Source: LINCOLN UNIVERSITY submitted to NRP
ASSESSMENT OF NUTRITIONAL, BIOACTIVE COMPONENTS AND HUMAN HEALTH BENEFITS OF GREEN LEAVES OF QUINOA
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1028544
Grant No.
2022-38821-37301
Cumulative Award Amt.
$499,335.00
Proposal No.
2021-12868
Multistate No.
(N/A)
Project Start Date
May 1, 2022
Project End Date
Apr 30, 2026
Grant Year
2022
Program Code
[EQ]- Research Project
Recipient Organization
LINCOLN UNIVERSITY
820 CHESTNUT ST
JEFFERSON CITY,MO 651023537
Performing Department
Agriculture and Env Sciences
Non Technical Summary
Quinoa grain is eaten worldwide as a healthy food, but consumption of quinoa leaves is uncommon. The fresh young leaves and tender shoots of quinoa can be eaten in salads or cooked as vegetables. The traditional knowledge suggests that daily intake of at least one serving of leafy greens can prevent chronic diseases, including heart diseases, cancer, diabetes, obesity, and micronutrients deficiencies; however, scientifically, very little is known on the nutritional, phytochemical compositions, and human health benefits of green leaves of quinoa. We hypothesize that consumption of quinoa leaves provides nutrients that prevent and/or treat chronic disease. We will test our hypothesis by following the objectives to (1) assesses nutritional compounds, (2) characterize the bioactive components for antioxidation and health benefits, (3) explore the growth and development of beneficial bacteria in the human intestine, and (4) educate and train students in crop production. In addition, we will disseminate this information to farmers, consumers and scientific community through demonstration plots, field days, workshops, and publications. Our collaborative, integrated-multidisciplinary approach addresses the CBG program priority area "Human health, obesity as it relates to nutrition and human sciences." Our proposal is innovative because characterization of nutritional and health benefits of quinoa leaves will help us to introduce a nutrient-rich vegetable in American food chain and bring economic benefits to the farmers.
Animal Health Component
40%
Research Effort Categories
Basic
30%
Applied
40%
Developmental
30%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70114301010100%
Goals / Objectives
The overall goal of the project is to evaluate the nutritional, bioactive components and human health benefits of green leaves of quinoa. The following objectives have been planned to achieve the goal:Assessment of nutritional compounds such as protein, fat, amino acid, fiber, carbohydrate, minerals, and vitamins.The evaluation of functional bioactive compounds (phytochemicals) includes total phenolic compounds, flavonoid content, antinutritive factors, antioxidation capacity, anticancer and antidiabetic activities.Explore the influence of non-cellulosic polysaccharides and their derivatives on the growth and development of beneficial bacteria in the human intestine/gut.Dissemination of information to vegetable growers and consumers through demonstration plots, field days, and workshops to encourage adaption as a leafy green vegetable.Train students in crop science, especially the analysis of nutritional and phytochemical research.
Project Methods
Plan of Operation and Methodology?Plant materials: Three quinoa lines (e.g., Ames 13724, PI 614885, and PI 665275 will be used for this study. These lines will be grown in the high tunnel (March - April) and field (May - June) at the Carver farm of Lincoln University, Jefferson City, MO, for leaf tissue collection. The experiment will be set up in a randomized complete block design (RCBD) with three replications over two years. Four-week-old young leaves of three lines, three replications, two environments (high tunnel and field), and over two years will be collected for the study.Objective 1: Assessment of nutritional compounds of young leaves of quinoa- protein, fat, amino acid, fiber, carbohydrate, minerals, and vitamins.1.1: Proximate analysis: Protein, fat, fiber, moisture, and ash of dry leaves of quinoa will be determined following the procedure described by the Association of Official Analytical Chemists (AOAC, 2006).1.2: Amino acid analysis: The complete amino acid composition of dry leaves of quinoa will be analyzed using AOAC (2006) official method 982.30 E (a, b, c).1.3: Mineral analysis: Powder of dried leaves of quinoa will be used for mineral analysis. The mineral elements will be analyzed using the Agilent 5110 inductively coupled plasma - optical emission spectrometer (ICP-OES) instrument.1.4: Vitamins analysis: The different vitamins (A, B1, B2, B3, C, and E) in quinoa leaves will be determined using AOAC (1990).Objective 2: Evaluation of bioactive compounds (phytochemicals) in quinoa leaves- total phenolics, flavonoids content, antinutritive factors, antioxidation capacity, anticancer, and antidiabetic activities in green leaves of quinoa (VSU). 