Tissue Culture and Rapid Propagation System of Roselle(Hibiscus sabdariffa L.)

[Objectives]This study was conducted to establish a tissue culture and rapid propagation system of roselle(Hibiscus sabdariffa L.)[Methods]A tissue culture and rapid propagation experiment was carried out with roselle plants as materials to study the suitable explants,the best disinfection method,the best growth medium,the best rooting medium and the transplanting and domestication method of roselle.[Results]The lateral buds of roselle were the best explants.Sterilizing with 0.1%mercuric chloride for 7 min showed a contamination rate of 5%and achieved a survival rate of 90%.With MS as the basic medium,adding 1.0 mg/L 6-BA and 0.5 mg/L IBA could obtain the best effect of bud induction.The medium with the highest proliferation rate was MS+6-BA 0.5 mg/L+BA 0.1 mg/L.On the basis of 1/2MS,adding 0.5 mg/L NAA+0.5 mg/L IBA could make adventitious buds root fastest and most,and greatly improve the propagation coefficient.And 1∶1 perlite rock:peat soil was the best transplanting substrate,with which the transplanting survival rate reached 95%.[Conclusions]This study provides technical reference for rapid cultivation and large-scale planting of roselle.

Comparative Effect of Foliar Application of Cow Dung, Wood Ash and Benlate on the Disease Initiation and Development of Roselle (Hibiscus sabdariffa L.) Leaf Spot Disease Caused by Coniella musaiensis Var. Hibisci. in Makurdi, Central Nigeria

A study on the comparative effect of cow dung, wood ash and benlate for the control of leaf spot disease of Roselle (Hibiscus sabdariffa L.) was carried out following the preparation and foliar application of the materials on selected Roselle plants of the green and red accessions (Acc1 & Acc3) in the field. The cow dung was fermented for 14 days while sour milk was added to wood ash, dissolved and all filtered and labeled. The spraying commenced soon after the plants were thinned to one and was done weekly until after 50% of the plants had flowered. Results showed significant (p = 0.05) difference between benlate, the synthetic fungicide and the bio-fertilizers (cow dung and wood ash) for the two years of the study. Disease severity was significantly lower in the benlate than in the bio-fertilizers. There was no significant difference in yield between the treatments although higher yield was recorded in benlate. Though the bio-fertilizers could not suppress infection, they aided the growth of the plants by boosting luxuriant growth. The result showed that these bio-fertilizers cannot be used as substitute to the synthetic fungicide for the control of this leaf spot disease caused by Coniella musaiensis var. hibisci. However, since they give equally good yield the bio-fertilizers can still be used to minimize the reliance upon use of chemical fungicides.

Production of Cashew Apple Wine Enriched with Hibiscus sabdariffa Extracts

This study focuses on enhancing the value of agricultural products by developing a process to produce wine from cashew apples enriched with extracts from Hibiscus sabdariffa. The formulation consisted of a blend of cashew apple juice and Hibiscus calyxes in a ratio of 90:10 (w/w). The Hibiscus calyxes were added at three different stages: before, during, and after fermentation. The physico-chemical analysis of the resulting wines revealed a pH range of 3.073 ± 0.005 to 3.583 ± 0.015 and acidity levels ranging from 4.018 ± 0.028 to 5.628 ± 0.059 g/L. The alcoholic strength ranged from 13.54% ± 0.036% to 13.86% ± 0.04%, with a soluble dry extract of 7.2 ± 0.25 to 8.1 ± 0.28 °B. Regardless of the stage of Hibiscus calyx addition, the fermentation kinetics and physico-chemical parameters met the standards set for wines.

