Cho, B. G.
(KT&G Central Research Institute)
,
Nho, K. B.
(KT&G Central Research Institute)
,
Shon, H. J.
(KT&G Central Research Institute)
,
Choi, K. J.
(KT&G Central Research Institute)
,
Lee, S. K.
(KT&G Central Research Institute)
,
Kim, S. C
(KT&G Central Research Institute)
,
Ko, S. R.
(KT&G Central Research Institute)
,
Xie, P. S.
(Guangzhou Institute for Drug Control)
,
Yan, Y. Z.
(Guangzhou Institute for Drug Control)
,
Yang, J. W.
(KT&G Central Research Institute)
A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginse...
A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.
A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.
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가설 설정
When the standard mixtures of ginsenoside-Rg1, ginsenoside-Re and ginsenoside-Rb1 were analyzed by using an ELSD of ELSD 2000 (KGTRTs method), the calibration graph of either ginsenoside-Rg1, -Re or -Rb1 was quadratic as shown in Fig. 2. The regression coefficient (r) for each graph of ginsenoside was 0.9999 or higher. In addition, there were good linear relationships between the detector responses of ginsenosides and their amount injected to the HPLC (r = 0.
제안 방법
A cross-examination of validation on the extraction and separation method of ginsenosides was carried out between KGTRI and GIDC. The reproducibility for the contents of ginsenoside-Rg1, -Re and -Rb1 and the recovery rates of the three ginsenosides were examined.
Ginsenosides were analyzed by the established HPLC/ELSD method and their peak resolution, repeatability, linearity and detection limit were examined.
In order to establish a quantitative analytical method of ginsenosides in red ginseng, several HPLC/ELSD conditions suggested by KGTRI and GIDC for the determination of ginsenosides were examined. A standard mixture containing ginsenoside-Rh1, -Rh2, -Rg2, -Rg1, -Rg3, -Rf, -Re, -Rd, -Rc, -Rb2 and -Rb1 was analyzed by HPLC/ELSD.
The extraction conditions of ginsenosides and their separation conditions using a SepPak C18 cartiidge suggested by KGTRI and GIDC were compare to establish a better method.
대상 데이터
All reagents used in this study were GR or HPLC grades.
Korean red ginseng in root-size of 30 Ji (=38 roots per 600g) was used as a ginseng sample. Rhizome of the ginseng sample was removed, and then its main body and lateral roots (=big tails) were ground to 60 mesh-size just before the analysis.
성능/효과
These results indicate that the linearities of both the HPLC/ELSD methods would be good. The detection limits of ginsenoside-Rg1 -Re and -Rb1 were 9 ng, 28 ng and 108 ng when analyzed by using an ELSD of ELSD 2000 model but the detection limits of the three ginsenosides were 80 ng ail when analyzed by using another ELSD of SEDEX 55, as shown in Table 3. These results suggest that linearity of ELSD for the analysis of ginsenosides and its sensitivity may be different from the HPLC conditions such as the mobile phase and the operating temperature of ELSD. Park et aL [11] reported that the there were linear relationships between the ginsenoside amounts of 0.
1 shows HPLC patterns of ginsenosides analyzed by KGTRIs method and GIDCs method. Such HPLC/ELSD analytical conditions as mobile phases, detector model and detection temperature were different between KGTRTs method and GIDC's method, but all the ginsenoside peaks except for ginsenoside-Rh1 and -Rh2 were well separated not only by KGTRI's method but by GIDC's method. This result indicates that the peak resolutions of most ginsenosides may be very good as long as ginsenosides are analyzed under the HPLC conditions using a Lichrosorb NH2 column and an ELSD.
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