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Simultaneous analysis of 12 kind maker components in Banhabaekchulchhonma-tang by RP-HPLC

RP-HPLC를 이용한 반하백출천마탕에서 12종 지표성분의 동시분석

  • Lee, Kwang Jin (Korea Institute of Oriental Medicine(KIOM), KM-Based Herbal Drug Development Group) ;
  • Yang, Hye Jin (Korea Institute of Oriental Medicine(KIOM), KM-Based Herbal Drug Development Group) ;
  • Ma, Jin Yeul (Korea Institute of Oriental Medicine(KIOM), KM-Based Herbal Drug Development Group)
  • 이광진 (한국한의학연구원 한의신약개발그룹) ;
  • 양혜진 (한국한의학연구원 한의신약개발그룹) ;
  • 마진열 (한국한의학연구원 한의신약개발그룹)
  • Received : 2014.03.21
  • Accepted : 2014.07.10
  • Published : 2014.07.31

Abstract

Banhabaekchulchhonma-tang is a traditional Korean herbal prescription with a range of pharmacological activities. In this study, the simultaneous analysis of 12 kinds of marker components, Homogentisic acid, Hesperidin, Naringin, Alisol A, Atractylenolide II, Atractylenolide III, Ginsenoside Rg1, Formononetin, Gastrodin, Berberine, Palmatine and 6-gingerol, in Banhabaekchulchhonma-tang was performed using high-performance liquid chromatography (RP-HPLC). The standard sample of commercial $C_{18}$ reversed phase-column using water (0.1% TFA) and acetonitrile as the mobile phase with a step gradient elution mode. The flow rate (1.0mL/min), injection volume ($10{\mu}L$) and column oven temperature ($40^{\circ}C$) at a 200, 220, 280 and 340nm wavelength was conducted. All calibration curves of the standard components showed good linearity ($r^2$ >0.999). In addition, the limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.012 to $0.878{\mu}g/mL$ and 0.009 to $0.290{\mu}g/mL$. The precision intra-day and inter-day were ranged from 0.07 to 1.21% and 0.20 to 0.90%, respectively. The recoveries ranged from 97.17 to 108.40%. Each sample amount showed a very small change. These results highlight the efficient quality evaluation of Banhabaekchulchhonma-tang.

반하백출천마탕은 다양한 약리활성을 가진 전통한방처방이다. 본 연구는 한방처방의 품질관리를 위한 기초자료로서, 고성능액체 크로마토그래피 (RP-HPLC)을 이용한 반하백출천마탕에서 12종 지표성분 Homogentisic acid, Hesperidin, Naringin, Alisol A, Atractylenolide II, Atractylenolide III, Ginsenoside Rg1, Formononetin, Gastrodin, Berberine, Palmatine, 6-gingerol를 동시분석 하였다. 표준액은 상업용 $C_{18}$ 역상 컬럼과 이동상으로 물(0.1% TFA)과 아세토나이트릴을 사용하여 기울기용매조성법인 유속은 1.0mL/min, 주입부피는 $10{\mu}L$, 컬럼오븐 온도 $40^{\circ}C$, 파장 200, 220, 280, 340nm에서 수행되었다. 이 결과 모든 표준성분의 검량선은 우수한 직선성 $r^2$ >0.999으로 나타내었다. 또한 검출한계(LOD)와 정량한계(LOQ)는 $0.012{\sim}0.878{\mu}g/mL$$0.009{\sim}0.290{\mu}g/mL$, 정밀성 intra-day와 inter-day는 0.07~1.21%및 0.20~0.90%, 회수율은 97.17~108.40%, 각각의 함량은 매우 미미한 변화를 타나내었다. 이 연구결과는 실험적으로 입증된 동시분석법의 확립에 따라 반하백출천마탕의 품질평가를 효율적으로 보여주었다.

Keywords

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