三七接骨丸中三七的质量控制标准研究

目的建立三七接骨丸中三七的快速检测方法及含量测定方法。方法采用薄层色谱法快速检测三七接骨丸中的三七药材,采用HPLC法测定三七中主要成分人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1的含量：采用Hibar ODS C18（4.6×250mm,5μm）色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0mL·min-1,柱温30℃,检测波长203nm。结果人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1分别在0.01270mg·mL-1~0.4064mg·mL-1（r=0.9997）、0.01416mg·mL-1~0.4530mg·mL-1（r=0.9998）、0.0054mg·mL-1~0.1742mg·mL-1（r=0.9999）范围内线性关系良好,平均回收率分别为人参皂苷Rb195.41%（RSD=0.78%,n=6）、人参皂苷Rg195.32（RSD=0.39%,n=6）、三七皂苷R194.74%（RSD=0.82%,n=6）。结论该方法准确有效、重复性好,可作为三七接骨丸中三七的质量控制方法。     

HPLC法测定活血通络片中三七皂苷R_1、人参皂苷Rg_1和Re的含量

目的:建立以高效液相色谱法测定活血通络片中三七有效成分含量的方法。方法:色谱柱为Agilent C18(150mm×4.6mm,5μm),流动相为乙腈-0.05%磷酸水溶液(梯度洗脱),流速为1mL.min-1,检测波长为203nm。结果:三七皂苷R1、人参皂苷Rg1、人参皂苷Re的进样量分别在0.38～3.80μg(r=0.9996)、0.94～9.40μg(r=0.9995)、0.15～1.50μg(r=0.9998)范围内与各自峰面积积分值呈良好线性关系;三者的平均回收率分别为99.23%、97.56%、107.32%,RSD分别为4.35%、4.94%、4.77%。结论:本方法简便、可行、重现性好,可用于活血通络片的质量控制。     

高效液相色谱法测定珍黄片的含量

目的：建立高效液相色谱法测定珍黄片的含量。方法：采用高效液相色谱法对制剂中人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1的含量进行测定。结果：人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1的线性范围分别为1．144～11．440μg（r=0．9999）、0．948～9．480μg（r=0．9999）和0．434-4．340μg（r=0．9998）,平均回收率分别为96．46％（RSD=1．01％）、98．17％（RSD=1．02％）和96．27％（RSD=1．16％）。结论：该方法简便、灵敏、准确,适用于珍黄片的含量测定。     

HPLC 法同时测定复方血栓通胶囊中5种皂苷类成分

目的：建立 HPLC 法同时测定复方血栓通胶囊中人参皂苷 Rg1、Re、Rb1、Rd 和三七皂苷 R1含量的方法。方法采用YMC －Pack Pro C18柱（250 mm ×4．6 mm,5μm）,以乙腈为流动相 A,水为流动相 B 进行梯度洗脱,检测波长为203 nm,流速为1．0 mL·min －1,柱温为35℃。结果测得三七皂苷 R1在50．10～501．00 ng、人参皂苷 Rg1在151．02～1510．20 ng、人参皂苷Re 在32．01～302．10 ng、人参皂苷 Rb1在102．23～1022．30 ng、人参皂苷 Rd 在16．32～163．20 ng 范围内线性关系良好（r ＝0．9999）;精密度、稳定性、重复性 RSD 均小于3％;平均加样收回率在95％～105％之间。结论该法简便、准确、重复性好,可用于复方血栓通胶囊的质量控制。     

RP-HPLC法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量

目的：建立反相高效液相色谱法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量测定方法。方法：采用Hibar ODS C18（4.6mm×250mm,5μm）色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0ml·min-1,柱温30℃,检测波长203nm。结果：人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1分别在0.5246～5.2464μg（r=0.9997）、0.6792～6.7920μg（r=0.9998）和0.2542～2.5424μg（r=0.9997）范围内线性关系良好,平均回收率分别为人参皂苷Rb199.12%（RSD=0.61%,n=6）、人参皂苷Rg197.50%（RSD=1.63%,n=6）、三七皂苷R197.43%（RSD=1.04%,n=6）。结论：该方法定量简单、准确、重复性好,可作为三七丹胶囊的质量控制方法。     

