Source: SOUTHERN UNIV submitted to NRP
ALTERNATIVE UTILIZATION OF ROSELLE HIBISCUS AS SMALL FARM AND NICHE MARKET CROP
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
0230455
Grant No.
2012-38821-20092
Cumulative Award Amt.
$500,000.00
Proposal No.
2012-02470
Multistate No.
(N/A)
Project Start Date
Sep 1, 2012
Project End Date
Aug 31, 2017
Grant Year
2012
Program Code
[EQ]- Research Project
Recipient Organization
SOUTHERN UNIV
(N/A)
BATON ROUGE,LA 70813
Performing Department
Plant & Soil Sciences
Non Technical Summary
The emergence of global economy for traditional horticultural crops has compelled Southern University to undertake research to embrace NIFA priority areas in sustainable agriculture and expanded opportunities for small farms to explore alternative crops to enhance farm sustainability. Roselle hibiscus, Hibiscus sabdariffa was selected and evaluated for its adaptability, flavonoid properties and value-addition in Phase I. The outcomes of Phase I activities warrant the continuation of this proposal to further validate some research findings and to find new uses to enhance profitability. The multi-institutional and multi-disciplinary research approach includes (i) developing appropriate cultural practices, (ii) evaluating it as a container flowering plant for the Christmas market, (iii) utilizing its anthelmintic foliage as forage supplement for goat production, (iv) evaluating the refined calyce and leaf extracts on apoptosis of cancer cells, and (v) developing alternative but marketable value-added products such as drinks, health supplements, antimicrobial food seasonings to strengthen niche marketing and commercialization in Louisiana. This proposal provides supportive information and niches to nurture entrepreneurship development. Results will enhance science-based knowledge and education in specialty crops and natural plant products database for USDA/ARS. It enhances SUAREC research capability and competitiveness by building human capacity in natural product science, and equipping analytical, bioassay and food safety laboratories with the state-of-the-art facilities. The acquired collaborative research experience and participation strengthen students' experiential learning. Funds provided by this project can be used as leverage to secure external funding and impact the livelihood of more than 50 niche market producers and 1,184 sustainable agriculture farmers.
Animal Health Component
40%
Research Effort Categories
Basic
20%
Applied
40%
Developmental
40%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2011099108010%
2052220102010%
2052220101040%
2052129106010%
3073820106010%
3133820111010%
5022220101010%
Knowledge Area
205 - Plant Management Systems; 313 - Internal Parasites in Animals; 201 - Plant Genome, Genetics, and Genetic Mechanisms; 307 - Animal Management Systems; 502 - New and Improved Food Products;

Subject Of Investigation
2220 - Medicinal crops, non-narcotic; 1099 - Tropical/subtropical fruit, general/other; 3820 - Goats, meat, and mohair; 2129 - Florist crops, other;

Field Of Science
1060 - Biology (whole systems); 1010 - Nutrition and metabolism; 1110 - Parasitology; 1080 - Genetics; 1020 - Physiology;
Goals / Objectives
1: To validate the performance of selected roselle hibiscus accessions from Phase I and to develop appropriate cultural practices for the production of calyces: (i) Genetic identification of roselle hibiscus accessions, (ii) Evaluation of fertilizers and (iii) Evaluation of the effects on dates of planting. 2: To produce container-grown flowering roselle hibiscus plants for Christmas. 3: To develop alternative niche market for roselle hibiscus value-added products to promote small-scale commercial market. 4: To assess roselle hibiscus as a nutritive and natural deworming forage for goat production. 5: To further evaluate the refined extracts of roselle hibiscus calyces and leaves for their antagonistic and anti- proliferation effects on selected human cancer cells.
Project Methods
(1) In collaboration with Thad Cochran Southern Horticultural Laboratory of USDA/ARS, simple sequence repeat (SSR) markers applications will be used to determine the genetic identity of four field-selected roselle hibiscus accessions. Using a complete block randomized design with a split-split plot arrangement of treatments, various levels of organic 8-5-5 fertilizer will be evaluated for their effects on the performance of a selected roselle hibiscus accession and its calyce and leaf quality in the field. Phenolic acids, anthocyanins and antioxidant activity as affected by fertilizer treatments will be determined by Folin-Ciocalteu, pH differential spectrophotometry and ABTS method, respectively. A field experiment will be conducted to assess whether the dates of seed germination and transplanting dates have any effect on earliness of flowering and maturity. Various planting dates will be assessed on their effects on fruit maturity, calyce production and plant performance. The horticultural performance of the four selected roselle hibiscus accessions will be continuously evaluated in conjuction with the above-mentioned experiments. (2) In the research to explore the possibility of using roselle hibiscus plants as a container-grown florist crop, stem cuttings harvested at various dates will be rooted and planted in the containers. Pinching , topping and the application of plant growth retardants will be used to manipulate plant growth. Finished products will be evaluated for their acceptability in the market. (3) In the development of alternative niche for roselle hibiscus value-added products to promote small-scale commercial market, several products will be targeted for development. Roselle hibiscus product development include roselle hibiscus concoctions, encapsulated hibiscus calyce powder, roselle hibiscus tea lead sachets, roselle hibiscus calyce powder seasoning mix in addition to cookies and chutney. Panel taste tests and marketability will be conducted to assess economic feasibility. In addition, the derived seasoning mixes will be assessed for their antimicrobial activities and their effects on meat tenderness and quality. Nutritive values of the developed products will be determined. (4) In the study to assess the roselle hibiscus foliage as a nutritive and natural deworming forage for goat production, the harvested foliage which is nutritive and anthelmintic ,will be fed to goats as a forage supplement. Chemical composition, acid detergent fiber and energy of the roselle hibiscus will be determined. Animal weight gain from roselle hibiscus foliage feeding will be compared to that of the traditional feeding regimen practiced in the field. Fecal samples of experimental goats will be weekly collected and parasitic egg count will be measured using modified McMaster's techniques. (5) To further ascertain the antagonistic and anti-proliferation effects of the roselle hibiscus extracts on selected human cancer cells, refined extracts of roselle hibiscus calyce and leaves will be prepared and evaluated in vitro.

