A feasibility study on organic lotus pollen tea as consumer product

R E G U L A R A R T I C L E Hongsamoot and Bumroongsook: Feasibility Study on Organic Lotus Pollen Tea 442 Emir. J. Food Agric ● Vol 27 ● Issue 5 ● 2015 content and antioxidant activity are parameters of the tea’s antioxidative quality (Anesini et al., 2008). The Department of Thai Foreign Trade Affair has found that imported consumer household goods including drinks and tea are growing steadily, causing an unbalanced trade. Therefore, this is a good opportunity for lotus growers to offer lotus tea, full of benefi cial fl avonoids, as an alternative to imported beverages that offer no health benefi t. In addition, healthy lotus tea fi ttingly caters to the need for healthy food products currently in high demand by the top-tiered consumer markets with high purchasing power such as those in the European Community, Japan, and the United States (Newswit, 2007). Therefore, organic lotus pollen tea is a good value-added product that can bring in more steady income to lotus growers. MATERIALS AND METHODS Pollen production and sample preparation Four varieties of lotus: Sattabutt, Sattabongkutt, Buntharik, and Patum were grown in a farm at the KMITL campus in Bangkok. The lotuses were grown organically, without any chemicals, by techniques such as cutting full bloom fl owers and leaves above the water to prevent thrip and caterpillar infestations and cleaning up of weeds and grasses to prevent insects from laying eggs and harboring in the area around the farm (Bumroongsook, 2003). Green manure and compost were added to prevent nutrient defi ciency (Malhi, 2009). At the age of 10 days, the fl owers were hand-picked in the morning and their sizes, weights, and the amounts of pollens collected were recorded. Thrip contamination was also examined. The collected pollens were dried in a shade for two days and removed of any pest contamination before they were ground into powder. The powder was then packaged into tea bags: 1.0 g in each 5×5 cm tea bag. Hedonic test Thirty people who have drunk lotus tea before were recruited as testers. Lotus pollen tea made from Sattabutt, Sattabongkutt, Buntharik, and Patum varieties were evaluated on their fl avor, aroma and color. The testers’ preferences of these attributes and their overall preference were graded on a 9 point hedonic scale, from extremely dislike to extremely like. Analysis of variance were conducted on the sample mean scores of these preferences. Statistically signifi cant attributes were further analyzed for the mean differences using Duncan Multiple Range test at P<0.05. Antioxidant activity The DPPH scavenging test used was the method of BrandWilliam et al. (1995). One bag of tea was soaked in 120 ml of hot water for 2 minutes. Then, 1 ml of the brewed tea was added to 1 ml of DPPH solution. Next, the mixture was incubated for 30 minutes. The absorbance of the mixture was measured at 517 nm with a Spekol 2000 UVvis spectrophotometer (AnalytikGena, Germany). Distilled water was used as blank and trolox as standard. Percentage of inhibition was calculated using the following equation: % inhibition=(Abst=0-Abst=30)/Abst=0* 100,


