高效液相色谱-二极管阵列检测/质谱法分析生脉饮煎剂中的人参皂甙类成分

采用高效液相色谱-二极管阵列检测/质谱（HPLC-DAD/MS）联用技术,以10 mmol/L醋酸铵和乙腈混合溶液梯度洗脱 系统为流动相,应用C18色谱柱对生脉饮煎剂中人参皂甙类成分进行分离鉴定。分析结果表明:生脉饮煎剂中主要含有17个 人参皂甙类成分,即20(R)-人参皂甙Rh1、Rh2、Rg3、Rg2,20(S)-人参皂甙Rh1、Rh2、Rg3、Rg2,人参皂甙Rf、Rg6、Rg5 、F4、Rk1、Rk3、Rh4,20(S)-和20(R)-原人参三醇。人参皂甙成分在煎煮过程中发生了很大变化,主要变成了一些中低 极性产物,这是因为煎煮过程中发生了水解、差向异构、脱水等反应。该方法简便、精确、灵敏度高,可以用来分析生脉 饮煎剂中人参皂甙的变化。     

高效液相色谱-飞行时间质谱联用法分析人参及人参皂苷与山楂配伍过程中的水解行为

利用高效液相色谱-飞行时间质谱联用的方法,分别对人参配伍山楂前后人参皂苷的变化进行分析,同时对人参皂苷Re、Rg1、Rb1、Rd与山楂配伍的水解规律进行系统研究,并与单独煎煮液、仿山楂配伍pH值煎煮液的水解产物进行比较,结果发现人参与山楂配伍后人参皂苷Rg1、Rb1含量明显减少,而人参皂苷Re、Rd、Rg2、Rg3、F2、Rh1含量明显增加,其中人参皂苷Re与山楂配伍后水解产物为人参皂苷20(R)-Rg2、20(S)-Rg2,仿山楂配伍pH值水解产物为人参皂苷20(R)-Rg2、20(S)-Rg2、Rg4、Rg6;人参皂苷Rg1与山楂配伍后水解产物为20(S)-Rh1、20(R)-Rh1,仿山楂pH值水解产物为20(S)-Rh1、20(R)-Rh1、Rh4、Rk3;人参皂苷Rb1与山楂配伍后水解产物为Rd、20(S)-Rg3,仿山楂pH值水解产物为F2、20(S)-Rg3;人参皂苷Rd与山楂配伍后水解产物为F2、20(S)-Rg3、20(R)-Rg3,仿山楂pH值水解产物为20(S)-Rg3、20(R)-Rg3。研究表明,不同人参皂苷和山楂配伍后与仿山楂pH值的水解产物并不相同,人参与山楂配伍改变了人参皂苷成分的种类及含量。本研究为临床方剂中人参与山楂配伍后成分的变化提供物质基础数据。     

Analysis of Low-polar Ginsenosides in Steamed Panax Ginseng at High-temperature by HPLC-ESI-MS/MS

A high performance liquid chromatography coupled with electrospray ionization-tandem mass spectrometry( HPLC-ESI-MS/MS) method was developed for the analysis and identification of ginsenosides in the extracts of raw Panax ginseng(RPG) and steamed Panax ginseng at high temperatures(SPGHT). A total of 25 ginsenosides were extracted include of which 10 low-polar ginsenosides, such as ginsenosides F4, Rk3, Rh4, 20S-Rg3, 20R-Rg3 and so on, were identified according to their HPLC retention time and MS/MS data. The results indicated that the low polar ginsenosides were seldom found in RPG. For the exploration of the transformation pattern of the ginsenosides in steam processing, the standards of ginsenosides Re, Rg1, Rb1, Rc, Rb2, Rb3 and Rd were selected and hydrolyzed at a temperature of 120 ℃. The results show that these polar ginsenosides can be converted to low-polar ginsenosides such as Rg2, Rg6, F4, Rk3 and Rg5 by hydrolyzing the sugar chains.     

Major ginsenoside contents in rhizomes of Panax sokpayensis and Panax bipinnatifidus

This study compared eight major ginsenosides (Rg1, Rg2, Rf, Re, Rd, Rc, Rb1 and Rb2) between Panax sokpayensis and Panax bipinnatifidus collected from Sikkim Himalaya, India. High-performance liquid chromatographic analysis revealed that all major ginsenosides were present in the rhizomes of P. sokpayensis except ginsenoside Rc, whereas ginsenoside Rf, Rc and Rb2 were not detected in P. bipinnatifidus.     

