Saikosaponin Content Research Articles

Quantitative and Differential Analysis between Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. Using HPLC-MS and GC-MS Coupled with Multivariate Statistical Analysis.

Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. have different clinical efficacies, with the former typically used to treat typhoid fever and the latter mainly used to clear liver heat. The differences in their clinical efficacy are closely related to their complex chemical composition, especially the active components. In this study, the saponins and volatile oils in two varieties of Radix Bupleuri grown in different regions were extracted and analyzed using high-performance liquid chromatography (HPLC) and gas chromatography coupled with mass spectrometry (MS), and the absolute contents of five saikosaponins were accurately quantified using an established HPLC-MS method in the multiple reaction monitoring mode. Multivariate statistical analysis was performed to reveal the difference in the active components between the two varieties. The saikosaponin content was significantly affected by variety and growing region, with all five saikosaponins being significantly higher in Bupleurum chinense DC. than in Bupleurum scorzonerifolium Willd. The results of principal component analysis and hierarchical cluster analysis show a clear distinction between the two varieties in terms of both saponins and volatile oils. Twenty-one saponins, including saikosaponin b2 and b1, and fifty-two volatile oils, including 2-tetradecyloxirane and chloromethyl cyanide, were screened and identified as differential compounds contributing to the significant difference between the two varieties. These compounds may also be responsible for the difference in clinical efficacy between Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. All the results suggest that the accumulation and diversity of active components in Radix Bupleuri are significantly affected by the variety. In contrast to previous reports, this study provides the absolute contents of five saikosaponins in Radix Bupleuri of different varieties and reduces the influence of the growing region on the analytical results by collecting samples from different regions. The results of this study may provide a reference for the identification and quality evaluation of different varieties of Radix Bupleuri.

Assessment of Genetic Diversity in Bupleurum spp. Basing Agronomic Traits, Medicinal Components and ISSR Markers.

Radix bupleuri is one of the bulk medicinal materials in China and it is widely adopted in clinical applications and drug discovery. The investigation of agronomic traits, active component content and genetic diversity in diverse Radix bupleuri germplasms may provide evidence to promote the selection of better strains. In this research, 13 germplasms from various sources were used to investigate the variations between different Radix bupleuri germplasms. Nine biological characteristics were noted in the field, and the levels of the two primary active ingredients were determined using high performance liquid chromatography (HPLC). Moreover, the molecular marker technique of inter-simple sequence repeat (ISSR) and the unweighted pair group method with arithmetic means (UPGMA) were employed to evaluate the molecular genetic diversity. The findings showed that there was a wide range of variation among the many varieties of Radix bupleuri, with coefficients of variation for agronomic traits and active component content ranging from 7.62% to 41.54% and 36.47% to 53.70%, respectively. Moreover, there are different degrees of relationship between the two. Since there was a significant correlation between root weight and saikosaponin content, it was possible to classify a plant based on its weight and anticipate its saikosaponin content. The 13 species were divided into four groups based on their germplasm by genetic markers-based cluster analysis. This indicated the possibility that the component content would not necessarily be related to germplasm and might easily be influenced by environmental factors. The use of ISSR marker technology made it possible to precisely identify the various Radix bupleuri provenances and its counterfeit products. There may be a way to prevent the misunderstandings caused by the appearance and composition of Chinese medicinal substances. In our study, the germplasm of Radix bupleuri that was widely circulated in the market was comprehensively evaluated in terms of agronomic traits, active components and molecular level, and identified by simple means, to provide a theoretical basis for the evaluation and screening of fine germplasms of Radix bupleuri.

Transcriptome and metabolome analysis to reveal major genes of saikosaponin biosynthesis in Bupleurum chinense

