UPLC-QTOF-MS/MS Analysis for Steaming Times-dependent Profiling of Steamed Panax quinquefolius and its Ginsenosides Transfromations induced by Repetitious Steaming

Bai-Shen Sun,Ming-Yang Xu,Zheng Li,Yi-Bo Wanag,Chang-Keun Sung 고려인삼학회 2012 Journal of Ginseng Research Vol.36 No.3

The metabolic profiles of Panax quinquefolius and its associated therapeutic values are critically affected by the repetitious steaming times. The times-dependent steaming effect of P. quinquefolius is not well-characterized and there is also no official guidelilne on its times of steaming, In this paper, a UPLC-Q-TOF-MS/MS method was developed for the qualitative profiling of multi-parametric metabolic changes of raw P. quinquefolius durilng the repetitious steaming process. Our method was successful in discriminating the differentially multi-steamed herbs. Meantime, the repetitious steaming-inducing chemical transformations in the preparation of black American ginseng (American ginseng that was subjected to 9 cycles of steaming treatment) were evaluated by this UPLC-Q-TOF-MS/MS based chemical profiling method. Under the optimized UPLC-Q-TOF-MS/MS conditions, 29 major ginsenosides were unambiguously identified and/or tentatively assigned in both raw and multi-steamed P. quinquefolius within 19 min, among them 18 ginsenosides were detected to bo newly generated during the preparatory process of black American ginseng. The mechanisms involved were further deduced to be hydrolysis, dehydration, decarboxylation and addition reactions of the original ginsenosiders in raw P. quinquefolius through analyzing 9 cycles of steaming extracts of 14 pure reference ginsenosides. Our novel steaming times-dependent metabolic profiling represents the paradigm shift in the global quality control of multi-steamed P. quinquefolius products.

Determination of 11 Ginsenosides in Black Ginseng Developed from Panax ginseng by High Performance Liquid Chromatography

Bai-Shen Sun,Li-Juan Gu,Zhe-Ming Fang,Chun Yan Wang,Zhen Wang,Chang-Keun Sung 한국식품과학회 2009 Food Science and Biotechnology Vol.18 No.2

A high performance liquid chromatography (HPLC) method has been developed for determination of 11 ginsenosides in black ginseng (BG, white ginseng that is subjected to 9 cycles of 95℃ for 3 hr). After eluted by gradient elution of water-acetonitrile without buffer in 70 min, 11 ginsenosides in BG were identified. The proposed method provided good linearity (R²>0.9995), accuracy (92.2-106.6%), and intra- and interday precision (RSD<2.6%). In addition, ginsenosides compositions in white, red, and black ginsengs were investigated using this method, respectively. Interestingly, in BG, the content of ginsenoside Rg3 which does not existed in white ginseng was 7.51 ㎎/g, approximately 20 times than that in red ginseng.

Ginseng extracts attenuates neurotoxicity and ameliorates cognitive impairment induced by D-galactose in rats

Bai-Shen Sun,Mi-Ra Lee,Beom-Sik Yun,Lei Lui,Zheng Li,Chun-Yan Wang,Zhen Wang,Dong-Liang Zhang,Jing-Jie Li,Chang-Keun Sung(성창근) 고려인삼학회 2009 고려인삼학회 학술대회 Vol.2009 No.5

Repetitious Steaming-induced Chemical Transformations and Global Quality of Black Ginseng Derived from Panax ginseng by HPLC-ESI- MS/MS^n Based Chemical Profiling Approach

Bai-Shen Sun,Fu-You Pan,성창근 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.5

A high performance liquid chromatography–electrospray tandem mass spectrometry (HPLC-ESI-MS/MSn) based chemical profiling method was developed to evaluate repetitious steaming-induced chemical transformations in black ginseng (BG and Korean white ginseng subjected to nine cycles of steam treatment). Under the optimized HPLC and ESI-MS/MS^n conditions, more than 13 and 17 peaks were separated and detected in white ginseng (WG) and BG within 85 min, respectively. The components were identified by comparing the mass spectrum and/or matching the empirical molecular formula with that of known published compounds. In total, 17major ginsenosides were identified in BG, 16 of which were determined to be newly generated during the BG preparatory process. The mechanisms involved were further deduced to be hydrolysis, dehydration, isomerization, and decarboxylation reactions of the original ginsenosides in WG by analyzing nine mimic cycles of steaming extracts of seven pure reference ginsenosides. A significant difference in chemical profiles between BGs developed from two batches of WG suggested that storage duration significantly influenced the quality consistency of not only the crude drug but also the BG derived from WG.