INTRODUCTION
Sacred lotus (Nelumbonucifera (Gaertn.))has long been honored the queen of aquatic plants in Thai culture.In Thailand, there are 4 common varieties of lotus: Sattabutt (white double), Sattabongkot (pink double), Buntharik (white single), and Patum (pink single).It is a widespread ornamental plant used especially in religious ceremony.Lotus cultivation is frequently plagued by thrip infestation, especially by the species Frankliniella schultzei (Trybom).Both the adults and larvae of this insect species ferociously attack lotus petal and pollen, forcing growers to use insecticide heavily.Even so, thrips are very diffi cult to control due to their prevalence and diffi cultto-locate habitat (Mungnimitr and Bumroongsook, 2007).In international trade, where export of lotus fl owers requires them to be pest free, thrips are a major cause for quarantine (Walsh et al., 2005).Consequently, growers have to strictly control them, both before and after harvest, yet it is still very diffi cult to meet the regulations of the major importing countries.An alternative to export whole lotus fl owers is to export them as pest-free herbal tea.
Lotus, all parts of it, is well known for its medical benefi ts.A wide range of pharmacological effects from different parts of this plant has been reported.According to Department of Medical Science (2004), the pollen protects against DNA mutation, promotes enzyme activity, destroys toxic substances, mildly decreases blood sugar, and nurtures the heart.The brownish yellow fi lament of the pollen has various therapeutic benefi ts to the heart, spleen, and liver.Hence, numerous traditional medicines have lotus pollen as an essential ingredient in their formularies.The pollen also contains several flavonoids such as quercetin, luteolin, isoquercitrin, and luteolin-glucoside.Flavoniods are found in edible plants (Miean and Mohamed, 2001).These substances are reported to benefi t health (Tokuşoğlu et al., 2003).Nowadays, lotus pollens are increasingly being used as natural health supplements.They possess antioxidant properties when brewed in hot water.Total polyphenol Lotus pollen tea, in particular, contains potent antioxidants.The fi rst step of this study was the screening for 3 desired qualities of 4 varieties of lotus (Nelumbonucifera Gaertn): Sattabongkutt, Buntharik, Sattabutt, and Patum.The 3 desired qualities were as follows: High pollen yield; potent antioxidant activity; and favorable aroma, fl avor, and color to a panel of testers.The results of the screening were the following: one, the pollen productions of a fl ower of each of the 4 varieties stated above were 1.28, 3.11, 0.43, and 1.83 gm, respectively; two, Buntharik had the highest antioxidant capacity at a trolox equivalent of 499.48 μg/ml or 59.94 mg/serving and at a total polyphenol equivalent of gallic acid of 63.26 μg/ml or 7.59 mg/serving; and three, the highest-rated variety in a hedonic test of overall preference was Buntharik.Thus, the Buntharik tea was chosen for the consumer acceptance studies conducted with 90 tea testers with equal number of males and females.The fi nding was found that males preferred this tea over females; panelists over 30 years old gave the highest score on overall preference; their level of acceptance was fairly acceptable, at 50%, but their purchase intention was lower, at 27.78%.
content and antioxidant activity are parameters of the tea's antioxidative quality (Anesini et al., 2008).
The Department of Thai Foreign Trade Affair has found that imported consumer household goods including drinks and tea are growing steadily, causing an unbalanced trade.Therefore, this is a good opportunity for lotus growers to offer lotus tea, full of benefi cial fl avonoids, as an alternative to imported beverages that offer no health benefi t.In addition, healthy lotus tea fi ttingly caters to the need for healthy food products currently in high demand by the top-tiered consumer markets with high purchasing power such as those in the European Community, Japan, and the United States (Newswit, 2007).Therefore, organic lotus pollen tea is a good value-added product that can bring in more steady income to lotus growers.

Pollen production and sample preparation
Four varieties of lotus: Sattabutt, Sattabongkutt, Buntharik, and Patum were grown in a farm at the KMITL campus in Bangkok.The lotuses were grown organically, without any chemicals, by techniques such as cutting full bloom fl owers and leaves above the water to prevent thrip and caterpillar infestations and cleaning up of weeds and grasses to prevent insects from laying eggs and harboring in the area around the farm (Bumroongsook, 2003).Green manure and compost were added to prevent nutrient defi ciency (Malhi, 2009).At the age of 10 days, the fl owers were hand-picked in the morning and their sizes, weights, and the amounts of pollens collected were recorded.Thrip contamination was also examined.The collected pollens were dried in a shade for two days and removed of any pest contamination before they were ground into powder.The powder was then packaged into tea bags: 1.0 g in each 5×5 cm tea bag.

Hedonic test
Thirty people who have drunk lotus tea before were recruited as testers.Lotus pollen tea made from Sattabutt, Sattabongkutt, Buntharik, and Patum varieties were evaluated on their fl avor, aroma and color.The testers' preferences of these attributes and their overall preference were graded on a 9 point hedonic scale, from extremely dislike to extremely like.Analysis of variance were conducted on the sample mean scores of these preferences.Statistically signifi cant attributes were further analyzed for the mean differences using Duncan Multiple Range test at P<0.05.

Antioxidant activity
The DPPH scavenging test used was the method of Brand-William et al. (1995).One bag of tea was soaked in 120 ml of hot water for 2 minutes.Then, 1 ml of the brewed tea was added to 1 ml of DPPH solution.Next, the mixture was incubated for 30 minutes.The absorbance of the mixture was measured at 517 nm with a Spekol 2000 UVvis spectrophotometer (AnalytikGena, Germany).Distilled water was used as blank and trolox as standard.Percentage of inhibition was calculated using the following equation: % inhibition=(Abs t=0 -Abs t=30 )/Abs t=0 * 100, where Abs t=0 was the absorbance of DPPH at time = 0 min, Abs t=30 was the absorbance of DPPH after 30 minutes of incubation.
Antioxidant activity was expressed as μg of trolox equivalent per ml or mg of trolox equivalent per serving.

