Antioxidant and Antimelanogenic Effects of Stevia rebaudiana Flower Extract

소경섭,이성률,김성혁,하창우,박윤아,장소희,박종필,구현정,손은화 한국자원식물학회 2019 한국자원식물학회지 Vol.32 No.3

Stevia rebaudiana (Asteraceae), a perennial plant, has been used as a low-calorie sweetener and is being developed as a therapeutic agent for diabetes, hypertension, myocardial diseases, and microbial infections. Despite the common use of its leaves and stem, the bioavailability of the components present in S. rebaudiana flowers, when used as ingredients of cosmetics, has not been well investigated. Herein, we investigated the antioxidative and antimelanogenic effects of an aqueous extract of S. rebaudiana flowers (Stevia-F). Total flavonoid and phenolic content in Stevia-F were determined to be 8.64 ± 0.23 ㎎ of quercetin equivalents/100 g and 631.5 ± 2.01 ㎎ of gallic acid equivalents/100 g, respectively. The IC50 values of Stevia-F for reducing power, and 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical, hydrogen peroxide, and nitric oxide scavenging activities were 5541.96, 131.39, 466.34, and 10.44 ㎍/mL, respectively. Stevia-F showed inhibitory effects on the tyrosinase (IC50 = 134.74 ㎍/mL) and α-glucosidase (IC50 = 114.81 ㎍/mL) activities. No significant cytotoxicity of Stevia-F was observed in B16F10 cells, treated with up to 100 ㎍/mL of the extract for 24 and 48 h (p > 0.05). Stevia-F (1–100 ㎍/mL) suppressed α-melanocyte stimulating hormone-induced melanin production in B16F10 cells (p < 0.05) and also inhibited the cellular tyrosinase activity (p < 0.05). Overall, our results show that Stevia-F possesses potential for inhibiting tyrosinase and α-glucosidase activities and has significant antioxidant capacity. The antimelanogenic potential of Stevia-F should extend the usage of S. rebaudiana flowers in the development of skinwhitening products.

Improving productivity of steviol glycosides in  Stevia rebaudiana via induced polyploidy

Daryush Talei,Mojtaba Khayam Nekouei,Mohsen Mardi,Saeid Kadkhodaei 한국작물학회 2020 Journal of crop science and biotechnology Vol.23 No.4

The polyploidy induction using mutagenic chemicals is one of the breeding approaches to improve enhancing productivity of yield and secondary metabolites in medicinal plants. In the present research, to induce polyploidy in Stevia rebaudiana plants, the seeds were treated with four concentrations of colchicine (control, 0.05%, 0.1% and 0.2%) for 12, 24, 36 and 48 h in a factorial experiment based on randomized complete block design with three replicates. The morphological and phytochemical traits of plants were measured before the flowering stage. The results indicated that different concentrations of colchicine had a significant effect on the plant height, number of leaves, number of branches, mean of leaf length, stomata size and stomata density. The exposure times of colchicine also caused signifi cant changes in the plant height, number of branches, leaf width and length (P ≤ 0.01). Enhancement of colchicine concentration, significantly increased the above mentioned morphological traits, along with stomata size and rebaudioside-A, while significantly decreased stomata density in comparison to the control (P ≤ 0.01). In contrast, the interaction of colchicine concentration and exposure time had a significant effect on leaf length. The findings of this study indicated that estimation of physiological changes and secondary metabolites amount is one of the effective methods in primary screening of polyploid plants in polyploidisation breeding program and flow cytometry is recommended to be used for accurate identification of ploidy level in Stevia rebaudiana.