UPLC-Q-TOF-MS/MS Analysis for Steaming Times-dependent Profiling of Steamed Panax quinquefolius and Its Ginsenosides Transformations Induced by Repetitious Steaming

Sun, Bai-Shen,Xu, Ming-Yang,Li, Zheng,Wang, Yi-Bo,Sung, Chang-Keun The Korean Society of Ginseng 2012 Journal of Ginseng Research Vol.36 No.3

The metabolic profiles of Panax quinquefolius and its associated therapeutic values are critically affected by the repetitious steaming times. The times-dependent steaming effect of P. quinquefolius is not well-characterized and there is also no official guideline on its times of steaming. In this paper, a UPLC-Q-TOF-MS/MS method was developed for the qualitative profiling of multi-parametric metabolic changes of raw P. quinquefolius during the repetitious steaming process. Our method was successful in discriminating the differentially multi-steamed herbs. Meantime, the repetitious steaming-inducing chemical transformations in the preparation of black American ginseng (American ginseng that was subjected to 9 cycles of steaming treatment) were evaluated by this UPLC-Q-TOF-MS/MS based chemical profiling method. Under the optimized UPLC-Q-TOF-MS/MS conditions, 29 major ginsenosides were unambiguously identified and/or tentatively assigned in both raw and multi-steamed P. quinquefolius within 19 min, among them 18 ginsenosides were detected to be newly generated during the preparatory process of black American ginseng. The mechanisms involved were further deduced to be hydrolysis, dehydration, decarboxylation and addition reactions of the original ginsenosides in raw P. quinquefolius through analyzing mimic 9 cycles of steaming extracts of 14 pure reference ginsenosides. Our novel steaming times-dependent metabolic profiling approach represents the paradigm shift in the global quality control of multi-steamed P. quinquefolius products.

Antioxidant activity and metabolic regulation of sodium salicylate on goat sperm at low temperature

Shen Wenzheng,Fu Yu,Bai Haiyu,Zhang Zhiyu,Cao Zhikun,Liu Zibo,Yang Chao,Sun Shixin,Wang Lei,Ren Chunhuan,Ling Ying-hui,Zhang Zi Jun,Cao Hongguo 아세아·태평양축산학회 2024 Animal Bioscience Vol.37 No.4

Objective: The purpose of this study was to explore the effect of sodium salicylate (SS) on semen preservation and metabolic regulation in goats. Methods: Under the condition of low temperature, SS was added to goat semen diluent to detect goat sperm motility, plasma membrane, acrosome, antioxidant capacity, mitochondrial membrane potential (MMP) and metabonomics. Results: The results show that at the 8th day of low-temperature storage, the sperm motility of the 20 μM SS group was 66.64%, and the integrity rates of the plasma membrane and acrosome were both above 60%, significantly higher than those of the other groups. The activities of catalase and superoxide dismutase in the sperm of the 20 μM SS group were significantly higher than those of the control group, the contents of reactive oxygen species and malondialdehyde were significantly lower than those in the control group, the MMP was significantly higher than that in the control group, and the contents of Ca2+ and total cholesterol were significantly higher than those in the control group. Through metabonomics analysis, there were significant metabolic differences between the control group and the 20 μM SS group. Twenty of the most significant metabolic markers were screened, mainly involving five metabolic pathways, of which nicotinic acid and nicotinamide metabolic pathways were the most significant. Conclusion: The results indicate that SS can effectively improve the low-temperature preservation quality of goat sperm.