Total phenolic content analysis
Total phenolic content was determined by the Folin-Ciocalteau method.Each bag of tea was extracted with 120 ml of water at 80 o C for 2 minutes.Its total polyphenol content was determined by a modifi ed method of Singleton et al. (1999).The total phenolic content of the tea was measured by using Folin-Ciocalteau reagent.Dilute reagent was mixed with a tea sample and left sitting at room temperature for 5 minutes.Then, 2% sodium carbonate was added, and the mixture incubated for 15 minutes at room temperature.The absorbance of the tea sample was measured at 765 nm with a Spekol 2000 UV-vis spectrophotometer (Analytik Gena, Germany).Distilled water was used as blank and gallic acid as standard.The results were reported as μg of gallic acid equivalent/ml and mg of gallic acid equivalent/serving.

Consumer acceptance test
A panel of 90 tea testers was recruited from the Ladkrabang community, Bangkok, to participate in this study.These testers were chosen according to the following three criteria: they were between 18-65 years old; they drank tea at least once a week; and the number of male and female testers in each age group should be equal.The panelists were divided into 3 age groups (less than 20, 20-30, and more than 30 years of age).Each group consisted of 30 persons with equal number of males and females.The Buntharik pollen tea was evaluated on the testers' preferences for its aroma, fl avor, color, nutritional value, and on their overall preference, using a 9 point intensity scale anchoring at extremely dislike to extremely like.These panelists were also asked to respond to items of a questionnaire at the end of the test.The items covered demographic and socioeconomic information, tea drinking behavior, and tea product acceptance.
Analyses of variance were conducted on the sample mean scores for aroma, fl avor, color, nutritional value, and overall preference.Statistically signifi cant attributes were further analyzed for the mean differences using Duncan Multiple Range Test at P=0.05.The analysis of the responses to the questionnaire only focused on obtaining frequencies and descriptive statistics.Cross tabulations were used to explore whether special relationships existed.

RESULTS AND DISCUSSION
This study shows that Sattabongkutt flower was the heaviest, followed by Sattabutt, Buntharik, and Patum.The widths of Sattabongkutt, Buntharik, Sattabutt, and Patum fl owers were 10.03, 7.12, 5.75 and 5.17 cm, respectively.Buntharik fl ower was the longest at 12.95 cm (p<0.05),followed by Sattabongkutt, Patum, and Sattabutt.The amounts of collected pollens were signifi cantly different among the four lotus varieties (p< 0.05); Buntharik had the highest amount of them, weighing 3.1 gm while Patum, Sattabongkutt, and Sattabutt had 1.83, 1.28 and 0.43 g, respectively.Thrip contamination on pollens, of adults and larvae of F. schultzei, was monitored, and Buntharik pollens were found to be most contaminated at 59.80 (p < 0.05), followed by Patum, Sattabongkutt, and Sattabutt at 8.40, 4.13 and 2.67, respectively (Table 1).Pleansri and Bumroongsook (2013) found heavy infestation of thrips in the middle part of a lotus fl owers where lotus pollen was located, thus pollen was easily contaminated by thrips population.Moreover, F. schultzei preferred Buntharik to other varieties.Laborious work of picking up dead thrips from dried pollens was needed.Minor damage of an organic product by pests should be perceived positively by consumers (Hadyanyah, 2009).
The highest antioxidant capacity among those of the four pollen teas was found in Buntharik tea.It was equal to a trolox equivalent of 499.48 g/ml or 59.94 mg/serving.This fi nding confi rmed with Thongrungroj (2007) that Butharik pollen had antioxidant capacity more than the other varieties.The antioxidant activity is to scavenge free radicals and benefi t human health, and be associated with the prevention of cardiovascular diseases, anticancer and anti-aging (Verlangieri et al., 1985).Moreover, its total polyphenol content was an equivalent of gallic acid 63.26 μg/ml or 7.59 mg/serving (Table 2).The antioxidant capacity and total phenolic content of the Buntharik tea was much higher than those of other varieties.The chemical composition of lotus pollen is complex includes polyphenol, quercitin, luteolin, isoquercitin, luteolin glucoside, volatile oil, alkaloid, and unidentifi ed compounds (Deparment of Medical Science, 2004;Thongrungroj 2007).Quercithin, a flavonoid glycoside, had antioxidant properties (Zhang et al, 2011).It is considered as one of the most prominent dietary antioxidants and it was found in fruits and vegetables and excerted benefi cial health benefi t (Boot et al., 2008).Polyphenol is an interesting group and some plant phenolic contents has strong antioxidant capacity (Cabrera et al., 2003;Karori et al., 2007).The plant crude extract could prevent the occurrence of most degenerative diseases due to its high phenolic content, and, its high antioxidant activity (Beltrán-Orozco1, 2013).Nevertheless, natural antioxidants are the result of multiple factors and its antioxidant capacity cannot only be associated to the phenolic content (Arazo et al., 2011) The results of the consumer hedonic test of 4 different lotus teas showed that, in the category of overall preference, most testers preferred ones made from Buntharik and Patum pollens.Most of them also preferred the aroma and the fl avor of these two teas (Table 3).There were no signifi cant differences among preferences on color (p>0.05).Based on the above information, we conclude that Buntharik pollen tea is the healthiest because of its highest antioxidant capacity and total phenolic content.Buntharik fl owers also gave the highest pollen yield, and Buntharik tea achieved the highest hedonic rating.

