Accepted Manuscript

Analytical methods

Intake of selected bioactive compounds from plant food supplements containing

fennel (Foeniculum vulgare) among Finnish consumers

Liisa Uusitalo, Maija Salmenhaara, Merja Isoniemi, Alicia Garcia-Alvarez,

Llus Serra-Majem, Lourdes Ribas-Barba, Paul Finglas, Jenny Plumb, Pirkko
Tuominen, Kirsti Savela, on behalf of the PlantLIBRA Projects Plant Food
Supplement Consumer SurveyePlantLIBRA Database groups

PII: S0308-8146(15)01255-8
DOI: http://dx.doi.org/10.1016/j.foodchem.2015.08.057
Reference: FOCH 18005

To appear in: Food Chemistry

Received Date: 19 June 2014

Accepted Date: 17 August 2015

Please cite this article as: Uusitalo, L., Salmenhaara, M., Isoniemi, M., Garcia-Alvarez, A., Serra-Majem, L., Ribas-
Barba, L., Finglas, P., Plumb, J., Tuominen, P., Savela, K., on behalf of the PlantLIBRA Projects Plant Food
Supplement Consumer Survey, ePlantLIBRA Database groups, Intake of selected bioactive compounds from plant
food supplements containing fennel (Foeniculum vulgare) among Finnish consumers, Food Chemistry (2015), doi:
http://dx.doi.org/10.1016/j.foodchem.2015.08.057

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Intake of selected bioactive compounds from plant food supplements containing fennel

(Foeniculum vulgare) among Finnish consumers

Running title: Bioactive compounds in fennel-containing plant food supplements

Liisa Uusitaloa1, Maija Salmenhaaraa, Merja Isoniemia, Alicia Garcia-Alvarezb, Llus Serra-

Majemb,c,d, Lourdes Ribas-Barbab,c, Paul Finglase, Jenny Plumbe, Pirkko Tuominena and Kirsti

Savelaa; on behalf of the PlantLIBRA Projects Plant Food Supplement Consumer Surveyf and

ePlantLIBRA Databaseg groups

a
Finnish Food Safety Authority Evira, Risk Assessment Research Unit, Mustialankatu 3, FI-

00790 Helsinki, Finland

b
Fundacin para la Investigacin Nutricional, Barcelona Science Park, University of Barcelona,

Barcelona, Spain
c
CIBER Obn of Physiopathology of Obesity and Nutrition, Institute of Health Carlos III, Madrid,

Spain.
d
Research Institute of Biomedical and Health Sciences. University of Las Palmas de Gran

Canaria, Spain.
e
Institute of Food Research, Norwich Research Park, Norwich, NR4 7UA. United Kingdom
f
The PlantLIBRA PFS Consumer Survey working group comprises: Llus Serra-Majem -

responsible for the survey, Alicia Garcia-Alvarez and Lourdes Ribas-Barba (Fundacin para la

Investigacin Nutricional, Barcelona Science Park, University of Barcelona, Barcelona, SPAIN;

and CIBER of Physiopathology of Obesity and Nutrition, Madrid, Spain); Liisa Uusitalo, Maija

1
Corresponding author at Finnish Food Safety Authority Evira, Risk Assessment Research Unit,

Mustialankatu 3, FI-00790 Helsinki, Finland. Tel. +358 50 570 9637, Fax +358 207724353, email

liisa.uusitalo@evira.fi

1
Salmenhaara and Merja Isoniemi (Finnish Food Safety Authority Evira, Helsinki, FINLAND);

Simone de Klein and Eva Melanie Meissner (PhytoLab GmbH & Co KG, Vestenbergsgreuth,

GERMANY); Bernadette Egan and Monique M Raats (Food, Consumer Behaviour and Health

Research Centre, University of Surrey, Guildford, Surrey, The UNITED KINGDOM); Franco M

Maggi, Flavia Bruno and Patrizia Restani (Dipartimento di Scienze Farmacologiche e

Biomolecolari, Universit degli Studi di Milano, Milano, ITALY); Mihaela Badea, Angela

Marculescu and Lorena Dima (Transilvania University of Brasov, Brasov, ROMANIA).

g
The ePlantLIBRA Database working group comprises: Jenny Plumb and Paul Finglas (Institute

of Food Research, Norwich, UK) responsible for the composition data within the database,

Mairead Kiely and Jacqueline Lyons (University College Cork, Ireland) responsible for beneficial

bioeffects data, Karin Norby (Danish Technical University, Copenhagen Denmark) responsible

for adverse effects data and Paul Finglas and Carlos Ramos (EuroFIR AISBL, Brussels, Belgium)

responsible for ePlantLIBRA sustainability.

