Using Flavonoid and Polyphenol Testing of Honey to Improve Consumer Education

Progress report for FW21-387

Project Type: Farmer/Rancher
Funds awarded in 2021: $25,000.00
Projected End Date: 09/30/2022
Host Institution Award ID: G308-21-W8613
Grant Recipient: Rock Front Ranch
Region: Western
State: California
Principal Investigator:
Alisha Taff
Rock Front Ranch
Expand All

Project Information

Abstract:

The American honey market is flooded with adulterated honey and consumers don't know how to determine which honey to buy. Until recently, there has been little technology for testing of honey to determine each honey's unique polyphenolic characteristics. 

Rock Front Ranch's honey is produced in a geographically isolated area with a unique diversity of flora for bees to forage, including some plants with recognized healing components.

Rock Front Ranch proposes to use state-of-the-art  Sweetwater Science Labs to determine the content and makeup of its two varieties of honey: Sage Honey and Wildflower Honey. The lab will use Liquid Chromatography Mass Spectroscopy to determine that pesticides and  pollutants are not present, in addition to listing each flavonoid contained in the honey.

Rock Front Ranch will apply this research by describing the healing qualities of these flavonoids (for example, antioxidants and antiseptic compounds) and promoting the honey for its unique characteristics.

Results will be published in a peer-reviewed scientific paper led by Dr. Michael Greenlief, who is the director of the Chemistry Mass Spectrometry Facility at the University of Missouri.

Workshop and presentations will  be held in CA for beekeepers and consumers. to describe the results of the testing and the potential effect on consumer health of the identified compounds. The benefit of using the results of the study to enhance honey marketing will also be discussed.

In addition, an informative brochure will be prepared  to educate beekeepers and consumers on the health-giving qualities of honey.

PROGRESS REPORT as per timeline dates:

Collected and submitted sample Aug. 2021. Received analysis Nov. 30, 2021. Created NMR handouts and new NMR page on website Fall of 2021.

PROCESSING AND BOTTLING WAS DRASTICALLY IMPACTED BY THE DROUGHT.

Education continued, with online interviews, online presentations and a video. Due to Covid 19, no in-person classes or workshops were given except for one presentation to Rotary Club, sponsored by Allan Hancock College in Santa Maria, CA.  attended by 25 people, 6 of whom were farmers, 4 were educators and 15 were members of the community.

 

Project Objectives:

Rock Front Ranch (RFR) proposes to utilize state-of-the-art testing from Sweetwater Science Lab in MO to use liquid chromatography mass spectroscopy to determine and analyze a wide range of flavonoids and polyphenols in RFR honey samples. Scientific studies show that it is the phenolic acids and flavonoids in honey which exhibit a wide range of biological effects and act as natural antioxidants. An article in Bee Culture, by Tianna Kolody, April 23, 2018 cites a study by Pyrzynska and Biesaga, in 2009, which states that it is the flavonoids and phenolic acids which give honey its unique flavors and antibiotic characteristics.

RFR's isolated geography and diversity of flora provide a rich source of certified glyphosate-free honey. Using Sweetwater's testing, we seek to show a direct connection between the components that go into the honey and its unique character, and to demonstrate a high degree of antioxidant properties which could prove the honey to be an excellent immune-system booster. During the time of COVID 19, this research becomes increasingly important as consumers are looking for natural foods that will assist them in staying healthy.

RFR will educate consumers on its findings by publishing a scientific paper in conjunction with Sweetwater Science labs, presenting at a symposium at the University of MO,  creating consumer-friendly marketing brochures, and by presenting 2 workshops and a Zoom presentation for beekeepers and consumers, detailing the findings and the implications for better consumer health.

Workshops for farmers were presented via Zoom.

Informative articles were shared on social media. Bee gold: Honey as a superfood (knowablemagazine.org)

 

Timeline:

Farmer Rancher Timeline

The timeline for the project is an Xcel sheet that includes dates, people responsible for each activity, milestones, and impacts on farmer/beekeepers, the market, and knowledge of the international community. For more details on each activity, please refer to Research Plan and Education Plan sections in this proposal.

