Progress 11/19/15 to 09/30/18
Outputs
Target Audience:The target audiences include other researchers in food science and nutrition either in academia or in industry, as well asfood product developers in industry that might use the information provided to them in the form of publications andpresentations to develop and market new functional food products with health-beneficial properties based on developedprototypes of functional foods containing compounds and bio-activities identified in native and non-native plants. Changes/Problems:The largest issue is that one the PhD students who has been working on the project, specifically the elderberry project, fell ill with a serious eye disease that has prevented him from making any significant progress over the last 1.5-2 years, specifically in the data analysis and report / dissertation writing aspect. What opportunities for training and professional development has the project provided?The three PhD students who have been working on this project, only one of which (Abhinandya Datta) has finshed his degree, received training in the use of chromatographic equipment and statistical analysis. How have the results been disseminated to communities of interest?Results have been reported in the form of presentations and publications. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Neem (Azadirachta indica) is an evergreen tree cultivated in various parts of the Indian sub-continent. It has been in use over centuries in the Indian folk medicine for its therapeutic value. Given the prominent role it has played in curing diseases of the villagers over centuries, it has been hailed as a "divine" tree, a "village dispensary" and " nature's drugstore" .Today, extensive research has shown that it may have anti-cancer, anti- diabetic, anti-inflammatory, anti-ulcerogenic, and anti-microbial effects. However, there are significant gaps in the scientific literature about the compounds contributing to its medicinal potential. In this study, select flavonoids, namely flavonols: myricetin, quercetin and kaempferol are quantified by a HPLC method in Neem powder and leaves as well as green and black tea leaves for comparison. It was observed that Neem dry powder had significantly higher (p< 0.05) flavonol content at 11.146 ±1.09 mg/g than green and black tea 5.782 ± 0.723 mg/g and 5.599 ± 0.484 mg/g respectively. The tea infusions of the plant materials also yielded similar results. The total phenolics were found to be the highest in green tea infusion because of its high flavan-3-ol content, followed by black tea, Neem powder and Neem tea cut leaves with values of 174.55 ± 12.85, 106.85 ± 6.511, 55.60 ± 7.90 and 87. 35 ± 7.42 mg/g respectively. DPPH and FRAP anti-oxidant assays yielded contradictory results which can be explained by the very principle of these assays. It was also observed that the particle size of ground tea leaves significantly influenced the yield of flavonols, phenolics and anti-oxidant assay. In addition, neem is extremely bitter, in large part due to its limonoid content, making it unpalatable and limiting its potential use in dietary supplements or foods alike. Bitterness reduction, especially of foods and beverages containing phytonutrients, is one of the biggest challenges in the food industry. The objective of this experiment of our overall study was to apply two adsorbent based strategies, namely solid phase extraction (SPE) and Amberlite XAD-16 (AMB) resin, to achieve de-bittering of neem tea and to determine the effects of the de-bittering on the bio-active, color, and volatile properties. The solid SPE treatment completely removed the flavonol, quercetin, from neem tea while in Amberlite XAD-16 treated tea (AMB) it was only insignificantly (p > 0.05) reduced. We also observed decreases in total phenolic content and consequently anti-oxidant activities after de-bittering. A 62% mean reduction of limonoid aglycones indicated diminished levels of bitterness. The loss of phenolics lead to a visually appreciable color change in the treated teas. The de-bittering also leads to a loss of sesquiterpenes, ketones and acids from neem tea. In conclusion, we found that while SPE cartridges were more efficient in removing bitterness, they caused a greater reduction in bio-active compounds than AMB XAD-16 resins, which may ultimately affect the health properties of neem tea. Over the last year, in the context of this project, we initiated an investigation into the bioactive compounds and the resulting bitterness of cacao and chocolate, specifically our goal is to identify and quantify the complex mixture of important bitter compounds within the three compound classes that are known to be important to