Chemical characteristics for different parts of Panax notoginseng using pressurized liquid extraction and HPLC-ELSD

The chemical characteristics for different parts of Panax notoginseng, including root, fibre root, rhizome, stem, leaf, flower and seed, were determined using high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) and pressurized liquid extraction (PLE). Eight major saponins, namely notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3 and Rd were also quantitatively compared among the different parts of P. notoginseng. The chromatograms showed that there was significant difference between underground (root, fibre root, rhizome) and aerial (leaf and flower) parts from P. notoginseng, though the similarities of entire chromatographic patterns among tested samples from underground (0.965 ± 0.029, n = 12) and aerial parts (0.987 ± 0.014, n = 5) were similar, respectively. Especially, no saponin was detected in the seed of P. notoginseng. Hierarchical clustering analysis based on eight investigated saponins or the ratios of contents for ginsenoside Rg1/Rb1 and ginsenoside Rb3/Rb1 showed that the samples from different parts of P. notoginseng were divided into three main clusters. One cluster was underground parts, which contained rich protopanaxatriol and protopanaxadiol types saponins. The leaf and flower were in the same cluster, which contained protopanaxadiol type saponins only. Especially, ginsenoside Rc, Rb2 and Rb3, rare in the underground parts, were rich in aerial parts of P. notoginseng. The stem of P. notoginseng was another cluster. Based on the cluster analysis, the chemical characteristics for different parts of P. notoginseng were revealed. They are composite cluster (underground parts), protopanaxadiol cluster (aerial parts) and interim (stem) cluster, which was the one between the two typical clusters, respectively. The result shows that chemical characteristics of underground parts and aerial parts from P. notoginseng are obviously different, which is helpful for pharmacological evaluation and quality control of P. notoginseng.     

A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography

A rapid ultra performance liquid chromatography coupled with photo diode array detection method (UPLC-PDA) was developed for the simultaneous determination of 11 saponins, namely notoginsenoside R1, ginsenoside Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, Rd and Rg3 in Panax notoginseng. The analysis was performed on Acquity UPLC system with Acquity UPLC BEH C(18) column and gradient elution of water and acetonitrile in 12 min. The high correlation coefficient (r(2)>0.9968) values indicated good correlations between the investigated compounds' concentrations and their peak areas within the test ranges. The LOQ and LOD were lower to 0.2-2.4 and 0.1-1.8 ng on column, respectively. The overall intra- and inter-day variations (R.S.D.) of 11 saponins were lower than 3.1%. The developed method was successfully used for the analysis of saponins in P. notoginseng with overall recovery of 93.0-101.6% for the analytes. The results show that UPLC is a powerful tool for analysis of components in Chinese medicines.     

HPLC法同时测定杜仲壮骨胶囊中3种皂苷的含量

目的：建立高效液相色谱法同时测定杜仲壮骨胶囊中三七皂苷 R1（C47 H80 O18）、人参皂苷 Rg1（C42 H72 O14）和人参皂苷 Re（C48 H82 O18）的含量测定方法,旨在进一步提高对杜仲壮骨胶囊的质量控制。方法采用 Cap-cell Pak －C18（4．6 mm ×250 mm,5μm）色谱柱,流动相为乙腈－1％磷酸（18.8∶81．2）,流速为1．5 mL·min －1,柱温为30℃,检测波长为203 nm。结果三七皂苷 R1线性范围为4．99～99．7μg·mL －1;人参皂苷 Rg1线性范围为5．76～115．1μg·mL －1;人参皂苷 Re 线性范围为54．05～1081μg·mL －1。结论该方法简便、精确、灵敏度高且重复性好,可反映本制剂中多个药材组分的质量,可用于杜仲壮骨胶囊的质量控制。     

HPLC法测定云南白药胶囊中三七皂苷R_1和人参皂苷Rg_1的含量

目的采用高效液相色谱法测定云南白药胶囊中三七皂苷R1和人参皂苷Rg1含量。方法色谱条件:Phenomenex C18柱(100mm×4.6mm,2.6μm);流动相:乙腈-水(19∶81);流速:1mL.min-1;柱温:30℃;检测波长:203nm。结果三七皂苷R1和人参皂苷Rg1的线性范围分别为0.53～3.17μg(r=0.999 8),2.27～13.62μg(r=0.999 8);回收率(n=6)分别为100.6%(RSD=2.1%),97.7%(RSD=1.8%)。结论该方法准确可靠,重复性好,可用于测定云南白药胶囊中三七皂苷R1和人参皂苷Rg1含量。     

NP-HPLC法测定三七药材中总皂苷含量

目的:建立正相高效液相色谱法同时测定三七药材中人参皂苷Rg1,Re,Rb1,三七皂苷R14种成分的含量测定方法,并对不同规格三七药材中三七总皂苷的含量进行比较.方法:采用正相高效液相色谱法,色谱柱用Phenomenex NH2色谱柱(4.6mm×250 mm,5μm),流动相为乙腈—水(82:18),等度洗脱,检测波长...     