Progress 09/01/12 to 08/31/17

Outputs
Target Audience:This project is designed to produce applied technical information for small and limited resource farmers and agricultural entrepreneurs who have desire to use alternative crops to start and/or to enhance their farm sustainability. It also targeted home gardeners, rural and urban residents who have desire to grow and consume health-promoting natural products. The educational focus of this project was to utilize the research activities as a means to provide experiential learning to students on roselle hibiscus cultivation, the use of natural product of hibiscus as alternative medicinal supplements and functional foods. and research methodologies. The project team also worked closely with and jointly with Southern University Agricultural Research and Extension county agents to deliver the know-how to clienteles of all walks of life. Roselle hibiscus production and utilization workshops and seminars were presented to more than 160 farmers and gardeners. About 1,100 community members were exposed to roselle production and research via north Louisiana Agri-business Council Ag Expose, "This week in Louisiana Agriculture" broadcast, various exhibitions throughout Louisiana. Targeted audiences also included scientific communities in natural products, USDA/ARS data base and molecular biology. Changes/Problems:No major changes had occurred on this project because appropriately trained scientists were on board for specific assigned task of the project. Problem encountered: Our sub-awardee, West Virginia State University submitted reimbursement invoice for $11,500.56 after August 25, 2017 which was the last day for final drawdown of the fund from this project (which ended August 31, 2017). What opportunities for training and professional development has the project provided?The projectoffered scientific research exposure to more than 50 undergraduate students and about 100 school students from various parishes in the state of Louisiana. It supported four M.S. degree and three Ph.D. degree candidates who conducted research activities relating to the objectives of the project. It provided an opportunity for two graduate students to be trained in the analytical laboratory protocol for phytochemicals. It had offered resources and opportunities for the project director(s) to make various seminars/ workshops to train farmers and home owners of various parishes on roselle hibiscus production and value-added products development. The project director attended and presented numerous papers in the scientific meetings ranging from domestic to international venues.A great deal of knowledge and skills have been acquired via scientific interactions and visitations. It has offered opportunity for the project to establish collaborative work with other institutions. How have the results been disseminated to communities of interest?Roselle hibiscus research findings had been presented to medicinal plant and natural products communities domestically, nationally and internationally. Domestically, research findings were presented to farmers and home owners of various parishes in Louisiana, school students and communities via workshops, field tours, Agri Expos and presentations to SUAREC faculty and staff. Nationally, roselle hibiscus production and value addition research were broadcasted via "This Week In Louisiana Agriculture", exhibited in the National Farm Bureau Convention held in New Orleans, ARD symposia, annual meeting of the American Council on medicinal plants and natural products. Internationally, the research findings were presentedat the 3rd World Congress on Medicinal Plants and Natural Products Research, 7th Global Summit on Agriculture and Horticulture, 3rd International Conference on Mass Spectrometry, International Conference and Exhibition on Food Processing& Technology, annual meeting of the American Society of Horticultural Science, and International Horticultural Congress and International Society for Horticultural Sciences. Scientific publications included Proceedings of Agriculture and Horticulture, Mass Spectrometry, Food Processing and Technology, ACTA Horticulture, Molecules, Food Chemistry, Planar Chromatography, International Innovation. What do you plan to do during the next reporting period to accomplish the goals?This project has officially ended. The project director plans to develop new proposal to validate these research findings and to advance further research activities to improve roselle hibiscus utilization and its contributions toward food and agriculture.

