm;流速为1.0 ml/min;柱温为35℃;进样量为20μg.结果 芦丁在5.0～50.0μg/ml浓度范围内与峰面积呈良好线性关系,回归方程为Y=99384X-22521,r=0.9999.平均回收率为99.3%,RSD为0.70%(n=6).结论 本法快速简便,准确,重现性好.     

RP-HPLC法测定枸杞中芦丁含量

目的建立反相高效液相色谱法测定枸杞中芦丁含量的方法。方法色谱分析条件Shim-packVP-ODS色谱柱(250mm×4.6mm,5μm),以乙腈∶水∶冰乙酸(5∶94.5∶0.5)为流动相A,乙腈∶水∶冰乙酸(70∶29.5∶0.5)为流动相B,采用梯度洗脱,流速0.5ml/min,检测波长342nm,柱温30℃。结果芦丁线性范围为10～100μg/ml(相关系数为r=0.9998),平均回收率(n=5)为99.0%;不同产地枸杞样品中芦丁含量存在差异,随保存时间的延长,芦丁含量明显下降。结论本方法重现性好,适用于测定枸杞中芦丁的含量。     

HPLC法测定痔速宁片中芦丁与没食子酸的含量

目的建立HPLC法测定痔速宁片中芦丁与没食子酸含量的方法。方法采用Thermo Acclaim TM 120C18色谱柱(150mm×4.6mm,5μm);流动相为甲醇-0.4mL·L~(-1)磷酸溶液,梯度洗脱;流速为1.0mL·min~(-1);检测波长为257和273nm;进样量5μL;柱温35℃。结果芦丁与没食子酸的线性范围分别为10.54~105.4μg·mL~(-1)(r=0.999 9,n=6)和51.075~510.75μg·mL~(-1)(r=0.999 9,n=6);提取回收率分别为99.2%(RSD=0.7%,n=6)和99.4%(RSD=0.9%,n=6)。结论该方法简便、可靠、准确,可用于该制剂的质量控制。     

桑叶中芦丁含量的动态变化研究

目的研究不同时期桑叶中芦丁的含量。方法 Kromasil C18柱(4.6 mm×250 mm,5μm);流动相:甲醇-0.5%磷酸水溶液(梯度洗脱),检测波长:358 nm;柱温:30℃;结果芦丁在0.126 2².02μg范围内有良好的线性关系,r=0.999 9,平均回收率为97.88%,桑叶中芦丁含量最高在霜降前后。结论初步表明了桑叶中芦丁含量随季节变化的规律。     

高效液相色谱法测定吉如心片中芦丁的含量

目的:建立HPLC法测定吉如心片中芦丁含量的方法。方法:色谱柱为HypersilC₁₈柱（250mm×4.6mm,5μm）;以乙腈-0.1%磷酸溶液（15:85）为流动相,流速为1.0ml·min^-1,检测波长257nm,柱温:35℃。结果:芦丁在18.64～93.18ng范围内线性关系良好。平均回收率为99.81%,RSD=0.62%。结论:该方法准确、可靠,可用于吉如心片的质量控制。     

HPLC测定复方芦丁片中芦丁的含量

目的:建立高效液相色谱法测定复方芦丁片中芦丁的含量.方法:以Wafters Nova-pak C18(3.9×150mm,4um)为色谱柱;以乙腈-四氢呋喃-0.2%枸橼酸溶液(14:1:85)为流动相;流速为1.0ml·min-1;检测波长为360nm.结果:芦丁在0.8372～5.0232μg(r=0.9999)范围内与峰面积呈良好的线性关系;平均回收率为99.05%,RSD为0.71%.结论:本法快速、准确,可用于测定复方芦丁片中芦丁的含量.     