2
Abstract

The purpose of this study was to estimate the intake of selected bioactive compounds from

fennel-containing plant food supplements (PFS) among Finnish consumers. The estimated

average intake of estragole was 0.20 mg/d, of trans-anethole 1.15 mg/d, of rosmarinic acid

0.09 mg/d, of p-coumaric acid 0.0068 mg/d, of kaempferol 0.0034 mg/d, of luteolin 0.0525

g/d, of quercetin 0.0246 mg/d, of matairesinol 0.0066 g/d and of lignans 0.0412 g/d. The

intakes of kaempferol, quercetin, luteolin, matairesinol and lignans from PFS were low in

comparison with their dietary supply. The intake of estragole was usually moderate, but a

heavy consumption of PFS may lead to a high intake of estragole. The intake of trans-anethole

did not exceed the acceptable daily intake, but PFS should be taken into account when

assessing the total exposure. To our knowledge, this study provided the first intake estimates

of trans-anethole, p-coumaric acid and rosmarinic acid in human populations.

Keywords Dietary supplements Plants Botanicals Fennel Bioactive compounds Survey

3
 1. Introduction

Food supplements are concentrated sources of nutrients or other substances with a nutritional

or physiological effect, the purpose of which is to supplement the normal diet. They are

marketed in dose form, for instance as pills, tablets, capsules and liquids in measured doses.

Plant food supplements (PFS) contain one or more botanical ingredients. They have a long

tradition of use, and are widely consumed in many European countries. However, comparable

data on the consumption of PFS on the European level are scarce, and little is known about the

risks and benefits associated with their consumption. The PlantLIBRA project aims to develop,

validate and disseminate data and methodologies for the risk and benefit assessment of PFS,

and to implement sustainable international cooperation related to PFS (PlantLIBRA , 2010).

Fennel (Foeniculum vulgare) is a perennial aromatic herb. In pharmacopoeias and well

established documents fennel has been described as being used in the symptomatic treatment

of dyspepsia (Weiss, 1991), as an expectorant for mild inflammation of the upper respiratory

tract (Weiss, 1991) and in the treatment of dysmenorrhea and pain in scrotal hernia

(Pharmacopoeia of the Peoples Republic of China, 2000). In traditional medicine, fennel is

used in the treatment of many types of symptoms (Hare, Caspari, & Rusby, 1916).

According to the results of the PlantLIBRA PFS Consumer Survey conducted in Finland,

Germany, Italy, Romania, Spain and the UK, fennel ranked sixth in the pooled list of most

consumed botanicals (Garcia-Alvarez et al., 2014). Among the Finnish subsample, fennel

ranked 31th. The essential oil of fennel, extracted from fennel seeds, contains trans-anethole

and estragole, among other compounds. Both have exhibited antimicrobial activity (Friedman,

Henika and Mandrell, 2002), but trans-anethole has also displayed tumorigenic properties in

laboratory animals (Joint FAO/WHO expert committee on food additives, 2000), and estragole

is carcinogenic and mutagenic at high doses (Committee on herbal medicinal products, 2005).

4
The objectives of the present article are as follows: to describe the consumption of PFS

containing fennel among a subsample of Finnish PFS consumers from the PlantLIBRA PFS

Consumer Survey; to present the consumer reasons for PFS usage, perception of the

usefulness of the products and the adverse effects experienced after usage; to describe the

socio-demographic and lifestyle-related characteristics of the consumers; and to estimate the

intake of selected bioactive compounds, including estragole and trans-anethole, from the

fennel-containing PFS consumed by these Finnish consumers. To our knowledge, this article

presents the first intake estimates of some of the bioactive compounds namely trans-

anethole, p-coumaric acid and rosmarinic acid in human populations.