Impacts
What was accomplished under these goals? More than 37 roselle hibiscus accessions from USDA/ARS Germplasm Depository, Universities and researchers in the USA were screened. Four accessions (of Jamaica, Nigeria, Senegal and South Africa origins) were selected based on horticultural characteristics and earliness of production. A new accession from a Florida grower was discovered to be another accession that has early maturity at the end of the project period. Such accession warrants further evaluation. The fertilizer applications of organic fertilizer 4-2-2 and 8-5-5 at 4 kg/ 30 m row at planting time did not significantly increase fruit yields. Preliminary data indicated that side-dressing of organic fertilizer 8-5-5 at 4 kg/30 m row increased plant height, chlorophyll content of leaves and fruit yield as the frequency of side-dressing increased. There appeared to be a trend that increased frequency of fertilizer application reduced the antioxidant content of extracts. Diseases that affected production were Phytopthera and Rhizoctonia root rot, and Alternaria leaf spot, and powdery mildew. No major insect pests were observed but the incidence of black head web-worm was observed in 2016 growing season. Elemental composition of the calyce and leaf were analyzed using Arco ICP and LECO CN analyzer and compared among selected roselle accessions. There were significant variations in elemental composition of the calyces when the analytical data from five accessions were compared. In general, all accessions studied exhibited higher amount of N, K, CA, P, Mg and Mn. The extracts of roselle calyces were found to exhibit antimicrobial activity against E. coli, Salmonella enterica and to some extent Listeria monocytogenes basing on in vitro study as well as vegetable wash which was conducted by former collaborating scientist of the project. In collaboration with Rutgers University, phytochemical profiles and antioxidant contentsof the calyces and leaves of 25 roselle hibiscus accessions using Folin Ciocalteu's reagent assay and ABTS assay, respectively were analyzed.. Anthocyanin components were evaluated and compared among calyces of different pigmentation. Delphinidin sambubioside and cyaniding sambubioside were the major components In general, extracts of darker pigmented calyces showed higher content of anthocyanin and antioxidant activities than those of lighter pigmented calyces. Chlorogenic acid and it isomers and neochlorogenic acid were the key phenolic acids presented in the leaves while quercetin rutinoside, quercetin glucoside, kaempferol-rutinoside and kaempferol glucoside, Quercetin and Kaempferol were the major flavonoid present in leaves of all roselle accessions. Trace amount of delphinidin sambubioside and cyaniding sambubioside were found in three accessions. In collaboration with West Virginia State University,the extracts of different pigmented calyces were evaluated for their effect on inhibiting human tumor cell growth. Darker pigmented extracts appeared to exhibit higher amounts of inhibition of cancer tumor cell growth than lighter pigmented calyces. Higher amounts of anthocyanin-enriched extracts exhibited greater effectiveness in inhibiting human cancer cell growth than lower amounts of anthocyanin-enriched extracts. Extracts of roselle hibiscus leaves also showed great degree in inhibiting human cancer cell growth. In vitro evaluation of human cancer cell growth was conducted using MTT. WST-8, crystal violet and CyQUANT assays. A dissertation study in the evaluation of the roselle extracts on the viability of prostate cancer cells and the determination of androgenic effects of roselle extracts in the presence of anti-androgenic compounds indicated roselle hibiscus extracts exhibited inhibition of LNCap-GFP cells. Various value-added products such as tea, roselle jelly and chutney, wine, and calyce powder capsules have been formulated and will be required for quality assurance and control in the future.

Publications

  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2017 Citation: Chin, Kit L. et. al. 2017. In vitro evaluation of anti-tumor activity of extracts of two wide-collected mushrooms. 3rd world Congress on Medicinal plants and Natural Product research. Kuala Lumpur, Malaysia. Oct, 2-3, 2017
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jing Zhen, Thomas S. Villani, Yue Guo, Yadong Qi, Kit Chin, Ming-Hsiung Pan, Chi-Tang Ho, James E. Simon and QingLi Wu. 2015. Food Chemistry 190 (2016):673-680. http://dx.doi.org/10.1016/j.Foodchem.2015.06.006.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jin Wang, X.S. Cao, Y. Qi, V.Ferchaud, K.L. Chin and F. Tang. 2015. Variation in chemical fingerprints and major flavonoid contents from the leaves of thirty-one accessions of Hibiscus sabdariffa L. J. of Planar Chromatography 28 (4):1-6.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Chin, K.L., J. Zhen, Y. Qi, S.L. Chin, M. Breithaupt, Q.L. Wu, J. Simon, J. Henson, and V. Ferchaud. 2015. A comparative evaluation: Phytochemical composition and antioxidant capacity of three roselle (Hibiscus sabdariffa L.) accessions. Proceeding of the 5th WOCMAP, ACTA Horticulture (in Press) 8 p.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Jin Wang, Xian Shuang Cao, Hao Jiang, Yadong Qi, Kit L. Chin, Yongde Yue. 2014. Antioxidant activity of leaf extracts from different Hibiscus sabdariffa accessions and simultaneous determination Five Major antioxidant compounds by LC-Q-TOF-MS. Molecules 19 (12): 21226-21238.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Chin, Kit L., Y. Qi, R. Payne. 2017. Phytochemistry of the roselle leaves. 18th Biennial Research Symposium of 1890 Research Directors. P.268.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Chin, S.L., Y. Qi, V.A. Ferchaud, R. Payne, J.F. Henson and K.l. Chin. 2017. Effects of fertilizer treatments on nutrient elements, total polyphenol and antioxidant contents of Hibiscus sabdariffa leaves. 18th Biennial Research Symposium of 1890 Research Directors. P.238.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Breithaupt, M.D., Y. Qi, K.L. Chin, V.A. Ferchaud, R. Payne and J.F. Henson. 2017. 18th Biennial Research Symposium of 1890 Research Directors. P.112
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Chin, Kit L., S. L. Chin, Y. Qi and V.A. Ferchaud. 2016. Organic fertilizer effect on nutritional elements, total polyphenol and antioxidant content of roselle (hibiscus sabdariffa L.) leaves. Proceeding of 7th Global Summit on Agriculture and Horticulture. P. 23.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Chin, Kit L., Y. Qi, A. Pertuz, J. Coppin, G-L Wu and J. Simon, 2016. The use of LC/UV/MS method for qualitative and quantitative analysis of phytochemicals in roselle (Hibiscus sabdariffa L.) leaves, Proceeding of 3rd International conference on Mass Spectrometry. P. 23.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Chin, K.L. 2013. The Healing Power of Hibiscus. International Innovation, August issue. P.50-52.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Chin, K.L., Y. Qi, QL Wu, R. Juliani, J. Simon & G. Hankins. 2013. Comparative Study on Roselle Hibiscus Varieties for the Development of Natural Products. BITs Annual Global Health Conference. P.039.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Chin, Kit, Y. Qi, D. Jaroni, G. Hankins and J. Simon. 2012. Roselle hibiscus (H. sabdariffa) as a as a niche market crop for product development. J. of Food Procession & Technol. 3 (10): 80.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Qi, Y., K.L. Chin, Q.L. Wu, J. Coppin, R.C. Nicholas and J. Simon. 2012. Total polyphenols, antioxidant content and chemical profiling of leaves of roselle (Hibiscus sabdariffa) accessions. The 2nd Annual Conference of the American Council for Medicinally active plants (ACMAP) J. of Medicinally active plants 1(5):20