Cooperators

Click linked name(s) to expand/collapse or show everyone's info
  • Dr. James Gawenis, PhD - Technical Advisor (Researcher)
  • Gary Troester - Technical Advisor

Research

Materials and methods:

Bees on Sage flowers.
Sage is still green and producing flowers in a very bad drought year.
Purple Sage in 2020.
Bees forage on purple sage during good years and bad.

Research Objective 1) Research on how foraged plant characteristics may affect the therapeutic value and taste of the two types of honey that we market. The testing will be performed by Sweetwater Science Labs using modern statistical software in combination with advanced analytical instrumentation [Liquid Chromatography Mass Spectroscopy (LCMS)]. Research began in 2021 by establishing a baseline for comparison of the more comprehensive study to be completed in summer 2022.

 

Research Objective 2) Tie the results of Research Objective 1 to an observational study of what plants are blooming at Rock Front Ranch at the time the honey was produced. Take special note of any plants with recognized therapeutic value. Correlate the honey characteristics determined by the testing with observed bloom times and correlate that to existing research on the therapeutic characteristics of the plants.

Black Sage and Purple Sage blooms were identified on the ranch in March of 2022.

Potential healing characteristics of Black Sage and Purple Sage have been identified.

  The healing characteristics of the plants found in our research will be compared with the honey, based on polyphenol and flavonoid content of the honey as identified by Sweetwater Science Labs.

Research Objective 3) Determine what therapeutic benefits (e.g. wound healing and cough suppression) could be derived from the identified flavonoids and polyphenols.

Sweetwater Science Labs specializes in food authenticity testing, with an emphasis on honey testing. They use a combination of liquid chromatography mass spectroscopy (LCMS) and nuclear magnetic resonance (NMR) for the most complete results in flavonoid and phenolic testing. This methodology is so new, and this advanced equipment is so necessary for it, that there is very little information available on these components of honey. We have found  studies in China and Poland, but, in the US, our study may be the first of its kind. It is important because it can potentially prove that honey has antiseptic and antibiotic actions, as well as a host of other potentially beneficial effects. See studies below.

Dr. James Gawenis, PhD, Sweetwater's co-founder,  chemist and CEO will work closely with Alisha Taff and the WIHPA team led by Nancy Smith on the analysis and dissemination of results. Sweetwater Science Labs has scheduled a presentation in 2022 at the Midwest Regional Chapter of the American Chemical Society convention in Iowa. The abstract of the presentation will be finished in July of 2022 and will be attached to this grant. The abstract will include a comparison of the honey from 4 different hives as well as a validation of the plant source. The presentation will be by Dr. Michael Greenlief and Dr. Jim Gawenis. 

In addition, Dr. Gawenis and Dr. Greenlief, who also teaches and collaborates at the University of Missouri, will oversee the publication of a scientific study detailing the methods used for honey analysis, the effect of the isolated geography of Rock Front Ranch and its diverse flora on the scientific components of the honey, and the outcomes of the comparison between the honey from 4 different hives.

Due to the effects of the drought that Rock Front Ranch is experiencing, the initial 2021 overall wildflower sampling did not occur but samples of Black Sage and Purple Sage were obtained. Because Black Sage has been found blooming in March 2022 at Rock Front Ranch, it is assumed that  samples of Black Sage Honey and Purple Sage Honey will be collected in summer of 2022 and sent to Sweet Science Labs for analysis, the results of which will be included in the presentation at the aforementioned Midwest Regional Chapter of. The samples will be of honey that is extracted and centrifuged, but not heated or adulterated.

A record of flower bloom time that corresponds with bee foraging will be kept by the Principal Investigator, Alisha Taff. That record will be correlated with the final results of testing to inform which flowers seem to contribute to the most flavonoid and polyphenol content.

Timing of testing will correspond to the collection of samples.

a) Tested initial samples of stored 2020 honey to establish a baseline. 

b) Tested samples of 2021 Black Sage and Purple Sage honey.

c) Harvest and process honey samples from 2022 crop of honey in July and August 2022. Send to Sweetwater Science Labs.

e) Receive analysis from lab and compare to observed bloom times on the ranch.