cocoa--methylxanthines, flavan-3-ols, and diketopiperazines. Chocolate is made from the fermented, dried, and roasted seeds of the Theobroma cacao tree. It is now understood that cacao, also known in English-speaking countries as cocoa, contains a variety of compounds, including well-known flavonoid polyphenols, the consumption of which has positive impacts on human heart health and blood pressure, cancer reduction, LDL cholesterol reduction, and insulin resistance improvements, as numerous in vitro, in vivo, and observational studies have confirmed. In important research published in 2017, performed, in part, by Harvard's Chan School of Public Health, a participant observational study, composed of men and women between the ages of 50 and 64, led to the determination that chocolate consumption is inversely correlated with atrial fibrillation (AR). This is an important finding, as AR is associated with multiple significant health problems such as "higher risk of stroke, heart failure, cognitive decline, dementia and mortality." A persuasive case has, therefore, been made that cacao is a healthful addition to a balanced diet. However, despite the strong evidence for the healthful qualities of cacao, which has led to an overall increase in consumer purchases of more high-cacao-content chocolate than in previous decades, sales figures still show that there is an unwillingness by many Americans to consume higher-cacao-content chocolate, which tends to be more bitter, a taste modality not readily appreciated by most. Therefore, if the bitterness of cacao could be minimized, higher-cacao-content and lower-sugar chocolate confection sales could capture an even larger segment of the conventional and healthy snack-food market by achieving the elusive combination of being both tasty and healthy. The key is to better grasp the causes of bitterness in cacao, which are complex and still not well understood. What has long been known is that the methylxanthine, theobromine, is an important contributing bitter compound in cacao, with its name even being derived from the genus of the cacao tree, Theobroma. Similarly, the related methylxanthine, caffeine, which is well known as an important bitter compound in coffee, has also been found to impart bitterness in cacao and chocolate. Certain flavan-3-ols, which are a subclass of the aforementioned healthy flavonoids, and which are found in black tea, are also bitter and present in cacao at levels above the detection threshold, especially the compound (-)-epicatechin. After project initiation, cocoa beans were obtained from Ghana, Madagascar, and Peru and have been roasted according to a standard roasting profile by chocolate manufacturer Patric Chocolate, LLC. Methods have been developed and validated, and standard curves have been established for the cocoa bitter compounds: Caffeine, Theobromine, Catechin, Epicatechin, and selected Diketopiperazines (DKPs), including cyclo(L-Pro-L-Val) which according to the most recent literature available is considered the most important DKP in regards to cocoa bitterness. The bitter compounds caffeine, Theobromine, Catechin, and Epicatechin were found in all samples. While a preliminary analysis of the data showed, as expected, some variation, we still need to analyze the data statistically. We were not able to replicate the finding reported by Stark (Stark T, Bareuther S, Hofmann T. Molecular definition of the taste of roasted cocoa nibs (Theobroma cacao) by means of quantitative studies and sensory experiments. J Agr Food Chem. 2006;54(15):5530-9) that the diketopiperazine cyclo(L-Pro-L-Val) is an important bitter compound! While we were able to positively identify cyclo(L-Pro-L-Val) in a very dark roasted cocoa using LC-MS-MS, which then allowed us to also detect and quantify this diketopiperazine in other roasts, the concentration of cyclo(L-Pro-L-Val) is apparently considerably lower, i.e. only about 5-10% of that reported by Stark, which put its concentration at or below its sensory threshold. However, since Stark's sample was not well characterized, i.e. no information was given on roast profile or how many samples were analyzed, a direct comparison of results is difficult. On the other hand, we were able to detect this diketopiperazine even in unroasted samples, indicating that DKPs are formed already during the fermentation stage of the beans.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Gedikoglu A, Clarke AD, Gruen, IU. 2018. Determination of flavonoids in citrus fiber and the impact of processing method on total phenolic content, total flavonoid content, total carotenoid content and oxygen radical absorbance capacity. GIDA: The Journal of Food 43 (6): 917-924.
