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Validation of Simultaneous Analysis Method of Standard Compounds in Fermented Kalopanax pictus Nakai by Bioconversion

생물전환을 통한 음나무발효물의 지표성분 설정 및 동시분석법 검증

  • 장원희 ((주)에스티알바이오텍 중앙연구소) ;
  • 이화영 ((주)에스티알바이오텍 중앙연구소) ;
  • 이봉진 ((주)에스티알바이오텍 중앙연구소) ;
  • 김진만 ((주)에스티알바이오텍 중앙연구소) ;
  • 박선주 ((주)에스티알바이오텍 중앙연구소)
  • Received : 2019.03.08
  • Accepted : 2019.03.18
  • Published : 2019.04.30

Abstract

The aim of this study was to select compounds for the standardization of fermented Kalopanax pictus Nakai (KP-F), to develop the analysis method using HPLC-PDA and to perform method validation. KP-F is a fermented powder developed to improve the original physiological activities and create a new functionality. Eleutheroside E, Acanthoside B, and Syringaresinol were selected as the standard compounds and developed our own method for simultaneous analysis. The analyte was isolated using C18 column with a gradient elution of 0.05 M phosphoric acid in water and methanol as the mobile phase at a flow rate of 1 mL/min and detected at 210 nm. As a result, all standard compounds showed good linearity with an $R^2$ (coefficient of correlation) of 1.000 and for the limit of detection range of $0.710{\sim}0.831{\mu}g/mL$, and the limit of quantification as $2.150{\sim}2.520{\mu}g/mL$. The precision was RSD (%) of less than 4.80%, while the accuracy was 4.70%>RSD (%) for the range 102.44~110.48%. In conclusion, the developed analysis method is suitable for the detection of Eleutheroside E, Acanthoside B, and Syringaresinol in KP-F.

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Fig. 2. Chromatogram of (A) standards mixture of Eleutheroside E, Acanthoside B, and Syringaresinol, (B) Kalopanax pictus Nakai (KP), (C) Fermented Kalopanax pictus (KP-F).

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Fig. 1. Chromatogram of (A) standards mixture of Kalopanaxsaponin B and Kalopanaxsaponin A, (B) Kalopanax pictus Nakai(KP), (C) Fermented Kalopanax pictus Nakai (KP-F).

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Fig. 3. Compound structures of Kalopanax pictus Nakai(KP) converted through Bioconversion.

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Fig. 4. Content graph of (A-1) Kalopanaxsaponin B and Kalopanaxsaponin A in Kalopanax pictus Nakai (KP), (A-2) Kalopanaxsaponin B and Kalopanaxsaponin A in fermented Kalopanax pictus Nakai (KP-F), (B-1) Eelutheroside E, Acanthoside B, and Syriangaresinol in Kalopanax pictus Nakai (KP), (B-2) Eelutheroside E, Acanthoside B, and Syriangaresinol in fermented Kalopanax pictus Nakai (KP-F).

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Fig. 5. UV spectrum of (A) Eleutheroside E, (B) Acanthoside B, and (C) Syringaresinol.

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Fig. 6. Chromatogram of (A) Blank (70% EtOH), (B) Standards mixture of Eleutheroside E, Acanthoside B, and Syringaresinol, (C) Fermented Kalopanax pictus (KP-F).

Table 1. Content of Kalopanaxsaponins and Liganan glycosides in fermented Kalopanax pictus Nakai (KP-F)

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Table 2. Calibration curves, detection limits (LOD) and quantification limits (LOQ) of Eleutheroside E, Acanthoside B, and Syringaresinol

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Table 3. Precision of fermented Kalopanax pictus Nakai (KP-F)

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Table 4. Accuracy of Eleutheroside E, Acanthoside B, and Syringaresinol

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Acknowledgement

Supported by : 중소기업청

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