Comparative Effect of Adding Synthetic Citric Acid to Natural Lemon Juice (Citrus aurantiifolia) on the Stability of Hibiscus Drinks Stored at 4°C and 37°C

The general objective of the work is to compare the effect of the addition of synthetic citric acid compared to the addition of natural fruit juice of Citrus aurantiifolia on the conservation of drink based on red chalice H. sabdariffa. The tests were carried out over a period of 5 weeks at 4°C and 37°C with seven batches of beverage samples prepared at the rate of a calyx/water ratio of 1/40 (kg·kg-1) and added respectively citric acid at 1, 2 and 4 g·L-1 and lemon juice at 10, 20 and 40 mL·L-1. The characterization of the different batches of beverages was carried out from day one. A follow-up of the residual anthocyanin content and the intensity of the red coloring were carried out over five weeks. The anthocyanin concentration was determined by the pH-differential method. The red color degradation index is determined based on the CIELAB color system (L*, a*, b* and L*). R and Minitab 18 software was used for data processing. The results of the monitoring of the parameters showed that the concentration of 1 g AC L-1 retains 2.7 mg more of the anthocyanins than adding 10 mL JC L-1 and longer maintains red color when stored at 4°C and 37°C/5 weeks. The 2 g AC L-1 and 4 g AC L-1 ratios have the same effects as the addition of 20 and 40 mL of lemon juice, all accelerating the degradation of anthocyanins and the red color. After 5 weeks of storage at 37°C, the effect of the temperature combined with the increase in the acidity of the samples (2 to 4 g AC L-1 and 20 to 40 ml JC L-1), have accelerated the total disappearance of the red color on all samples.

Effects of Hibiscus sabdariffa Linn. fruit extracts on α-glucosidase enzyme,glucose diffusion and wound healing activities

Objective:To provide in vitro evidence for antidiabetic activity through potential inhibition of α-glucosidase enzyme,glucose diffusion and enhancement in the wound healing using methanolic extract and fractions from Hibiscus sabdariffa Linn.fruit.Methods:The inhibitory action of methanolic extract and fractions of such fruit on aglucosidase enzyme and glucose movement through in vitro assay assessment was reported.Their activities on wound healing were tested using the scratch assay.Results:Ethyl acetate fraction at 50 mg/m L concentration exhibited significant aglucosidase inhibition(95.79 mg/m L) with P < 0.05.At the same concentration,the methanolic extract as well as other fractions revealed lower α-glucosidase inhibition and higher glucose diffusion retardation across the dialysis tube than the control.Ethyl acetate and butanol fractions displayed notably higher glucose diffusion inhibitory activity of 5.21 mmol/L and 5.2 mmol/L,respectively as compared to methanolic extract and n-hexane fraction of 6.58 mmol/L and 6.49 mmol/L,respectively.Conversely,compared to other fractions the methanolic extract and ethyl acetate fraction manifested proliferative effect at the incubation time of 6 h during the wound healing study.Conclusions:It is established that methanolic extract and fractions from H.sabdariffa Linn.fruit can inhibit the α-glucosidase enzyme and glucose movement as well as influence the wound healing activity positively.

Study on Quality Standard of Hibiscus trionum L.

[Objectives]To establish the quality standard for Hibiscus trionum L.[Methods]The moisture,total ash,acid insoluble ash and extract of H.trionum L.were determined by the method of Chinese Pharmacopoeia 2020 edition.The medicinal materials were qualitatively identified by microscopic identification and TLC,and the content of quercetin was determined by HPLC.[Results]The moisture,total ash,acid insoluble ash and extract contents of the 12 batches of samples were 7.69%-12.94%,10.44%-14.62%,1.89%-3.64%,16.56%-21.81%,microscopic characteristic was obvious,and a method for TLC and content determination of H.trionum L.was established.[Conclusions]A perfect quality standard was established,which can be used for the quality control of H.trionum L.

