본 연구는 인삼 잎의 이용증대를 위해 마이크로웨이브에 의한 인삼 잎의 잔류농약 추출효과와 발효 인삼 잎의 ginsenoside 유용 유도체의 전환 검토 및 품질 특성을 분석 하였다. 인삼 잎에 잔류되어 있는 tolclofos-methyl와 azoxystrobin을 microwave로 추출하기 위한 용매는 hexane이 가장 효율적 이었다. tolclofos-methyl와 azoxystrobin이 잔류되어 있는 인삼 잎에서의 microwave를 이용한 추출 최적 조건은 power 50∼95 watts, 추출용매는 hexane, 추출시간은 3분으로 나타났다. 인삼 잎 추출물의 발효에서 발효 전과 비교하여 Rg1과 Rb1은 감소한 반면 Rh1, Rg3, Rk1 및 Rh2는 발효 후 모두 증가한 것으로 나타났다. 특히 홍삼에서 대표적인 성분으로 알려져 있는 Rg3의 경우 발효전 2.77 ㎍/g에서 발효 후 균주의 종류에 따라 70.62∼77.61 ㎍/g으로 증가하였다. 7일간 발효 후 인삼 잎의 총 페놀성 화합물 및 전자공여능은 일부 균주에서는 발효전과 비교하여 감소하다가 다시 증가하는 경향을 나타내었으나, 발효가 진행됨에 따라 전반적으로 감소되는 경향을 나타내었다.
This study was carried out to extract ginsenosides in by-products from honeyed red ginseng. Response surface methodology (RSM) was used to optimize the extraction conditions. Based on D-optimal design, independent variables were ethanol (extraction solvent) concentration (30-90%, v/v), extraction temperature (25-70oC), and extraction time (5-11 h). Extraction yield (Y1) and total ginsenosides (Y2) in the extract were analyzed as dependent variables. Results found that extraction yield increased with increasing extraction temperature and time, whereas it was decreased with increasing ethanol concentration. Similar trends were found for the content of ginsenosides in the extracts, except for ethanol concentration, which was increased with increasing ethanol concentration. Regression equations derived from RSM were suggested to coincide well with the results from the experiments. The optimal extraction conditions for extraction yield and total ginsenosides were an extraction temperature of 56.94oC, ethanol concentration of 57.90%, and extraction time of 11 h. Under these conditions, extraction yield and total ginsenoside contents were predicted to be 84.52% and 9.54 mg/g, respectively.
Microwave-assisted extraction (MAE) is known as a more environmental-friendly process with economic advantages in terms of less time, less solvent, less energy and less waste than the current reflux method that is time-consuming. MAE was applied to extract three pesticides (quintozene, tolclofos-methyl and procymidone) from ginseng in a pesticide residue analysis and eliminate pesticide contaminants from ginseng prior to production of the ginseng extracts. The optimal conditions for pesticide extraction from ginseng were 50% in power (150 watts), 2 minutes, 20 mL hexane and 2.0 g sample. The results of optimal conditions were compared with those of Soxhlet method (7 hours, 150 mL hexane). The recoveries between two methods were almost same while time and amount of solvent used in MAE were significantly decreased in comparing with those in the Soxhlet methods. When the 45% ethyl alcohol as a extraction solvent was applied to eliminate pesticides from ginseng prior to the production of the ginseng extracts, it was found that 100% quintozene, 69% tolclofos-methyl and 83% procymidone were eliminated from ginseng. The data showed that MAE may be used to eliminate pesticide contaminants in ginseng prior to making the ginseng extracts.
Background: Hot steaming is known to be effective in improving the biological activities of plant extracts by breaking down useful compounds to low molecular weight ones.
Methods and Results: This study aimed to develop an optimal extraction and steam processing method for enhancing the low molecular ginsenoside contents of the adventitious roots culture of wild mountain ginseng. The total ginsenoside was optimally extracted when 70% EtOH was used at 50℃, whereas low molecule ginsenoside such as Rg2, Rh1, Rh4 and Rk1 could be extracted using 70% EtOH at 70℃. The adventitious roots culture of wild mountain ginseng is known to contain four major ginsenosides, i.e., Rb2, Rb1, Rg1 and Rd, however new ginsenosides Rg6, Rh4, Rg3, Rk1 and Rg5 were new abundantly obtaind after steam processing method was applied. The contents of total ginsenosides were the highest when thermal steam processing was conducted at 120℃ for 120 min. Unlike ginsenosides such as Rg1, Re, Rb1, Rc, Rb2, and Rh1, which decreased after steam processing, Rg3, Rk1, and Rg5 increased after thermal processing. Steam processing significanltly reduced the content of Rb1, increased that of Rg6 by about ten times than that in the adventitious roots culture of wild mountain ginseng.
