一测多评法测定五味子中4种木脂素类成分的含量

建立五味子中4个木脂素类化合物一测多评方法学考察模式,验证该方法在五味子质量控制中应用的可行性和技术适应性。采用HPLC以五味子醇甲为内参物,建立五味子酯甲、五味子甲素和五味子乙素与五味子醇甲的相对校正因子,并进行含量计算,实现一测多评。同时采用外标法测定13批五味子药材中4个成分的含量,比较计算值与实测值的差异,验证一测多评法的准确性。结果表明,建立的校正因子重现性良好,采用校正因子计算的含量值与外标法实测值之间无显著差异。表明在缺少对照品的情况下,以外标法测定五味子醇甲含量,利用相对校正因子实现对五味子酯甲、五味子甲素和五味子乙素的测定准确可行,此方法可以用于五味子中木脂素类成分的定量评价研究。     

枣仁安神方配伍变化对五味子醇甲及五味子甲素药动学的影响

目的：考察枣仁安神方配伍变化对五味子醇甲和五味子甲素药动学影响。方法：SD大鼠分为五味子组、五味子-丹参组、五味子-酸枣仁组及枣仁安神方组,灌胃给药,采用高效液相色谱法（HPLC）对0~24h内大鼠血浆中五味子醇甲和五味子甲素测定,DAS 3.0软件计算药动学参数。结果：与五味子组相比,五味子-丹参组,五味子醇甲CL/F明显增加,五味子甲素V_1/F显著减小;五味子-酸枣仁组,五味子醇甲AUC_（0-t）显著降低,五味子甲素t_1/2K_a值明显减小,而枣仁安神方组五味子醇甲和五味子甲素t_1/2K_a值均明显增加,五味子醇甲AUC_（0-t）、AUC_（0-∞）显著增加,CL/F、V_1/F显著降低。结论：枣仁安神方的不同配伍对五味子醇甲和五味子甲素的药动学存在不同影响,与五味子组相比,复方显著减缓五味子醇甲和五味子甲素的消除,延长了作用时间。     

五味子配方颗粒中木脂素成分分析

目的:建立测定五味子配方颗粒中主要木脂素类成分(五味子醇甲、五味子酯甲、五味子甲素、五味子乙素)含量的方法,分析不同厂家的五味子配方颗粒中木脂素成分的差异。方法:采用RP-HPLC法,并结合二极管阵列检测器(PDA)进行光谱分析。色谱柱:Hypersil ODS C18柱(4.6 mm×250 mm,10μm),流动相:A为乙腈,B为水,梯度洗脱,流速1.0 mL/min,柱温25℃,检测波长254 nm。结果:不同厂家五味子配方颗粒中五味子醇甲含量为0~1.292 3 mg/包、五味子酯甲含量为0~0.560 8 mg/包、五味子甲素含量为0~0.168 8 mg/包、五味子乙素均未检测出。结论:不同厂家产品五味子醇甲、五味子酯甲、五味子甲素含量差异显著。     

五味子甲素的体内药动学及药理活性研究进展

五味子甲素是中药五味子的主要生物活性成分,属于联苯环辛烯型木脂素,具有保护肝功能、抗溃疡性结肠炎、神经保护、抗肿瘤等多种药理活性.五味子甲素在体内吸收迅速,可广泛分布于各组织,以肝脏、肾脏分布为主.本文系统综述了五味子甲素的药代动力学及其药理活性,为研究开发新型给药系统,提高五味子甲素的生物利用度提供理论依据.     

HPLC法测定南五味子软胶囊中五味子酯甲和五味子甲素的含量

目的研究并建立南五味子软胶囊的含量测定方法。方法采用高效液相-紫外检测法,以四氢呋喃-甲醇-水(10∶30∶60)为流动相,检测波长为280 nm,测定五味子酯甲和五味子甲素的含量。结果五味子酯甲在0.72～7.2μg范围内呈良好的线性关系,五味子甲素在0.56～5.6μg范围内呈良好的线性关系;加样回收率五味子酯甲为100.13%,RSD为1.05%,五味子甲素为99.53%,RSD为0.81%;精密度良好,五味子酯甲和五味子甲素的RSD分别为1.20%、1.02%;重复性也很好,五味子酯甲和五味子甲素的RSD分别为0.95%、1.06%。结论本方法稳定可靠,灵敏简便,重现性好,可作为南五味子软胶囊的的质量控制方法。     