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2016
Citation:
Datta, A. 2016. FUNCTIONALITY OF AZADIRACHTA INDICA A. JUSS (NEEM) IN BEVERAGES. Dissertation, University of Missouri.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Datta A., Gr�n IU, Kwasniewski, MT, and Fernando, LN. 2017. Comparison of Two Adsorbent Based de-Bittering Procedures for Neem (Azadirachta indica A. Juss) Tea- Effect on Polyphenols, Anti-Oxidant Capacity, Color and Volatile Profile. Plant Foods for Human Nutrition 72(1):88-95;
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Datta, A., Fernando, L.N. and Gruen, I.U. 2015. Effects of two de-bittering strategies on flavonols, phenolic compounds and anti-oxidant activity of Neem tea. Institute of Food Technologists Annual Meeting, Chicago, IL, July 11-14, Pres. #098-058
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:The target audiences include other researchers in food science and nutrition either in academia or in industry, as well asfood product developers in industry that might use the information provided to them in the form of publications andpresentations to develop and market new functional food products with health-beneficial properties based on developedprototypes of functional foods containing compounds and bio-activities identified in native and non-native plants. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The Ph.D. student, who worked on the project, in additon to (as previously reported) acquiring chromatograhy skills, specifically high performance liquidchromoiatography, as well as learning about the interpretation of mass spectrometric data, he also attained a considerable amount of skills in statitics, specifically experimental design as it relates to sensory analysis. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?In the next year, we intend to finalize the analytical methodologies and work with the Professional Manufacturing Candy Association (PMCA) on acquiring the cocoa samples for the study.
Impacts
What was accomplished under these goals?
In the context of this project, we initiated an investigation into the bioactive compounds and the resulting bitterness of cacao and chocolate, specifically our goal is to identify and quantify the complex mixture of important bitter compounds within the three compound classes that are known to be important to cocoa--methylxanthines, flavan-3-ols, and diketopiperazines. Chocolate is made from the fermented, dried, and roasted seeds of the Theobroma cacao tree. It is now understood that cacao, also known in English-speaking countries as cocoa, contains a variety of compounds, including well-known flavonoid polyphenols, the consumption of which has positive impacts on human heart health and blood pressure, cancer reduction, LDL cholesterol reduction, and insulin resistance improvements, as numerous in vitro, in vivo, and observational studies have confirmed. In important research published in 2017, performed, in part, by Harvard's Chan School of Public Health, a participant observational study, composed of men and women between the ages of 50 and 64, led to the determination that chocolate consumption is inversely correlated with atrial fibrillation (AR). This is an important finding, as AR is associated with multiple significant health problems such as "higher risk of stroke, heart failure, cognitive decline, dementia and mortality." A persuasive case has, therefore, been made that cacao is a healthful addition to a balanced diet. However, despite the strong evidence for the healthful qualities of cacao, which has led to an overall increase in consumer purchases of more high-cacao-content chocolate than in previous decades, sales figures still show that there is an unwillingness by many Americans to consume higher-cacao-content chocolate, which tends to be more bitter, a taste modality not readily appreciated by most. Therefore, if the bitterness of cacao could be minimized, higher-cacao-content and lower-sugar chocolate confection sales could capture an even larger segment of the conventional and healthy snack-food market by achieving the elusive combination of being both tasty and healthy. The key is to better grasp the causes of bitterness in cacao, which are complex and still not well understood. What has long been known is that the methylxanthine, theobromine, is an important contributing bitter compound in cacao, with its name even being derived from the genus of the cacao tree, Theobroma. Similarly, the related methylxanthine, caffeine, which is well known as an important bitter compound in coffee, has also been found to impart bitterness in cacao and chocolate. Certain flavan-3-ols, which are a subclass of the aforementioned healthy flavonoids, and which are found in black tea, are also bitter and present in cacao at levels above the detection threshold, especially the compound (-)-epicatechin. After project initiation, cocoa beans were obtained from Ghana, Madagascar, and Peru and have been roasted according to a standard roasting profile by chocolate manufacturer Patric Chocolate, LLC. Methods have been developed and validated, and standard curves have been established for the cocoa bitter compounds: Caffeine, Theobromine, Catechin, Epicatechin, and selected Diketopiperazines (DKPs), including cyclo(L-Pro-L-Val) which according to the most recent literature available is considered the most important DKP in regards to cocoa bitterness. The bitter compounds caffeine, Theobromine, Catechin, and Epicatechin were found in all samples. While a preliminary analysis of the data showed, as expected, some variation, we still need to analyze the data statistically. We were not able to replicate the finding reported by Stark (Stark T, Bareuther S, Hofmann T. Molecular definition of the taste of roasted cocoa nibs (Theobroma cacao) by means of quantitative studies and sensory experiments. J Agr Food Chem. 2006;54(15):5530-9) that the diketopiperazine cyclo(L-Pro-L-Val) is an important bitter compound! While we were able to positively identify cyclo(L-Pro-L-Val) in a very dark roasted cocoa using LC-MS-MS, which then allowed us to also detect and quantify this diketopiperazine in other roasts, the concentration of cyclo(L-Pro-L-Val) is apparently considerably lower, i.e. only about 5-10% of that reported by Stark, which put its concentration at or below its sensory threshold. However, since Stark's sample was not well characterized, i.e. no information was given on roast profile or how many samples were analyzed, a direct comparison of results is difficult. On the other hand, we were able to detect this diketopiperazine even in unroasted samples, indicating that DKPs are formed already during the fermentation stage of the beans.