Progress 09/01/15 to 08/31/16

Outputs
Target Audience:This project is designed to produce technical information forsmall and limited farmers and agricultural enterpreneurs who haveadesire to usealternative crops to start and/or enhance their farm sustainability.It also targeted home gardeners, rural and urban residents who have the desire to grow and consume health-promoting natural products. It providedexperiential learning in roselle hibiscus research and enhancedknowledge of students on roselle hibiscus plant, the use of natural products as alternative medicinal supplements and functional foods. This project also workedclosely and jointly with Southern University Ag Center to reach out to clienteles of all walks of life through various parish county agents. Roselle production and utilization workshop was presented to 36farmers/home gardeners of St Helena and Tangipahoa. Roselle hibiscus tea was featured in the North Louisiana Agri-Business Council AgExpo withabout 650 general attendees.One hundred and seventy-fivehibiscus seed packages were given out at this Expo. Three undergraduate students, three Ph.D. studentsparticipated in the research work. More than 28attendees of the SUAREC Fall Garden Workshophave visited and gained knowledgeat the Roselle hibiscus research plots. Changes/Problems:A minor budget revision to acquire equipment for in-vitro study,value-added product development and financial assistance to students is needed. Such modification does not alter the scope and objectives of the research project. What opportunities for training and professional development has the project provided?The project has provided an opportunity for the project director to attend project-related conferences and to interact with professionals. Increased knowledge and skills in lab analytical protocol and roselle hibiscus tea development was acquiredvia the attendance of Worldtea Expo and Emerald conference... How have the results been disseminated to communities of interest?Roselle production potential for St. Helena and Tangipahoa parishes was presented to the agriculture advisory committee of these twoparishes. A field tour was given to 28attendees of the SUAREC Fall Garden Workshop. Value-added productsuch as hibiscus tea was presented and served to about 180general public and agricultural community attending the North LouisianaAgri-Business Council AgExpo. Value-added products such as hibiscus seasoning, wine, canned drink and capsule products were show cased at the SU Ag Center building. Research findings have been periodically disseminated to the public. What do you plan to do during the next reporting period to accomplish the goals?Further testing and selection to validate the performance of roselle hibiscus accessions will be continued to ascertain adaptability performance. Genetic verification of the four selected accessions will be conducted. Phytochemical profiles, total phenol and antioxidant activity of potential roselle hibiscus accessions are to be assessed and completed. There is a need to continue the in-vitro evaluation of the hibiscus extracts on selected cancer cell apoptosis, the developmental research on value-added hibiscus products for the QA/QC,and the evaluation of roselle hibiscus as a container flowering plant.