스테비아로부터 분리한 Austroinulin의 RAW264.7 세포에 대한 항염증 효과

변명우(Myung-Woo Byun) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.4

본 연구에서는 스테비아는 이용성이 높은 식물임에도 불구하고 diterpene 성분에 대한 생리활성이 과학적으로 주로 연구된 바 없어 active-guided fractionation 방법을 이용하여 항염증 효과를 나타내는 지표화합물을 규명하기 위해 실리카겔 칼럼크로마토그래피 방법을 이용하여 분리 및 정제한 후, <SUP>1</SUP>H와 <SUP>13</SUP>C-NMR, COSY, DEPT, HMQC, HMBC spectrum(500 MHz, CDCl3), MS, IR 분석을 통하여 분자량 322의 astroinulin임을 구조 동정하였다. 특히 HMBC spectrum을 통해 분리된 화합물이 decalin system에서 C-9에 3-me-thylpenta-2,4-dienyl 치환체를 가지고 있음을 알 수 있었다. 분리된 astroinulin을 RAW264.7 세포에 2.5, 5, 10 μg/mL 처리한 결과 농도 의존적으로 NO의 생성을 억제하였다. 마찬가지로 iNOS protein의 발현에서도 농도 의존적으로 저해하였으며, 화합물을 2.5, 5, 10 μg/mL 처리한 결과 각각 11.4, 26.8, 45.1% 감소하였다. Astroinulin 화합물을 10 μg/mL의 낮은 농도에서 NO와 iNOS를 각각 67.9, 45.1%를 저해하는 결과를 나타내었다. 이와 같은 결과는 스테비아에서 분리된 astroinulin 화합물은 RAW264.7 세포에서 LPS에 의해 유도된 NO와 iNOS 단백질을 유의적으로 억제하는 것으로 확인되어 이를 이용한 기능성식품 및 의약품 개발 가능성을 지닌 약용 식물자원인 것으로 판단된다. The leaves of Stevia rebaudiana are well-known in Japan, Korea, and China as a natural sweetener. Medicinal uses of this plant originated in Paraguay and Brazil in the form of aqueous decoctions of the leaves used as a contraceptive agent and for the treatment of hyperglycemia. In the present study, the antioxidant, anti-hyper-tension, and anti-inflammatory activities of S. rebaudiana extracts are investigated for their use in food. The biologically-active compound was isolated and purified from S. rebaudiana. The isolated compound was identified as austroinulin (C20H34O3; molecular weight 322) by mass, IR spectrophotometry, 1D, and 2D-NMR. Austroinulin was characterized as a diterpenoid possessing a 3-methylpenta-2,4-dienyl at C-9. When subjected to an inflammatory mediator inhibitory assay from lipopolysaccharide (LPS)-activated macrophages, the austroinulin inhibited the enhanced production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) expression (10 μg/mL=67.9 and 45.1%, respectively). This was significant and dose-dependent. The results suggest that austroinulin from S. rebaudiana inhibited the NO and iNOS in RAW 264.7 cells.

Optimization of supercritical fluid extraction of steviol glycosides and total phenolic content from <i>Stevia rebaudiana</i> (Bertoni) leaves using response surface methodology and artificial neural network modeling

Ameer, Kashif,Chun, Byung-Soo,Kwon, Joong-Ho Elsevier 2017 Industrial crops and products Vol.109 No.-

<P><B>Abstract</B></P> <P>Stevia leaves include natural, non-caloric sweetening compounds known as steviol glycosides (SGs)–mainly stevioside (ST) and rebaudioside-A (Reb-A). Along with sweetness, stevia generates interest because of its dietetic and therapeutic significance related to the presence of other bioactive compounds in its leaves, such as phenolics, which in association with SGs may contribute to human health by exerting anti-inflammatory, antihyperglycemic, anticarries, chemopreventive, insulinotropic and diuretic properties. In this study, response surface methodology (RSM) and artificial neural network (ANN) modeling were compared in terms of their estimation capabilities for building effective models with maximum response values. A supercritical fluid extraction (SFE) process was optimized by employing a 5-level-3-factor central composite design to achieve maximum target response values for total extract yield, ST yield, Reb-A yield and total phenolic content of 15.85%, 95.76mg/g, 62.95mg/g and 25.76mg GAE/g, respectively. The optimized SFE parameters included a modifier concentration of 40%, an extraction temperature of 45°C, and a pressure of 225bar. The ANN model proved an attractive alternative to RSM owing to its improved estimation and predictive capabilities. SFE yielded higher target response values than conventional maceration extraction (24h) and was a faster, lower energy, and greener extraction method with reduced CO<SUB>2</SUB> emissions and lower solvent consumption.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Stevia leaves consist of bioactive compounds with therapeutic benefits. </LI> <LI> Supercritical fluid extraction was optimized for target responses. </LI> <LI> Response surface methodology and artificial neural network modeling were compared. </LI> <LI> Artificial neural network modeling proved to be superior. </LI> <LI> Supercritical fluid extraction was more efficient with less resource consumption. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Stevia rebaudiana의 항산화 효과

