Product Optimization, Storage Quality and Sensory Acceptance of Low Calorie Beverage Developed from Bitter Gourd and Kiwifruit

Abstract Bitter gourd (Momordica charantia L.) fruit is good source of many nutraceutical compounds and possess antioxidant, anti-diabetic and hypoglycaemic activities. However, its utilization in the preparation of beverages is limited due to its bitter after taste. Therefore, to realize the functional and therapeutic benefits of bitter gourd, an attempt was made to optimize nutritious and low caloriebitter gourd based beverage by blending with kiwifruit (Actinidia deliciosa), a store house of bioactive compounds and substituting sugar with stevioside (steviol glycoside). The standard (sugar sweetened) bitter gourd (BG)-kiwifruit (K) blended beverage was developed by utilizing 30% fruit part of BG:K blended juice (80: 20) with 40oB TSS and 1.3% acidity. Further, to develop the low calorie beverage, sucrose (table sugar) was replaced with 25, 50, 75 and 100% equi-sweetness level of stevioside (steviol glycoside). Results revealed that 75% substitution of sucrose with stevioside resulted in shelf stable beverage with identical taste, good antioxidant potential (68.80%) and strong antimicrobial activity (26 mm ZOI) with reduced calorie values (28.5 Kcal/100g) compared to the sugar sweetened control sample (150.60 Kcal/100g). Hence, the developed beverage can be commercialized as low calorie beverage with additional health benefits of natural compounds of bitter gourd and kiwifruit with highest bioactivity.

Turbo-extraction of glycosides from Stevia rebaudiana using a fractional factorial design

ABSTRACT Stevia rebaudiana (Bertoni) Bertoni, Asteraceae, leaf extract has recently called the attention of food industry as a proposal for natural sweetener. The sweet flavor is attributed to the glycosides, in especial stevioside and rebaudioside A, which are the plant main chemical markers. The aim of the work reported here was to optimize the turbo-extraction of stevia leaves using water, ethanol 70% and 90% (w/w) as green solvents. A 25-2 factorial design was applied to study the linear effects of the drug size, solvent to drug ratio, temperature, time and also the turbolysis speed on the extraction of glycosides. The glycosides exhaustive extraction showed that ethanol 70% gave better results and was used for turbo-extraction. The stevioside and rebaudioside A contents were quantified by a validated method by high performance liquid chromatographic with photodiode array detector. The contents of stevioside and rebaudioside A in fluid extract increased with the drug size, but decreased at high shearing speeds and solvent to drug ratio, while their yields decreased at higher temperature and were not affected by turbo speed. An increase in solvent to drug ratio reduced significantly the glycosides percent in dried extract. Optimal solution for S. rebaudiana leaves turbo-extraction was determined by desirability functions. The optimal extraction condition corresponded to drug size of 780 µm, solvent to drug ratio of 10, extraction time of 18 min; temperature of 23 ºC and turbo speed of 20,000 rpm, resulting in yields of 4.98% and 2.70%, for stevioside and rebaudioside A, respectively. These yields are comparable to the ones recently published for dynamic maceration, but with the advantage of shorter extraction times. This work demonstrates that turbolysis is promising for S. rebaudiana glycosides extraction and stimulate new research on the purification of these extracts, which may become an interesting source of income for developing countries such as India and Brazil.

Micropropagaión de Stevia rebaudiana Bertoni, un endulzante natural a través de explantes con meristemos pre existentes / Micropropagation of Stevia rebaudiana Bertoni, a natural sweetener, through pre existing meristem explants

Las hojas de Stevia rebaudiana son fuente de esteviosidos y rebaudiosidos, sustancias endulzantes con bajo contenido calórico. La propagación sexual y clonal de estevia es difícil debido a la calidad de la semilla y el tamaño reducido de la planta. Para evaluar la multiplicación, brotes establecidos in vitro fueron cultivados en ½ MS con cinco concentraciones de BAP (0.0, 2.22, 4.44, 8.88 y 17.6 µM). Posteriormente, los tallos multiplicados se subcultivaron en presencia de cinco concentraciones de ANA (0.0, 2.69, 5.37, 10.74 de 21.48 µM) para evaluar enraizamiento. Finalmente, tallos multiplicados sin enraizar, tratados o no con 0.4% de ANA, y otros enraizados in vitro fueron transferidos a condiciones ex vitro. Todos los experimentos fueron distribuidos usando un DCA. Los resultados indicaron que el medio 1/2MS adicionado con BAP indujo una mayor tasa de multiplicación. 10.74 µM de ANA indujo el mejor enraizamiento; sin embargo, los tallos sin enraizamiento resultaron en la mayor supervivencia ex vitro.