RP-HPLC法同时测定丹七片中三七皂苷R_1和人参皂苷Rg_1的含量

目的:建立以反相高效液相色谱法同时测定丹七片中三七皂苷R1和人参皂苷Rg1含量的方法。方法:色谱柱为KromasilC18(250mm×4.6mm,5μm),流动相为乙腈-水-磷酸(20.5∶79.5∶0.02),流速为1.0mL.min-1,柱温为室温,检测波长为203nm。结果:三七皂苷R1、人参皂苷Rg1的检测浓度分别在4.67~46.68(r=0.999 0)、4.62~115.50μg.mL-1(r=0.999 9)范围内与各自峰面积积分值呈良好线性关系;平均回收率分别为97.93%和97.98%,RSD分别为1.31%(n=6)和1.38%(n=6)。结论:本方法简便、快速、准确,可用于丹七片的质量控制。     

HPLC法测定益心复脉颗粒中人参皂苷Rg_1、Re、Rb_1的含量

目的:建立以高效液相色谱法测定益心复脉颗粒中人参皂苷Rg1、Re、Rb1含量的方法。方法:色谱柱为Diamond C18(150mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),流速为1mL·min-1,检测波长为203nm,柱温为25℃,进样量为10μL。结果:人参皂苷Rg1、Re和Rb1分别在0.645～6.450μg(r=0.999 8)、0.54～5.40μg(r=0.999 7)和0.605～6.050μg(r=0.999 9)范围内与各自峰面积积分值呈良好线性关系;三者平均加样回收率分别为100.59%(RSD=2.03%)、98.70%(RSD=1.46%)和98.99%(RSD=1.19%)。结论:本方法灵敏度高、简便、准确,可用于益心复脉颗粒的质量控制。     

HPLC法同时测定肠肽胶囊中三七皂苷R1、人参皂苷Rg1及Rb1含量

目的:建立肠肽胶囊中三七有效成分的HPLC含量测定方法。方法:色谱柱为SinoChrom ODS-BP（150 mm×4.6mm,5μm）,以乙腈（A）和水（B）为流动相,梯度洗脱,检测波长为203 nm,流速为1.0 ml·min^-1,柱温为室温,进样量为20μl。结果:三七皂苷R₁、人参皂苷Rg₁及人参皂苷Rb₁的线性范围依次为63.9～319.5μg·ml^-1、112.6～563.2μg·ml^-1、102.5～512.8μg·ml^-1,线性关系良好（r值分别为0.999 7、0.999 4、0.999 9）;三种皂苷的平均回收率分别为96.47%、98.75%,97.55%,RSD分别为1.52%、1.73%、1.81%（n=6）。结论:本方法简便可行、准确、灵敏度高、专属性强,可用于肠肽胶囊的质量控制。     

HPLC-ELSD法测定愈伤灵胶囊中5个皂苷类成分

目的建立愈伤灵胶囊中皂苷类成分的含量测定方法。方法用HPLC-ELSD法测定愈伤灵胶囊中的三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的含量。色谱条件为色谱柱C18柱,乙腈-水为流动相梯度洗脱:0²0 min,25%乙腈;2020 min,25%乙腈;20³0 min,25%30 min,25%³0%乙腈;3030%乙腈;30⁴5 min,30%45 min,30%⁵0%乙腈;4550%乙腈;45⁴8 min,50%48 min,50%⁹0%乙腈,柱温25℃,流速1 mL·min-1。ELSD漂移管温度110℃;载气流速2.5 L·min-1。结果三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd分别在0.097 490%乙腈,柱温25℃,流速1 mL·min-1。ELSD漂移管温度110℃;载气流速2.5 L·min-1。结果三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd分别在0.097 4⁰.974 2、0.379 30.974 2、0.379 3³.793、0.033 53.793、0.033 5⁰.334 8、0.330 90.334 8、0.330 9³.310和0.097 23.310和0.097 2⁰.972 4μg与峰面积线性关系良好,相关系数r均>0.999 3,平均回收率分别为100.1%、99.9%、100.2%、100.2%、100.5%。结论本方法可行,专属性强、重复性好,能有效地控制该制剂的质量。     