HPLC-ELSD法同时测定复方三七漱口液中三七皂苷R_1、人参皂苷Rg_1、Rb_1的含量

目的建立同时测定复方三七漱口液中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的HPLCELSD法。方法采用DIKMA Diamonsil C18(2)(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-水梯度洗脱,流速1.0 mL/min,柱温30℃,蒸发光散射检测器漂移管温度47℃,载气流速1.5 L/min。结果三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1分别在0.41¹.53、1.551.53、1.55⁵.83、1.585.83、1.58⁵.92μg呈良好线性关系;复方三七漱口液中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的平均回收率(n=9)分别为99.31%、100.30%、99.98%,RSD分别为1.60%、0.56%、0.97%。结论该方法简便、准确,专属性、重复性好,可用于复方三七漱口液的质量控制。     

HPLC法测定跌打巴布剂中三七皂苷R1和人参皂苷Rg1含量

目的建立跌打巴布剂中三七皂苷R1和人参皂苷Rg1的含量测定方法。方法采用HPLC法,以Agilent TC-C18ODS柱为色谱柱,以乙腈-水(24:76)为流动相,检测波长203nm,流速1mL/min。结果三七皂苷R1线性范围为0.583~29.15μg(r=0.9999,n=5),回收率为99.9%~100.3%。人参皂苷Rg1线性范围为0.336~16.8μg(r=0.9999,n=5),回收率为99.8%~100.2%。结论本方法准确、可靠、灵敏度高,可用于跌打巴布剂的定量检测。     

三七总皂苷和灯盏花素单用与联用时主要成分的药动学比较

目的考察三七总皂苷(主要成分为三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd)和灯盏花素(主要成分为野黄芩苷)复方粉针制剂在健康比格犬体内的药动学,并与单独使用三七总皂苷粉针或灯盏花素粉针时的药动学参数进行比较。方法建立检测给药后不同时间点的比格犬血样中三七总皂苷各组分及野黄芩苷浓度的液相色谱-串联质谱联用(LC-MS/MS)方法,计算各成分的药动学参数。结果单次给予复方粉针后,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及野黄芩苷的血浆消除半衰期(t1/2)分别为(1.08±0.30),(0.95±0.16),(1.40±0.39),(51.08±10.42),(64.84±17.70)及(2.00±0.88)h;峰浓度(Cmax)分别为(4641.00±758.84),(11325.00±1418.62),(1822.00±253.37),(39380.00±5644.03),(12964.00±2738.41)及(2669.00±841.79)ng·mL-1;血药浓度-时间曲线下面积(AUC0-∞)分别为(2832.31±308.38),(3454.00±473.08),(1210.80±161.06),(1360410.90±277244.88),(320529.65±101345.47),(450.68±90.50)ng·mL-1·h。与三七总皂苷粉针或灯盏花素粉针单次给药相比,复方粉针单次给药后,血药浓度-时间曲线相似;三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rd的Cmax显著升高(P<0.05);人参皂苷Rb1和野黄芩苷Cmax差异无统计学意义(P>0.05);t1/2、AUC0-∞差异无统计学意义(P>0.05)。结论三七总皂苷粉针和灯盏花素粉针联合用药可提高三七总皂苷部分组分的Cmax,对野黄芩苷的药动学没有明显影响。     

HPLC-ELSD测定心可舒胶囊中三七皂苷R1、人参皂苷Rg1的含量

目的建立心可舒胶囊中三七皂苷R1、人参皂苷Rg1的含量测定方法。方法以COSMOSIL5C18-PAQPackedColumn（250mm×4．6mm,5μm）为色谱柱,以乙腈．水（27：73）为流动相,流速为1mL·min-1,ELSD为检测器,漂移管温度：41℃;雾化器（氮气）压力：350kPa。结果三七皂苷R1、人参皂苷Rg1的线性范围分别为0．1232--6．16,0．3272～16．36μg,r值分别为0．9999,0．9997;回收率分别为98．7％,99．2％,RSD分别为2．72％,1．54％。结论该方法简便,准确,灵敏度高,重复性好,可作为心可舒胶囊的含量测定方法。     