정은혜(Eun Hye Jung),서혜림(Hye Lim Seo),김민규(Min Gyu Kim),김영우(Young Woo Kim),조일제(Il Je Cho) 한의병리학회 2013 동의생리병리학회지 Vol.27 No.6

스테비아(Stevia rebaudiana)잎 첨가 쌀쿠키의 품질 특성

김다솔 ( Dah-sol Kim ),신지훈 ( Jihun Shin ),주나미 ( Nami Joo ) 대한영양사협회 2017 대한영양사협회 학술지 Vol.23 No.1

The purpose of this study was to determine the optimal recipe of rice cookies with two different amounts of Stevia rebaudiana leaf and grape seed oil, using a central composite design (CCD). In addition, mixing conditions of rice cookies were optimized by sensory evaluation and mechanical and physicochemical analysis using response surface methodology (RSM). RSM was used to obtain 10 experimental points (including two replicates of Stevia rebaudiana leaf and Grape seed oil), and the formulation of Stevia rebaudiana leaf added rice cookies was optimized using rheology. The results of mechanical and physicochemical analysis showed significant values for lightness, redness, yellowness, hardness, spread factor, loss rate, leavening rate, sweetness, moisture, pH, and density (P<0.001), results of the sensory evaluation showed significant values for color, flavor, taste, texture, appearance, and overall quality (P<0.05). As a results, optimal sensory ratio was found to be 1.98 g of Stevia rebaudiana leaf and 37.94 g of Grape seed oil.

스테비아잎 분말을 첨가한 두유의 품질 특성

최순남 ( Soon Nam Choi ),주미경 ( Mi Kyoung Joo ),정남용 ( Nam Yong Chung ) 대한영양사협회 2014 대한영양사협회 학술지 Vol.20 No.2