Stevia rebaudiana Bertoni leaves are source of stevioside and rebaudioside, non-caloric sweetener substances. Seed and cutting estevia propagation is difficult due to seed sterility and small size plant, respectively. To evaluate shoot proliferation, in vitro-established estevia shoots were cultured in ½ MS with five (0.0, 2.22, 4.44, 8.88 and 17.6 µM) BAP levels. Thereafter, proliferated shoots were cultured on ½ MS with five NAA levels (0.0, 2.69, 5.37, 10.74 and 21.48 µM) to evaluate shoot rooting. Finally, non-rooted shoots, in vitro-rooted shoots and non-rooted shoots treated with a 0.4% NAA powder were transferred to ex vitro conditions. All experiments were distributed using a complete randomized design. The data indicated that BAP treated shoots showed a higher rate of shoot proliferation. An 87% of rooting and higher number of roots per explant was achieved with 10.74 µM of NAA. Non-rooted shoots transferred directly from Stage II showed the best survival rate.

Acid and Alkaline Hydrolysis Studies of Stevioside and Rebaudioside A.

Hydrolysis studies of the two diterpene glycosides stevioside and rebaudioside A that were isolated from Stevia rebaudiana were performed using acid and alkaline conditions. Acid hydrolysis was carried out using H2SO4 whereas alkaline hydrolysis was performed using NaOH. Using these methods, partial hydrolyzed glycosides of stevioside and rebaudioside A and their sugar residues were identified from the alkaline and acid hydrolysis studies respectively. The structures of the acid and alkaline hydrolysis products were achieved on the basis of extensive NMR and high resolution mass spectral data, and in comparison with the data reported in the literature as well as TLC comparison with their corresponding standard compounds.

Evaluación del contenido de esteviósido y rebaudiósido A en una población de Stevia rebaudiana Bertoni (kaâ heê) cultivada comercialmente: estudio preliminar / Evaluation of the content of stevioside and rebaudioside A in a population of Stevia rebaudiana Bertoni (kaâ heê) commercially cultivated: a preliminary study

Stevia rebaudiana Bertoni (kaâ heê) es un arbusto originario de la Región Oriental de Paraguay. Sus hojas contienen glucósidos diterpénicos que producen un sabor dulce, perosin valor calórico y son de 150 a 300 veces más dulces que la sacarosa (solución al 0,4%). Durante las temporadas agrícolas 1998 y 1999 se establecieron sembrados deselección individual, en el Instituto Agronómico Nacional (Caacupé, Paraguay). El objetivo de este trabajo fue determinar el contenido de esteviósido y rebaudiósido A en plantasseleccionadas en campo a fin de evaluar la variabilidad existente en la población e identificar clones con alto contenido de dichos glucósidos. De una población original de 2000 individuos, se seleccionaron fenotípicamente 140 plantas por características agronómicas superiores y se cosecharon en etapa de prefloración y libres de polinización.En los Laboratorios del Instituto Nacional de Tecnología Normalización y Metrología (INTN) fueron evaluados los contenidos de esteviósido y rebaudiósido A por detección por cromatografía liquida de alta resolución. El contenido de esteviósido fluctuó entre 0 y 21 % concentrándose el 92 % de la población entre valores del 5 al 15 % .En cuanto al contenido de rebaudiósido A se obtuvieron valores entre 0 y 12 % correspondiendo al 68% de la población un contenido entre 3 y 9 %. La demanda actual de stevia se halla en creciente aumento, por lo que las acciones orientadas a establecer líneas clonales con alto contenido de principios edulcorantes permitirá a los productores primarios la obtención decultivos más productivos e incrementara el rendimiento de obtención de glucósidos.