用大孔树脂提取和纯化五味子总木脂素

目的以五味子中有效成分五味子醇甲、五味子甲素及五味子乙素含量为考察指标,筛选分离纯化五味子总木脂素的最佳树脂;研究不同大孔吸附树脂及其不同的工艺条件对总木脂素分离纯化的影响;建立五味子总木脂素的纯化方法。方法大孔吸附树脂与阴离子交换树脂(脱色树脂)联合使用,采用静态与动态的吸附-解吸两种模式进行纯化;通过动态吸附曲线观察吸附量及洗脱能力;利用HPLC法验证纯化后五味子醇甲、五味子甲素及五味子乙素的含量;上样吸附后,用5倍量柱床体积的水洗除杂,再用体积分数为95%的乙醇洗脱,用量为6倍量柱床体积。结果弱极性树脂AB-8适合用于五味子总木脂素的纯化;纯化后,五味子醇甲、五味子甲素、五味子乙素3种有效成分含量的质量分数之和由1.197%提高到14.92%,总转移率为72.3%。结论用大孔树脂提取和纯化五味子总木脂素的方法在保留五味子中有效成分的同时,显著降低固形物得率,可用于工业推广。     

多指标优选五味子种子中木脂素类成分提取工艺

目的优选五味子中木脂素类成分提取工艺。方法以五味子醇甲、五味子醇乙、五味子酯甲、五味子甲素、五味子乙素、五味子丙素及总木脂素提取量为指标,采用正交试验考察回流次数、回流时间、乙醇浓度、乙醇用量对该指标的影响。结果优选的工艺为5倍量95%乙醇,回流提取3次,每次60 min。结论优选的提取工艺稳定可行,可用于为五味子种子中木脂素类成分的提取。     

综合评分结合响应面法筛选北五味子木脂素提取工艺

目的筛选北五味子木脂素的提取工艺。方法以五味子醇甲、五味子醇乙、五味子酯甲、五味子甲素、五味子乙素和五味子丙素含量为评价指标,乙醇浓度、提取次数、提取时间和溶媒用量为考察因素,采用响应面Box-Behnken设计试验,筛选提取工艺参数。结果通过回归模型的预测,得到北五味子木脂素类成分最佳提取工艺为乙醇浓度88.24%,料液比10.37倍,提取时间34.70 min,提取次数2.76次。根据工业化生产需要,将提取工艺参数调整为11倍量90%的乙醇提取3次,每次35 min。结论采用多指标综合评价法能够更客观优化北五味子木脂素的提取工艺,为北五味子木脂素成分的分离纯化研究提供依据。     

高效液相色谱法测定舒神胶嚢中五味子甲素的含量

舒神胶囊主要由百合、郁金、五味子等十余味中药组成,其中五味子为方中主要组成药物之一,五味子中主要含五味子甲素、五味子乙素、五味子酯甲等木脂素类化合物,故选择五味子甲素作为控制本品质量的指标性成分.     

五味子甲素的研究进展

本文综述了五味子甲素的药理作用、提取工艺及测定方法,并对其提取工艺及测定方法进行了比较研究,旨在更有效地为五味子甲素的现代研究提供依据.     

Determination of deoxyschizandrin in rat plasma by LC-MS

A simple, rapid and sensitive LC-MS method was developed for quantification of deoxyschizandrin in rat plasma. A 50 miccrol plasma sample was extracted by ether and performed on Elite Hypersil C(18) column (200 mm x 4.6 mm, 5 microm) with the mobile phase of methanol-water (84:16, v/v) in a run time of 6.5 min. The analyte was monitored with positive atmospheric pressure chemical ionization (APCI) by selected ion monitoring (SIM) mode. A good linear relationship was obtained over the range of 1.0-50.0 ng/ml and the validated method was successfully applied for the pharmacokinetic studies of deoxyschizandrin in rat. After oral administration of 4 mg/kg deoxyschizandrin and Schisandra extract which contained the same dose of deoxyschizandrin to male rats, the C(max) of deoxyschizandrin were 15.8+/-3.1 and 34.3+/-16.8 ng/ml, T(max) were 0.51+/-0.13 and 3.83+/-1.83 h, T(1/2) were 5.3+/-2.2 and 6.5+/-3.4h.     