2.1: Total Phenolic Compounds: The total phenolic contents of each extracted sample will be determined using the Folin-Ciocalteu method as we described previously using Gallic acid as a standard (Al-Shaya et al., 2020).2.2: Flavonoid content: Flavonoid represents a diverse class of compounds, and their determination is often based on the use of appropriate standard and spectrophotometric procedures involving the reaction of AlCl3 in the presence or absence of sodium Nitrite (NaNO2). We will employ two different procedures to determine flavonoids as described previously (Al-Shaya et al., 2020).2.3: Antinutritive factors: Antinutritive activity will be determined by assaying the concentration of total saponins, phytic acid, oxalate, nitrate, and nitrite, and chymotrypsin/trypsin inhibitors. Saponins: The amounts of total saponins will be determined using a spectrophotometric method described by Benyong et al. (2014). Two grams of quinoa leaves powder will be extracted in ethanol, petroleum ether, and butanol as described and used for spectrophotometric analysis using Oleanolic acid as a reference standard.Phytic acid: The total phytic acid will be determined using the procedure described by Agostinho et al. (2016).Nitrate: The total Nitrate in quinoa leaves extracts will be determined using a spectrophotometric method as described by Lastra (2003).Oxalate: The total and soluble oxalate content in all the samples will be analyzed by following the titration method using KMnO4 as described in AOAC (1990) and modified by Madhu-Babu et al. (2018).Chymotrypsin/Trypsin inhibitors: The inhibitory activity in the leaves extract of quinoa will be determined against chymotrypsin and trypsin using the substrate BTpNA and BApNA, respectively, in Tris (2-Amino-2-hydroxymethyl-propane-1,3-diol) buffer, pH 8.0 at 37 °C in 30 min as described by Pesoti et al. (2015).2.4: Antioxidation Capacity: A comparative analysis of antioxidation activity in each quinoa leaves sample will be determined using chemical assays including 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) method, and 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+) method for the oxygen scavenging capacity, and Ferric ion reducing antioxidation power (FRAP) assay.2.5: Anticancer activity: We will determine the anticancer activity of quinoa levees extract in colon cancer cells following Al-Shaya et al., (2020).2.6: Antidiabetic activity: Antidiabetic properties will be determined by assaying inhibition of alpha-amylase and alpha-glucosidase activities by quinoa extracts as described by Sekhon-Loodu and Rupasinghe (2019).Objective 3: Influence of non-cellulosic polysaccharides and their derivatives on the growth and development of beneficial bacteria in the human intestine, thereby boosting immunity. (MSU- KH)Non-cellulosic polysaccharides arabinoxylans (AX) will be extracted from freeze-dried quinoa leaf following a method of Lopez et al. (1999) with slight modification as described by Sarker et al. (2020). Arabinoxylan oligosaccharides (AXOS) will be synthesized enzymatic hydrolysis of AX using AX degrading enzyme. Extraction of ferulic acid from freeze-dried quinoa leaf will be performed by alkaline hydrolysis method (Arabi et al., 2016), and Short-chain fatty acid (SCFA) will be extracted using the Reverse Phase HPLC method (Vargas et al., 2020).In the colon, the main types of bacteria are obligate anaerobes. They are members of the genus Bacteroides, anaerobic gram-positive cocci, such as Peptostreptococcus sp., Eubacterium sp., Lactobacillus sp., and Clostridium sp. We will culture the selected bacteria of these species using different concentrations of AX, AXOS, ferulic acid, and SCFA. Comparing with the respective control, we will determine the influence of these compounds on the growth and development of these compounds' beneficial bacteria in our intestine.Objective 4: Dissemination of information to vegetable growers and consumers through demonstration plots, field days, and workshops to encourage the adaption of quinoa as a leafy green vegetable.Three demonstration plots will be set-up in each year for the second and third year of the project, one in farmers' high tunnel and two others in the field (total 6 plots). Similar demo plots will be arranged at Lincoln University Carver farm high tunnel and field. An annual quinoa field day and workshop will be organized at LU's Carver Farm to disseminate information on quinoa production. Farmers/vegetable growers, extension, and marketing people will be invited to attend the event. An economic analysis will be conducted to evaluate the feasibility and net benefit of vegetable quinoa production by estimation all the relevant costs and yields associated with vegetable quinoa production. Information on vegetable quinoa production will be disseminated through the production and distribution of fact/guide sheets, LU extension programs, newsletters, and departmental websites. In addition, findings will be presented at scientific meetings and conferences and published in peer-reviewed journals.Objective 5: Train graduate students in crop science, especially the analysis of nutritional and phytochemical research. A total of one graduate from LU and three undergraduate students, one each from LU, VSU, and MU, will work on this project. These students will be intricately involved with all project activities and will increase their knowledge and strengthen their research capabilities in the areas of advanced sciences and technologies.Data analysis: Statistical analyses for all data (data generated in objectives 1, 2 and 3) will be analyzed using SAS® version 9.3 packages (SAS Institute, 2011). Mean differences will be determined using Fisher's least significant difference (LSD); a p-value of <0.05 is considered to be significant.