Publications
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:The target audiences include other researchers in food science and nutrition either in academia or in industry, as well as food product developers in industry that might use the information provided to them in the form of publications and presentations to develop and market new functional food products with health-beneficial properties based on developed prototypes of functional foods containing compounds and bio-activities identified in native and non-native plants. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The Ph.D. student, who worked on the project, acquired chromatograhy skills, specifically high performance liquid chromoiatography, as well as learned about the interpretation of mass spectrometric data. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?As of 2018, it is still the case that much is to be learned about the variation that exists in bitter-compound composition of cacao, especially across multiple cacao origins and roast profiles, even considering that the three primary classes of bitter compounds mentioned above (DKPs, flavonoids, and methylxanthines) are well known. We intend to decrease chocolate bitterness by involving process-based bitter compound minimization, thereby leaving the naturally occuring healthy polyphenols "unharmed". Specifically the post-importation processing steps of roasting, alkalization, formulation, and conching are worth consideration in the reduction of bitterness in cacao given their potential to increase some bitter compounds while reducing others. However, considering literature and our preliminary data above, which suggests that roast profile likely has a significant impact on the bitter compound profile of cacao, and considering that roasting is the first post-importation process to be carried out on cacao with the goal of flavor development, while also being recognized as the most important processing step overall, we will first consider roasting optimization while the potential of alkalization, formulation, and conching may be studied at a later date.
Impacts
What was accomplished under these goals?
In the context of this project, we initiated an investigation into the bioactive compounds and the resulting bitterness of cacao and chocolate, specifically our goal is to identify and quantify the complex mixture of important bitter compounds within the three compound classes that are known to be important to cocoa--methylxanthines, flavan-3-ols, and diketopiperazines. Chocolate is made from the fermented, dried, and roasted seeds of the Theobroma cacao tree. It is now understood that cacao, also known in English-speaking countries as cocoa, contains a variety of compounds, including well-known flavonoid polyphenols, the consumption of which has positive impacts on human heart health and blood pressure, cancer reduction, LDL cholesterol reduction, and insulin resistance improvements, as numerous in vitro, in vivo, and observational studies have confirmed. In important research published in 2017, performed, in part, by Harvard's Chan School of Public Health, a 55,502 participant observational study, composed of men and women between the ages of 50 and 64, led to the determination that chocolate consumption is inversely correlated with atrial fibrillation (AR). This is an important finding, as AR is associated with multiple significant health problems such as "higher risk of stroke, heart failure, cognitive decline, dementia and mortality." A persuasive case has, therefore, been made that cacao is a healthful addition to a balanced diet. However, despite the strong evidence for the healthful qualities of cacao, which has led to an overall increase in consumer purchases of more high-cacao-content chocolate than in previous decades, sales figures still show that there is an unwillingness by many Americans to consume higher-cacao-content chocolate, which tends to be more bitter, a taste modality not readily appreciated by most. Therefore, if the bitterness of cacao could be minimized, higher-cacao-content and lower-sugar chocolate confection sales could capture an even larger segment of the conventional and healthy snack-food market by achieving the elusive combination of being both tasty and healthy. The key is to better grasp the causes of bitterness in cacao, which are complex and still not well understood. What has long been known is that the methylxanthine, theobromine, is an important contributing bitter compound in cacao, with its name even being derived from the genus of the cacao tree, Theobroma. Similarly, the related methylxanthine, caffeine, which is well known as an important bitter compound in coffee, has also been found to impart bitterness in cacao and chocolate. Certain flavan-3-ols, which are a subclass of the aforementioned healthy flavonoids, and which are found in black tea, are also bitter and present in cacao at levels above the detection threshold, especially the compound (-)-epicatechin. After project initiation, cocoa