두유에 첨가하는 당을 스테비아잎 분말로 대체하여 단맛을 유지하면서도 새로운 맛과 향을 부여한 스테비아잎 분말 첨가 두유를 제조하여 그 품질 및 관능 특성을 조사한 결과는 다음과 같다. 1. 외관을 관찰한 결과 대조군은 시판 두유 특유의 농후한 황색을 나타내는 반면, 첨가군은 스테비아잎 분말이 지니는 고유의 색으로 인해 녹색을 띠었으며, 스테비아잎 분말 첨가량이 증가할수록 녹색이 점차 진해졌다. 각 시료의 당도는 대조군13.0%, 스테비아잎 분말 첨가군은 12.1∼13.5%의 범위를 보였다. 제조 당일 pH는 대조군 7.88, 스테비아잎 분말 첨가군은 7.90∼7.98로(P<0.05), 스테비아잎 분말이 증가할수록 pH가 증가하였다. 대조군은 제조당일 pH 7.88, 4일 저장 시료는 pH 7.82로, 첨가군은 제조 당일 7.90∼7.98, 4일 시료7.83∼7.90으로 감소(P<0.05)하였다. 2. 제조 당일 대조군의 점도는 11.09 cp, 스테비아잎분말 첨가군은 11.13∼12.71 cp이었고 스테비아잎분말 첨가량이 증가할수록 점도가 증가하였다(P<0.05). 4일 시료는 대조군 10.53 cp, 첨가군은 10.91∼12.05 cp이었고 대조군에 비해 스테비아잎분말 첨가군의 점도가 더 높았다(P<0.05). 저장기간 중 점도의 변화는 대조군의 경우 제조 당일 11.09 cp, 4일째는 10.53으로 점차 감소하였다. 스테비아잎 분말 첨가군에서는 SP4군을 제외하고 감소하였으나 유의적이지 않았다. 3. 제조 당일 L값은 대조군 67.45, 스테비아잎 분말첨가군은 64.94에서 59.40으로 점차 감소하였으며 (P<0.05), 저장기간에 따라 모든 시료에서 L값이 감소하였다. a값은 제조 당일 대조군 -4.74, 스테비아잎 분말 첨가량이 증가할수록 -7.63∼-6.45로 유의적으로 감소하였다(P<0.05). 저장일수 증가에 따라 a값은 대조군의 경우 제조 당일 -4.74에서 -4.52로 증가하였고, 스테비아잎 분말 첨가군에서도 저장일수가 증가할수록 a값이 증가하였다. b값은 제조 당일 대조군 20.13, 스테비아잎 분말첨가군은 18.71∼19.63의 값을 보여 황색이 점차감소하였다. 4. 맛은 대조군 4.09, SP3군 4.10으로 다른 군에 비해 다소 높았으며 대조군과 유의적 차이를 보이지 않았다. 색은 대조군 3.87, SP3군이 3.94로 대조군과 비교하였을 때 유의적으로 높았다(P<0.05). 향은 SP3군 3.85로, 대조군 3.53 등 다른군에 비해 높았는데(P<0.05), 색, 맛 및 향에 대한 관능검사 결과로 보았을 때 SP3의 경우 비교적 좋은 관능평가 결과를 보였다. 선호도는 대조군 3.91, SP3군 4.01로 다른 시료보다 높았다(P<0.05). 따라서 맛, 향, 색 및 선호도를 종합적으로 평가하였을 때 SP3군이 스테비아잎 분말을 첨가한 두유로 좋은 평가를 보였다. 위의 결과를 볼 때 두유에 스테비아잎 분말을 0.06% 첨가한 SP3군의 경우 두유의 관능적 품질을 유지하면서 스테비아잎 특유의 맛과 향을 부여한 저칼로리 식품으로서 가능성이 있을 것으로 생각되었으며, 다양한 식품분야에서의 스테비아잎 분말의 활용도가 더 높아질 것으로 생각된다. 또한 본 연구에서 첨가한 스테비아잎 분말의 양이 비교적 적어 기능성이 크게 반영되지 못하였지만 그 특유의 맛에 익숙해진다면 첨가량을 증가시켜 기능성과 저열량 특성이 부여된 건강식품의 하나로 자리 잡을 수 있을 것으로 생각된다. This study was carried out to evaluate the effects of stevia (Stevia rebaudiana Bertoni) leaf powder on soybean milk quality. Soybean milk was prepared with the addition of 0.1 g (SP1), 0.2 g (SP2), 0.3 g (SP3), 0.4 g (SP4) of stevia leaf powder. The pH of soybean milk added with stevia leaf powders ranged from 7.90 to 7.98, whereas that of control was 7.88. The viscosity of soybean milk added with stevia leaf powder was 11.13¡­12.71 cp while that of control was 11.09 cp. Soymilk added with stevia leaf powder yielded values of Hunter L (59.40¡­64.94), a (￡­7.63¡­￡­6.45), and b (18.71¡­19.63), whereas that of control were 67.45, ￡­4.74 and 20.13, respectively. There was decrease upon addition of stevia leaf powder. During storage, the pH, viscosity, L, and b values were decreased continuously. According to sensory evaluation, taste, and flavor of SP3 were better than the other groups. Quality characteristics between control and SP3 showed no significant differences. Therefore, this study showed the possibility of substituting 0.3 g of stevia leaf powder instead of sugar for creating a standard quality soybean milk with low calories. Upon the results of this study, it may be assumed that there are consequences positive responses to health-oriented consumers when using stevia leaf powder.