Selective Cytotoxicity and Apoptogenic Activity of Hibiscus Sabdariffa Aqueous Extract Against MCF-7 Human Breast Cancer Cell Line

Background: Herbal compounds are attractive anticancer candidates due to their low toxicity. Pervious studies have demonstrated that Hibiscus sabdariffa is promising as an anticancer agent against several cancer types;however, its potential therapeutic role in breast cancer remains to be investigated. Materials and Methods: In the present study, the cytotoxic effects of Hibiscus sabdariffa aqueous extract (HSE) on a human breast adenocarcinoma cell line (MCF-7) and fetal foreskin fibroblast (HFFF) were investigated. Different concentrations of water extract of calyces were added and the percentage of cell survival was determined after 24, 48, and 72 hours using an MTT assay. Apoptosis induction was assessed by DNA fragmentation. Results: At the concentration of 0.5 mg/ml of the extract and following 72 hours of incubation, the number of viable MCF-7 cells was less than 50%. The extract was not cytotoxic against normal HFFF cells in all tested concentrations. Also, HSE induced apoptosis only in MCF-7 cells. Conclusions: These results suggest that HSE inhibits the growth of MCF-7 cells selectively in a concentration- and time-dependent manner. As this herbal substance has been shown to be nontoxic at very high doses in experimental animals, it might be a good anticancer drug candidate for breast cancer treatment.

Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections

Objective:To explore the prevention of rerurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa(H.sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H.sabdariffa extract.Methods:In this particular study,six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used.The susceptibility of fungal isolates,time-kill curves and biofilm forming capacity in the present of the H.sabdariffa extract were determined.Results:Various levels minimum inhibitory concentration of the extract were observed against all the isolates.Minimum inhibitory concentration values ranged from 0.5 to 2.0 mg/mL Timekill experiment demonstrated that the effect was fungistatic.The biofilm inhibition assay results showed that H.sabdariffa extract inhibited biofilm production of all the isolates.Conclusions:The results of the study support the potential effect of H.sabdariffa extract for preventing recurrent candiduria and emphasize the significance of the plant extract approach as a potential antifungal agent.

Mechanism of the decreased food consumption and weight gain in rats following consumption of aqueous extract of the calyx of Hibiscus sabdariffa during pregnancy

Objective:To investigate the possible mechanisms of the decreasing fluid and food consumption following Hibiscus sabdariffa(HS) consumption.Methods:On the 1st day of pregnancy,rats were randomly divided into three groups with six animals per each group.One group was given tap water,one was given with extract at 0.6 g/100 mL while the third group was given with extract at 1.8 g/100 mL as their drinking solution.All groups received normal rat chow and drinking solution ad libitum.Fluid& food intake and weight were measured daily throughout pregnancy and Na^+ concentration in plasma was determined on the 18th day of pregnancy.Results:Results showed decreased fluid and food consumption,decreased weight gain and increased sodium ion concentration in plasma of rats with HS extract compared with the control group.Conclusions: Consumption of aqueous extract of the calyx of HS during pregnancy decreases food consumption and weight gain through mechanisms that may depend on Na^+ in HS content and elevating Na^+ concentration.

In Vitro Studies: Potensial Use of Dyospiros kaky, Avium grafeolens, Guazuma ulmifolia, Hibiscus sabdariffa as an Agent of Anti-cholesterol

God has made man and has prepared everything for human life.This universe was created for humans to be healthy.The lifestyle of humans in the millenial era has made circulatory disorders in the form of hypercholesterolemia.This study aims to determine the efficacy of herbal plants to reduce blood cholesterol levels.The method used is the in vitro method.Herbal plant used in this experiment Dyospiros kaky 50%,Avium grafeolens 20%,Guazuma ulmifolia 20%,Hibiscus sabdariffa 10%.This study showed that cholesterol level decreased from 200 mg/dL to 102 mg/100 g at dose 20 mg/dL,64.5 mg/100 g,at dose 40 mg/dL,and 94.2 mg/100 g,at dose 60 mg/dL.The best dose to decrease blood cholesterol was 40 mg/dL.This study indicated that Dyospiros kaki is an agent of anti-cholesterol.