Effects of Schisandra lignans on P-glycoprotein-mediated drug efflux in human intestinal Caco-2

Schisandra fruits (Schisandraceae) are often used in traditional medicine and can be taken concomitantly with conventional medicine. In this study, the effects of dibenzocyclooctadiene lignans from Schizandra chinensis on P-gp-mediated efflux were examined to investigate a possible interaction with P-gp substrates. The cellular accumulation of rhodamine-123 in Caco-2 cells was measured with 12 Schisandra lignans. Most compounds resulted in slight or moderate increases of rhodamin-123 cellular uptake, indicating their P-gp inhibitory activity. Among them, deoxyschizandrin exhibited the most potent effect on the accumulation of rhodamine-123. Subsequently, bidirectional transports of digoxin and rhodamine-123 in Caco-2 cells were determined with deoxyschizandrin, the most active compound for the rhodamine-123 assay. In the bidirectional transport study, apical-to-basal (A-to-B) transports of digoxin and rhodamine-123 were increased, whereas basal-to-apical (B-to-A) transports were decreased by deoxyschizandrin in concentration- and time-dependent manners. At 50 microM of deoxyschizandrin, the transport ratios (B-A/A-B) for digoxin and rhodamine-123 were 2.2 and 2.1 compared with the control ratios of 15.2 and 12.2, respectively. These results demonstrated that deoxyschizandrin effectively inhibited the P-gp-mediated efflux in Caco-2 cells, suggesting they could potentially increase the absorption of drugs that can act as a P-gp substrate.     

Molecular mechanism of apoptosis induced by schizandrae-derived lignans in human leukemia HL-60 cells.

Schizandrae chinensis, a traditional Chinese medicine herb, has been used to treat hepatitis B disease in Chinese hospital clinic. We have isolated two bioactive compounds, deoxyschizandrin and gamma-schizandrin, from S. chinensis. In the present, we reported that deoxyschizandrin and gamma-schizandrin could induce apoptosis in human promyelocytic leukemia cells (HL-60), as characterized by DNA fragmentation and poly (ADP) ribose polymerase (PARP) cleavage. Further molecular analysis showed that deoxyschizandrin and gamma-schizandrin caused the loss of mitochondrial membrane potential (DeltaPsim), cytochrome c release from mitochondrion to cytosol, truncation of Bid protein, and activation of caspase-3 and -9. However, they did not increase the intracellular level of reactive oxygen species (ROS). Antioxidants such as N-acetyl cysteine (NAC) and catalase did not block the apoptosis induced by deoxyschizandrin or gamma-schizandrin. These findings suggest that deoxyschizandrin and gamma-schizandrin-induced apoptosis in HL-60 cells involved ROS-independent mitochondrial dysfunction pathway.     

高效液相色谱法测定舒宁胶囊中五味子甲素的含量研究

目的建立高效液相色谱法,对舒宁胶囊中五味子甲素的含量进行测定,为今后保健食品的研究工作提供可靠的参考依据。方法采用SunFireTM-C 18色谱柱(250mm×4.6mm×5μm),流动相为甲醇∶水=(75∶25),检测波长采取254nm,流速为1mL·min^-1,柱温为35℃。结果五味子甲素在4.95~131.95μg·mL^-1浓度范围内具有良好的线性关系,r=0.9999,平均加样回收率为100.95%,RSD为1.93%。结论所建立的高效液相色谱法操作简便、灵敏度高、重现性好,适用于舒宁胶囊中五味子甲素的含量测定。     