BackgroundBupleurum chinense DC. is a widely used traditional Chinese medicinal plant. Saikosaponins are the major bioactive constituents of B. chinense, but relatively little is known about saikosaponin biosynthesis. In the present study, we performed an integrated analysis of metabolic composition and the expressed genes involved in saikosaponin biosynthetic pathways among four organs (the root, flower, stem, and leaf) of B. chinense to discover the genes related to the saikosaponin biosynthetic pathway.ResultsTranscript and metabolite profiles were generated through high-throughput RNA-sequencing (RNA-seq) data analysis and liquid chromatography tandem mass spectrometry, respectively. Evaluation of saikosaponin contents and transcriptional changes showed 152 strong correlations (P < 0.05) over 3 compounds and 77 unigenes. These unigenes belonged to eight gene families: the acetoacetyl CoA transferase (AACT) (6), HMG-CoA synthase (HMGS) (2), HMG-CoA reductase (HMGR) (2), mevalonate diphosphate decarboxylase (MVD) (1), 1-deoxy-D-xylulose-5-phosphate synthase (DXS) (3), farnesyl diphosphate synthase (FPPS) (11), β-amyrin synthase (β-AS) (13) and cytochrome P450 enzymes (P450s) (39) families.ConclusionsOur results investigated the diversity of the saikosaponin triterpene biosynthetic pathway in the roots, stems, leaves and flowers of B. chinese by integrated transcriptomic and metabolomic analysis, implying that manipulation of P450s genes such as Bc95697 and Bc35434 might improve saikosaponin biosynthesis. This is a good candidate for the genetic improvement of this important medicinal plant.

Phylogenetic position of Bupleurum sikangense inferred from the complete chloroplast genome sequence.

Bupleurum sikangense is an endemic species to China distributed in Xizang (Tibet), which has high saikosaponin content and potential medicinal value. Morphologically, it extremely resembles B. commelynoideum. In order to get a better understanding of the relationship between B. sikangense and B. commelynoideum, and on the phylogenetic status of the two species in the genus, the complete chloroplast (cp) genomes of them were sequenced. The genome organization, repeat sequences, codon usage, RNA-editing sites, and variation of their cp genomes revealed high similarity between the species. Some highly variable regions like trnK-UUU_rps16, rps16_trnQ-UUG, ndhC_trnV-UAC, petA_psbJ, accD_psaI, and petL_psbE were identified, providing potential molecular markers for differentiating the two species. Phylogenetic analysis indicated that B. commelynoideum has a closer relationship to B. chinese than that to B. sikangense. Overall, this study will not only improve our knowledge about cp genomes of these two species, and but also provide data for further research on species identification, safe medical application, conservation genetics, etc., of Bupleurum plants.

Exogenous abscisic acid reduces saikosaponin accumulation by inhibiting saikosaponin synthesis pathway gene expression under drought stress in Bupleurum chinense DC.

Abscisic acid (ABA) plays an important role in stress resistance and secondary metabolism of plants under drought stress. Bupleurum chinense DC. is a medicinal plant cultivated in arid areas in China. In this study, the effects of progressive drought stress (22 days) and exogenous ABA (5 mg L–1) treatment on plant physiological characteristics and saikosaponin secondary metabolism were investigated in 2-year-old B. chinense. The samples were collected on 6, 10, 14, 18 and 22 days after drought stress (DADS). In drought stressed B. chinense plants treated with ABA, root weight loss and malondialdehyde accumulation was reduced, while drought resistance increased, as indicated by the accumulation of soluble protein and soluble sugar content, and by the increased activities of superoxide dismutase and peroxidase. At the early stage of drought stress (first 6 days), saikosaponin increased more sharply in plants exposed to drought stress and its accumulation indicated increased sensitivity to this condition. At this time, saikosaponin content reached 31.83 mg g−1, which was 33.6 % higher than that in control plants. Using principal component analysis and fuzzy membership function calculation, comprehensive evaluation values of gene expression in the drought (D) and D + ABA treated groups at different drought exposure time were evaluated. Exogenous ABA inhibited the expression of saikosaponin biosynthesis-related genes from 6 to 14 DADS of drought stress. However, gene expression was up-regulated at later stages of drought stress (the 22 DADS), while genes involved in saikosaponin content were down-regulated in the D group compared with those in the D + ABA group. Indeed, saikosaponin content in both of these groups was significantly higher than that in the control group. Correlation analysis suggested that the expression of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) gene was positively correlated with total saikosaponin (SSs) (p < 0.01), and saikosaponin a (SS-a) and saikosaponin d (SS-d) (p < 0.05) content, while that of farnesyl pyrophosphate synthase (FPS) gene was positively correlated with SSs (p < 0.05), and SS-a and SS-d (p < 0.01) content in the D group. These results indicated that HMGR and FPS play a key role in saikosaponin biosynthesis during drought stress. Moreover, the negative correlations between multiple genes and saikosaponin after ABA treatment [particularly that of cytochrome P450−7 with SS-d (p < 0.01) and glycosyl transferase-8 with SS-d (p < 0.05)], indicated that exogenous ABA reduced saikosaponin accumulation by inhibiting the expression of saikosaponin biosynthesis-related genes in B. chinense exposed to drought stress. This study provides new insight into the effect of ABA on secondary metabolism and it will be of great importance to cultivators and researchers aiming to understand how ABA application affects medicinal plant cultivation.