Conclusions: Our study showed that the optimal extraction and steam processing method increased the content of total ginsenosides and allowed the extraction of minor ginsenosides from major ones.
Background : The study about ginseng cultured roots have been reported mainly ginsenosides in saponins family. Other phytochemical such as non-saponins of fatty acid has been revealed its bioactive activity including anti-oxidation, whitening, anti-cancer. Supercritical extraction (SE) process mainly refer to the extraction with CO2, is usually from a solid matrix, is a sample preparation step for analytical purposes. SE produce no residual solvent and possess high stability of the extract component, which is advantageous for fatty acid analysis. Methods and Results : Fermented ginseng cultured roots used in the experiment were used for fermentation using Pediococcus pentosaceus. SE performed at different temperature, pressure and extraction time using non-fermented and fermented ginseng roots. Further we fractionated from fermented ginseng using Methanol, Hexane, Ethanol, Ethyl acetate and Butanol. We compared fatty acids contents ginseng extractions by GC analysis. Methyl linoleate contents was 44% of fatty acids supercritical extraction contained. The contents of Methyl linoleate was the most dominant component among 37 types of fatty acids by SE and other extractions solvent. Total fatty acids contents obtained by SE process from fermented ginseng (1325.61ppm) was twice than from non-fermented ginseng (618.47ppm). Conclusion : Fatty acids contents by SE was increased at high pressure. The best condition for fatty acids contents extraction was 60℃, 350bar and 3h.
Background : From 2000 years ago, Panax ginseng is identified as precious pharmaceutical plant. Depend on growing environment, the name would be vary. For instance, it is called "mountain cultured ginseng (jangnoesam)" which is artificially grown ginseng, "Cultured ginseng (jaebaesam)" which refer to the ginseng grown in the forest, and lastly "Wild ginseng (sansam)" which inhabits in deep mountain. The main active compounds in the Panax ginseng is called ginsenoside and many researches have been performing in biological field. However, most studies focus on functional ability of ginseng. In this study, to seek the suitable extraction condition and antioxidant activity, cell cultured Panax ginseng was extracted according to different ethanol concentration and extraction time. Methods and Results : To establish the optimal extraction condition, the sample was pulverized into 500 μm and added 10% (v/v), 30% (v/v), 50% (v/v), 70% (v/v) and, 90% (v/v) EtOH. After that, the samples are extracted in different time by ultrasonic bath (Power sonic 520, Hwashin Co., Korea). The extracts was filtered by Whatman No. 2 filtering paper. Eventually, the saponin was separated by n-butanol as the ginsenoside, the combination of terpenoid and sugar. The extraction yield of 90% cell cultured panax ginseng EtOH extract was 7.36±0.33%, which was the lowest extraction yield and simultaneously, 10% EtOH extract showed 1.8 times more yield that of 90% EtOH extract. The saponin extraction yield revealed 10% and 70% EtOH extract showed 1.64±0.06% and 3.13±0.08%, respectively. Conclusion : The suitable extraction yield in cell cultured panax ginseng and saponin were evaluated by different extraction condition such as ethanol concentration and extraction time. As a result, when 10% EtOH was applied as solvent, the yield was doubles of 90% EtOH extract. As ethanol became high concentrations, the extraction yield was gradually increased. Among them, crude saponin, the main active compounds in Panax ginseng was extracted the most by 70% EtOH and that value was 3.13±0.08%.
A new extraction method-heated ultrasonic extraction was qualitatively and quantitatively analyzed for the extraction of major ginsenosides from ginseng extract; this new high-performance liquid chromatography (HPLC) method was compared with the official extraction method of Korean industrial standards and standard for health functional food. Methods and Results : Ginsenoside compounds were analyzed for 35 minutes by the new HPLC analysis method using a Halo® RP-Amide column. The new HPLC analysis method was validated by the measurement of intra-day and inter-day precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) of each ginsenoside. The correlation coefficients (r2) for the calibration curves of the ginsenoside compounds were over 0.9997 in terms of linearity. The heated ultrasonic extraction method using ultrasonication for 30 minutes at 50℃ yielded higher amount of ginsenosides than the extraction method of the Korean industrial standards owing to the enhancement of extraction efficiency. Conclusions : Compared to the other extraction methods, the heated ultrasonic extraction method yielded a higher amount of ginsenoside Rb1 than Rg1 index compounds for the quality evaluation of ginseng roots.