Clinical, Hematological and Biochemical Health Benefit Effects of Hibiscus sabdariffa Lin Dried Calyces Beverage in Human

Background/Objective: This study was carried out to investigate the health benefit effects of Hibiscus sabdariffa Lin (H. sabdariffa L.) dried calyces beverage on some clinical, biochemical and hematological parameters in humans. Methods: The dried calyces were harvested in the two regions (Adamaoua and West) of Cameroon. The proximate, mineral composition and phytochemical screening were evaluated. A standardized extraction procedure was set up and from the calyces;we prepared a drink for 32 male volunteers’ subjects aged from 21 to 32 years, specially recruited for the experiment. Each participant consumed 500 mL twice a day (in the morning and in the evening) as supplement beverage during two weeks. The anthropometrics (age, height, weight, body mass index (BMI)), clinical (systolic and diastolic blood pressure), hematological (RBC, Hb, PCV, MCV, MCH, MCHC, WBC, Lymphocytes, MID cells, Granulocytes, platelet and MPV) and biochemical (TC, HDL-C, LDL-C, TG, serum iron, blood glucose, creatinine, urea, ASAT and ALAT) parameters were determined in the blood on days 0 and at the end of each week. Results: Crude protein, lipid, fiber and ash content of calyx ranged respectively from 4.57 - 5.98, 10.10 - 11.33, 20.39 - 22.30 and 9.15% - 10.38% while the levels of minerals were ranged from 512.0 - 740.6, 77.8 - 177.7, 52.84 - 52.85, 1.10 - 2.10, 41.2 - 119.5, 3.25 - 8.20 and 0.56 - 17.5 mg/100g respectively for Ca, Mg, K, Na, P, Fe and Zn. The phytochemical screening tests revealed the presence of alkaloids, flavonoids, tannins, phenols and anthocyanins on methanol and aqueous extracts. A significant increase of RBC, Hb, PCV, MPV, HDL-C, TG and creatinine and a significant decrease of WBC, MID cells, LDL-C and TC (p < 0.05) were observed during the study period. Furthermore, there was no significant change on BMI, MCV, MCH, MCHC, lymphocyte, granulocyte, platelet, serum iron, blood glucose, ASAT, ALAT and urea levels. Conclusion: H. sabdariffa L. dried calyces from Cameroon are rich sources of crude fibers and minerals. The H. Sabdariffa L. dried calyces drink can be safely used for people suffering for anemia. It also revealed good cholesterol lowering potential. No hepatoxicity and no kidney damage have been observed as far as serum enzymes were concerned.

Influence of Polyphenolic Contents on the Antioxidant Properties of Hibiscus sabdariffa Extract (HSE), Aged Garlic Extract (AGE) and Garlic Tablet (GT) in Vitro

This study compared the antioxidant properties of Hibiscus sabdariffa extract (HSE), commercial aged garlic extract (AGE, Kyolic) and garlic tablet (GT). The results indicate that HSE, AGE and GT are effective antioxidant as xanthine/xanthine oxidase generated superoxide ions are significantly inhibited in the presence of 20% (v/v) diethyl ether extract of HSE, AGE and GT by 100%, 66.39% and 18.61% respectively, while uric acid production is not affected byAGEand GT. However, at 2.5% and 5.0% (v/v) reaction volumes,HSEsignificantly inhibits uric acid production by 9.5% and 33.0% respectively, and this suggests that components ofHSEinhibit xanthine oxidase activity while AGE and GT scavenge superoxide. In addition, it is also found that the three extracts scavenge ABTS radical cations in dose-dependent manner. In all cell free assays, HSE is found to be more effective as an antioxidant when compared with AGE and GT under the same experimental conditions. Chromatographic and colorimetric analyses suggest thatHSEhas numerous different types of phenolic compounds with higher amount of phenolic compounds (14.9 mg/g) when compared withAGE(2.8 mg/g) and GT (3.6 mg/g) catechin equivalent respectively. Therefore, the effectiveness of antioxidant activities of these three extracts may be related to their phenolic content.