Product acceptance
According to gender, the results showed that, overall, males preferred lotus pollen tea more than females did; however, their preferences for aroma, fl avor, or color of the tea were not different.According to age, testers 30 years old and over preferred this tea the most, followed by the ones between 20 and 30 years old and those less than 20 year old, in that order (Table 4).Out of the total of 90 purposedly chosen panelists, 45 were females and 45 were males.They were grouped according to their ages into 3 groups (<aged 20, 20-30, and > 30 or over) with equal representation from both genders (Table 5).They were also grouped according to their income into 3 groups.The biggest group, at 44.45% of the total number of testers, had members whose incomes were in the range of 10,000-30,000 baht; the next biggest group, at 31.11%, had members with more than 30,000 baht income; and, the smallest group, at 22.44%, was those with income between 5,000-10,000 baht (Table 5).
The testers were asked to specify the type of tea that they have been drinking.Of all the panelists, 45.56% responded that they drank green tea, 31.11%drank Chinese tea, 16.67% drank instant tea, and the remaining 6.66% drank other types of tea.Tea-drinking time was classifi ed into 4 categories (shown with the percentage of drinkers):before 10:00 (16.67%), 10:00-13:00 (41.11%), 13:01-17:00 (31.78%), and 17:01-20:00 (4.44%).As for how many testers drank tea at what frequency, 13.33 % of the panelists drank it 1-2times a week, 51.11% drank between 3-5 times, and 35.56 % drank more than 5 times.The panelists' responses indicated that 80% of them bought their tea by themselves and 20.00% of them got their tea by other means.The panelists'responses to the question of at what place they bought their tea were the following: mostly at supermarkets (67.78%), convenient stores (13.33%), co-op food stores (8.89%), grocery stores (5.56%), and other places (4.44%) (Table 6).As for the level of acceptance indicated by the panelists, half of them responded that the tea was fairly acceptable, 16.67% very acceptable, 24.44% just acceptable, and 8.89% not acceptable.On the question whether they would buy the tea if a box of 20 tea bags was on the market for 50 baht, 27.78% of the panelists indicated that they would buy the product, 31.11% would not, and 41.11% was uncertain whether they would (Table 7).Purchase intention is one of the components of consumer cognitive behavior on how an individual intends to buy a specifi c product.The organic lotus pollen tea products is a healthy drink and less harmful to the environment than their non-organic equivalents.
It is assumed that rational considerations will favor the  purchase of organic tea.However, many factors effect the purchase behavior of consumers and consumption of organic tea (Sakthirama et al., 2013).Pual and Rana (2012) indicated that healthy attributes, availability of products and education had positively infl uence the consumer's attitude towards buying organic food.Income, age and willing to pay at a premium price for safe food had minor effect on the purchase of organic food (Lockie et al., 2004).Sheng-Hun et al. ( 2010) stated that the negative relationship between price and purchasing intention whereas respondents from Taiwan preferred tea drinking products with health benefi t claims.The consumer goes through different buying decision process before he makes a decision to purchase a product.Further research should be conducted on various types of buying infl uences to develope market strategies to target consumers (Sumi and Kabir, 2010).

CONCLUSIONS
Growing lotus for pollen production organically is laborious and time consuming, but the growers can save the costs of pesticides and synthetic fertilizers.In this study, we found that there were thrips infesting the pollens, but they were easily removed.Based on its pollen yield, antioxidant capacity, total polyphenol content, and 9-point hedonic rating, the Buntharik organic pollen tea was chosen as a viable consumer product.The product acceptance study gained basic information for further development of organic lotus pollen tea into a truly marketable product.If fully developed, the tea may be preferred by people over 30 years of age.Campaigns on health benefi ts of the lotus pollen tea should be staged, offering the tea as a healthy substitute for unhealthy beverages.Comprehensive market research and development for product diversity should also be conducted.