Study of the noncovalent interactions of ginsenosides and amyloid‐β‐peptide by CSI‐MS and molecular docking

Noncovalent interactions between drugs and proteins play significant roles for drug metabolisms and drug discoveries. Mass spectrometry has been a commonly used method for studying noncovalent interactions. However, the harsh ionization process in electrospray ionization mass spectrometry (ESI‐MS) is not conducive to the preservation of noncovalent and unstable biomolecular complexes compared with the cold spray ionization mass spectrometry (CSI‐MS). A cold spray ionization providing a stable solvation‐ionization at low temperature is milder than ESI, which was more suitable for studying noncovalent drug‐protein complexes with exact stoichiometries. In this paper, we apply CSI‐MS to explore the interactions of ginsenosides toward amyloid‐β‐peptide (Aβ) and clarify the therapeutic effect of ginsenosides on Alzheimer's disease (AD) at the molecular level for the first time. The interactions of ginsenosides with Aβ were performed by CSI‐MS and ESI‐MS, respectively. The ginsenosides Rg1 bounded to Aβ at the stoichiometries of 1:1 to 5:1 could be characterized by CSI‐MS, while dehydration products are more readily available by ESI‐MS. The binding force depends on the number of glycosyls and the type of ginsenosides. The relative binding affinities were sorted in order as follows: Rg1 ≈ Re > Rd ≈ Rg2 > Rh2, protopanaxatriol by competition experiments, which were supported by molecular docking experiment. CSI‐MS is expected to be a more appropriate approach to determine the weak but specific interactions of proteins with other natural products especially polyhydroxy compounds.     

Retention behavior of ginsenosides on a poly(vinyl alcohol)-bonded stationary phase in hydrophilic interaction chromatography

The influences of the organic component of the mobile phase and the column temperature on the retention of ginsenosides on a poly(vinyl alcohol) (PVA) bonded stationary phase operated under hydrophilic interaction chromatographic mode were investigated. The retention of the ginsenosides was found to increase with increasing amount of acetonitrile (MeCN) in the mobile phase, which is typical of hydrophilic interaction chromatographic behavior. It was also found that the retention of the analytes was highly affected by the type of the organic modifier used. Aqueous MeCN (75–90%) gave the most satisfactory retention and separation of ginsenosides Rf, Rg1, Rd, Re, Rc, Rb2 and Rb1 compared with aqueous methanol, isopropyl alcohol or tetrahydrofuran at the same composition levels. The effects of the different types of organic modifiers on the retention of the analytes were attributed to their solvent strength and hydrogen-bond accepting/donating properties. The effect of temperature on the retention of ginsenoside on the PVA-bonded phase was assessed by constructing van’t Hoff plots for two temperature ranges: subambient (273–293 K) and ambient-elevated (298–333 K) temperatures. van’t Hoff plots for all analytes were linear at the two temperature intervals; however, the slopes of the lines corresponding to ginsenosides Rg1 and Re were completely different from those for the rest of the analytes especially in the subambient temperature range. Enthalpy-entropy compensation (EEC) studies were conducted to verify the difference in thermodynamics observed for ginsenosides Rg1 and Re compared with the other analytes. EEC plots showed that Rf, Rd, Rc, Rb2 and Rb1 were possibly retained by the same retention mechanism, which was completely different from that of Rg1 and Re at subambient temperatures. Retention prediction models were derived using multiple linear regression to identify solute attributes that affected the retention of the analytes on the PVA-bonded phase. The mathematical models derived revealed that the number of hydrogen-bond donors and the ovality of the molecules are important molecular properties that govern the retention of the compounds on the chromatographic system.     