舒胸片中人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1含量测定

目的：建立舒胸片中人参皂苷Rg1、人参皂苷Rb,和三七皂苷R1含量的HPLCI]定方法。方法：采用DiamosilC18柱（4．6X250mm,5gm）,流动相乙腈一水,梯度洗脱,柱温30℃,流速lmL／min,203nm波长下检测。结果：人参皂苷Rg1、人参皂苷Rb1、三七皂苷R,分别在0．442～11．050gg（r=0．9999）、0．344～8．600μg（P=0．9999）、0．208～5．200gg（r=0．9995）范围内线性关系良好,平均回收率分别为98．93％（RSD为1．7％1、98．88％（RSD为1．4％）、98．98％（RSD为1．4％）。结论：以HPLC法检测舒胸片中人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1,方法简便、准确、灵敏度高、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。     

心脑治胶囊中人参皂苷Rg_1、Rb_1及三七皂苷R_1的检测方法

目的建立心脑治胶囊中人参皂苷及三七皂苷含量的检测方法。方法采用高效液相色谱法,色谱柱为ZorbaxC18柱,乙腈-水梯度洗脱(0~3min,20%~25%乙腈;3~15min,25%~45%乙腈);流速1.0mL.min-1;检测波长203nm。结果人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1线性关系良好(r≥0.999),平均回收率(n=5)分别为105.68%(RSD=1.52%)、104.34%(RSD=0.73%)、102.93%(RSD=1.26%)。结论本含量测定方法灵敏、简便、快速、准确,适用于心脑治胶囊内在质量的控制。     

高效液相色谱梯度洗脱法同时测定三七总皂苷中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1含量

目的 采用高效液相色谱梯度洗脱法测定三七总皂苷中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量。方法 色谱柱为C18柱（250 mm×4.6 mm,3.5μm）,流动相为乙腈-水、梯度洗脱,检测波长为203nm,流速1.0 mL/min,柱温为室温。结果 人参皂苷Rg1进样量线性范围是6.0~18μg（r=0.996 7）,平均回收率为98.79%,RSD=0.33%（n=6）;人参皂苷Rb1进样量线性范围是6.0~18μg（r=0.996 4）,平均回收率为98.64%,RSD=0.46%（n=6）;三七皂苷R1进样量线性范围是1.6~4.8μg（r=0.997 1）,平均回收率为98.84%,RSD=0.56%（n=6）。结论 该方法准确、灵敏度高,可用于三七总皂苷的质量控制。     

高效液相色谱法同时测定三七总皂苷中人参皂苷Rg_1、Re、Rb_1与三七皂苷R_1含量

目的建立高效液相色谱法同时测定三七总皂苷中人参皂苷Rg1、Re、Rb1与三七皂苷R1的方法。方法采用高效液相色谱法 ,固定相为氨基键合相 ,流动相为乙腈 水 ( 81∶1 9,V∶V) ,检测波长2 0 3nm。结果三七总皂苷中人参皂苷Rg1、Re、Rb1和三七皂苷R1与其他成分分离良好 ,保留时间分别约为 5 7min、8 9min、2 5 1min和 9 9min。人参皂苷Rg1在 80～ 2 80mg/L(r =0 9992 )、Re在 2 0～ 1 80mg/L(r=0 9993 )、Rb1在 95～ 2 85mg/L(r=0 9991 )、三七皂苷R1在1 8～ 1 4 6mg/L(r=0 9991 )内线性关系良好 ,人参皂苷Rg1、Re、Rb1和三七皂苷R1的回收率分别为 99 1 %、98 4 %、98 6%和 97 1 % ,RSD分别为 2 1 %、2 0 %、2 2 %、2 8%。结论本法可用于三七的质量控制     

苦碟子注射液对血栓通主要皂苷成分药物动力学的影响

目的研究苦碟子注射液对血栓通(冻干)中三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd在大鼠体内药动学特征的影响。方法 12只SD大鼠随机分为2组,分别为血栓通组、苦碟子注射液-血栓通配伍组,尾静脉注射,于给药后不同时间采集血样,血样经乙腈沉淀蛋白后取上清,采用超高效液相色谱串联三重四级杆质谱(UPLC-MS/MS)测定5个皂苷成分的血药浓度。结果与苦碟子注射液配伍后,血栓通中5个化合物的t1/2、AUC0^t、AUC0t、AUC0^∞、CL、MRT0∞、CL、MRT0^t、MRT0t、MRT0^∞、Vd等主要药物动力学参数有不同程度的改变。结论苦碟子注射液在一定程度上能影响血栓通中主要皂苷成分在大鼠体内的药动学特征。     