Publications will include a scientific study published with the assistance of Dr. James Gawenis and associates from the University of Missouri in late summer/early fall of 2022.

In a 2009 study by Krystyn Pyrynska and Magdalena Biesaga (Dept. of Chemistry, University of Warsaw, Poland), Analysis of phenolic acids and flavonoids in honey, the abstract states: "Honey is rich in phenolic acids and flavonoids, which exhibit  a wide range of biological effects and act as natural antioxidants. The analysis of polyphenols has been regarded as a very promising way of studying floral and geographical origins of honeys. . .We pay particular attention to. . .high-performance liquid chromatography and electrophoresis)." this study gave us confidence that we are on the right track with our proposal. As mentioned, we are using a private laboratory, Sweetwater Science Labs, to use high-performance liquid chromatography as well as nuclear magnetic resonance to determine the number of types of phenolic acids and flavonoids to enable us to educate consumers as to the therapeutic nature of our honey in particular and all pure, unadulterated honey in general.

Additional studies include: 

Analysis of Polyphenols in Honey: Extraction, Separation and Quantification Procedures. Pascual-Mate et al. 13 July 2017. Spain.

The abstract states: "High performance liquid chromatography with ultraviolet or mass spectrometry detectors is by far, the most commonly employed instrumental procedure to separate and quantify polyphenols in honey."

This study gives us confidence that we are on the right track with our testing program.

Phenolic acids and flavonoids profiles of commercial honey from different floral sources and geographic sources. Yiuchung Cheung et al. March 4, 2019. China.

The abstract states: "Honey is well-known for its nutritional and medicinal values since prehistoric times. In the present study, 40 honey samples were collected from supermarkets in China. .  .These samples were explored for the detection of 16 phenolic acids and 14 flavonoids by employing high-performance liquid chromatography (HPLC). . .it was observed that certain phenolic compounds significantly contributed towards the classification of honey samples. . .Thus, this systematic study provides a fundamental knowledge of honey quality for therapeutic product development."

Honey for Nutrition and Health: A Review. Stefan Bogdanov et al. Article, Journal of the American College of Nutrition. 2013

The abstract states: "Due to the variation of botanical origin honey differs in appearance, sensory perception and composition. The main nutritional and health relevant components are carbohydrates. . .but also about 25 different oligosaccharides. It contains small amounts of proteins, enzymes, amino acids, minerals, trace elements, vitamins, aroma compounds and polyphenols. The review . . . shows that honey has a variety of positive nutritional and health effects, if consumed at higher doses of 50 to 80 g intake."

 

Research results and discussion:

Nov 30 2021 Sweetwater Analysis

   Two major challenges in 2021, the pandemic and an extreme drought, affected our ability to achieve the level of results we had hoped for in terms of the variety of wildflower floral sources. However, we were able to harvest honey samples of Black Sage Honey and Purple Sage Honey that were sent to Sweetwater Laboratories for analysis using Nuclear Magnetic Resonance to obtain full honey profiles. (Attached above.)

projects.sare.org/…/Analysis_of_phenolic_acids_and_flavonoid.pdf

   Research is ongoing into the healing properties of honey in general. Our study will build on this research and add to the body of knowledge around the healing properties of honey and specific plant properties that contribute to the makeup of the honey.

   The attached scientific study by Krystyna Pyrzynska and Magdalena Biesoga, published in Trends in Analytical Chemistry, Vol. 28, No. 7, 2009 will be used to build our research upon. The advanced equipment available from Sweetwater Science Labs and the University of Missouri chemistry department under Dr. Michael Greenlief serve to enable us to obtain more detailed and specific information about the composition of our honey in relationship to the flowers that are available for the bees to forage than the 2009 study was able to provide. Sweetwater Science Labs will also compare the study produced in conjunction with this project with a 2020 report they prepared previously for Rock Front Ranch.

The results of a November 30, 2021 Sweetwater Analysis which are attached, show no discernible pesticides or antibiotics in Rock Front Ranch honey.