Impacts
What was accomplished under these goals? An experiment to assess the effects of the application frequency of organic fertilizer 8-5-5 on roselle hibiscus performance was conducted in the field of silty clay loam soil with soil pH of 7.0. The application rate was 4 kg/30 m row of organic fertilizer 8-5-5. The between-rowspacing was 2.13 m. The treatments were control, one application of organic fertilizer at planting, one application of organic fertilizer at planting + one subsequent application one month after first application and one application of organic fertilizer at planting + two subsequent applications at monthly interval following the first application, respectively. Ten individual plants were randomly selected for yield, plant height and SPAD data. The fruit yield/plant basis increased significantlywith increased number of organic fertilizer applications consistently across four roselle accessions (Jamaica, Nigeria, Senegal and South Africa). The increased number ofapplications of organic fertilizer exhibited asignificant linear increase in yield correspondingly. There was no significant fertilizer application and accession interaction indicating that the application of fertilizer increased yield consistently across all accessions.Likewise, plant height and the chlorophyll content measured as SPAD data increased correspondingly with increased number of fertilizer application. The data onthe effectof fertilizer applications on the total phenol content and antioxidant activitywill be collected using Folin-Ciocalteu assay and ABTS radical cation assay. In the accession screening, four new accessions were evaluated. Theperformances of these four accessions were compared to the "Jamaica" accession, a recommended accession for Louisiana growing. A new earliest maturing accession known as "Flor-Ted" was identified while "Thai Red", "Malaysia" accessions were identified as late maturing accessions. "Flor-Ted" accession will need to bescreened for its horticultural characteristics and yield performance to ascertain adaptability. Because of the exhibited antimicrobial activity of the roselle hibiscus extract, a new study was conducted to evaluate its cosmeticuse. Anew value-added infused hibiscus product, soap was developed from this study. Seven soap prototypes were made using whole and/or ground calyces extracts with either glycerin or lard bases. Antibacterial analysis was conducted using the cup plate agar method before performing consumer acceptability analysis. A panel of 32 men and women evaluated the soaps on color, appearance, texture, fragrance and lathering ability. Lower concentrations of infused hibiscus soap showed no significant effectiveness against E. coli while higher concentrations infused soap showed potential antimicrobial effect. A literature review indicated that androgens exerted their effects by binding androgen nuclear receptors causing changes in transcriptional and post-transcriptional activity resulting in modulation of cell growth and cancer cell development. A dissertation study was conducted to evaluate the roselle hibiscus extracts on the viability of prostate cancer cells and to determine the androgenic effects of roselle hibiscus extracts in the presence anti-androgenic compounds. Preliminary result indicated that the roselle hibiscus extracts exhibited inhibition of in-vitro LNCaP-GFP cells. This project also workedclosely and jointly with Southern University Ag Center to reach out to clienteles of all walks of life through various parish county agents. Roselle production and utilization workshop was presented to 36farmers/home gardeners of St Helena and Tangipahoa. Roselle hibiscus tea was featured in the North Louisiana Agri-Business Council AgExpo withabout 650 general attendees.One hundred and seventy-fivehibiscus seed packages were given out at this Expo. Three undergraduate students, three Ph.D. studentsparticipated in the research work. More than 28attendees of the SUAREC Fall Garden Workshophave visited and gained knowledgeat the Roselle hibiscus research plots.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Lakeisha Y. Butler, Melisa Morgan, K. Chin, J. Snowden and W. Grey. 2016. Analysis of the calyces from the Jamaica species of Hibiscus sabdariffa for androgenic compounds. Experimental Biology. April 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: M. Morgan, L. Butler, W. Grey, K. Chin and J. Snowden. 2015. Investigating the androgenic potential of Jamaica species, H. sabdariffa. Annual meeting of the Society of Toxicology. South Central Chapter, Houston, TX. October 2015
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Morgan,, M.. and Butler, L. 2015. Investigating the androgenic potential of Jamaica species, H. sabdariffa. Wilbur B. Clarke Chemistry Symposium. Baton Rouge, LA. October, 2015
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jin Wang, Xianshuang Cao, Vanessa Ferchaud, Yadong Qi, Hao Jiang, Feng Tang, Yongde Yue and Kit L. Chin. 2015. Variations in chemical fingerprints and major flavonoid contents from the leaves of thirty-one accessions of Hibiscus sabdariffa L. Biomedical Chromatography. (Wileyonlinelibrary.com) DOI 10.1002/bmc.3623.


Progress 09/01/14 to 08/31/15

Outputs
Target Audience:This project targeted small and limited resource farmers, potential agricultural entrepreneurs, home gardeners, rural and urban residents who have desire for helath-promoting natural products,Also targeted were graduate and undergraduate students, scientists, extension agents and those with alternative medicine and functional food interests. This project worked closely and jointly with SUAREC cooperative extension service to reach out to clienteles of all walks of life especially in the area of functional food. Changes/Problems:A minor budget modification may be undertaken. What opportunities for training and professional development has the project provided?Twenty-eight small farmers attended a workshop in St. Helena/Tangipahoawhere production pointer, insect and disease control and techniques of hibiscus drinks, jelly, chutney and hot sauce preparations were featured. One undergraduate student in plant andsoil sciencefrom Southern University's agricultural program was engaged in summer internship participating inhibiscus production and product development research activities. How have the results been disseminated to communities of interest?Roselle hibiscus research at Southern University was presented to an agricultural committee in St Helena and Tangipahoa Parishes via workshop. Field tours and presentationswere coducted to farmers and general public via Louisiana Small Farmers Conference, north Louisiana Agri-Business Council Ag Expo, National Ag Day Celebration, Improving Information Access Conference. Research findings have been published in scientific journals. What do you plan to do during the next reporting period to accomplish the goals?Further testing to validate the performance of selected H. sabdariffa accessions will be needed to ascertain adaptability performance. Genetic verifiication of the four selected roselle hibiscus will be conducted. Phytochemical analysis and determination including antioxidant activities for various plant parts of all potential H. sabdariffa accessions are to be completed and evaluated. To continue the in-vitro bioassays to confirm preliminary testedresults on cancer cell proliferation effected by the hibiscus calyce and leaf extracts.To continue the developmental research on value-added hibiscus products for the purpose of refining the quality and acceptability of the products. To further evaluation the potential use of roselle hibiscus plants as container flowering plants.