荷花中芦丁含量测定方法研究

目的建立荷花中芦丁的含量测定方法。方法采用反相高效液相色谱法,对样品处理方法、色谱柱和流动相等影响分离效果的主要因素进行了优化。选用Thermo BDS Hypersil C18色谱柱(4.6 mm×250 mm,5μm);甲醇-1%冰乙酸溶液(32∶68)为流动相;检测波长为360 nm。结果芦丁的线性范围0.119 9⁰.839 3μg,r=0.999 998;平均回收率97.93%,RSD为1.8%(n=6)。结论该方法简便、实用、结果可靠,分离效果好。     

高效液相色谱法测定曲克芦丁注射剂的含量

目的 :建立以高效液相色谱法测定曲克芦丁注射剂含量的方法。方法 :色谱柱为HypersilBDSC18(150mm×5mm ) ,流动相为乙腈 -四氢呋喃 -0 05mol/L磷酸二氢钠溶液 (15∶8∶77 ,pH=4 0) ,柱温为30℃ ,流速为0 8ml/min ,检测波长为254nm。结果 :曲克芦丁标准溶液浓度在20～200μg/ml范围内线性关系良好 (r=0 9998) ,平均回收率为100 7 %;高、中、低3种浓度曲克芦丁样品溶液日内精密度为1 61 %,日间精密度为2 46 %。结论 :该方法简便 ,结果准确 ,适用于曲克芦丁注射剂的质量控制。     

HPLC法测定莲子心中芦丁的含量

目的 建立高效液相色谱(HPLC)法测定莲子心中芦丁的含量.方法 以Ultimate C18 (250 mm×4.6 mm, 5μm)为色谱柱,流动相为甲醇-0.1%磷酸溶液(40:60),流速为1.0 ml/min,检测波长为256 nm,柱温为30 ℃.结果 芦丁进样量在0.122～1.094 μg 范围内线性关系良好(r=0.999 9),平均回收率为100.5%,RSD为1.72 % (n=6).结论 本法简便、准确、可靠,可作为莲子心中芦丁的定量分析方法.     

HPLC法测定肾复康片中芦丁、槲皮素、山柰素的含量

目的：建立以高效液相色谱法测定肾复康片中芦丁、槲皮素、山柰素含量的方法。方法：采用高效液相色谱法,色谱柱为CAPCELL Pak MG C18色谱柱(4.6×150mm,5um),流动相：甲醇-0.4%磷酸溶液,梯度洗脱;检测波长：360nm;流速：1ml.min-1。结果：芦丁在0.01583～0.1425mg/ml范围内线性关系良好(r=0.9999),平均加样回收率为99.60%,RSD=1.07%(n=6);槲皮素在0.00805～0.0725mg/ml范围内线性关系良好(r=0.9997),平均加样回收率为98.56%,RSD=1.31%(n=6);山柰素在0.00387～0.0349mg/ml范围内线性关系良好(r=0.9996),平均加样回收率为97.20%,RSD=1.46%(n=6)。结论：本方法简便可靠,重现性好,可用于肾复康片中芦丁、槲皮素、山柰素的含量测定。     

芸香苷对实验性急性胰腺炎肺损伤的保护作用及机制

目的探讨芸香苷(rutoside,Ru)对实验性急性胰腺炎(AP)肺损伤的保护作用及机制。方法牛磺胆酸钠(STC)逆行胆胰管注射诱发大鼠AP模型后,立即按Ru15、30、60mg.kg-1.h-13个剂量持续静脉输注6h,观察AP大鼠动脉血气分析、肺湿重/干重比值、肺组织病理学、肺TNF-α、ICAM-1、NF-κB表达的变化。结果60mg.kg-1剂量组可升高AP大鼠降低的动脉PO2;Ru30、60mg.kg-1剂量组可降低肺湿/干重比值,同时可以改善肺病理损伤情况;Ru15、30、60mg.kg-1剂量组均可降低肺TNF-α、ICAM-1、NF-κB的表达。结论静脉输注Ru对实验性AP胰外肺损伤具有一定的保护作用,其发挥保护作用的机制可能与降低TNF-α、ICAM-1、NF-κB的表达有关。     

偏最小二乘紫外分光光度法同时测定复方芦丁片中2组分的含量

目的:将偏最小二乘法与紫外分光光度法结合,建立一种同时测定复方芦丁片中芦丁和维生素C含量的新方法。方法:在234～300nm波长范围内取33个波长点测定16组标准混合溶液的吸光度值并建立校正集,结合残差平方和确定主因子数,运用偏最小二乘法预报待测样品中2组分的含量。结果:模拟样品中芦丁和维生素C平均回收率分别为102%、92.7%,均方根误差为0.04879、0.3410,预测相对误差为2.259%、6.315%;将此模型用于复方芦丁片中2组分的测定,芦丁和维生素C平均加样回收率分别为102%～103%、96.3%～104%。结论:该方法不需分离样品,具有操作简便、结果准确等优点。     