2. Materials and Methods

2.1. PlantLIBRA PFS Consumer Survey

A cross-sectional 12-month retrospective PFS consumption survey was conducted in Finland

and five other European countries. Before initiating the fieldwork, an ethical statement was

obtained from the Coordinating Ethics Committee, Hospital District of Helsinki and Uusimaa,

Finland. The details of the PlantLIBRA PFS Consumer Survey and its data collection procedures

have been described elsewhere (Garcia-Alvarez et al., 2014). In this survey, the sample size in

Finland was 401 PFS consumers. They came from four cities located in different parts of the

country; Helsinki, Turku, Kuopio and Oulu. The gender and age group quotas were set as

follows: 75% adults (18 to 59 years) and 25% elderly adults (60 years and over) with 50% males

and 50% females.

The survey participants were regular PFS consumers, who were identified using a short

screening questionnaire. The respondents were considered eligible for inclusion if they met

either of the following specified criteria (Garcia-Alvarez et al., 2014)):

1) They had taken at least 1 PFS in the previous 12 months, in an appropriate dose

form at a minimum frequency of:

5
 a) 1 daily dose for at least 2 consecutive or non-consecutive weeks, or

b) 1 or more doses per week for at least 3 consecutive weeks, or

c) 1 or more doses per week for at least 4 consecutive or non-consecutive

weeks;

2) They had taken 2 or more different PFS, in an appropriate dose form, at a

minimum frequency of 1 or more doses per week, with the sum of the usage

period for the 2 or more products being equal to at least 4 weeks.

Eligible consumers subsequently completed a more detailed questionnaire on their PFS usage

habits in the preceding 12 months, providing details of plant and product names, dose forms,

the frequency of use, reasons for use, adverse effects, places and patterns of purchase and

information sources on products. These questions were asked for each of up to a maximum of

five different PFS used. In addition, respondents were asked to provide socio-demographic

data, including age, gender, the level of education and employment status, self-reported

height and weight, and health-related lifestyle information.

Fieldwork and data collection for the PlantLIBRA cross-sectional survey were conducted by an

international market research company, European Fieldwork Group, from May 2011 to

September 2012. The duration of the fieldwork ensured that any seasonal variability in the

usage of products was captured.

All data from the completed surveys were entered into the statistical package SPSS for

Windows version 18 (IBM Corporation, Somers, NY, USA), which was also used for data

analysis.

2.2. Fennel content of plant food supplements

A database of botanical ingredients was compiled for all reported PFS products. Each product

was coded according to its botanical ingredients in local, scientific and English names.

6
Botanicals were also coded. Respondent data (i.e. consumption habits and socio-demographic

information) were recorded in a separate database.

PFS containing fennel were identified from the Finnish survey data. Fennel was identified in 6

different PFS products reported by the consumers. Information on the botanical concentration

in each product was obtained from the internet or from the producer/distributor. The

amounts of PFS consumed were calculated from the data. The outcome variable was the

average daily consumption of each product that contained fennel, both during the actual

consumption period reported by the consumer, and during the preceding 12 months. The

calculation process is presented in Figure 1. For PFS in liquid form, the size of the dose usually

taken was not asked for in the questionnaire, but some consumers reported it in an open field.

If not, we used the amount recommended by the manufacturer as the default dose. If the

recommended dose was expressed as a range, we used the average, maximum and minimum

values, respectively, for the average, maximum and minimum intake scenario of bioactive

compounds.

2.3. Intake of bioactive compounds from fennel-containing PFS

Bioactive compounds found in fennel were reported using the ePlantLIBRA database (EuroFIR

AISBL and PlantLIBRA, 2013). ePlantLIBRA is an extensive database containing biological effects

data as well as quality evaluated composition data from peer reviewed publications of

bioactive compounds analysed in botanicals. The bioactive compounds were selected for the

analysis based on 1) prior knowledge of the biological effects of the compounds and 2) the

number of the analysis results available in the database for each compound. For fennel, 70

analytical results were available for 36 different bioactive compounds, of which 13 were

selected for this study.

Information on the content of bioactive components in each botanical was extracted from the

ePlantLIBRA database (EuroFIR AISBL and PlantLIBRA, 2013). The data were saved in Excel

7
format. For each bioactive component, all the analysis results were checked, and

concentration information from appropriate studies was selected for use in the calculations.