Impacts
What was accomplished under these goals? Situation: Roselle hibiscus, is one of the highest volume specialty natural and botanical products in international commerce. The international market for herbal products is estimated to be US$ 62 billion (2011) which is poised to grow to US $5 trillion by the year 2050 (The 4th Global summit on Medicinal and Aromatic Plants program announcement, 2009). The production of H. sabdariffa in Louisiana and the southern region can potentially play a part in reducing our dependency on importation of calyces from Africa and other countries, and provide forage for goats as nutritive and natural dewormer herbage. Roselle hibiscus can become an important niche market crop for functional food producers and forage crop that can potentially improve the livelihood of at least 1,184 limited resource farmers in Louisiana. No locally grown fresh and dried calyces are available in the commercial market. The Red Stick farmers market in Baton Rouge, Louisiana could not adequately supply calyces to meet consumers' demand. Experiment/Accomplishments: In the experiment on the evaluation of organic fertilizer 8-5-5 applied on plant performance of four hibiscus accessions at planting, it was found that the amount of anthocyanin content in calyx was 8.73 mg/g, 9.16 mg/g, 9.89 mg/g and 17.37 mg/g, for Jamaica accession, Nigeria accession, Senegal accession and South Africa accession, respectively. Data indicated darker pigmented calyx was associated with higher content of anthocyanins. However, weak association was observed between total phenol/total antioxidant with anthocyanin content. Delphinidin-sambubioside and cyaniding-sambubioside were the major components of the anthocyanin. Delphinidin was only observed in the darker-pigmented calyx. Leaf and calyx samples were collected, dried and grounded from each treatment of the experiment studying the effect of organic fertilizer applications on phenolic compounds and antioxidant activities. The seeds of five accessions reflecting different country-origin (Sudan PI496938, Senegal T, South Africa Trevael, Jamaica and Taiwan PI 273389) were germinated in the greenhouse for container flowering plant study along with pruning. In the value-addition study, a trial effort was carried to incorporate various components of the hibiscus plant to develop products with most health-benefiting factors. A preliminary HPLC/MS analysis showed that the leaf alone has the highest total phenols followed by leaf and calyx combined, calyx, whole shoot (fruits & shoot with no leaves), and shoot stem alone. These data suggest versatility of hibiscus plant parts for product development. Hibiscus beverage made from dried calyx and dried whole shoot (whole fruit + stem) were preliminarily taste tested for their comparison and acceptability. Based on this taste test evaluation, new formulation will be developed for further testing. Leaf samples from plants receiving organic fertilizer 8-5-5 treatments were prepared for HPLC/MS analysis. Following analysis, hibiscus extracts effected by fertilizer application will be assessed for their bioactivities such as inhibition of in vitro cancer cell proliferation. Roselle hibiscus research findings were presented to a scientific community of aromatic and medicinal plants via the World Horticultural Congress and published journal articles. About 300 high school students visited the Roselle hibiscus exhibitand value-added products displayvia the celebration of National AG Day at Southern University. Roselle hibiscus production research findings were presented to about 15 small farmers during the 5th Annual Louisiana Small Farmers Conference. Thirty people attended the presentationon the use of medicinal crops including Roselle hibiscus via "Improving Information Access Conference". Hibiscus value-added products were featured and exposed tomore than 100 attendees of the annual North Louisiana Agri-business Council Ag Expo. Twenty-eight small farmers attended the St. Helena/Tangipahoa Hibiscus workshop where production pointer, insect and disease control and hibiscus drinks, jelly, chutney and hot sauce were featured. One plant andsoil science student from Southern University's agricultural program was engaged in summer internship participating inhibiscus production and product development research activities. Four graduate students (two M.S. and two Ph.D.) wereoffered graduate assistantships and tuition stipends to conduct research on hibiscus plant performance research and value-addition studies.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Jing zhen, Thomas S. Villani, Yue Guo, Yadong Qi, Kit Chin, Min-Hsiung Pan, Chi-Tang Ho, James E. Simon and QingLi Wu. 2014. Ohytochemistry, antioxidant capacity, total phenolic content and anti-inflammatory activity of Hibiscus sabdariffa leaves. Food Chemistry 190 (2016) 673-680
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jin Wang, Xianshuang Cao, Yadong Qi, Vanessa Ferchaud, Kit L. Chin and Feng Tang. 2015. High performance thin-layer chromatographic method for screening antioxidant compounds and discrimination of Hibiscus sabdariffa L. by principal component analysis. J. of Planar Chromatography 28 (4): 1-6.


Progress 09/01/13 to 08/31/14

Outputs
Target Audience: This project targeted small and limited resource businesses, farmers' market vendors, farmers, rural and urban residents, producers who could grow alternative crops for additional income. Also targeted were graduate and undergraduate students, scientists, extension agents and those with alternative medicine and functional food interests. This project worked closely and jointly with NIFA-Evans-Allen project entitled " Evaluation of roselle (Hibiscus sabdariffa) accessions on disease resistance, horticultural and antimicrobisl pprperties". Hibiscus value-added products were exhibited at the Louisiana State Legislative Day where more than 100 general publics and legislative staff and students at the State capital lobby perusedthe exhibition and sampled products. Eight Africa American small farmersattended the St. Helena parish meeting where hibiscus production as a niche market crop for under-served farmers was presented. One plant and soil science student from Southern University's agricultural program was engaged in summer internshipparticipating in hibiscus production and product development research activities. Four graduate students (two M.S. and two Ph.D.)were offered graduate assistantships and tuition stipends to conduct research on hibiscus plant performance study. A significant number of particpants (estimated to be around 85) viewed the digital poster and the presentation at theSymposium on Aromatic and Medicinal Plants of International Horticultural Congress held in Brisbane, Australia in August, 2014.A brief discussion of roselle hibiscus research was presented to32 high school students who also toured the value-added roselle hibiscus processing facility. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? PD attended the International Horticultural Congress held in Brisbane, Australia anddelivered a presentation on " A cimparative evaluation::phytochemical compositionand AntioxidantCapacities of Three Roselle (Hibiscus sabdariffa) Accessions". The PD also took the opportunity to visit andstudy the various horticulturalresearch and production sites ofinterest as a stopover visit in New Zealand. How have the results been disseminated to communities of interest? Roselle hibiscus research at Southern University was presented to an agricultural committee in St Helena Parish. Workshop and field tour were coducted to vegetable gardening and church groups. The project also host a tour of the roselle hibiscus processing facilityto demonstrate product development activities to a group of 32 high school students. Three articles on roselle hibiscus have been submitted for publication. What do you plan to do during the next reporting period to accomplish the goals? The project will reach and provide research findings to both traditional and non-traditional producers across the state, graduate and undergraduate students, scientists, extension agents and those with alternative medicine and functional food interests. We will also implement aspects of the study that are still pending. We will continue to encourage the adoption of the new techniques created and encourage new business ventures relating to roselle hibiscus.