This study was performed to enhance contents of low molecular weight ginsenoside Rh2 and Rg3 using an ultra high pressure and steaming process in wild cultured-Root in wild ginseng. For selective increase in contents of Rg3 and Rh2 in cultured wild ginseng roots, an ultra high extraction was applied at 500MPa for 20 min which was followed by steaming process at 90℃ for 12 hr. It was revealed that contents of ginsenosides, Rb1, Rb2, Rc and Rd, were decreased with the complex process described above, whereas contents of ginsenoside Rh2 and Rg3 were increased up to 4.918 mg/g and 6.115 mg/g, respectively. In addition, concentration of benzo[α]pyrene in extracts of the cultured wild ginseng roots treated by the complex process was 0.64 ppm but it was 0.78 ppm when it was treated with the steaming process. From the results, it was strongly suggested that low molecular weight ginsenosides, Rh2 and Rg3, are converted from Rb1, Rb2, Rc, and Rd which are easily broken down by an ultra high pressure and steaming process. This results indicate that an ultra high pressure and steaming process can selectively increase in contents of Rg3 and Rh2 in cultured wild ginseng roots and this process might enhance the utilization and values of cultured wild ginseng roots.
In this study, ginseng flower water extracts were analyzed to set up the ginsenoside content and quality optimization condition. The highest total ginsenoside content among the ginseng flower water extracts was 67.44mg/g which was extracted at 85℃ for 3 hours. In addition, the ginsenoside content decreased according to the increased extraction temperature and time. The highest total content of Rb2 and Re was 37.42mg/g at 75℃ for 6 hours. Total content of Rb2 and Re decreased according to the increased extraction temperature and time. The highest prosapogenin (Rg2 + Rg3 + Rh1) content among the total of ginseng flower water extracts was 18.58mg/g which was extracted at 95℃ for 12 hours. The sweetness, absorbance were increased according to the increased extraction temperature and time. But pH was decreased according to the increased extraction time.
본 연구는 홍삼의 가능성 식품개발을 위한 기초 자료로 활용하기 위하여 추출 시간에 따른 홍삼 부위별 ginsenoside의 함량 변화를 비교하였다. 동체, 지근 및 세근의 총 ginsenoside 최고 함량은 동제 21시간, 지근 18시간, 세근 12시간 추출하였을 때 각각 23.04, 65.68 295.92 mg/100 ml이었고, 추출시간이 증가할수록 ginsenoside의 총량은 감소하였다. Ginsenoside Rg1 과 Rb1 의 최고 함량은 동제 21시간, 지근 15시간, 세근 12시간 추출하였을 때 각각 5.76, 28.39, 117.83 mg/100 ml 이었고, Rb2 와 Re의 함량은 동체 21시간, 지근 18시간, 세근 9시간 추출하였을 때 각각 5.76, 28.39, 117.83 mg/100 ml이었다. 홍삼으로부터의 총 ginsenoside의 추출비율은 동체 21.3%, 지근 21.1%, 세근 67.1%이었다.
본 연구는 백삼 추출액의 사포닌 함량을 높이고 품질과 관련된 특성을 증진 시킬수 있는 최적 추출 시간 및 온도를 밝히기 위하여 실시되었다. 백삼의 물 추출액의 총 ginsenosides 함량은 75℃ 에서 18시간 추출액 8.32 mg/10ml, 85℃ 에서 6시간 추출액 5.93 mg/10 ml, 95℃ 에서 6시간 추출액 4.92 mg/10 ml으로 최고의 함량을 나타냈다. 항당뇨 작용을 나타내는 ginsenoside Rb2 와 Re을 합한 최고 함량은 75℃ 에서 18시간 추출액 1.76 mg/10 ml, 85℃ 6시간 추출액 1.34 mg/10 ml, 95℃ 6시간 추출액 0.56 mg/10 ml이었으며, 항암 작용을 나타내는 Rg3 의 함량은 75℃ 에서 36시간 추출액 1.67 mg/10 ml, 85℃ 에서 36시간 추출액 3.13 mg/10 ml, 95℃ 에서 24시간 추출액이 3.56 mg/10 ml이었다. 추출온도와 시간에 따른 이화학성의 변화는 추출온도가 높을수록, 추출시간이 길어질수록 pH는 낮아 졌으나 환원당, 탁도, 당도는 증가하였다. 따라서 백삼추출액을 제조할때 1차 추출에서는 사포닌 최고 함량 추출 온도와 시간으로 추출한 다음 다시 온도를 높게 2차 추출 하여 맛을 증진시켜 혼합하는 방법이 좋을 것으로 생각된다.