HPLC-VWD法同时测定丹参及三七混合物中8种有效成分的含量

目的建立HPLC-VWD法同时测定丹参、三七混合物中8种有效成分的含量。方法采用Eclipse XDB-C18(150mm×4.6mm,5μm)色谱柱;流动相:乙腈-0.1%磷酸水梯度洗脱;流速:1.0mL.min-1;柱温:15℃;采用切换波长的方法:在203nm下检测三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1;在286nm下检测丹酚酸B;在270nm下检测隐丹参酮、丹参酮ⅡA、丹参酮Ⅰ。结果三七皂苷R1、人参皂苷Rg1、人参皂苷Re、丹酚酸B、人参皂苷Rb1、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA在相应的浓度范围内与峰面积呈现良好的线性关系,相关系数均大于0.9995;平均回收率97.56%~103.0%,RSD均小于3.0%。结论该方法采用波长切换法同时测定不同吸收波长的化合物,方法简便、重复性好。     

HPLC法测定蓝玉簪颗粒中三七皂苷R1、人参皂苷Rg1、Rb1的含量

目的：建立HPLC法测定蓝玉簪颗粒中三七皂苷R1与人参皂苷Rg1、Rb1含量的方法。方法：色谱柱为Agilent C18（250mm×416mm,5μm）,以乙腈-水为流动相,梯度洗脱;检测波长为203nm;流速为1．0ml·min^-1;柱温为35℃。结果：三七皂苷R1在0．56—3．54峙、人参皂苷Rg1在0．41—8．12μg、人参皂苷Rb1在0．40～5．31μg范围内线性关系良好,相关系数r均为0．9999。三七皂苷R1与人参皂苷Rg1、Rb1的平均回收率分别是99．45％、99．11％、99．84％;RSD值分别为0．97％、0．56％、0．24％（n=6）。结论：本方法操作简便、可靠,重复性好,可作为蓝簪这颗粒中三七皂苷R1、人参皂苷Rg1、Rb1的含量测定方法。     

The blood-brain barrier permeability of 20(S) and 20(R)-protopanaxatriol epimers and dammar-20(22)E,24-diene-3β,6α,12β-triol in MDCK-pHaMDR cell monolayer model

The blood-brain barrier permeability of 20(S) and 20(R)-protopanaxatriol epimers and dammar-20(22)E,24-diene-3β,6α,12β-triol were investigated using the MDCK-pHaMDR cell monolayer model. The bidirectional permeability tests were carried out, and the apparent permeability coefficients (P_app) were calculated. The two protopanaxatriol epimers showed good permeability with P_app values of ~10^–5 cm/s, whereas dammar-20(22)E,24-diene-3β,6α,12β-triol showed poor permeability with P_app of <1×10^–7 cm/s. The three compounds showed differences in intracellular accumulations due to their different structures. Inhibition of P-gp with verapamil showed that the transport mechanisms in MDCK-pHaMDR cell monolayer for compounds 1and 2 epimers were not only simple passive diffusion but also involving an efflux way mediated by P-gp. These findings provided new basis for the further study of compounds 1 and 2 acting on the brain.     

HPLC法测定大鼠血清中三七皂苷R1、人参皂苷Rg1、Rb1的含量

目的 建立测定大鼠血清中三七皂苷R1、人参皂苷Rg1、Rb1含量的方法.方法 采用Kromasil 100-5C18色谱柱(250 mm×4.6mm,5μm),流动相为乙睛-0.2％磷酸水溶液,梯度洗脱;紫外检测波长203 nm,流速1 ml/min,柱温25℃.结果 三七皂苷R1、人参皂苷Rg1、Rb1均在0.2～5...     