Impacts
What was accomplished under these goals? The demonstrated medicinal properties and increased global market demand for healthy natural products have made roselle hibiscus a lucrative alternative functional crop to supplement small farm income. Research has been undertaken to develop marketable products for enterprise development in Louisiana. Various roselle hibiscus value-added productswhich haveeconomicviability in the market and can be commercialized were potentially developed. An emergence of new entrepreneurs in roselle hibiscus value-added products is envisioned in the near future. This could mean that a new roselle hibiscus industry worthmillions of dollars can be potentially established from the research findings at Southern University Agricultural Research& Extension Center. A newbusiness opportunity will have a significant impact on Louisiana economy and the well-being of Louisiana small and limited resource farmers. Accomplishments: Objective 1:To validate the performance of selected roselle hibiscus accessions and to develop appropriate cultural practices for the production of calyces. A repeated experiment to evaluate the effect of organic fertilizer applications on the performance of four selected roselle hibiscus accessions was conducted in the field with silty clay loam soil. The origins of these roselle hibiscus accessions wereJamaica, Nigeria, Senegal and South Africa. The treatments were Control,4 kg.of organic fertilizer 8-5-5/20 m. at planting, 4 kg.of organic fertilizer 8-5-5/20 m at planting + 4 kgone month after first application, 4 kg.of organic fertilizer 8-5-5/20 m. at planting followed by two subsequent application of 4 kg/20m. at one month interval.It was observed that the application of ferilizer increased total fruit yield when their effects were compared to that of the Control. In general, increased yields were associated with increased amount of fertilizer applications. Higher amount of organic fertilizer applications increased plant height and chlorophyll content of the leaf but did not delay flowering. In a seperate inorganic fertilizer application study, high nitrogen application enhanced vegetative growth and delayed flowering. Experimental plants performed well without the occurrence of any major diseases that impacted the plants. Objective 2. To produce container-grown flowering roselle hibiscus for Christmas plants. Initial greenhouse trial of the selected roselle hibiscus accessions indicated that these plants produced fruits much earlier prior to Christmas, andwere susceptible to powdery mildew infestationunder the cold and humid condition during the months of October and November. However, field grown roselle hibiscus cutting made good flower/fruit cuttings. Future trials on the late maturing roselle hibiscus accessions may be needed to assess the viability of container-grown flowering hibiscus. Objective 3. To develop alternative niche market for roselle hibiscus value-added products to promote small-scale commercial market. Preliminarily, several roselle Hibiscus value-added products have been developed. Such products included hibiscus beverage, hibiscus jelly and chutney, hibiscus seasoning mix, health supplement capsules, hibiscus hot sauce and mix, hibiscus cookies, granola bar with hibiscus calyce. All these potential products are needed to evaluate for their nutritive values, acceptability study and labelling requirements. Objective 4. To assess roselle hibiscus as a nutritive and natural deworming forage for goat production. Prelimiary observation trial indicated that roselle hibiscus plant (leaves, fruits and stems) was palatable to the experimental goats. Initial trial was conducted to evaluate the effect of roselle hibiscus feeding in addition to regular diet on intestinal parasites. Six goats were used as a control treatment which provided regular goat diet feeding. Another six goats were fed with roselle hibiscus plants in addition to regular goat diet. After five days of feeding, fecal samples were collected and stored for laboratory parasitive egg counts. Objective 5.To further evaluate the refined extracts of roselle hibiscus calyces and leaves for their antagonistic and anti- proliferation effects on selected human cancer cells. Four different-pigmented roselle hibiscus calyces were evaluated for their total anthocyanin, phenol and antioxidant contents using HPLC/UV/MS, Folin Ciocalteu's reagent and ABTS method, respectively. The in-vitro inhibition of selected human cancer cells (U87 glioma, Pan-C1 pancreatic and CH157-MN mengloma) by the extracts of these calyces were used to correlate the bioactivity with anthocyanin, total phenol and antioxidant contents. In the cancer cell proliferation assay, crystal violet and WST8 assaay methodswere used to examine the effects of leaf extracts on cell proliferation of four human tumor lines, namely U87 cells, A172 cells, PanC-1 cells and Ch157-MN cells according to manufacturer's protocol. In this study, 96 well plates were used here. The crystal violet was less consistent than the WST8 assay. initial results show little difference in cell proliferation inhibition by the whole leaf extracts among accessions. The extract of leaf blade were more effective in inhibition of cell proliferation than that of the leaf petiole.