The low quality fresh ginseng was extracted by water at 80℃ and 240bar for 20min (HPE, High pressureextraction process). The cytotoxicity on human normal kidney cell (HEK293) and human normal lung cell (HEL299) of theextracts from HPE showed 28.43% and 21.78% lower than that from conventional water extraction at 100℃ in adding themaximum concentration of 1.0㎎/㎖. The human breast carcinoma cell and lung adenocarcinoma cell growth were inhib-ited up to about 86%, in adding 1.0㎎/㎖ of extracts from HPE. This values were 9-12% higher than those from conven-tional water extraction. On in vivo experiment using ICR mice, the variation of body weight of mice group treated freshginseng extracts from HPE of 100㎎/㎏/day concentration was very lower than control and other group. The extracts fromHPE was showed longer survival times as 35.65% than that of the control group, and showed the highest tumor inhibitionactivities compared with other group, which were 70.64% on Sarcoma-180 solid tumor cells. On the high performance liq-uid chromatogram (HPLC), amount of ginsenoside-Rg2, Rg3, Rh1 and Rh2 on fresh ginseng were increased up to 43-183%by HPE, compared with conventional water extracts. These data indicate that HPE definitely plays an important role ineffectively extracting ginsenoside, which could result in improving anticancer activities. It can be concluded that low qualityfresh ginseng associated with this process has more biologically compound and better anticancer activities than that fromnormal extraction process.
In this study, raw ginseng water extract solutions were analyzed to set up the functional saponin content and quality optimization condition. The highest saponin content among the total raw ginseng water extracts was 74.6 mg/100 ml which was extracted at 75℃ for 24 hours. In addition, the saponin content decreased according to the increased extraction temperature and time. The highest total content of Rb2 and Re was 19.9 mg/100 ml at 75℃ for 12 hours which decreased according to the increased extracted temperature and time. The highest prosapogenin (Rg2 + Rg3 + Rh1) content among the total raw ginseng water extracts was 28.6 mg/100 ml which was extracted at 85℃ for 36 hours. The reducing sugar content, sweetness and turbidity were increased according to the increased extraction temperature and time. But pH were decreased according to the increased extracted time.
The low quality fresh ginseng was fermented by Phelinus linteus or Hericium erinaceum mycelium. This fermented ginseng was extracted by water at 100℃ or water with ultrasonification at 60℃. Total phenolic compounds was improved by ultrasonification extraction process, compare to conventional water extraction. All extracts enhanced the growth of human B and T cells, showing 2.68 times and 3.43 times higher, respectively, than the control. The secretion of TNF-α and IL-6 from human immune cells was enhanced as 3.53×10-4 pg/cell, 3.40×10-4 pg/cell by adding H. erinaceum mycelium fermented ginseng. H. erinaceum mycelium fermented ginseng yielded higher nitric oxide production from macrophage than Lipopolysaccharides (LPS). The cytotoxicity on human normal kidney cell (HEK293) was as low as 20.5% in adding the maximum concentration of 1.0 mg/ml of fermented ginseng. Generally, the extracts from ultrasonification extraction process showed 10% lower toxicity than that by conventional process. H. erinaceum mycelium fermented ginseng had the highest anticancer activity on human lung cancer and stomach cancer cells as 69.33% and 75.32%, respectively at 1.0 mg/ml. It can be concluded that, in general, H. erinaceum mycelium fermented ginseng has relatively better immune and anticancer activities than P. linteus fermented ginseng. Expecially, the extracts treated with ultrasonification had higher activities than that from conventional extraction process.
An advanced extraction method by ultrasonic extraction with applied solid phase extraction (SPE) has been developed for the determination of simultaneous eight major ginsenosides, namely ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, and Rd in the root of Panax ginseng. Four extraction methods including n-BuOH reflux extraction (Method A), 70% EtOH reflux extraction (Method B), 50% MeOH reflux extraction with SPE (Method C), and 50% MeOH ultrasonication with SPE clean-up process (Method D) were investigated for the determination of eight major ginsenosides. Total contents of ginsenosides were highest by extraction of Method C as 2.408±0.011%. However, Method D was evaluated as relatively simpler and more efficient method due to short extraction time, small solvent consumption and less expensive, compared to conservative reflux method. Ginsenosides were also satisfactorily separated with good resolution and the accuracy range was between 1.05 and 4.06% as relative standard deviation (RSD) by Method D. SPE condition and HPLC condition were further optimized for determination of eight major ginsenosides by the ultrasonic extraction method. Conclusively, ultrasonic extraction of 2 g sample of ginseng using ultrasonic bath and 1 loading for SPE was evaluated as proper condition for extraction of ginseng.