The lengthy 2022 report which will be prepared using samples from the 2022 honey crop (or limited honey samples if drought continues into 2022) will show whether there are any discernible levels of antibiotics and/or pesticides and also will provide validation of floral sources in the tested honey.

A challenge caused by the ongoing drought at Rock Front Ranch is that the number of floral sources will be far fewer than they were in 202o. So far, only Black Sage and Purple Sage are blooming. This is being met by increased research that we are doing currently into the possible medicinal benefits of Black Sage and Purple Sage. We are researching these two herbs more deeply than originally planned because they are currently blooming, and it is assumed that we will have fewer herbs overall to research if the drought continues. The overall results of the project will be served by the deeper research, and our use of NMR testing in our marketing will continue with more emphasis on these two herbs that are over 51% present in our "Black and Purple Sage Honey". It is required that a floral source be 51% or over in order for a honey to be named for that source. In the past, we moved the bees into the Black Sage and Purple Sage fields and then removed them in order to preserve the validity of our floral source. Our "Wildflower Honey" is more threatened by the drought, as the multifloral nuances we have obtained in the past may be threatened by a dearth of wildflowers. Sweetwater Science Labs will test for and identify the floral sources in the wildflower honey compared to the wildflower sources in our 2020 honey. So far, 2020 has had the largest variety of floral sources of any year we have bottled honey.

projects.sare.org/…/Analysis_of_phenolic_acids_and_flavonoid.pdf

Flowers identified by Rock Front Ranch that have reputed medicinal qualities to be verified in the honey profile by Sweetwater Science Labs:

Yerba Buena: (will most likely only bloom if extensive rainfall occurs.) Full of antioxidants. Also contains caffeine. Reputed to give energy boost. Also reputed to improve skin quality, soothe inflammation, soothe stomach issues (inhaling scent), relieve asthma (crush leaves and inhale scent),  build immunity (contains catechins and polyphenolics that could be identified in the honey with NMR testing.

Manzanita: (Rock Front Ranch staff is currently watching for manzanita to bloom this year.) Used to treat a variety of digestive issues and promote relaxation. Considered relatively safe, but it is recommended that you check with a health practitioner before using, especially if you have serious health issues or are taking prescribed medication.

Black and Purple Sage-artemisia nova, salvia mellifera:  (is currently blooming on the ranch as of March 2022.)Used externally to relieve sore muscles and joints.

MARKETING RESEARCH:

Polls will be conducted on social media and in person in stores asking customers whether their interest in purity and potentially healing components is low, medium or high. We will then compare sales pre- and post- NMR testing advertising campaign to determine whether testing is helpful in marketing honey. There has been very little research done in this area and the results will inform the marketing efforts of other farmers. Results will be shared with other farmers, universities, and with presentations.

 

 

 

Participation Summary
1 Farmer participating in research

Educational & Outreach Activities

4 Consultations
2 Curricula, factsheets or educational tools
2 Online trainings
3 Published press articles, newsletters
1 Tours
4 Webinars / talks / presentations
4 Other educational activities: To meet the challenges of the pandemic, presentations and consultations were mostly via Zoom. In winter 2021, Alisha Taff participated in a round table discussion through local Allan Hancock College. The presentation involved some national level experts such as Dr. Tim LaSalle and Hunter Francis as well as staff from the college. The roundtable discussion was attended via Zoom by dozens of local residents, including college staff, farmers and interested consumers.
On World Bee Day in 2021, Alisha Taff participated in a facebook live presentation describing the risks of honey contamination and the value of NMR testing.
On February 22, Alisha Taff did a Zoom presentation through Allan Hancock College, on beekeeping in general, including the importance of testing for purity.
Plans are underway for a facebook live presentation for farmers and consumers through SLO Food Coop, using facebook live. SLO Food Coop will bring many participants to the table and will handle the logistics of the presentation. Alisha Taff will make the presentation.
2 presentations were given to middle school children at Central Coast Unified School District.

Participation Summary:

25 Farmers
6 Ag professionals participated
Education/outreach description:
The attached page from our website describes the importance of NMR Testing.
Our website was updated to include educational material about NMR testing, presented in a way that the average consumer can understand. This page can be seen on our website: www.rockfrontranchhoney.com

RFR Honey Sell Sheet

RFR's Honey Sell Sheet was updated to feature artistic renderings of Cuyama Valley wildflowers as well as information about NMR testing and it emphasizes the characteristic of glyphosate free honey.