Total saikosaponin content in some species of Bupleurum L

The aim of this study was to determine the total saikosaponin content in some species of Bupleurum L. genus. For the first time, the saikosaponin content was investigated in roots of B. scorzonerifolium from Buryatia and Mongolia. The differences in the quantitative content of saikosaponins in the B. scorzonerifolium roots can be explained by the possible polymorphism of this plant. Dynamic change of the total saikosaponin content in roots of B. scorzonerifolium in different phenophases didn’t undergo significant changes. The total saikosaponin content was at the highest level in the roots of B. bicaule.

What degree of light deficiency is suitable for saikosaponin accumulation by Bupleurum chinense DC.?

Total saikosaponin yield in Bupleurum chinense DC. roots depends on the root biomass production and the saikosaponin content in roots, which are both affected by abiotic factors. Bupleurum chinense is relatively shade-tolerant. To test how total saikosaponin yield responds to light conditions, different light intensities (500, 200, and 50 μmol m−2 s−1) were established in artificial climate incubators. Responses of plant morphological and physiological characteristics and of saikosaponin synthesis to different light intensities were measured to find suitable light conditions for improving total saikosaponin yield. Among these light conditions, moderately low light intensity (MLI, 200 μmol m−2 s−1) produced plants with the highest leaf area, chlorophyll content, effective quantum yield of PSII (ΦPSII), photochemical quenching coefficient (qP), electron transport rate (ETR), maximum carboxylation efficiency (Vcmax), maximum electron transport rate (Jmax), and triose phosphate utilization rate (VTPU), which suggests a well-functioning photosynthetic apparatus capable of fully utilizing the limited light energy. Moreover, the plants had the highest root–shoot ratio, saikosaponin content, and total saikosaponin yield, which indicates that they allocated more resources to storage and defensive adaptations. Under extremely low light intensity (ELI, 50 μmol m−2 s−1), more energy was partitioned to photoprotection, and strong oxidation resistance developed, but saikosaponin accumulation and root biomass were lowest due to greatly reduced carbon assimilation. The MLI improved both the quantity and the quality of herbal extracts from B. chinense: the total saikosaponin yield was 1.3 times the corresponding value of the control (500 μmol m−2 s−1), and lower illumination levels were counterproductive. Our results suggest that B. chinense would be suitable for introducing to an agroforestry system.

Effects of light deficiency on the accumulation of saikosaponins and the ecophysiological characteristics of wild Bupleurum chinense DC. in China

Bupleurum chinense DC. is one of the most important traditional Chinese medicines. Among its primary pharmacologically active components, saikosaponins play a vital role. We investigated the ecophysiological characteristics, accumulation of secondary metabolites, and antioxidant system of wild plants at low light intensity in the forest understory at shady and sunny sites. Compared with plants at sunny sites, plants at the shady site were more abundant, had higher specific leaf area (SLA), a lower maximum electron transport rate (Jmax), higher triose phosphate utilization (TPU), and higher chlorophyll b content (Chlb), but lower net photosynthesis (Pn), root and aboveground biomass, dark respiration rate (Rd), light compensation point (LCP), chlorophyll a to b ratio, carotenoid content, and nonphotochemical quenching (NPQ). These features suggest that under low light conditions, the species developed larger leaves to catch more light energy through light-harvesting complex II to increase its photosynthetic efficiency (its ability to use weak light) and photosynthetic electron transport, thereby improving carbon metabolism and survival. The maximum quantum efficiency of photosystem II was about 0.8 at both sites; thus, photoinhibition was not a major problem. Contents of saikosaponins a and d and the total saikosaponin content (SSa, SSd, and SStotal) were much higher at the shady site, accompanied by increased superoxide dismutase (SOD), catalase(CAT), and peroxidases (POD) activity and decreased malondialdehyde (MDA). This suggested that low light activates antioxidant stress-response pathways, leading to increased SSa, SSd, and SStotal. Our results suggest that B. chinense has strong shade tolerance and would be suitable for cultivation in an agroforestry system.

