• Journal of Pharmaceutical Investigation
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• 제38권1호
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• pp.57-61
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• 2008

A HPLC-ELSD method was developed to determine saikosaponin derivatives from Bupleuri Radix. Eight saikosaponins, saikosaponin c, i, h, a, $b_2$, g, $b_1$ and d, were analyzed under optimized HPLC conditions [column: Eclipse XDB $C_{18}$ ($150{\times}4.6mm$ i.d., $5{\mu}m$; mobile phase: $H_2O$ with 0.1% $CH_3$COOH (v/v) for solvent A and AcCN with 0.1% $CH_3$COOH (v/v) for solvent B, gradient elution; flow rate: 1mL/min; injection volume: $20{\mu}L$]. Good linearity was achieved in the range from 62.5 to $250{\mu}g/mL$ for each compound, and intra-day precision and accuracy at each concentration level varied between 0.05 and 5.45% and between 93.9 and 109.6%, respectively, whereas those for inter-day variations were between 0.91 to 2.73% and 94.3 to 106.1%. This HPLC-ELSD method was applied for the determination of sakosaponins from Bupleuri Radix samples, and saikosaponin a $(0.79{\pm}0.20mg/g)$, c $(0.33{\pm}0.06mg/g)$ and d $(0.48{\pm}0.15mg/g)$ were observed as major compounds. The other saikosaponins were shown under limit of quantification level thus couldn't be quantified. The present study suggested that the introduced HPLC-ELSD method is selective and reliable, and not only saikosaponin a, but also saikosaponin c and d should be employed as the standard markers for Bupleuri Radix.

• Molecules and Cells
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• 제22권3호
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• pp.269-274
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• 2006

In order to characterize saikosaponin biosynthesis in Bupleurum falcatum, the expression of five isoprenoid pathway genes and their relationship to saikosaponin accumulation in the hairy roots were analyzed. The hairy roots exhibited a rapid accumulation of saikosaponins when incubated in a root culture medium (3XRCM). Homology-based RT-PCR was used to isolate core fragments of five genes, HMGR, IPPI, FPS, SS, and OSC, from the hairy roots. The deduced amino acid sequences exhibited amino acid identities of more than 85% to previously reported genes. Using the fragments as probes, the expression of these five genes in the hairy roots during incubation in 3XRCM medium was examined. Expression of all five genes in the hairy roots increased soon after incubation. In particular, the SS and OSC genes were coordinately induced at 8 days of incubation, and their expression persisted throughout the incubation period. A quantitative HPLC analysis showed that the saikosaponin content of the hairy root culture also began to increase at 8 days of culture. The correlation between SS transcript level and saikosaponin content in the hairy roots suggests that transcriptional regulation plays a regulatory role in saikosaponin biosynthesis.

• 생약학회지
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• 제24권2호
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• pp.153-158
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• 1993

As a part of pharmacological studies of saikosaponins, which were reported to exhibit diverse biological activities especially concerning with liver function, effects of saikosaponin on metabolizing enzymes and lipid peroxide contents in liver were examined. As the result, UDP-glucose dehydrogenase activity and lipid peroxidation which were due to acetaminophen were inhibited by saikosaponin treatment. But other metabolizing enzyme activities were not modified.

• 생약학회지
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• 제21권3호
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• pp.205-209
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• 1990

The study for the quantitative analysis of saikosaponin of Bupleurum falcatum root produced by tissue culture was attempted. The optimal concentration of 2,4-D was 0.1 mg/l for inducing the callus. The induction and growth of the adventitious root from the callus was obtained in the suspension culture containing MS basal medium supplemented with no plant growth regulators. The results of the quantitative analysis of saikosaponins by high performance liquid chromatography (HPLC) showed that the content of saikosaponin a and c was high in the one-year-old root from somatic embryos and that of saikosaponin d was remarkably high in three-months-old adventitious root from callus.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.121-126
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• 2006

Saikosaponin productivity was examined in a Bupleurum falcatum L. BFHR2 hairy root culture in response to changes in the sucrose content $(2{\sim}8%)$, nitrogen content $(0{\sim}250mM\;NH_4NO_3)$, phosphate content $(0{\sim}12mM\;NaH_2PO_4)$, and the potassium content $(0{\sim}87.2 mM\; KCl)$ of the culture media. We found that the conditions for maximal saikosaponin production differed from those for optimal root growth. Highest saikosaponin yield was achieved for 8% sucrose, 62mM $NH_4NO_3$ 1.2 mM $NaH_2PO_4$, and 0.5mM KCl.

