The main active components of Panax ginseng are ginsenosides. Ginsenoside Rb1 and Rg1 are accepted as marker substances for quality control worldwide. The analytical methods currently used to detect these two compounds unfairly penalize steamed and dried (red) P. ginseng preparations, because it has...
The main active components of Panax ginseng are ginsenosides. Ginsenoside Rb1 and Rg1 are accepted as marker substances for quality control worldwide. The analytical methods currently used to detect these two compounds unfairly penalize steamed and dried (red) P. ginseng preparations, because it has a lower content of those ginsenosides than white ginseng. To manufacture red ginseng products from fresh ginseng, the ginseng roots are exposed to high temperatures for many hours. This heating process converts the naturally occurring ginsenoside Rb1 and Rg1 into artifact ginsenosides such as ginsenoside Rg3, Rg5, Rh1, and Rh2, among others. This study highlights the absurdity of the current analytical practice by investigating the time-dependent changes in the crude saponin and the major natural and artifact ginsenosides contents during simmering. The results lead us to recommend (20S)- and (20R)-ginsenoside Rg3 as new reference materials to complement the current P. ginseng preparation reference materials ginsenoside Rb1 and Rg1. An attempt has also been made to establish validated qualitative and quantitative analytical procedures for these four compounds that meet International Conference of Harmonization (ICH) guidelines for specificity, linearity, range, accuracy, precision, detection limit, quantitation limit, robustness and system suitability. Based on these results, we suggest a validated analytical procedure which conforms to ICH guidelines and equally values the contents of ginsenosides in white and red ginseng preparations.
The main active components of Panax ginseng are ginsenosides. Ginsenoside Rb1 and Rg1 are accepted as marker substances for quality control worldwide. The analytical methods currently used to detect these two compounds unfairly penalize steamed and dried (red) P. ginseng preparations, because it has a lower content of those ginsenosides than white ginseng. To manufacture red ginseng products from fresh ginseng, the ginseng roots are exposed to high temperatures for many hours. This heating process converts the naturally occurring ginsenoside Rb1 and Rg1 into artifact ginsenosides such as ginsenoside Rg3, Rg5, Rh1, and Rh2, among others. This study highlights the absurdity of the current analytical practice by investigating the time-dependent changes in the crude saponin and the major natural and artifact ginsenosides contents during simmering. The results lead us to recommend (20S)- and (20R)-ginsenoside Rg3 as new reference materials to complement the current P. ginseng preparation reference materials ginsenoside Rb1 and Rg1. An attempt has also been made to establish validated qualitative and quantitative analytical procedures for these four compounds that meet International Conference of Harmonization (ICH) guidelines for specificity, linearity, range, accuracy, precision, detection limit, quantitation limit, robustness and system suitability. Based on these results, we suggest a validated analytical procedure which conforms to ICH guidelines and equally values the contents of ginsenosides in white and red ginseng preparations.
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가설 설정
1) Concentration of all samples was 0.5 mg/mL.
In this study, we determined the ginsenoside Rb1 and Rg1 content of commercially available ginseng extract. We also measured the crude saponin content, the concentration change of the major naturally occurring ginsenoside Rb1 or Rg1 and the generation of artifact ginsenosides as a function of time at 80°C.
제안 방법
Change in ginsenosides content during simmering at 80°C (mg%, w/v). Aliquots (100 mL) of the simmering ginseng root were sampled at 3, 6, 12, 24, 36, 48, 60 and 72 h, and the samples were subjected to MCI gel adsorption chromatography to determine the crude saponin content. The content of the (20S)-ginsenoside Rg3 and (20R)-ginsenoside Rg3 increased to a greater extent.
A glass vessel containing 250 g of fresh ginseng root and 2,500 mL of water was simmered (approximately 80°C) for 72 h. Aliquots (sample A, 100 mL) of the simmering ginseng root were removed at 3, 6, 12, 24, 36, 48, 60, and 72 h, and the samples were subjected to gel adsorption chromatography (MCI; Supelco, Bellefonte, PA, USA) or butanol partition to determine the crude saponin content. The crude saponin fraction was dissolved to a concentration of 20 mg/mL, and a HPLC quantitative analysis of the 11 ginsenosides was performed.
UPLC equipment reduces recycling time and increases resolution, but the equipment is not prevalent. Consequently, the authors recommend a newly validated HPLC analytical method for ginsenoside Rb1 and Rg1 that includes (20S)-ginsenoside Rg3 and (20R)-ginsenoside Rg3 for the qualitative and quantitative determination of reference materials for P. ginseng preparations.
The linear relationship was evaluated for the full range of the analytical procedure by visual inspection of the signals as a function of analyzed concentration. Five sample concentrations were employed to establish linearity, and the correlation coefficient, y-intercept, and the slope of the regression line were measured to quantify the same. The range was derived from both the content of the four naturally occurring ginsenosides in the red ginseng sample and the linearity data.
Two HPLC systems were employed to determine purity and content: The HPLC analysis of the ginsenosides for preliminary validation tests was performed using 3 different HPLC configurations. The 1st HPLC setup: Discovery HS C18 material (Supelco, Bellefonte, PA, USA; 250×4.
대상 데이터
The four reference ginsenosides employed for this validation study were 100% pure as determined by HPLC (ODS, 4.6×200 mm, 203 nm, aqueous CH3CN gradient).
데이터처리
Statistical analyses were performed using Student’s t-test in Excel (Microsoft office professional plus 2010; Microsoft, Seattle, WA, USA).
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