HPLC法切换波长同时测定六味五灵片中五味子甲素和乙素的含量

目的通过切换波长HPLC法同时测定六味五灵片中五味子甲素和乙素的含量。方法采用HPLC法,使用Shim-pack VP-ODS色谱柱,以甲醇-水（70∶30）为流动相,五味子甲素检测波长为254nm,五味子乙素检测波长为220nm,流速为1.0mL·min^-1,柱温为40℃。结果五味子甲素在1.22-24.40μg范围内呈良好的线性关系（r=0.9999）,五味子甲素平均回收率为99.52%,RSD=0.79%;五味子乙素在0.2528-5.056μg范围内线性关系良好（r=0.9999）,五味子乙素平均回收率为100.43%,RSD=1.4%。结论本法灵敏、准确、专属性强,重复性好,可用于同时测定六味五灵片中五味子甲素和乙素含量。     

HPLC切换波长法同时测定参芪五味子片中五味子酯甲和五味子甲素的含量

目的通过切换波长高效液相色谱法同时测定参芪五味子片中五味子酯甲和五味子甲素的含量。方法采用高效液相色谱法,使用Agilent Zorbax SB-C18(250 mm×416 mm,5μm)色谱柱,以甲醇-水(69∶31)为流动相,流速:1.0 mL.min-1,柱温:40℃,五味子酯甲检测波长为222 nm,五味子甲素检测波长为251 nm。结果五味子酯甲、五味子甲素分别在0.171~3.420μg(Y=24 107 309X+2.717 84×105,r=0.999 9),0.218~5.460μg(Y=2 069 343X+1.041 10×105,r=0.999 9)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99.73%,99.21%,RSD(n=6)分别为1.3%,1.4%。结论本法灵敏、准确、简便、快速,可用于同时测定参芪五味子片中五味子酯甲和五味子甲素含量。     

RP-HPLC法测定复方五味子胶囊中五味子甲素的含量

目的建立测定复方五味子胶囊中五味子甲素含量的高效液相色谱法。方法采用Kro-masilC18（250mm×4.6mm,5μm）的色谱柱;以甲醇-0.1%冰醋酸水溶液（82∶18）为流动相;流速：1ml/min;检测波长：254nm;柱温为30℃。结果五味子甲素在0.1137～0.4547mg/ml范围内与峰面积呈良好的线性关系（r=0.9992,n=6）,平均加样回收率为99.01%（RSD=1.1%,n=6）。结论HPLC法测定复方五味子胶囊中五味子甲素含量方法简便、准确、可靠。     

高效液相色谱法测定抗风湿液中染料木苷、染料木素、去氢二异丁香酚、五味子甲素的含量

目的采用多波长HPLC梯度洗脱法建立同时测定抗风湿液中染料木苷、染料木素、去氢二异丁香酚和五味子甲素含量的方法。方法采用C18色谱柱(250 mm×4.6 mm,5μm);流速:1.0 mL·min-1;流动相A为乙腈,流动相B为0.5%醋酸溶液,梯度洗脱;检测波长:染料木苷和染料木素为263 nm,去氢二异丁香酚为274 nm,五味子甲素为250 nm;柱温为35℃。结果染料木苷、染料木素、去氢二异丁香酚和五味子甲素质量浓度分别在9.65¹93.00μg·mL-1(r=0.999 9)、7.25193.00μg·mL-1(r=0.999 9)、7.25¹45.00μg·mL-1(r=0.999 7)、5.90145.00μg·mL-1(r=0.999 7)、5.90¹18.00μg·mL-1(r=0.999 2)、5.30118.00μg·mL-1(r=0.999 2)、5.30¹06.00μg·mL-1(r=0.999 6)时与其峰面积呈良好的线性关系;染料木苷、染料木素、去氢二异丁香酚和五味子甲素的平均加样回收率分别为98.8%、99.1%、96.9%、98.1%,RSD分别为1.0%、1.3%、1.3%、1.5%(n=6)。结论该方法是一种快速、灵敏、准确的分析方法,可作为抗风湿液的含量控制方法。     

HPLC法测定五酯胶囊中五味子甲素的含量

目的：建立HPLC法测定五酯胶囊中五味子甲素含量的方法。方法：色谱柱为C18,以甲醇-乙腈-0.4%磷酸溶液（70：10：20）为流动相,检测nm。结果：平均回收率98.9%RSD：1.86%。（n=6)结论：该法简便,灵敏,准确,重现性好。