Determination of marker constituents in radix Glycyrrhizae and radix Notoginseng by near infrared spectroscopy

High-performance liquid chromatographic (HPLC) methods were developed for the determination of glycyrrhizin in radix Glycyrrhizae and ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf and Rg, in radix Notoginseng. These methods were used as reference methods for near-infrared (NIR) spectroscopy. Spectroscopic calibrations were developed for the determination of glycyrrhizin, the total content of ginsenosides and the individual major ginsenosides Rb1, Rd, Re and Rg1. Standard errors of cross validation (SECV) were 1.22 mg g(-1) for glycyrrhizin (concentration range 21.3-34.1 mg g(-1)) and 0.99 mg g(-1) for the sum of ginsenosides (concentration range 55.3-71.1 mg g(-1)). The corresponding coefficients of determination (R2) were 0.94 and 0.98, respectively. The SECVs were generally less than a factor of 2.5 of the repeatability standard deviation of the HPLC methods.     

Development of a rapid and convenient method to separate eight ginsenosides from Panax ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detection

Ginsenosides exhibit diverse biological activities and are major well-known components isolated from the radix of Panax ginseng C.A. Meyer. In the present work, a rapid and facile method for the separation and purification of eight ginsenosides from P. ginseng by high-speed counter-current chromatography coupled with evaporative light scattering detector (HSCCC-ELSD) was successfully developed. The crude samples for HSCCC separation were first purified from ginseng extract using a macroporous resin; the extract was loaded onto a Diaion-HP20 column and fractionated by methanol and water gradient elution. The ginsenosides-protopanaxadiol (PPD) and protopanaxatriol (PPT) fractions were subsequently eluted with 65 and 80% methanol and water gradient elution, respectively. Furthermore, these two fractions were separated by HSCCC-ELSD. The two-phase solvent system used for separation was composed of chloroform/methanol/water/isopropanol at a volume ratio of 4:3:2:1. Each fraction obtained was collected and dried, yielding the following eight ginsenosides: Rg(1), Re, Rf, Rh(1), Rb(1), Rc Rb(2) and Rd. The purity of these ginsenosides was greater than 97% as assessed by HPLC-ELSD, and their structures were characterized by electrospray-ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance spectroscopy. This is the first report regarding the separation of the ginsenosides Rh(1), Rb(2) and Rc from P. ginseng by HSCCC.     

Marked production of ginsenosides Rd, F2, Rg3, and compound K by enzymatic method

The hydrolysis of protopanaxadiol-type saponin mixture by various glycoside hydrolases was examined. Among these enzymes, crude preparations of lactase from Aspergillus oryzae, beta-galactosidase from A. oryzae, and cellulase from Trichoderma viride were found to produce ginsenoside F(2) [3-O-(beta-D-glucopyranosyl)-20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], compound K [20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], and ginsenoside Rd {3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol}, respectively, from protopanaxadiol-type saponin mixture in large quantities. Moreover, the crude preparation of lactase from Penicillium sp. having a high producing activity of ginsenoside Rh(1) (6-O-beta-D-glucopyranosyl-20(S)-protopanaxatriol) from protopanaxatriol-type saponin mixture gave ginsenoside Rd as a main product, ginsenoside Rg(3) {3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol}, and compound K from protopanaxadiol-type saponin mixture. The hydrolytic pathways of ginsenosides Rb(1), Rb(2), and Rc to ginsenosides Rd, Rg(3), and F(2), and compound K by crude preparations of four glycoside hydrolases were also studied. This is the first report on the enzymatic preparation of an intestinal bacterial metabolite, ginsenoside F(2), in quantity, and a considerable amount of a minor saponin, ginsenoside Rg(3), from a protopanaxadiol-type saponin mixture.     

Determination of ginsenosides (ginseng saponins) in dry root powder from Panax ginseng, Panax quinquefolius, and selected commercial products by liquid chromatography: interlaboratory study

Twelve collaborating laboratories assayed 4 products, namely, Panax ginseng, Panax quinquefolius, and 2 ginseng products, for 6 ginsenosides: Rb1, Rb2, Rc, Rd, Re, and Rg1. Collaborators also received a negative control for the recovery study. Pure ginsenosides were provided as reference standards for the liquid chromatography (LC) analysis and the system suitability tests. The LC analyses were performed on the methanol extract using UV detection at 203 nm. For P. ginseng, individual ginsenosides were consistent in their means; repeatability standard deviations (RSDr) ranged from 4.17 to 5.09% and reproducibility standard deviations (RSDR) ranged from 7.27 to 11.3%. For P. quinquefolius, the Rb1 and Rb2 ginsenosides were higher and lower in concentration than P. ginseng, with RSDr values of 3.44 and 6.60% and RSDR values of 5.91 and 12.6% respectively, and other analytes at intermediate precisions. For ginseng commercial products, RSDr values ranged from 3.39 to 8.12%, and RSDR values ranged from 7.65 to 16.5%. A recovery study was also conducted for 3 ginsenosides: Rg1, Re, and Rb1. The average recoveries were 99.9, 96.2, and 92.3%, respectively. The method is not applicable for the determination of Rg1 and Re in ginseng product at levels <300 mg/kg.     