In order to provide a representative assessment of exposure, we calculated three separate

intake scenarios for each bioactive component: one using the average concentration, one

using the maximum and one using the minimum concentration. If a concentration range for

the bioactive compound was presented, we used the high end of the range for the maximum

scenario, and the low end of the range for the minimum scenario. If a standard deviation was

given, we added two standard deviations to calculate the maximum scenario, and subtracted

two standard deviations for the minimum scenario. If only the average concentration was

given, minimum and maximum scenarios were not calculated. Finally, If the concentration was

expressed as less than (<), we calculated only the maximum scenario.

When there were two or more appropriate analysis results for the concentration of a

particular bioactive compound, we used the average of the average values for the average

scenario. The analysis result with the highest average concentration was selected for the

maximum scenario, and that with the lowest average concentration for the minimum scenario.

Minimum and maximum scenarios were calculated as described above.

3. Results

3.1. Consumer perceptions on fennel-containing plant food supplements

According to the PlantLIBRA PFS Consumer Survey, the most common reasons for taking

fennel-containing PFS were to boost the defense system/immunity, enhance energy/tonics

and improve stomach/digestive function (each 22.7% of fennel-containing PFS), followed by

body weight loss and for anaemia/low haemoglobin/iron (each 18.2% of fennel-containing

PFS). Other reasons and conditions given by the consumer were to maintain hair/skin health,

to prevent flu/colds, to improve the mood, for healthy joints and bones, for the heart/blood

circulation and to maintain general health (each less than 10% of fennel-containing PFS).

8
The consumers mostly thought that the fennel-containing PFS always or sometimes helped

them (54.5 and 27.3% of fennel-containing PFS respectively). Less than 10% of the fennel-

containing PFS were thought to help rarely, not at all, or the consumer was not sure. The

majority of consumers of PFS containing fennel did not report adverse effects following their

consumption, but one of the products was experienced to cause stomach upset and one to

cause increased urination (each 4.5% of fennel-containing PFS).

3.2. Consumer and plant food supplement characteristics

Table 1 presents the characteristics of the Finnish PFS consumers. The majority of consumers

who had used fennel-containing PFS were women. Most had a medium or high level of

education. More than half had normal weight, while almost half were overweight or obese,

and none were underweight. Most of the consumers of fennel-containing PFS considered their

health status to be good or very good, even if regular use of medication during the previous

12 months was common. The majority had consumed other dietary supplements than PFS

during the previous year, and more than half had received alternative or complimentary health

care, such as from an acupuncturist, homeopath or massage therapist. One fifth of the

consumers of fennel preparations were current smokers, and a third of them followed a

special diet. The most common reasons for following a special diet were food allergies (6/21

respondents) and lactose intolerance (3/21 respondents).

Fennel was included in 6 of the PFS products mentioned by the Finnish users (Table 2). Two of

the preparations with fennel were in a tablet form, and 4 in a liquid form. The amount of

fennel ranged from 3 mg to 14.1 mg in one tablet, and from 1.36 to 9.6 mg per one ml liquid.

Each fennel-containing product was named by 1-6 consumers. Fennel-containing PFS were

consumed on the average on 65 days, or about two months, per year.

3.3. Intake of bioactive compounds

9
The intakes of selected bioactive components from PFS containing fennel are presented in

Table 3. Intakes of 11 single compounds and 2 group variables of similar compounds (i.e. total

isoflavones and a mixture of lignans) were calculated.

The differences between the minimum, average and maximum estimates mainly reflect the

uncertainty in the concentrations of the bioactive components in the plant. The variation was

highest for estragole, which showed a 78-fold difference between the minimum and maximum

intake estimate. The concentrations of four bioactive compounds (daidzein, genistein, apigenin

and total isoflavones) were only given in the database as the upper limit of the concentration,

and only maximum intake estimate could therefore be calculated for these compounds. For six

bioactive compounds (p-coumaric acid, kaempferol, luteolin, quercetin, matairesinol and

mixture of lignans) there was no information on the range of the concentrations, and only the

average intake estimates are presented.