High Infectivity of an Endoparasitic Fungus Strain, Esteya vermicola, against Nematodes

Chun Yan Wang,Zhe Ming Fang,Bai Shen Sun,Li Juan Gu,Ke Qin Zhang,성창근 한국미생물학회 2008 The journal of microbiology Vol.46 No.4

Esteya vermicola, as the first recorded endoparasitic fungus of pinewood nematodes, exhibits great potential as a biological agent against nematodes. However, only two strains of this species have been described so far. In this study, we identified a novel endoparasitic fungal strain, CNU 120806, isolated from infected nematodes in forest soil samples during a survey of nematophagous fungi in Korea. This strain showed similar morphological characteristics and infection mode with the two previously described strains of E. vermicola. All strains are characterized by the ability to produce two types of conidiogenous cells and conidia, and to parasitize nematodes with lunate adhesive conidia. Moreover, the CNU 120806 strain showed 100% identity with E. vermicola CBS 115803 when their partial sequences of 28S rRNA gene were compared. Molecular phylogenetic analysis further identified CNU 120806 as a strain of E. vermicola, by clustering CNU 120806 and E. vermicola CBS 115803 into a single subclade. Culture medium influenced the proportion of dimorphic CNU 120806 conidia, and further changed the adhesive and mortality rates of nematodes. The CNU 120806 strain exhibits high infection activity against nematodes on nutrient-rich PDA medium. Almost all tested nematodes were killed within 8~10 days after inoculation. This study provides justification for further research of E. vermicola, and the application and formulation of this fungus as a bio-control agent against nematodes.

Ginseng extracts ameliorates cognitive impairment and protects lipid peroxidation of brain induced by scopolamine in rats

Mi-Ra Lee,Beom-Sik Yun,Bai-Shen Sun,Lei Lui,Zheng Li,Chun-Yan Wang,Zhen Wang,Dong-Liang Zhang,Jing-Jie Li,Chang-Keun Sung(성창근) 고려인삼학회 2009 고려인삼학회 학술대회 Vol.2009 No.5

녹용 추출물이 치매 동물모델의 기억력 개선과 관련효소 활성에 미치는 효과

이미라(Mi-Ra Lee),손백신(Bai-Shen Sun),고리주안(Li-Juan Gu),왕춘년(Chun-Yan Wang),방철명(Zhe-Ming Fang),왕젠(Zhen Wang),모은경(Eun-Kyoung Mo),이선영(Sun-Young Ly),성창근(Chang-Keun Sung) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.4

본 연구는 녹용 추출물이 scopolamine으로 기억손상을 유발한 실험동물의 기억력 개선과 AChE, MAO-B의 활성억제, 신경전달 물질인 ACh 함량을 치매 치료제인 tacrine과 비교하여 분석하였다. 공간인지력을 평가하는 Morris water maze test에서 scopolamine(2 ㎎/㎏, i.p.)으로 기억손상을 유발한 경우 4일간의 인지훈련에도 불구하고 platform을 찾아가는 escape latency 시간이 증가되었으며, tacrine과 녹용추출물 투여군은 훈련 2일째부터 유의적으로 escape latency 시간이 감소하였다. 실험 5일째 platform을 제거한 probe test에서도 녹용 추출물 처리군은 scopolamine 처리군보다도 escape latency 시간이 유의적으로 감소하여 장기기억 개선효과가 있음을 보여주었다. 뇌의 신경전달물질인 ACh 생성은 대조군 대비 scopolamine 투여군은 84.7% 감소하였으나, 녹용 추출물 투여군(97.5%)은 치매 치료제인 tacrine(97.8%)과 비슷하게 정상적인 수치로 ACh 생성량을 증가시켰다. Tacrine 처리군은 scopolamine 투여로 상승한 뇌조직과 혈청의 AChE 활성을 유의적으로 감소시켰고, 녹용 추출물 처리군은 뇌조직의 AChE 활성을 유의적으로 감소시켰다. 뇌조직의 MAO-B 활성은 그룹 간 유의적인 차이는 없었으나, tacrine과 녹용 추출물 처리군이 scopolamine 투여군보다 MAO-B 활성을 감소시키는 경향을 보였다. 따라서 녹용 추출물은 scopolamine으로 기억손상을 유발한 치매 동물모델에서 신경전달물질을 분해하는 효소의 활성을 저해함으로써 장기기억 활성을 촉진하고 콜린성 신경계를 자극하여 기억 및 학습 증진에 효과적으로 작용하는 천연물 유래 기능성 물질로 사료된다. The aim of this study was to investigate the ameliorating effects of deer antler extract on the learning and memory impairments induced by the administration of scopolamine (2 ㎎/㎏, i.p.) in rats. Tacrine was used as a positive control agent for evaluating the cognition enhancing activity of deer antler extract in scopolamine- induced amnesia models. The results showed that the deer antler extract-treated group (200 ㎎/㎏, p.o.) and the tacrine-treated group (10 ㎎/㎏, p.o.) significantly ameliorated scopolamine-induced amnesia based on the Morris water maze test. Although there was no statistical significance of brain ACh contents among the experimental groups, the brain ACh contents of the deer antler extract-treated group was slightly higher than that of the scopolamine-treated group. The inhibitory effect of deer antler extract on the acetylcholinesterase activity in the brain was significantly lower than that of scopolamine-treated group. The tacrine- and the deer antler-treated groups reduced the MAO-B activity compared to the scopolamine-treated group, but not significantly. These results suggest that the deer antler extract could be an effective agent for the prevention of the cognitive impairment induced by cholinergic dysfunction.