The use of linear ion trap for qualitative analysis of phytochemicals in Korean ginseng tea

A new approach to qualitative analysis of ginsenosides in challenging matrices was developed on the basis of high‐performance liquid chromatography/tandem mass spectrometry. Using the extracts from samples of ginseng tea, the approach was validated. Analysis of extracts was carried out using a reversed‐phase chromatography with SB‐C₁₈ sorbent. For compound identification, electrospray ionization and a quadrupole/linear ion trap mass‐spectrometer in different modes were used. A meticulous study of the fragmentation of ginsenosides in the linear ion trap and its application for analysis of these compounds was performed in this work. The accuracy of the identification was proven with standards of ginsenosides Rb1, Rg1, Re, Rf, Rd, Rb2, Rb3 and Rc. Copyright © 2012 John Wiley & Sons, Ltd.     

Pharmacokinetic study of five ginsenosides using a sensitive and rapid liquid chromatography–tandem mass spectrometry method following single and multiple oral administration of Shexiang Baoxin pills to rats

Shexiang Baoxin pills (SBP) are a traditional Chinese medicine that are used for treating coronary heart disease. Ginsenosides are the main effective components of SBP, but a comprehensive and deep pharmacokinetic study of ginsenosides in SBP, including multiple dosing and linear or nonlinear properties, is lacking. This study was designed to investigate and compare the pharmacokinetic characteristics of ginsenosides in SBP at a single dose and in multiple doses. A liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method was developed for the simultaneous determination of the ginsenosides Rg1, Re, Rb3, Rc and Rb1 in rat plasma. Rats were randomly assigned to receive a single dose of 4, 8 or 12 g/kg and multiple doses (4 g/kg) of SBP for 8, 15 or 22 consecutive days. The results revealed that ginsenosides, following a single oral dose of 4 or 8 g/kg, were absorbed rapidly, with a T_max ranging from 0.250 to 1.08 h. The AUC_0–t and C_max of the ppd‐type ginsenosides Rb3, Rc and Rb1 were greater than those of the ppt‐type ginsenosides Rg1 and Re. Nondose‐dependent exposure was observed at doses of 4–12 g/kg for all of the ginsenosides. After multiple dosing, the plasma levels of the ppt‐type ginsenosides decreased, whereas those of the ppd‐type ginsenosides did not change significantly. In conclusion, the LC‐MS/MS method was successfully applied to investigate the pharmacokinetics of ginsenosides after single and multiple oral administrations of SBP. The ginsenosides did not accumulate after multiple dosing. The ppd‐type ginsenosides displayed more favorable pharmacokinetic properties compared with the ppt‐type ginsenosides. Copyright © 2014 John Wiley & Sons, Ltd.     

Purification and characterization of new special ginsenosidase hydrolyzing multi-glycisides of protopanaxadiol ginsenosides, ginsenosidase type I

In this paper, the new type ginsenosidase which hydrolyzing multi-glycosides of ginsenoside, named ginsenoside type I from Aspergillus sp.g48p strain was isolated, characterized and generally described. The enzyme molecular weight was about 80 kDa. Ginsenosidase type I can hydrolyze different glycoside of protopanaxadiol type ginsenosides (PPD); i.e., can hydrolyze the 3(carbon)-O-beta-glucoside of Rb1, Rb2, Rb3, Rc, Rd; can hydrolyze 20(carbon)-O-beta-glucoside of Rb1, 20-O-beta-xyloside of Rb3, 20-O-alpha-arabinoside(p) of Rb2 and 20-O-alpha-arabinoside(f) of Rc to produce mainly F2, compound-K (C-K) and small Rh2, but can not hydrolyze the glycosides of protopanaxatriol type ginsenoside (PPT) such as Re, Rf, Rg1. So, when the ginsenosidase type I hydrolyzed ginsenosides, the enzyme selected ginsenoside-aglycone type, can hydrolyze different glycosides of PPD type ginsenoside; however no selected glycoside type, can hydrolyze multi-glycosides of PPD type ginsenosides. These properties were novel properties, and differentiated with the other previously described glycosidases.     