연구논문 : 스테비아잎 분말을 첨가한 식빵의 품질특성

최순남 ( Soon Nam Choi ),주미경 ( Mi Kyoung Joo ),정남용 ( Nam Yong Chung ) 한국식품조리과학회(구 한국조리과학회) 2014 한국식품조리과학회지 Vol.30 No.4

This study was carried out to evaluate the effects of stevia leaf powder on bread quality. Different breads were prepared by the addition of 0%, 0.5%, 1.0%, 1.5% and 2.0% stevia leaf powder to wheat flour. The weight of the breads containing stevia leaf powder ranged from 525.0~543.5 g, while that of the control was 548.8 g. The volume of the breads prepared by adding stevia leaf powder was 2225.0~2278.8 mL, while that of the control was 2307.5 mL. The height and specific volume of the breads decreased with the addition of stevia leaf powder. The Hunter L values of the breads also decreased, whereas a and b values increased with the addition of stevia leaf powder. The L and b values of the bread decreased continuously during storage. The hardness, springiness, cohesiveness and chewiness all increased with the addition of stevia leaf powder. Overall the acceptability and taste determined by sensory evaluation of the bread with 1.5% stevia leaf powder were better than that of the control.

Genotypic characterization and steviol glycoside quantification in a population of  Stevia rebaudiana Bertoni from Paraguay

Liz Bogado-Villalba,Héctor Nakayama Nakashima,Rosanna Britos,Julio Cesar Masaru Iehisa,María Eugenia Flores Giubi 한국작물학회 2021 Journal of crop science and biotechnology Vol.24 No.2

Stevia rebaudiana (Bertoni) Bertoni is a widely grown species in various regions of the world, mainly due to its sweetening properties attributed to steviol glycosides. Despite the fact that Paraguay is the center of origin of S. rebaudiana , genetic characterization of local genotypes has not been studied. Considering that accumulation of steviol glycosides is a complex trait, molecular breeding could be an effective method for its improvement. Therefore, the identification of molecular markers associated with steviol glycoside accumulation is important. The aim of this study was to assess the genetic relationship among eight advanced lines of S. rebaudiana and two varieties selected from local germoplasm and quantify their steviol glycoside content. The stevioside and rebaudioside A content in methanolic extract was determined by high-performance liquid chromatography (HPLC), and the molecular characterization of the ten genotypes of S. rebaudiana was performed using SSR and ISSR markers. A wide range of variability was observed for the metabolites of interest under greenhouse conditions. The variety Eirete presented highest stevioside and rebausiode A content, followed by the variety Katupyry. One of the advanced line presented both metabolite content similar to that of Katupyry. Genotyping of these lines showed two main clusters. Cluster I contained the both varieties and advanced lines with higher stevioside and rebaudioside A content. Our results suggest that S. rebaudiana lines with genetic profi le similar to that of cluster I, determined mainly by genotype at the markers SUGMS 35 and ISSR 47, might present higher stevioside and rebaudioside A content.

NF-κB와 MAPK 억제를 통한 스테비아 잎의 항염증효과

김선영,조미정,황보민,백영두,정태영,조일제,지선영,Kim, Seon-Young,Jo, Mi-Jeong,Hwangbo, Min,Back, Young-Doo,Jeong, Tae-Young,Cho, Il-Je,Jee, Seon-Young 대한한방안이비인후피부과학회 2013 한방안이비인후피부과학회지 Vol.26 No.3

Objectives : Stevia rebaudiana is a well-known herbal sweetener in the Korea, Japan and China, and its medical uses were originated from countries in South America. Although it has been shown the various medical effects of S. rebaudiana including contraception and treatment of human diseases such as hyperglycemia, it has almost not been studied about the efficacy of S. rebaudiana methanolic extract (SRE) on the acute inflammation and its action mechanism. Methods : To investigate the anti-inflammatory effects of SRE, we treated SRE and examined the level of inflammatory mediators in LPS-stimulated Raw264.7 cells. Results : Treatment of macrophage with LPS markedly induced the production of NO, $PGE_2$ and pro-inflammatory cytokines. Pretreatment of SRE blocked the induction of inflammatory mediators and the expression of iNOS protein. More importantly, LPS-induced phosphorylation of $I{\kappa}B-{\alpha}$ was suppressed by the treatment of SRE, suggesting SRE inhibition of NF-${\kappa}B$ activation. Furthermore, SRE blocked LPS-induced phosphorylation of MAPKs. Conclusions : SRE inhibited the induction of NO, PGE2 and pro-inflammatory cytokines in Raw264.7 cells. SRE's effect may be mediated with its inhibition of NF-${\kappa}B$ activation and MAPK phosphorylation, which suggests its uses as an anti-inflammatory agents.