Progress 05/01/24 to 04/30/25

Outputs
Target Audience:The target audience (s) are: - small and marginal farmers/crop producers, - consumers, - extension personel, - scientific communities, - authentic food resturants, Changes/Problems:1. Research progress was temporarily slowed when one of the PIs ( Dr. Rafat Siddiqui of Virginia State University) suddenly died in July 2024. We are recovering now with minor adjustments by hiring one Post-doc. 2. We have requested a one-year, no-cost extension (May 2025 to April 2026). If it is granted, we can finish the project according to our plan during the extended period (May 2025 to April 2026). What opportunities for training and professional development has the project provided?1. A 'Quinoa Field Day' was organized on July 2024 at Lincoln University Carver Farm, near Jefferson City, MO. 2. Participation at the Missouri State Fair in August 2024 to promote quinoa production as well as the health benefits of both quinoa greens and grains. 3. Publishing articles and oral and poster presentations. How have the results been disseminated to communities of interest?1. Demonstration plots for quinoa greens and grains production were established at Lincoln University Carver Farm, Jefferson City, MO, and Columbia Center for Urban Agriculture (CCUA), Columbia, MO. 2. Publications, presentations at meetings, conferences, and state fairs, 3. Distribution of handouts. 4. Organizing a 'Quinoa field day." What do you plan to do during the next reporting period to accomplish the goals?1. Organizing a 'Quinoa Field Day' in July 2025 at LU Carver Farm, MO. 2. Presenting research findings at different meetings and conferences. 3. Participate as keynote speaker at the '7th International Nutrition Research Conference' to be held in Singapore in March 2025 and talk about 'Quinoa Green Leaves: A Promising Nutrient-rich Vegetable to Improve Human Health.' 4. Establish demonstration plots at Lincoln University Carver farm, Jefferson City, MO, and Columbia Center for Urban Agriculture (CCUA), Columbia, MO during summer 2025. 5. Manuscript preparation and publication.

Impacts
What was accomplished under these goals? It is the third year of the three-year project (May 2022 to April 2025). We have requested a one-year no-cost extension (May 2025to April 2026). The following are the accomplishments during the third-year project period (May 2024 to April 2025). 1. One manuscript entitled 'Nutritional composition of quinoa (Chenopodium quinoa Willd.) inflorescences: A comparison with quinoa green leaves and grains' has been published in 'Crops' journal, doi.org/10.3390/crops4010006, in 2024. 2. One international presentation in Bangladesh about the multipurpose use of quinoa in April 2024. Also,presented one poster at the tri-Socitey Meeting in San Antonio, TX, in November 2024. 3. Biochemical and bioactive components analysis of quinoa leaf tissues has been completed, and data analysis and manuscript preparation is in progress. 4. Quinoa field day was organized at the Carver Farm of Lincoln University in July 2024. About 70 participants attended, including vegetable growers, extension personnel, consumers, and research scientists. The President of LincolnUniversity, and the Dean and Research Director of the College of Agriculture and Environmental Sciences inaugurated theday-long event. 5. One graduate student will graduate in May 2024. 6. Established demonstration plots for quinoa greens and grains production, including using young quinoa whole plants as a forage for small ruminants.

Publications

  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Pathan, S.; Ndunguru, G.; Ayele, A. G. Comparison of the Nutritional Composition of Quinoa (Chenopodium quinoa Willd.) Inflorescences, Green Leaves, and Grains. Crops 2024, 4, 7281. https://doi.org/10.3390/crops4010006
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Ndunguru, G., Pathan, S., and Ayele, G. 2024. Maximizing quinoa harvest- yielding leafy greens and grains from a single plant. Poster presented at the Tri-Society Annual Conference, San Antonio, TX.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Pathan, S. 2024. Use of different plant parts of quinoa- nutrients rich entities. Paper presented as an invited speaker at the BSMR Agricultural University, Gazipur, Bangladesh in April, 2024.