Complete assignment of 1H and 13C NMR data for nine protopanaxatriol glycosides

Nine protopanaxatriol glycosides isolated from mild acid hydrolysis products of crude root saponins of Panax notoginseng were identified as 20(R)‐ginsenoside‐Rh1, 20(S)‐ginsenoside‐Rh1, ginsenoside‐Rg1, ‐Re and ‐Rg2, notoginsenoside‐R2 and ‐R1, a mixture of 25‐hydroxy‐20(S)‐ginsenoside‐Rh1 and its C‐20 (R) epimer, ginsenoside‐Rh4. The complete assignments of the ¹H and ¹³C NMR chemical shifts for these glycosides were obtained by means of 2D NMR techniques, including ¹H–¹H COSY, ROESY, HMQC, HMBC and HMQC‐TOCSY spectra. The glycosylation shift effect of protopanaxatriol and the differences in chemical shifts between 20(R)‐ and 20(S)‐protopanaxatriol isomers are also discussed. Except for ginsenoside‐Re and ‐Rg2, complete NMR assignments of the other seven glycosides are reported for the first time. Copyright © 2002 John Wiley & Sons, Ltd.     

人参中人参皂苷的直接高压微波辅助降解

采用高效液相色谱-电喷雾质谱联用法测定了人参提取液中的人参皂苷. 考察了天然人参皂苷发生降解的条件, 同时研究了单体人参皂苷Rg1, Re, Rb1, Rc, Rb2和Rd的降解, 并对降解产物进行了分析. 结果表明, 随着提取压力的升高, 提取液中天然人参皂苷的含量逐渐减少, 同时产生多种次级人参皂苷. 当微波提取压力达到600 kPa, 提取时间为10 min时, 提取液中的主要天然人参皂苷达到完全降解, 次级人参皂苷Rg3含量达到最高. 在单体人参皂苷Rb1, Rc, Rb2和Rd的降解产物中均得到人参皂苷Rg3.     

Determination of ginsenoside content in Asian and North American ginseng raw materials and finished products by high-performance liquid chromatography: single-laboratory validation

A single-laboratory validation study was conducted for the quantification of Rg1, Re, Rb1, Rc, Rb2, and Rd in Asian ginseng (Panax ginseng C.A. Meyer) and North American ginseng (Panax quinquefolius L.) raw materials and finished products by RP-HPLC. The extraction with aqueous methanol was optimized for whole root, powdered extract, and finished product (raw, tablet, and capsule matrixes) test articles. Root materials were treated with base to hydrolyze acidic malonyl ginsenosides to their neutral counterparts. Calibration curves for each ginsenoside were linear over the following ranges (microg/g): 5-394 for Rg1, 15-1188 for Re, 39-2981 for Rb1, 6-499 for Rc, 5-406 for Rb2, and 7-600 for Rd, all having a coefficient of determination (r2) of > or = 99.5%. The LOD for Rg1, Re, Rb1, Rc, Rb2, and Rd was determined to be 1.06, 1.25, 2.19, 1.24, 1.27, and 1.70 microg/mL, respectively. Quantitative determinations performed with eight test materials by two analysts over 3 days (n = 12) resulted in RSDr values that ranged from 1.11 to 7.61%.     

Rational development of a selection model for solvent gradients in single-step separation of ginsenosides from Panax ginseng using high-speed counter-current chromatography

A new model of solvent gradients selection was rationally developed for the preparative separation of target compounds. The solvent gradients were selected based on a three-stage screening process where stationary phase retention was ensured by introducing a new parameter termed as the phase ratio. The phase ratio was calculated after mixing the upper phase of a solvent system with the lower phase of a different solvent system (1:1, v/v). The developed model was applied to the one-step separation of eight ginsenosides from Panax ginseng. Three gradients were selected on the basis of new model and eight ginsenosides, Rb(1), Rb(2), Rc, Rd, Re, Rg(1), Rf, and Rh(1), were efficiently separated by high-speed counter-current chromatography coupled with evaporative light scattering detector. The structures of all compounds were characterized by electrospray-ionization mass spectrometry and nuclear magnetic resonance spectroscopy.     