Anti-melanoma activity of Bupleurum chinense, Bupleurum kaoi and nanoparticle formulation of their major bioactive compound saikosaponin-d

Ethnopharmacological relevanceBupleurum chinense is a traditional Chinese medicinal herb which has been used to treat various inflammatory and infectious diseases, while Bupleurum kaoi is an endemic plant in Taiwan. We determined whether B. chinense and B. kaoi and their biologically active saikosaponin compounds possess anti-melanoma activity. In addition, we developed a novel saikosaponin-d nanoparticle system to improve its solubility, and evaluated its antiproliferative effects and molecular mechanisms in melanoma cells. Materials and methodsEthanolic extracts from B. chinense and B. kaoi were prepared, and their saikosaponin contents were determined by high performance liquid chromatography analysis. Saikosaponin-d nanoparticles were synthesized, and their physicochemical properties were evaluated by particle size analyzer, transmission electron microscopy, differential scanning calorimetry, X-ray diffractometry, and Fourier transform infrared spectroscopy. Human A375.S2 melanoma cells were cultured, and cell viability determined by the MTT assay. Apoptosis was evaluated by determination of mitochondrial membrane potential, and signal transduction pathways investigated by Western blotting. ResultsEthanolic extracts from B. kaoi showed more potent antiproliferative effect on human A375.S2 melanoma cells compared to B. chinense. The saikosaponin-a, -c and -d contents were higher in B. kaoi compared to B. chinense. Saikosaponin-d was the most potent compound in terms of anti-melanoma activity, and saikosaponin-d nanoparticles exhibited increased water solubility due to lowered particle size, amorphous transformation and intermolecular hydrogen bond formation with the excipient. Furthermore, saikosaponin-d nanoparticles showed enhanced antiproliferative activity against melanoma cells, and induced apoptosis through the mitochondrial pathway. The anti-melanoma activity was mediated by phosphorylation of JNK and p38, phosphorylation of p53, increased level of cytochrome c, and activation of caspase 9. ConclusionsB. kaoi contains higher saikosaponin content and shows greater anti-melanoma activity than B. chinense. Saikosaponin-d nanoparticles have improved solubility, and may have potential use in the future as a form of treatment for melanoma.

Chemical and biological comparison of raw and vinegar-baked Radix Bupleuri

Ethnopharmacological relevanceRadix Bupleuri (RB) is a commonly used herbal drug in Traditional Chinese Medicine (TCM), and it can be baked with vinegar to afford vinegar-baked Radix Bupleuri (VBRB), which is used in TCM for liver diseases treatment. In this study, the chemical compositions and biological effects between raw and two processed RBs by different vinegars were systematically compared. Materials and methodsThe chemical compositions of raw and two processed RBs were analyzed by 1H NMR spectroscopy coupled with multivariate analysis. Two different extraction procedures were used, including direct extraction and liquid–liquid partition. Then HPLC was applied to determine the changes of saikosaponin contents. In addition, their liver protective effects against CCl4 induced liver injury were also investigated, and the biochemical parameters and histopathology were measured after treatment of mice with raw RB and two processed RBs (5g/kg/day) for 14 days. ResultsMultivariate analysis showed clear differences between the raw and the two processed RBs, and the vinegar-baking process induced elevated contents of ssb1, ssb2, acetic acid, malic acid, citric acid, 5-HMF, and ligustrazine, as well as the decreased contents of ssa, ssd, sucrose, glycine, succinic acid etc. In addition, both raw and processed RBs showed liver protective effects against CCl4 induced liver injury, and the vinegar-baked RBs showed better effects than that of raw RB. ConclusionsThe raw and vinegar-baked RBs differed not only in the chemical compositions but also in the pharmacological effects. And two processed RBs also showed chemical differences, suggesting that the type of vinegar had an important effect on vinegar baking. In order to ensure the therapeutic effect and safety of TCM, the effect of different vinegars on processing of herbal drugs should be further studied.