Stevia rebaudiana Bertoni (kaâ heê) is a bush native to the Eastern Region of Paraguay. Its leaves contain diterpene glycosides that produce a sweet taste but without any caloric value and are 150 to 300 times sweeter than sacharose (0, 4% solution). During 1998 and 1999 agricultural seasons, individual selection sown fields were established in the National Agronomical Institute (Caacupé, Paraguay). The objective of this work was todetermine the content of stevioside and rebaudioside A in these selected plants in the field in order to evaluate the variability existent in the population and identify clones with high content of those glycosides. Of an original population of 2,000 specimens, 140 plants were selected phenotipically by the superior agronomical characteristics and they were harvested in the period of pre-flowering and were pollination free. The content of stevioside and rebaudioside A were evaluated in the laboratories of the National Instituteof Technology, Standardization and Metrology (INTN in Spanish) by high resolution liquid chromatography. The content of stevioside ranged from 0 to 21% concentrating the 92% of the population between 5 to 15%. In relation to rebaudioside A, values between 0 and 12% were obtained being 68% of the population between a content of 3 to 9%. As the current demand of Stevia is increasingly rising, the actions oriented to establish clonallines with high content of sweetener principles will allow primary producers to obtain more productive cultivations and increase the output of glycoside obtainment.

Synthesis and antitumor activity of ent-kaurene diterpenoids / 中国药科大学学报

Aim: To synthesize the ent-kaurene diterpenoid and its derivatives from natural available stevioside.Methods: The crucial pharmacophore of exo-methylene ketone was designed and the carboxyl group at C-4 was modified.The cytotoxic inhibition activities in vitro of target compounds were evaluated against human cancer cells BEL-7402,HO-8910,MCF-7 and HL-60 by a MTT method.Results and Conclusion: The struc-tures of new target compounds were identified by ~1H NMR,FT-IR and EI-MS.The preliminary experimental re-sults showed that some derivatives of ent-kaurene diterpenoid possessed fair inhibitory activity against tumor cell lines compared with that of their parent steviol.

Novel agents for enzymatic and fungal hydrolysis of stevioside / Novos agentes para a hidrólise enzimática e fúngica do esteviosídeo

A comparative study on the potential of some biological agents to perform the hydrolysis of stevioside was carried out, aiming at establishing an alternative methodology to achieve the aglycon steviol or its rearranged derivative isosteviol, in high yields to be used in the preparation of novel bioactive compounds. Hydrolysis reactions were performed by using filamentous fungi (Aspergillus niger, Rhizopus stolonifer and Rhizopus arrhizus), a yeast (Saccharomyces cerevisiae) and enzymes (pancreatin and lipases PL250 and VFL 8000). Pancreatin showed the best hydrolytic activity, furnishing isosteviol at 93.9% of yield, at pH 4.0, using toluene as a co-solvent. Steviol was produced using both pancreatin at pH 7.0 (20.2% yield) and A. niger atpH 7 (20.8% yield).

Um estudo comparativo do potencial de alguns agentes biológicos capazes de hidrolisar o esteviosídeo foi realizado,objetivando-se estabelecer uma metodologia alternativa para a obtenção da aglicona esteviol ou seu produto de rearranjo, isoesteviol, em rendimentos elevados que permitam o uso destas agliconas para o preparo de novos compostos bioativos. As reações de hidrólise foram realizadas usando fungosfilamentosos (Aspergillus niger, Rhizopus stolonifer e Rhizopus arrhizus), uma levedura (Saccharomyces cerevisiae) e enzimas(pancreatina, lipase PL250 e lipase VFL 8000). A pancreatina mostrou a melhor atividade hidrolítica dentre os sistemastestados, fornecendo isoesteviol com rendimento de 93,9% em pH 4,0, usando tolueno como co-solvente. Esteviol foi produzido tanto usando pancreatina em pH 7,0 (20,2% derendimento) quanto usando A. niger em pH 7,0 (20,8% de rendimento).