10
 4. Discussion

In the present study, we estimated the intake of selected bioactive compounds in fennel. The

intake estimates were based on the consumption of plant food supplements (PFS) containing

fennel among 401 Finnish PFS consumers from the PlantLIBRA projects survey on PFS usage.

The concentration information on bioactive compounds in fennel was acquired from the

ePlantLIBRA database. Because of the large variation in the concentrations of the bioactive

compounds between the different analyses, we estimated the intake using three possible

scenarios for each bioactive compound: the first using the analysis result with the lowest or

minimum concentration; the second using the average of all suitable analysis results; and the

third using the analysis result with the highest or maximum concentration.

The present study population was a quota-based convenience sample recruited from four

cities, and the results are not representative of the Finnish population. In the pre-screening

phase, regular PFS consumers were selected to participate in the survey, and those who

tended to consume PFS on an irregular basis were excluded. Therefore, the intake figures were

overestimates of the intake among all consumers of PFS.

When assessing the benefits and adverse effects of bioactive compounds from PFS, one must

keep in mind that the same bioactive compounds are also supplied by other foods and

botanicals. For instance, estragole, which is present in PFS containing fennel, is supplied also

from fennel used in cooking and from fennel tea. In addition to fennel, other herbs such as

basil, anise, tarragon and oregano (Smith et al., 2002) also contain considerable amounts of

estragole. Estragole has additionally been used as a flavouring substance in foods, although its

use was prohibited in the EU in 2008 (Regulation (EC) no 1334/2008 ).

The concentrations of the bioactive compounds in botanicals vary according to many factors,

including the geographical origin, plant maturity, harvesting techniques, storage conditions,

processing and method of extraction (Smith et al., 2002), and the analysis results also

11
displayed large variation in some cases. This variation was reflected in the difference between

the minimum and maximum intake scenarios, and produced uncertainties in the intake

estimates. Of the bioactive compounds analysed in the present study, the difference the

between minimum and maximum intake estimate was 78-fold at the highest.

One major source of uncertainty in the intake estimates is related to the plant part consumed.

It was not always known which plant part was used in the PFS, and analysis results may only be

available for a certain plant part. Fennel provided a typical example on this: some bioactive

compounds had been analysed from the seeds, and some from the whole plant above the

surface. In addition, there is no documentation on the effect of the supplement manufacturing

process on the concentration of the bioactive compounds. Due to these uncertainties, a

validation study comparing the computed and analyzed concentrations of bioactive

compounds in PFS would be of high value.

Fennel-containing PFS were typically consumed periodically among the Finnish consumers,

which was reflected in the approximately ten-fold difference compared to the annual use. We

calculated the three intake scenarios for bioactive compounds average, minimum and

maximum- of bioactive compounds both for the consumption period reported by the

consumer, and for the consumption during the preceding 12 months, which was the reference

period of the survey.

The average intake estimate of estragole from fennel-containing PFS was 0.20 mg/d. This

figure was close to the estimated per capita intake of estragole from spices in the USA

population 0.166 mg/d (Joint FAO/WHO expert committee on food additives (JECFA)2009). The

estimated average intake of estragole from PFS is about half of the intake of herb and spice

eaters, i.e. consumers in the USA, whose proportion of the population was estimated to be 10

% (Smith et al., 2002).

The maximum estimate of estragole intake from PFS (0.78 mg/d) was one third higher than the

estimated intake from three cups of fennel tea per day (Raffo, Nicoli, and Leclercq, 2011). It

12
was one fifth lower than the estimated intake in the total population (1 mg/d), based on a

review (Active principles (constituents of concern) contained in natural sources of flavourings,

2005). A conservative estimate of estragole intake among consumers of foods with estragole

as a flavouring substance was 4.3 mg/d (Scientific Committee on Food, 2001), but its use as a

flavouring is now prohibited in the EU (Regulation (EC) no 1334/2008).