Determination of saikosaponins in threeBupleurumplants by HPLC analysis

In this study, saikosaponins were determined from roots of three Bupleurum plants collected in June 2009. Bupleuri Radix (Bupleurum spp. root) is one of the most important crude drugs in Korea, China and Japan. High performance liquid chromatography (HPLC) was used for the determination of saikosaponin ‘a’, ‘b’ and ‘c’ in three Bupleurum plants. The highest total saikosaponin content was found in the B. falcatum ‘Mishima’ (1281.94 mg%). Total saikosaponin contents of B. falcatum and B. latissimum were 669.67 and 489.95 mg%, respectively. B. falcatum ‘Mishima’ and B. falcatum contained more saikosaponin ‘a’ than other saikosaponins. The contents of saikosaponin ‘a’ of B. falcatum ‘Mishima’ and B. falcatum were 745.74 and 484.60 mg%, respectively. In contrast, B. latissimum, an endemic species of Korea had higher saikosaponin ‘c’ (351.90 mg%) content than the other two saikosaponins and the content was higher than B. falcatum (181.71 mg%). B. latissimum, a Korean endemic species, is a good source for extraction/production of saikosaponin ‘c’.

Analysis of saikosaponins in Bupleuri Radix and Caihu‐shugan‐san using reversed‐phase HPLC with pulsed amperometric detection

A simple and sensitive reversed-phase (RP) HPLC coupled with pulsed amperometric detection (PAD) method was developed to determine the saikosaponin content in Bupleuri Radix or Caihu-shugan-san. Four saikosaponins in Bupleuri Radix and Caihu-shugan-san were extracted with a 6:4 solution of 10 mM sodium phosphate buffer (pH 8)/100% ethanol. Pulsed amperometric detection of carbohydrates in four major saikosaponins was highly sensitive when used with a water-acetonitrile gradient on an alkaline RP column with a post-column delivery system. The limits of detection (S/N=3) and of quantification (S/N=10) of saikosaponins were 0.01-0.02 and 0.03-0.05 μg/mL, respectively. The intra- and inter-day precision (RSDs) were each <9.7% and the average recoveries were 95.0-97.6% in Bupleuri Radix. This method can be used to analyze saikosaponins in Bupleuri Radix and Caihu-shugan-san.

Breeding of new varieties "Zhongchai No. 2" and "Zhongchai No. 3" of Bupleurum chinense

To breed new varieties with better uniformity and commercial quality as well as higher saikosaponin contents. The excellent germplasm resources were selected from "zhongchai no. 1" population. Single plant method was applied to get better varieties. All the breeding material was investigated according to morphological characters, agronomic characters and the contents of saikosaponin a and saikosaponin d. The experiments of comparative test and varieties regional test were carried out. The bred new varieties of "zhongchai No. 2" and "zhongchai No. 3" had better uniformity. The dark brown roots ratios of the two varieties were 83.2%, 89.9%, respectively. The contents of saikosaponins (a + d) of the two varieties reached 1.31%, 1.02%, respectively. "zhongchai No. 2" and " zhongchai No. 3" both had the advantages of better uniformity, darker brown roots and higher saikosaponin contents.

Effects of Cultivation Temperature on the Growth and Saikosaponin Production of Bupleurum falcatum L. Cultivars

The effects of cultivar and cultivation temperatures on root extract, ash content and saikosaponin con- tent of Bupleurum falcatum L. were investigated using four Japanese and one Korean cultivars. Results showed that the Korean cultivar obtained higher root extract and saikosaponin contents but lower ash con- tent than the Japanese cultivars at cultivation temperatures of 16.5-17.6 °C. Higher cultivation tempera- ture (21.0°C) consistently yielded low values in all parameters for both cultivars. Results also showed inverse relationships between root extract and ash content, and between ash content and saikosaponin con- tent. On the other hand, root extract showed direct relationship with saikosaponin content. This study indicated the usefulness of component analysis of plant materials for traditional medicinal preparation since each plant material may have different concentration of their active ingredients.

Localization and Dynamic Change of Saikosaponin in Root of Bupleurum chinense

Anatomical, histochemical and phytochemical methods were used to investigate the structure, the localization and content changes of total saikosaponin and saikosaponin-a of the roots of Bupleurum chinense DC. at different developmental stages. Results showed that saikosaponin was mainly distributed in pericycle and primary phloem in the young root; but in the mature root, it was mainly distributed in vascular cambium and secondary phloem. During the whole growth period from the pre-blossom, blossom, fruit, and fruit mature periods until the pre-withering period, it was in the fruit mature period that both the total saikosaponin content and the saikosaponin-a content reached the highest level. So the last 20 d of October was considered as the right collecting season for the drug of B. chinense. In addition, the quality of 1-year-old drug was better than that of 2-year-old drug due to its higher saikosaponin content. On the other hand, judging from the external characteristics of the drug, the one with an acerose taproot and more lateral roots was of better quality. The results offered theoretical bases for selecting medicinal material of high quality and determining the most appropriate harvesting stage and part of B. chinense.