HPLC法同时测定疏血通脉胶囊中3种皂苷的含量

目的建立测定疏血通脉胶囊中三七有效成分含量的方法。方法采用高效液相色谱法,以Kromasil C18(4.6 mm×250 mm,5μm)为色谱柱,以乙腈-水为流动相进行梯度洗脱,流速1.0 mL·min-1,检测波长203nm。结果人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1进样量分别在1.02¹0.20μg、0.9510.20μg、0.95⁹.50μg和0.359.50μg和0.35³.50μg范围内与各自峰面积积分值呈良好线性关系(r=0.999 9),平均回收率分别为100.40%、98.90%和99.61%,RSD分别为1.71%、1.98%和1.69%(n=6)。结论所采用的方法操作简单,可靠,准确,可作为疏血通脉胶囊的质量控制方法。     

三七总皂苷对抗H_2O_2所致大鼠脑微血管内皮细胞损伤的物质基础研究

目的研究三七总皂苷对H2O2致大鼠脑微血管内皮细胞损伤产生保护作用的物质基础。方法分离并培养大鼠脑微血管内皮细胞(RBMEC),以MTT和LDH的释放为指标,观察三七总皂苷(TPNS)、三七皂苷R1、人参皂苷Rg1、Rd、Re和Rb1对RBMEC损伤的保护作用。结果500μmol·L-1的H2O2对原代培养的RBMEC产生明显的损伤。20～200mg·L-1的TPNS,16·0μmol·L-1的R1,37·5～75·0μmol·L-1的Rg1,8·0～16·0μmol·L-1的Rd及5·4～54·0μmol·L-1的Rb1对H2O2致RBMEC的损伤具有明显的保护作用(P<0·05或P<0·01),Re则无此作用。结论TPNS对H2O2所致原代培养的RBMEC损伤具有明显的保护作用,产生这一作用的主要物质基础可能为Rb1、Rg1和Rd。     

富集纯化前上样液参数影响三七皂苷类成分的研究

目的 考察富集纯化前上样液参数pH值、氯化钠浓度、时间等对三七皂苷类成分的影响.方法 用UV、TLSC分别测定3种不同上样液在不同时间段的三七总皂苷、人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1的含量.结果 3种上样液放置72 h后,人参皂苷Rg1含量均增加(P＜0.01);三七皂苷R1含量均减少(P＜0.05);人参皂苷Rb含量变化无差异;pH5.5上样液和pH 7.0上样液中的总皂苷含量均增加(P＜0.05),pH5.5且含4％氯化钠的上样液中总皂苷含量无明显变化.结论 在富集纯化过程中,上样液的pH值、无机盐浓度和时间对皂苷类成分含量变化有影响.     

Pharmacokinetic and absolute bioavailability study of total panax notoginsenoside, a typical multiple constituent traditional chinese medicine (TCM) in rats

LC/ESI/MS method was employed for the pharmacokinetic evaluation of total panax notoginsenoside (TPNS) in rats. After oral or intravenous administration of TPNS at the dosage of 300.0 or 10.0 mg kg(-1) to rats respectively, panax notoginsenoside R1, ginsenoside Rg1, Rd, Re and Rb1 were simultaneous determined in rat plasma. Pharmacokinetic parameters and absolute bioavailability of panax notoginsenoside R1, ginsenoside Rg1, Rd, Re and Rb1 were obtained by the Drug And Statistics for windows (DAS) pharmacokinetic software. The pharmacokinetic parameters of all analytes were different form each other. T(1/2) were changed from 0.72 to 22.16 h and AUC were changed from 1.03 to 98.94 mg/l.h after oral or intravenous administration TPNS or Xuesaitong (TPNS) injection. The absolute bioavailability of R1, Rg1, Rd, Re and Rb1 were of 9.29%, 6.06%, 2.36%, 7.06% and 1.18%, respectively.     