Effect of Different Sowing Dates on Disease Initiation and Development of Roselle (<i>Hibiscus sabdariffa</i>L.) Leaf Spot Disease Caused by <i>Coniella musaiensis</i>Var. <i>hibisci</i>in Makurdi, Central Nigeria

The experiment was conducted mainly to investigate the effect of change in sowing dates on disease severity of Roselle (Hibiscus sabdariffa L) leaf spot caused by Coniella musaiensis Var hibisci, plant height, number of branches, number of fruits and calyx yield of Roselle (H. sabdariffa L). Five dates were selected with an interval of fourteen days (14) between the dates for two years (2003 and 2004) to determine the most appropriate time to sow the crop using two Roselle accessions;green (Acc1) and red (Acc3).The first date was determined by the establishment of the rains in the season. It was observed that change in sowing dates had significant (p = 0.05) effect on disease severity but not on plant height, number of branches, number of fruits and calyx yield. Plants sown on July 18/27 had lowest disease indices;and are thus the best and therefore recommended for cropping of Roselle.

Physicochemical and Microbiological Properties of Bissap (Hibiscus sabdariffa) and Ginger (Zingiber officinale (L) Rose) Juices Sold in the Western Part of Abidjan (Côte d’Ivoire)

Juices made from the calyx of Hibiscus sabdariffa (Malvaceae), commonly called Bissap, and the rhizome of Zingiber officinale (Zingiberaceae), known as Gnamankoudji, are widely consumed in C&ocirc;te d'Ivoire. The artisanal preparation of these juices often makes their quality problematic. The aim of this study is to determine some physicochemical and microbiological properties of these drinks sold in the city of Abidjan. To do this, samples of Bissap and Gnamankoudji juices were collected near schools in the Mamie Adjoua (Yopougon) and Abobodoumé (Attécoubé) districts. Physico-chemical and microbiological analyses were performed on these collected samples. The results show that the Bissap and Gnamankoudji juices have an acid pH (2.47 and 3.71), a low protein content (0.78% and 2.11%) and a high water content (80.13% and 85.21%), respectively. The reducing sugar content ranged from 1.20% to 3.34%, with high total sugar (695 mg/mL and 812 mg/mL), low ash (0.70 ± 0.07;0.91 ± 0.01). On the other hand, these juices contain variable concentrations of minerals from one site to another (magnesium, calcium, phosphorus, potassium and iron). Potassium is the most abundant mineral element, followed by phosphorus and magnesium. Moreover, it is observed that the Gnamankoudji juices contain vitamin C (4.67 to 5.58 mg/100mL), contrary to the Bissap juice. The microbiological analysis indicates the presence of aerobic mesophilic germs (AMG) but a total absence of pathogenic germs in all juices. The important presence of nutrients and the total absence of pathogenic germs in these drinks justify their regular consumption by the population.

Sterilization of <i>Hibiscus rosa-sinensis</i>L. Vegetative Explants Sourced from Plants Grown in Open Environment and Influences of Organic Ingredients on <i>In Vitro</i>Direct Regeneration

This paper reports on the effects of organic ingredients in facilitating direct shoot regeneration from nodal explants of Hibiscus rosa-sinensis L. This paper also compares the sterilization conditions for 3 types of explants (node, internode, and shoot tip) harvested from an open field. The optimized sterilization conditions for the explants were 40% Clorox- 20 min exposure, 10% Clorox-15 min exposure, and 5% Clorox-40 min exposure for the node, internode and shoot tip, respectively. In the direct shoot regeneration using the nodal explants, we found MS medium containing 40 g/L sucrose, 0.3% (w/v) activated charcoal, and supplementations with myo-inositol, thiamine and nicotinic acid were suitable. The in vitro shoot survival rate was 30% with a mean leaf numbers of 2.68 produced, and a mean leaf length of 1.71 cm achieved after 5 weeks of culture on the modified medium.