beans were obtained from Ghana, Madagascar, and Peru and have been roasted according to a standard roasting profile by chocolate manufacturer Patric Chocolate, LLC. Methods have been developed and validated, and standard curves have been established for the cocoa bitter compounds: Caffeine, Theobromine, Catechin, Epicatechin, and selected Diketopiperazines (DKPs), including cyclo(L-Pro-L-Val) which according to the most recent literature available is considered the most important DKP in regards to cocoa bitterness. The bitter compounds Caffeine, Theobromine, Catechin, and Epicatechin were found in all samples. While a preliminary analysis of the data showed, as expected, some variation, we still need to analyze the data statistically. We were not able to replicate the finding reported by Stark (Stark T, Bareuther S, Hofmann T. Molecular definition of the taste of roasted cocoa nibs (Theobroma cacao) by means of quantitative studies and sensory experiments. J Agr Food Chem. 2006;54(15):5530-9) that the diketopiperazine cyclo(L-Pro-L-Val) is an important bitter compound! While we were able to positively identify cyclo(L-Pro-L-Val) in a very dark roasted cocoa using LC-MS-MS, which then allowed us to also detect and quantify this diketopiperazine in other roasts, the concentration of cyclo(L-Pro-L-Val) is apparently considerably lower, i.e. only about 5-10% of that reported by Stark, which put its concentration at or below its sensory threshold. However, since Stark's sample was not well characterized, i.e. no information was given on roast profile or how many samples were analyzed, a direct comparison of results is difficult. On the other hand, we were able to detect this diketopiperazine even in unroasted samples, indicating that DKPs are formed already during the fermentation stage of the beans.
Publications
|
Progress 11/19/15 to 09/30/16
Outputs
Target Audience:The target audiences include other researchers in food science and nutrition either in academia or in industry, as well as food product developers in industry that might use the information provided to them in the form of publications and presentations to develop and market new functional food products with health-beneficial properties based on developed prototypes of functional foods containing compounds and bio-activities identified in native and non-native plants. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project was part of a Ph.D. candidates research. How have the results been disseminated to communities of interest?Yes, via a publication. What do you plan to do during the next reporting period to accomplish the goals?Determine the volatile composition of neem.
Impacts
What was accomplished under these goals?
Neem (Azadirachta indica) is an evergreen tree cultivated in various parts of the Indian sub-continent. It has been in use over centuries in the Indian folk medicine for its therapeutic value. Given the prominent role it has played in curing diseases of the villagers over centuries, it has been hailed as a "divine" tree, a "village dispensary" and " nature's drugstore". Today, extensive research has shown that it may have anti-cancer, anti- diabetic, anti-inflammatory, anti- ulcerogenic, and anti-microbial effects. However, neem is extremely bitter, in large part due to its limonoid content, making it unpalatable and limiting its potential use in dietary supplements or foods alike. Bitterness reduction, especially of foods and beverages containing phytonutrients, is one of the biggest challenges in the food industry. The objective of this experiment of our overall study was to apply two adsorbent based strategies, namely solid phase extraction (SPE) and Amberlite XAD-16 (AMB) resin, to achieve de-bittering of neem tea and to determine the effects of the de-bittering on the bio-active, color, and volatile properties. The solid SPE treatment completely removed the flavonol, quercetin, from neem tea while in Amberlite XAD-16 treated tea (AMB) it was only insignificantly (p > 0.05) reduced. We also observed decreases in total phenolic content and consequently anti-oxidant activities after de-bittering. A 62% mean reduction of limonoid aglycones indicated diminished levels of bitterness. The loss of phenolics lead to a visually appreciable color change in the treated teas. The de-bittering also leads to a loss of sesquiterpenes, ketones and acids from neem tea. In conclusion, we found that while SPE cartridges were more efficient in removing bitterness, they caused a greater reduction in bio-active compounds than AMB XAD-16 resins, which may ultimately affect the health properties of neem tea.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Datta A., Gr�n IU, Kwasniewski, MT, and Fernando, LN. 2016. Comparison of Two Adsorbent Based de-Bittering Procedures for Neem (Azadirachta indica A. Juss) Tea- Effect on Polyphenols, Anti-Oxidant Capacity, Color and Volatile Profile. Plant Foods for Human Nutrition. DOI: 10.1007/s11130-016-0595-9
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