백삼과 홍삼추출물의 학습수행과 Acetylcholinesterase 억제에 미치는 효과

이미라(Mi-Ra Lee),손백신(Bai-Shen Sun),고리주안(Li-Juan Gu),왕춘년(Chun-Yan Wang),모은경(Eun-Kyoung Mo),양선아(Sun-Ah Yang),이선영(Sun-Young Ly),성창근(Chang-Keun Sung) 고려인삼학회 2008 Journal of Ginseng Research Vol.32 No.4

백삼과 홍삼추출물 투여가 scopolamine으로 기억을 손상시킨 실험동물의 학습수행 개선과 뇌조직의 AChE 억제 및 ACh 함량을 평가하여 다음과 같은 결론을 얻었다. 1. 백삼과 홍삼추출물(200 mg/kg, p.o.) 투여는 scopolamine (2 mg/kg, i.p.)으로 유발된 기억손상 동물모델의 Morris water maze 시험에서 tacrine 투여군과 대조군처럼 인지훈련이 반복될수록 escape latency 시간이 현저히 감소하였다. probetest에서도 scopolamine 투여군보다 유의적으로 감소하여 인삼추출물이 장기기억(Long-term memory)과 working memory의 개선에 효과가 있음을 확인하였다. 2. Y-maze test를 통한 실험동물의 자발적 변동행동력은 실험군간 차이가 없었으며, 총 출입회수도 차이가 없어 인삼 추출물은 실험동물의 일반적 운동성에 영향을 주지 않았다. 3. 해마조직의 AChE 활성은 인삼추출물 군에서 감소하는 경향을 보였으나 유의적 차이는 없었다. 대뇌피질과 혈청 AChE 활성은 홍삼추출물 투여군에서 유의적으로 감소하였다. 4. 인삼추출물 투여군의 뇌조직 ACh 함량은 대뇌피질과 해마조직에서 scopolamine 투여군보다 증가하는 경향을 보였으나 그룹간의 유의적인 차이는 없었다. 5. 이를 통하여 볼 때 인삼추출물 특히 홍삼추출물은 장기기억 활성촉진, AChE 억제, ACh 활성촉진으로 콜린성 신경계를 자극하여 기억 및 학습증진에 매우 효과적으로 작용하는 항치매 물질로 사료된다. In the present study, we assessed the effects of white ginseng and red ginseng extract on the learning and memory impairments induced by scopolamine. The cognition-enhancing effect of ginseng extracts was investigated using the Morris water maze and Y-maze test. Drug-induced amnesia was induced by treating animals with scopolamine (2 mg/kg, i.p.), an antagonist of muscarinic acetylcholine (ACh) receptor. Tacrine was used a positive control. Ginseng extract (200 mg/kg, p.o.), tacrine (10 mg/kg, p.o.) administration significantly reduced the escape latency during training in the Morris water maze (p<0.05). At the probe trial session, scopolamine significantly increased the escape latency on day 5 in comparison with control (p<0.01). The effect of ginseng extracts on spontaneous alternation in Y-maze was similar to that of scopolamine treated group. In addition, numbers of arm entracts were similar in all experimental groups. Moreover, red ginseng extract significantly inhibited acetylcholinesterase activity in the cortex and serum (p<0.05). Brain ACh contents of ginseng extract treated groups increased more than that of scopolamine group, which did not show statistically significant. These results suggest that ginseng extract may be useful for the treatment of cognitive impairment.