Optimization of Ultrasonic-assisted Extraction of Polysaccharide from Hibiscus syriacus L.

The ultrasonic-assisted extraction of polysaccharides from Hibiscus syriacus L. at 180 W low power was optimized by the single factor tests and orthogonal test. Extraction temperature,ultrasonic treatment time,extraction times,and material-to-liquid ratio were considered as independent variables. The results suggested that the best extraction condition were determined as the extraction temperature at 80 ℃,the ultrasonic treatment time of 40 min,the material-to-liquid ratio at1∶ 20（ g/ml）,and the extraction times of 3. Under the optimal conditions,the extraction rate of polysaccharides from Hibiscus syriacus was 3. 114%.

Effect of Roselle calyces extract on the chemical and sensory properties of functional cupcakes

Roselle calyces(RC)are a major crop for export and used to make a common drink in Egypt.Dried RC are commercially available and appreciated to obtain concentrated extracts which might be used in the food and pharmaceutical industries for color and heath benefits.The objective of this research was to determine the chemical and the sensory properties of cupcakes formulated with Roselle calyces extract(RCE).Proximate analysis,anthocyanins,ascorbic acid,titrable acidity,%retaining of anthocyanins,color and sensory evaluations were done.RC cupcakes had high sensory scores(P<0.05)compared to control cupcakes.The parameter a*was significantly red in the RC cupcakes compared to control cupcakes along with 77%retaining of anthocyanins.The consumption of 100 g of the RC cupcakes would provide 465 mg/100 g dry matter anthocyanin that is more than 2 folds of the minimum average of the daily intake of anthocyanins for Americans,along∼1/3 of the daily dietary fiber intake to achieve fiber adequacy according to the Scientific Advisory Committee on Nutrition.RC cupcake can be a functional food and would have a“clean”label with cost effective advantage.

基于UHPLC-LTQ-Orbitrap的发酵玫瑰茄化学成分筛查和研究

本研究采用高效液相色谱-线性离子阱-静电场轨道质谱(UHPLC-LTQ-Orbitrap MS)技术,在电喷雾负离子模式下,对玫瑰茄水提液及其发酵液中有机酸类、黄酮类、花青素类和木质素类化学成分进行分析鉴定。选取Agilent Zorbax SB C18色谱柱(250mm×4.6mm×5μm),以0.1%甲酸水溶液(A)-甲醇(B)为流动相,流速1mL/min,梯度洗脱分析。根据高分辨质谱提供的精确相对分子质量数据和多级质谱碎片离子,获取目标化合物的相对分子质量和结构信息。结合保留时间、对照品及相关参考文献等信息,共筛选鉴定出41种化合物,包括34种有机酸类、4种黄酮类、2种花青素类以及1种木脂素类。结果表明,玫瑰茄发酵前后,其活性成分的种类及含量均呈现不同程度的变化,该方法阐明了玫瑰茄发酵前后化学成分的变化,可为玫瑰茄的进一步研究提供参考。

玫瑰花 玫瑰茄 枸杞子复合发酵液化学成分的HPLC-HRMS分析

目的:研究玫瑰花、玫瑰茄及枸杞子3者复合物提取液经肠膜明串株菌肠膜亚种发酵前后化学成分的差异。方法:采用高效液相色谱-线性离子阱-静电场轨道阱质谱(HPLC-LTQ-Orbitrap MS)技术对其复合物提取液及发酵液中化学成分进行筛选及鉴定。结果:根据所获得的精确相对分子质量,同时结合色谱保留行为、质谱裂解规律、特征碎片离子、对照品比对以及相关文献报道,共筛选鉴定了48个化学成分,包括39个有机酸、6个黄酮及3个花青素类成分。结论:玫瑰花、玫瑰茄及枸杞子3者复合物提取液发酵前后化学成分的种类和含量均发生不同程度的变化,其中苷类成分可以转化为更利于人体吸收利用的苷元类成分,可为其他发酵中药的现代研究提供借鉴思路。