芸香苷静脉输注对大鼠急性胰腺炎腺泡细胞凋亡的影响及机制

目的探讨芸香苷(Rutoside,Ru)静脉输注给药对实验性急性胰腺炎(acute pancreatitis,AP)腺泡细胞凋亡的影响及作用机制。方法3%牛磺胆酸钠(sodium taurocholate,STC)逆行胆胰管注射诱导大鼠AP模型,取胰腺做病理学检查。应用末端脱氧核苷酸转移酶介导的末端标记(terminadeoxynucleotidyl transferase biotin-dUTP nick end labeling,TUNEL)方法检测胰腺组织中的腺泡细胞凋亡。用免疫组化法检测Fas、FasL蛋白的表达。结果Ru15、30、60mg·kg-1·h-13个静脉输注剂量组可改善胰腺的病理组织学变化,且凋亡指数(apoptosis index,AI)高于模型组〔6h Ru大、中、小剂量治疗组分别为(23·33%±3·49%)、(21·56%±3·81%)、(16·24%±2·83%),6h模型组(8·54%±1·28%),P<0·01〕;Fas蛋白阳性表达细胞的灰度值低于模型组〔6h Ru大、中剂量治疗组分别为(189·17±5·79)、(203·83±10·94),6h模型组(226·86±10·37),P<0·01〕;FasL在各组腺泡细胞中均有不同程度的表达,但各治疗组的阳性表达细胞的灰度值高于模型组〔6h Ru大、中、小剂量治疗组分别为(220·63±7·41)、(218·72±4·48)、(207·22±22·67),6h模型组(181·70±9·01),P<0·05,P<0·01〕,各组的AI在各时间点均与Fas阳性表达细胞的灰度值呈正相关,与FasL阳性表达细胞的灰度值和胰腺严重程度呈负相关(P<0·05,P<0·01)。结论Ru对实验性AP的保护作用可能与促腺泡细胞凋亡有关,Fas/FasL系统可能参与此过程而诱导细胞凋亡。     

Inhibition of Polymorphic Human Carbonyl Reductase 1 (CBR1) by the Cardioprotectant Flavonoid 7-monohydroxyethyl Rutoside (monoHER)

PURPOSE: Carbonyl reductase 1 (CBR1) reduces the anticancer anthracyclines doxorubicin and daunorubicin into the cardiotoxic metabolites doxorubicinol and daunorubicinol. We evaluated whether the cardioprotectant monoHER inhibits the activity of polymorphic CBR1. METHODS: We performed enzyme kinetic studies with monoHER, CBR1 (CBR1 V88 and CBR1 I88) and anthracycline substrates. We also characterized CBR1 inhibition by the related flavonoids triHER and quercetin. RESULTS: MonoHER inhibited the activity of CBR1 V88 and CBR1 I88 in a concentration-dependent manner. The IC(50) values of monoHER were lower for CBR1 I88 compared to CBR1 V88 for the substrates daunorubicin and doxorubicin (daunorubicin, IC(50)-CBR1 I88 = 164 microM vs. IC(50)-CBR1 V88 = 219 microM; doxorubicin, IC(50)-CBR1 I88 = 37 microM vs. IC(50)-CBR1 V88 = 59 microM; p < 0.001). Similarly, the flavonoids triHER and quercetin exhibited lower IC(50) values for CBR1 I88 compared to CBR1 V88 (p < 0.001). MonoHER acted as a competitive CBR1 inhibitor when using daunorubicin as a substrate Ki = 45 +/- 18 microM. MonoHER acted as an uncompetitive CBR1 inhibitor for the small quinone substrate menadione Ki = 33 +/- 17 microM. CONCLUSIONS: The cardioprotectant monoHER inhibits CBR1 activity. CBR1 V88I genotype status and the type of anthracycline substrate dictate the inhibition of CBR1 activity.     

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.