• 한국약용작물학회지
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• 제10권5호
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• pp.317-326
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• 2002

본 시험은 국내 재래종 정선시호와 일본에서 도입된 삼도시호의 품질과 생산성 향상을 위한 적정 파종시기 및 재식밀도의 구명과 saikosaponin 생산에 기여하는 요인들을 구명하여 시호의 안정적 생산을 위한 기초 자료로 이용하고자 실험을 수행하여 얻어진 결과를 요약하면 다음과 같다. 1. 파종시기와 재식거리에 따른 시호품종간에서 정선시호는 삼도시호보다 분지수는 적었고 초장은 길었다. 1년생의 정선시호는 삼도시호보다 지상부중은 적었고 생근중과 건근중은 높았으며, 2년생에서는 정선시호와 삼도시호간에 유의적인 차이는 없었다. Saikosapon함량은 1년생 또는 2년생의 정선시호가 삼도시호보다 saikosaponin a 및 total saikosaponln함량이 높았다. 2. 파종시기에 따른 생육특성은 1년생이나 2년생의 시호 모두 4월 30일 파종구에서 경태와 분지수가 가장 굵고 많았으며, 1년생 시호에 있어서 파종시기가 빠를수록 saikosaponin c 함량이 높게 나타났다. 3. 1년생 시호는 소식일수록 초장, 경태, 분지수, 지상부중, 그리고 근경, 생근중, 건근중, 지근수 등의 생육이 촉진되었으나 2년생 시호는 소식에서 주근장만 길었다. 4. 품종, 파종시기 및 재식거리에 있어서 상호작용은 1년생 시호만이 지상부중, 주근장, 근경에서 유의적인 차이를 보였다. 1년생의 정선시호는 4월10일에 파종하면서 재식밀도를 $30{\times}15\;cm$로 재배한 것에서 지상부중, 근경 및 TSS 함량이 가장 증가되었으며, 1년생 삼도시호는 3월20일에 재식밀도를 $30{\times}10\;cm$로 재배한 것에서 지상부중과 근경이 가장 높았다.

• 한국약용작물학회지
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• 제6권4호
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• pp.323-327
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• 1998

시호 재분화 체세포클론과 야생 개체의 뿌리에서 사이코사포닌 함량을 정량하여 비교 분석하였다. 사이코사포닌 함량은 1년생 체세포클론, 경상북도 영덕시호, 강원도 정선시호, 2년생 체세포클론 순으로 높게 나타났다. 사이코사포닌 함량 변이는 3개월간 배양한 캘러스에서 유도되어 2년간 재배된 체세포클론에서 2년간 배양한 캘러스에서 유도되어 1년간 재배된 체세포클론에서보다 높게 나타났다. 이러한 결과는 시호의 경우 사이코사포닌 변이가 캘러스 배양 기간에 별로 영향을 받지 않는다는 것을 보여주는 것이다. 1년생 체세포클론에서 야생 개체에서보다 사이코사포닌 함량이 2배 이상 높은 4개의 체세포클론이 발견되었는데, 이러한 결과는 체세포클론 변이를 이용한 우량 변이주 선발의 가능성을 보여 주는 것이다.

• 한국자원식물학회지
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• 제11권3호
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• pp.283-289
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• 1998

To determine the differences among cultivated , non-cultivated and imported bupleurum falcatum, contents and compositions of fatty acids and saikosaponins which are known as a major biologically acitive compounds in Bupleurum falcatum were meausred with GC and HPLC. Color of grinded powder of Bupleuri radix determined by colorimeter showed that the imported one has the highest white and yellow coloer Among the Bupleuri radix cultivated in China, Hong-siho(red-Bupleurum) produced in Yungil Province contained the highest red color. Cultivated Korean Bupleurum radix contained higher saikosaponi a, c, and than the imported ones which were cultivated in Yungil, Ankuk , and Gillium Province in China. Levels of saikosaponin c are 3 times higher in cultivated Korean Bupleuria radix compared the with imported ones. Furthermore, Bupleurum falcatum cultivated in Korea contained higher amount of palmitic and linoleic acids than those of improted and non-cultivated ones. Particularly they contain two to six times higher linoleic acid.

• 생약학회지
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• 제16권4호
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• pp.253-258
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• 1985

Various determination methods of saikosaponins in Bupleuri Radix are discussed, and the content of each saikosaponin which is original and/or artifact is rearranged. The changes of saikosaponin a and d into saikosaponin $b_1\;and\;b_2$, respectively, under gastric pH, were studied.

• Archives of Pharmacal Research
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• 제21권2호
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• pp.135-139
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• 1998

High titer rabbit polyclonal antibodies (pAbs) which show a specificity for saikosaponin a (SSA), have been generated. The immunogen used was a conjugate of SSA linked through its glucose moiety to bovine serum albumin by periodate oxidation method. The antibody titers obtained from two rabbits, innoculated with the immunogen, reached a plateau after the fourth and third booster injection, respectively. The specificity of the pAbs was determined by hapten inhibition assays using several SSA-like structures. SSA competitively inhibited the binding of the rabbit anti-SSA pAbs to SSA-ovalbumin on solid phase, a coated antigen on the well. The antibodies showed high specificity to SSA, exhibiting no significant cross-reactivity with any of SSA analogues tested.