Comparative Study on Saikosaponin Contents in Root of Unsexual Propagation and Sexual Reproduction in Bupleurum falcatum

Comparative Study on Saikosaponin Contents in Root of Unsexual Propagation and Sexual Reproduction in Bupleurum falcatum

High-performance liquid chromatographic analysis of saponin compounds in Bupleurum falcatum.

A mixture of saponin compounds (saikosaponin c, a, and d) in the 70% ethanol extract of a powdered sample of Bupleuri radix are analyzed by an Inertsil ODS-3 C(18) column at a flow rate of 1.0 mL/min and detection wavelength of 203 nm. Well resolved chromatograms of saikosaponin c, a, and d are obtained with a gradient elution of acetonitrile-water from 40:60 (v/v) to 50:50 (v/v). The total time required for a single analysis is approximately 20 min. Calibration curves for saikosaponin c, a, and d are linear up to 2.5 mg/mL. The coefficient of variability values for saikosaponins in the extract are below 4%, and the recoveries for saikosaponin c, a, and d are 95.2 +/- 1.1, 96.5 +/- 0.9, and 96.2 +/- 1.0%, respectively. The changes in saikosaponin contents for a two-year growth of Bupleurum falcatum are measured by the established high-performance liquid chromatography method.

Studies on Dry Matter Production and Efficiency for Solar Energy Utilization in Bupleurum falcatum L. at Different Plant Ages

Two Bupleurum falcatum cultivars, which originated from Jeongseon, Korea and Kumamoto, Japan were grown for two years to clarify the dry matter (DM) production, efficiency for solar energy utilization (Eu) and saikosaponin a,c and d contents in Fukuoka, Japan. Aerial parts, root and total DM production differed with the cultivar both in the first and second years. Generally, DM production correlated with leaf area index, and there was a significant relationship between total DM and Eu in the plants of the first year in both cultivars. High Eu was observed at the flowering stage in both cultivars in both years. The maxium Eu was 1.02-1.23% in the first year and 1.16-2.09% in the second year. Total saikosaponin content was higher in the first year than in the second year. Although the total saikosaponin content was higher in Jeongseon cultivar than in Kumamoto cultivar in the second year, no difference betweeen the two cultivars was observed in the first year. We concluded that annual cultivation with a high saikosaponin content was preferable for the high yield of saikosaponin.

Studies on the Agronomic Characteristics, Yield, and Saikosaponin Content of Two Bupleurum Species in Taiwan

Two Bupleurum species (Bupleurums spp.), i.e., B. falcatum L. cv. Tainung No. 1 and B. kaoi Liu, Chao, et Chuang, were compared for their differences in seed germination, plant characteristics and root yield, and saikosaponin content. Experimental data showed that the most suitable temperature for seed germination of the two species was 16 degrees C. Two treatments, cold stratification at 4 degrees C for 8 weeks and presoaking by running water for 2 days, resulted in higher germination rates. Tainung No. 1 possessed a higher 1,000-seed weight than B. kaoi. However, field survival rate was higher for B. kaoi than for Tainung No.1. Results from field experiments also revealed that stem diameter, leaf width and fresh weight of various plant parts except the root were superior for Tainung No. 1 to B. kaoi. On the contrary, tiller number and root diameter and weight of B. kaoi were higher than those of Tainung No. 1. Harvest data and elevation had significant effects on the agronomic performance of the two Bupleurum spp. Measurements of most traits of the 6 month-old plants were superior to those of the 3 and 10 month-old plants. Cultivation at higher elevation (850 m) favored the development of leaf weight and root length, while cultivation at lower elevation (85 m) facilitated the development of plant height, root diameter, and root weight. Analysis of saikosaponin concentration in the root tissue revealed that average contents of 3.19 and 3.80 mg/g, respectively, for plants grown at the elevations of 850 m and 85 m. Comparison between the two species showed no significant difference in saikosaponin content, ranging from 3.45 to 3.55 mg/g.