Progress 05/01/23 to 04/30/24

Outputs
Target Audience:The target audience (s) are: - small and marginal farmers/crop producers, - consumers, - extension personel, - scientific communities, - authentic food resturants, Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?1. Organized a 'Quinoa Field Day' on July 2023at Lincoln University Carver Farm, near Jefferson City, MO. 2. Publishing article, oral and poster presentation. How have the results been disseminated to communities of interest?1. Demonastration plots for quinoa greenswere established at 5different places, such as Lincoln University Carver Farm;Columbia Center for Urban Agriculture, Columbia, MO; Science Center at St. Louis, St. Louis, MO; Living Roots Urban Farmat St. Louis, MO; Alabama State University, Montgomery, Al. 2. Publications, presentations at meetings, conferences, and state fair, 3. Distribution of handouts. 4. Organizing a 'Quinoa field day." What do you plan to do during the next reporting period to accomplish the goals?- Organizing a 'Quinoa Field Day' in July 2024 at LU Carver Farm, MO. - Presenting research findings at different meetings and conferences. - Establish demonstration plots - Manuscript preparation and publication.

Impacts
What was accomplished under these goals? It is the secondyear of the three-year project. The following are the accomplishments during the second-year project period (May 2023to April 2024). 1. A comparative nutrional analysis of quinoa green leaves,inflorescences and grains have completed. A manuscript entitled 'Nutritional composition of quinoa (Chenopodium quinoaWilld.) inflorescences: A comparison with quinoa green leaves and grains' has submitted to 'Crops' journaland is under review. 2.Biochemical andbioactive componentsanalysis of quinoa leaf tissues has been completed and data analysis and manuscript preparation arein progress. 3. Quinoa field day was organized at the Carver Farm of Lincoln University in July2023. About60 participants, incuding vegetable growers, extension personnel, consumers, and research scientists were attended. The President of Lincoln University and the Dean and Research Director of the College of Agriculrure and Environmental Sciences inagurated the day-long event. 4. One graduate student has joined the project in August, 2023. 5. Demonastration plots for quinoa greens and grains production were established at 5different places, such as Lincoln University Carver Farm:Columbia Center for Urban Agriculture, Columbia, MO; Science Center at St. Louis, St. Louis, MO; Living Roots Urban Farmat St. Louis, MO; Alabama State University, Montgomery,Al.

Publications

  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Pathan, S., Ndunguru, G., and Ayele, A.G. 2023. Quinoa: A Super Crop with many uses. Paper presented at the Tri-Society Annual Meeting, St. Louis, MO.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: S. Pathan*, G. Ndunguru, M.K. Shahadat, A.G. Addissu. 2024. Use of Nutrient-rich Different Plant Parts of Quinoa. Paper presentation at the ARD Symposium, 6-10 April 2024, Nashville, TN.


Progress 05/01/22 to 04/30/23

Outputs
Target Audience:The target audience (s) of this project are crop growers/farmers, extension workers,leafy green consumers, and scientific community. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?1. Organized threequinoa greens demonstration plots. 2. Organizeda 'Quinoa field day' in May 2022. What do you plan to do during the next reporting period to accomplish the goals?1. Set up quinoa greens in five demonstration plots from May to September 2023. 2. Schedule a 'Quinoa field day' in July 2023. 3. Making oral and poster presentations at CSSA-ASA annual meeting in October 2023 and the ARD biennial research symposium in 2024.

Impacts
What was accomplished under these goals? It is the first year of the three-year project. The following are the accomplishments during the first-year project period (May 2022 to April 2023). 1. One article has been published, including data of proximate analysis (protein, fat. carbohydrate, fiber), amino acids, and minerals. (Pathan et al. 2023. Horticulturae, 9, 209. https://doi.org/10.3390/horticulturae9020209). 2. Leaf tissues were collected, and ground leaf powder distributed to all three participating Universities. Biochemical analysis at Virginia State University and nutritional analysis at Lincoln University are in progress. 3. Study of the 'influence of quinoa green's non-cellulosic polysaccharides and their derivatives on the growth and development of beneficial bacteria in the human intestine/gut' is in progress at Mayville State University, ND. 4. Quinoa greens field day was organized at the Carver Farm of Lincoln University in May 2022 with 30 participants, incuding vegetable growers, extension personnel, consumers, and research scientists. The President of Lincoln University and the Dean and Research Director of the College of Agriculrure also visited the quinoa greens plot in Carver farm high tunnel in May 2022. 5. One undergraduate and one graduate student have been recruited for the project.

Publications

  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Pathan, S. 2022. Quinoa: A nutrient-rich, climate smart dual-purpose new crop for grains and greens production in Bangladesh. Paper presented at the special conference at the Sher-e-Bangla Agricultural University, Dhaka, Bangladesh 11 December 2022.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Pathan, S., Ndunguru, G., Islam, M.R., Jhumur, S., and Ayele, A. 2023. Production of quinoa leafy greens in high tunnel for season extension in Missouri. Horticulturae, 9, 209. https://doi.org/10.3390/horticulturae9020209