Solid-Phase Extraction and High-Performance Liquid Chromatography for Simultaneous Determination of Important Bioactive Ginsenosides in Pharmaceutical Preparations

A robust method based on high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection has been developed for simultaneous determination of six important ginsenosides (Rg1, Re, Rb1, Rc, Rb2, and Rd) in pharmaceutical preparations. For sample preparation, simple and efficient extraction by ultrasonication, combined with solid-phase extraction (SPE) for clean-up, was effective without consuming large amounts of solvent. Chromatographic separation was performed on an ODS column with optimized gradient elution by means of a dual-solvent-pumping system. The validated method results in excellent separation, and quantitative determination is highly precise and accurate. The problem of co-elution of ginsenosides Rg1 and Re is also solved, with good resolution (R_S approx. 1.5). Intraday variation was between 0.2 and 4.4% and interday variation was between 0.4 and 6.5% (n=5 for both). The accuracy was satisfactory—in the range 93.9 to 103.4% from replicate evaluation at three different spiking concentrations. Overall limits of detection based on a typical injection volume of 5 μL were from 1.16 to 1.58 ng μL⁻¹. The validated method enabled complete assessment for quality control of ginseng samples. The technique may be performed with less sample preparation and, consequently, reduced possibility of sample loss.     

Determination of Seven Major Ginsenosides in Different Parts of Panax quinquefolius L.(American Ginseng) with Different Ages

Ginsenosides Rg1,Re,Rb1,Rc,Rb2,Rb3,and Rd in different parts of the American ginseng plant were investigated.The extraction process was a pressurized microwave-assisted extraction(PMAE).The seven ginsenosides were separated and determined by high-performance liquid chromatography(HPLC) with a ultraviolet(UV) detector,at 203 nm.The experiment results showed significant variations in the individual ginsenoside contents of the American ginseng in different parts and ages of the plant.The results demonstrated that the leaves,root hairs,and rhizomes of Panax quinquefolius L.contained higher ginsenoside contents,followed by the main roots and stems.The leaves contained dramatically higher levels of ginsenoside Rg1,Rb3,and Rd than the other four parts.Higher contents of Rb1 and Re were present in the main roots,root hairs,and rhizomes.The amount of ginsenoside content in the stems was the lowest.The total content of the seven ginsenosides in main roots,root hairs and rhizomes increased with the age of the plant.In contrast,the ginsenoside contents in the leaves and stems decreased with a year of growth.     

Validated quantification for selective cellular uptake of ginsenosides on MCF-7 human breast cancer cells by liquid chromatography–mass spectrometry

The cellular behavior of ginsenosides on cancer cells has not been measured directly despite their potent anticancer activities and biological actions. A liquid chromatography–mass spectrometry (LC-MS) method was developed to measure the selective cellular uptake of ginsenosides in both cell lysates and culture media. Fifteen ginsenosides were separated within 17 min with good peak shapes using a 2-μm sub-particle size C18 column. Quantification was performed by triple-quadrupole MS with electrospray ionization in negative ion mode. The sample preparation containing the solid-phase extraction was linear (correlation coefficient, r ² > 0.992) for all analytes, while the limit of quantification ranged from 0.5 to 2.0 ng/mL in both matrices. The assay precision (%CV) and accuracy (%bias) at three different concentrations (5, 20, and 100 ng/mL) were 1.4% to 11.6% and 94.9% to 106.4%, respectively. When this method was used to examine the selective cellular uptake of ginsenosides, the relative non-polar and protopanaxadiol class ginsenosides, such as Rg3, Rk1, Rg5, Rh2, compound-K, and protopanaxadiol (PPD), showed cellular uptake in the MCF-7 cells, but the relative polar and protopanaxatriol class of ginsenosides did not accumulate in the cells. The most non-polar ginsenoside PPD, which is an aglycone of the protopanaxadiol type, resulted in the highest uptake rate. These results show that the different anticancer activities are due to the selective uptake of ginsenosides based on their chemical structures. This LC-MS-based method can be used to estimate the biological activity of ginsenosides on cells from their structural diversity.