心灵丸中三七皂苷R1、人参皂苷Rg和Rb1的含量测定

目的建立同时测定心灵丸中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1含量的高效液相色谱（HPLC）法。方法色谱柱为Agilent Hypersil ODS柱（250mm×4．0mm,5μm）,流动相A为乙腈,B为水,梯度洗脱（0min 19％A→12min 19％A→60min 36％A）,流速为1．0mL／min,检测波长为203nm。结果三七皂苷R1进样量在0．1378～2．7560μg范围内与峰面积线性关系良好,r=0．9997,平均回收率为98．95％,RSD为0．67％;人参皂苷Rg1进样量在0．5672-11．3440μg范围内与峰面积线性关系良好,r=0．9999,平均回收率为99．10％,RSD为0．29％;人参皂苷Rb1进样量在0．4186～8．3720μg范围内与峰面积线性关系良好,r=0．9999,平均回收率为99．02％,RSD为0．42％。结论HPLC法简便准确、专属性强,可用于心灵丸的质量控制。     

Simultaneous quantification of six major active saponins of Panax notoginseng by high-performance liquid chromatography-UV method

A simple, sensitive and specific high-performance liquid chromatography-UV (HPLC-UV) method has been developed for the first time to simultaneously quantify the six major active saponins of Panax notoginseng, namely notoginsenoside R1, ginsenoside Rg1, Rb1, Rg2, Rh1 and Rd. Astragaloside IV is used as the internal standard. This HPLC assay was performed on a reversed-phase C18 column with gradient elution of acetonitrile and 0.01% formic acid in 30 min. The method provided good reproducibility and sensitivity for the quantification of six saponins with overall intra- and inter-day precision and accuracy of less than 4.0% and higher than 90%, respectively. This assay is successfully applied to the determination of the six saponins in 23 notoginseng samples. The results indicated that the developed HPLC assay can be readily utilized as a quality control method for P. notoginseng.     

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.     

A rapid and sensitive liquid chromatography–tandem mass spectrometric method for the determination of timosaponin B-II in blood plasma and a study of the pharmacokinetics of saponin in the rat

A rapid, sensitive and selective liquid chromatography–tandem mass spectrometric (LC–MS/MS) method was developed and validated for the determination of Timosaponin B-II (TB-II), a pharmacologically active constituent isolated from Anemarrhena asphodeloides. This method was used to examine the pharmacokinetics and bioavailability of TB-II in rats using ginsenoside Re as an internal standard. After simple protein precipitation of the plasma samples with acetonitrile, the analytes were separated on an ODS column (150 mm × 2.1 mm i.d., 5 μm) with the mobile phase of acetonitrile–water (35:65, v/v) containing 0.05% formic acid and detected by electrospray ionization mass spectrometry in the negative multiple reaction monitoring (MRM) mode with a chromatographic run time of 3.0 min. The calibration curves were linear over the range of 5–15,000 ng/ml and the lower limit of quantification (LLOQ) was 5 ng/ml in rat plasma. In this range, relative standard deviations (R.S.D.) were <7.4% for intra-day precision and <9.0% for inter-day precision. The accuracy was within the range of 97.7–107.3%. The method was successfully applied to assess the pharmacokinetics and oral bioavailability of TB-II after intravenous and oral administration in rats, with the oral bioavailability being only 1.1%.     

Simultaneous determination of 10 nucleosides and nucleobases from different cultivation years of Fritillaria unibracteata var. wabuensis by HPLC-DAD

Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis (FUW) due to their physiological and pharmacologicalactions. In the present study, we developed a sensitive and reliable HPLC-diode-array detection method to simultaneously determine ten nucleosides and nucleobases, including cytosine, uracil, cytidine, uridine, thymine, adenine, inosine, guanosine, thymidine and adenosine. Complete separation of all the analytes was achieved on a Zorbax 300 Å 300 Extend C₁₈ column with a gradient of methanol-ultrapure water at a flow rate of 1 mL/min in less than 30 min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. The optimized method provided good linearity (R²≥0.9993 for all the analytes), satisfactory precision (RSD≤3.715%), good repeatability (RSD≤3.748%) and good recovery (RSD from 97.688% to 102.923%). In addition, the developed method was successfully applied to simultaneous determination of ten nucleosides and nucleobases from FUW, and their content changes of various cultivation time (1-7 years) were further analyzed for the first time. Our findings were useful for ensuring the cultivation time choice of artificial cultivation, quality control, pharmaceutical studies and clinical efficacy of FUW.