Publications

  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2014 Citation: Chin, Kit L.,J. Zhen, Y. Qi, S.L. Chin, M. Breithaupt, Q.L. Wu, J. Simon, J. Henson and V. Ferchaud. 2014. A comparative evaluation: Phytochemical composition and antioxidant capacity of three roselle (Hibiscus sabdariffa) accessions. abstract. 29th International Horticultural Congress 2014.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Jin Wang, Xian Shuang Cao, Hao Jiang, Yadong Qi, Kit L. Chin and Yongde Yue.2014. Antioxidant Activity of Leaf Extracts from Different Hibiscus sabdariffa Accessions and Simultaneous Determination Five Major Antioxidant Componds by LC-Q-TOF_MS. Molecules 19: 21226-21238.
  • Type: Journal Articles Status: Under Review Year Published: 2014 Citation: Jin Wang, Xian Shuang Cao, Yadong Qi, Vanessa Ferchaud, Kit L. Chin and Fen Tang.2014. High-Performance Thin-Layer Chromatographic Method fro Screening Antioxidant Compounds and Discrimination of Hibiscus sabdariffa by Principal Component Analysis. J. of Planar Chromatography.
  • Type: Journal Articles Status: Under Review Year Published: 2014 Citation: Jin Wang,Vanessa Ferchaud, Yadong Qi, Kit L. Chin, Yongde Yue,Fen Tang, QingLi Wu and Jim Simon.2014. A Validated HPLC-DAD Method for Fingerprint Analysis and Simultaneous Determination five Flavonoids from the leaves of Thirty-one Accessions of Hibiscus sabdariffa. J. of Food Composition and Analysis.
  • Type: Other Status: Other Year Published: 2014 Citation: Eva Dunbar Davis. 2013. The Health Benefits of Edible Hibiscus. Southern University M.S. degree capstone project paper. 14pp.


Progress 09/01/12 to 08/31/13

Outputs
Target Audience: This project targeted small and limited resource businesses, farmers’ market vendors, farmers, rural and urban residents, producers who could grow alternative crops for additional income. Also targeted were graduate and undergraduate students, scientists, extension agents and those with alternative medicine and functional food interests. Other target audiences were administrators at 1890 and other land grant institutions, homemakers, families, and other interested businesses. In collaboration with NIFA funded project (2008-38814-04772), more than 80 internationalparticipants attended the presentation and interacted with the PD as speaker on "Roselle hibiscus (Hibiscus sabdariffa) as a niche market crop for product development" at the International Conference and Exhibition on Food Processing and technology. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? It has offered an opportunity for the project director to present roselle hibiscus collaborative research findings to a group of international professionals and attendee at the World Congress of Nutirtion and Health and acquired scientific inputs for current and future product development endeavors. It also offered opportunities to visit roselle hibiscus research sites and interact with scientists. How have the results been disseminated to communities of interest? The initial results were disseminated to communities via conference/workshops. What do you plan to do during the next reporting period to accomplish the goals? An effort to carry out research activities and experiments to accomplish the goal and objectives of the project, particularly on the screening of varieties, carryinggenetic identification of the four selected accessions andanalyzing the photochemical and antioxidant contents of roselle hbiscus foliage.. In vitro evaluation of the roselle leaf extracts on the inhibition of cancer cell proliferation will be carried out collaboratively with West Virginia State University. The plan also attempt to exhibit the developed products and make more scientific presentations

Impacts
What was accomplished under these goals? Four selected roselle hibiscus accessions from the previous project were planted in the field along with Jamaica accession derived from early maturing fruits (harvested in July 2012). The macro and micro nutrient elemental composition, total polyphenol and antioxidant contentsof the leaves and calyces of two roselle hibiscus accessions were quantified. A new Malaysian accession was added to the field selection . Requisition of materials and supplies and equipment necessary for the initiation of product development were initiated. In collaboration with NIFA funded project (2008-38814-04772), more than 80 international participants attended the presentation and interacted with the PD as speaker on "Roselle hibiscus (Hibiscus sabdariffa) as a niche market crop for product development" at the International Conference and Exhibition on Food Processing and technology. Roselle hibiscus research at Southern University and selected products were exhibited at the Southern University's Legislative Day at the State Capitol. The exhibits were viewed and the developed products were sampled by more than 120 people (legislators, business entity, professional, students and general public). Five students presented the conceptualized roselle hibiscus business plan at the Opportunity Funding Corporation annual conference held in Atlanta, GA. The presentation has sparked a great deal of interest by attending entrepreneurs for product development venture. More than 65 farmers, homemakers, retired church volunteers from various parishes attended the roselle hibiscus product development workshops and field days. Two graduate students completed MS thesis work based on this project. One graduate student presented a poster presentation at the annual Council for Medicinally Active Plant Cnference.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Chin, K.L., Y. Qi. QL Wu, H.R. Juliani, J. Simon and G. Hankins. 2013. Comparative study on roselle hibiscus varieties for the development of natural products. BIT's First World Congress of Nutrition and Health. Dalian, P.R. of China. p. 39.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Chin, K. L., Y. Qi, D. Jaroni, G. Hankins and J. Simon. 2012. J. of Food processing and Technology 3 (10): 80