人参皂苷 Rg1减轻大鼠离体心脏缺血再灌注损伤的作用研究

目的：探讨人参皂苷 Rg1对大鼠离体心脏缺血再灌注损伤的减轻作用及机制。方法制备大鼠离体心脏 I/ R 模型,将18只大鼠随机分为正常对照组、I/ R 组、人参皂苷 Rg1组。100μmol/ L 人参皂苷 Rg1进行预处理10 min,观察血流动力学指标左心室内压最大上升/下降速率和心率压力乘积的变化,检测灌流液中肌酸激酶（CK）、乳酸脱氢酶（LDH）活性。Western blotting 检测心肌组织 PI3K 下游激酶 Akt、ERK1/2及其磷酸化水平的变化,磷酸化GSK-3β的蛋白表达。结果与 I/ R 组比较,人参皂苷 Rg1预处理显著改善心脏功能,降低灌流液中 CK、LDH 活性（ P ﹤0.05）;升高 p-Akt 和 p-GSK-3β的蛋白表达（ P ﹤0.05）。结论人参皂苷 Rg1对大鼠离体心脏缺血再灌注损伤有减轻作用,其机制与激活 PI3K-Akt 通路有关。     

Simultaneous determination of nucleobases, nucleosides and saponins in Panax notoginseng using multiple columns high performance liquid chromatography

A new multiple columns HPLC method for simultaneous determination of 16 characteristic components, 5 nucleobases and nucleosides (uracil, cytidine, uridine, guanosine and adenosine), and 11 saponins (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, notoginsenoside R4, notoginsenoside Fa, ginsenoside Rb1, notoginsenoside R2, ginsenoside Rg2, ginsenoside Rh1, ginsenoside Rd and notoginsenoside K), in the root of Panax notoginseng, a valued traditional Chinese medicinal herb, were developed. Notoginsenoside R4, Fa and K were first quantitatively determined in P. notoginseng. The 5 nucleobases and nucleosides compounds were separated on a Zorbax SB-Aq column (150 × 4.6 mm, 5.0 μm) and 11 saponins were analyzed using a Zorbax Bonus-RP column (150 × 4.6 mm, 5.0 μm) with column switching. The column temperature was set at 30 °C. Mobile phase was composed of 5 mM ammonium acetate aqueous (A), water (B) and acetonitrile (C) using a gradient elution. The flow rate was 1.5 mL/min and detection wavelengths were set at 260 nm for nucleobases and nucleosides, and 203 nm for saponins. The developed method had good repeatability and sensitivity for quantification of 16 analytes with overall precision (including intra- and inter-day) less than 3% (RSD), and LOD and LOQ were less than 1.33 μg/mL and 5.12 μg/mL, respectively. The method was successfully applied to the simultaneous determination of 16 analytes in 15 samples of P. notoginseng collected from different places of China, which indicated that multiple columns HPLC can be used for comprehensive quality control of P. notoginseng.     

Pharmacokinetic study of ginsenoside Re with pure ginsenoside Re and ginseng berry extracts in mouse using ultra performance liquid chromatography/mass spectrometric method

Ginsenoside Re is the major ginsenoside in ginseng berry(GB) extract and its pharmacokinetics were studied following the intravenous and oral administration of pure Re or ginseng berry extract in mouse with doses of 10 and 50 mg/kg using ultra performance liquid chromatography mass spectrometric (UPLC/MS) method which can simultaneously determine ginsenoside Re, Rg1 and Rh1 in mouse serum. The serum samples were pretreated by protein precipitation and chromatographic separation was performed on AQUITY UPLC BEH C₁₈ column using gradient elution with the mobile phase of 5 mM ammonium formate and acetonitrile. Analytes and digoxin (I.S.) were analyzed and identified using an electrospray negative ionization mass spectrometry in the selected ion monitoring mode with the linear concentration range of 5.0–5000 ng/mL and lower limits of detection (LLOD) under 2.5 ng/mL. Ginsenoside Re was rapidly cleared from the body with a short half-life (0.2 ± 0.03 h for male and 0.5 ± 0.08 h for female mice after i.v.) and oral absorption was generally poor (F% 0.19–0.28). Notably, GB extract showed a superior oral absorption of ginsenoside Re (F% 0.33–0.75) at equivalent ginsenoside Re dose to pure ginsenoside Re, indicating that GB extract might be a good form for ginsenoside Re intake.