 

On May 20, 2021, Alisha Taff was interviewed on facebook live. The topic was broadly about beekeeping, but her emphasis was on the importance of bee health, the influence of glyphosate (the absence of which we identify using NRM testing of our honey), and the importance of bee health in general on consumers and on the environment.

On February 8, 2022, Alisha gave a presentation to the Rotary Club of Santa Maria, CA through Allan Hancock College. This presentation included the importance of bee health and the use of NMR testing to determine the purity of honey.

Two discussions/presentations were presented via Zoom through Allan Hancock College of Santa Maria, CA.

Two outdoor presentations were given to middle school children of Central Coast Unified School District.

Below is a link to our Rock Front Ranch facebook page World Bee Day interview.

(20+) Facebook 

 

 

Learning Outcomes

3 Farmers reported changes in knowledge, attitudes, skills and/or awareness as a result of their participation

Project Outcomes

2 Farmers changed or adopted a practice
1 Farmers intend/plan to change their practice(s)
2 New working collaborations
Project outcomes:

Nov 30 2021 Sweetwater Analysis

The purpose of the project was to contribute to beekeeping sustainability by researching the effects of adding the results of NMR testing to the marketing efforts of Rock Front Ranch Products LLC. The 2021 report from Sweetwater Labs in Columbia, MO is attached. This report shows a very high level of purity in the RFR honey. The results have been published on social media and in direct e-mail contacts with honey buyers from a number of supermarket chains and Whole Foods Markets. Response from the public has been favorable. 

Because of the extreme drought in the Cuyama Valley in 2021, and continuing into 2022, the honey we are marketing in 2021 and 2022 is actually from our 2020 crop. All our NMR testing in each year has shown the absence of pesticides or glyphosates. Response from the public is very positive to this information. We have some wholesale customers, including over 40 Whole Foods Markets, who tell us the NMR testing is a major reason why they choose our honey. The number one reason is the amazing taste of our very pure honey, and the number two reason is now the NMR testing.

This logo is added to our honey marketing and used on social media to show that our purity is documented.
NMR testing verified.
Success stories:

A beekeeper in Honoka'a Hawaii has become aware of the importance of honey testing. Her business is growing and she also gives workshops to beekeepers on the Big Island of Hawaii. She will be incorporating the results of our workshop into her classes and workshops and has applied for a grant to form a local marketing cooperative that will also emphasize testing and glyphosate-free honey. As of March, 2022, this Hawaii project has received notice of funding. 

 

Recommendations:

The challenge of extreme drought has rendered our study less robust than we originally intended. By increasing the number of presentations given, and increasing our NMR testing marketing efforts we hope to accomplish the same results of educating the public and increasing farmer income by using testing to verify the purity and qualities of our honey. 

The use of Sweetwater Labs to perform the testing gives us verifiable credentials and has sparked the interest of research scientists and academics, one of whom plans to write a scientific study of our project. The results of the study will be presented at a symposium (probably on Zoom because of Covid19) through the chemistry department at the University of Missouri at Columbia. This approach will improve consumer education and spark interest in the scientific community as well as among knowledgeable consumers. This is a very important part of our project.

Our approach relies heavily on our ability to reach consumers with the importance of honey purity and the confidence that NMR testing gives us that our honey is pure. The biggest challenge we have had is the inability to host on-farm workshops due to Covid. We are confident that the venues we have been able to use (Zoom, Facebook Live, and, more recently, a public presentation, have allowed us to achieve the same type of educational and marketing results that our original plan would have provided. In fact, we think our reach has actually increased due to our efforts to provide the information virtually that would have been limited to a smaller group of participants in classes and workshops.

An insight we have gained from that result is that we should continue to use online teaching even after the pandemic, in order to reach more farmers and consumers. In the future, we will increase our online educational and marketing efforts, and will advise other beekeepers to do the same.

Information Products

    Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture or SARE.