The intake of daidzein and genistein, two bioactive compounds that belong to the isoflavone

group, from the diet or the diet together with supplements has been estimated in the adult

populations in countries including Finland (Valsta et al., 2003), Italy (Bertoli et al., 2008), Japan

(Kimira, Arai, Shimoi and Watanabe, 1998), the Netherlands (Boker et al., 2002) and in native

and immigrants groups in the UK (Ward et al., 2010) and the USA (Bandera et al., 2011). The

average intake figures, based on 24-h recalls (Valsta et al., 2003), food frequency

questionnaires (Bandera et al., 2011; Bertoli et al., 2008; Boker et al., 2002) or food records

(Ward et al., 2010), were consistently higher than the maximum estimate in our study. The

mean intake of daidzein in dietary studies ranged from 0.073 mg/d (Bertoli et al., 2008) to 16.2

mg/d (Kimira et al., 1998), while the mean of the maximum intake scenario in the present

study was 0.0000037 mg/d. Similarly, the mean intake of genistein in previous studies ranged

from 0.098 mg/d (Bertoli et al., 2008) to 23.3 mg/d (Kimira et al., 1998), and in our study the

mean of the maximum intake scenario was 0.0000037 mg/d. The total dietary and/or

supplemental isoflavone intake (the sum of daidzein, genistein, glycitein, biochanin A and/or

formononetin) was also considerably higher in previous studies in Australia (Hanna, O'Neill, &

Lyons-Wall, 2010), Denmark (Zamora-Ros et al., 2012), France (Zamora-Ros et al., 2012),

Germany (Zamora-Ros et al., 2012), Greece (Zamora-Ros et al., 2012), Italy (Zamora-Ros et al.,

2012), Korea (Lee, Cho, Park, Shin, & Sung, 2013), the Netherlands (Zamora-Ros et al., 2012),

Norway (Zamora-Ros et al., 2012), Spain (Zamora-Ros et al., 2012), Sweden (Zamora-Ros et al.,

2012), the UK (Ward et al., 2010; Zamora-Ros et al., 2012) and the USA (Bandera et al., 2011)

13
(mean intake 0.11 (Zamora-Ros et al., 2012)-24.6 mg/d (Lee et al., 2013)) than the mean of the

maximum intake scenario from PFS in the present study (0.0000037 mg/d).

Kaempferol and quercetin belong to the flavonols group, and their dietary intake has been

estimated in adult populations in Australia (Johannot and Somerset, 2006), China (Cao, Zhang,

Chen, and Zhao, 2010), Finland (Bobe et al., 2008), Japan (Kimira et al., 1998), Korea (Lee et al.,

2013), the Netherlands (de Vries, Janssen, Hollman, van Staveren and Katan, 1997), Scotland

(Kyle, Sharp, Little, Duthie and McNeill, 2010) and the USA (Lin et al., 2006), using dietary

methods such as the food frequency questionnaire (Bobe et al., 2008; Lin et al., 2006), food

records (Cao et al., 2010; de Vries et al., 1997; Kimira et al., 1998) or the 24-h recall (Johannot

& Somerset, 2006; Lee et al., 2013) as the dietary method. As in the case of the isoflavones,

the intake of kaempferol and quercetin from fennel-containing PFS was also minimal

compared to the reported intake from the diet. The mean dietary intake of kaempferol ranged

from 0.501 mg/d (Bobe et al., 2008) to 14.97 mg/d (Cao et al., 2010), and the mean intake

from PFS in the present study was 0.0218 mg/d. The mean dietary intake of quercetin ranged

from 3.0 mg/d in a Korean population (Lee et al., 2013) to 28.5 mg/d in a Dutch study (de Vries

et al., 1997), while the average intake estimate in the present study was 0.16 mg/d.

The intake of flavones apigenin and luteolin has been investigated among adult populations in

Australia (Johannot and Somerset, 2006), China (Cao et al., 2010), Finland (Bobe et al., 2008),

Japan (Kimira et al., 1998), and the USA (Lin et al., 2006) using food frequency questionnaires

(Cao et al., 2010; de Vries et al., 1997), food records (Cao et al., 2010; Kimira et al., 1998) or

24-h recalls (Johannot & Somerset, 2006). Similarly to isoflavones and flavonols, the intakes of

apigenin and luteolin from PFS were negligible compared to the intake from foods. The

maximum intake estimate of apigenin in the present study was 0.000258 mg/d, while the

mean dietary intake in previous reports ranged from 0.089 mg/d in Finland (Bobe et al., 2008)

to 4.23 mg/d in China (Cao et al., 2010). For luteolin, the average intake estimate in the

14
present study was 0.000335 mg/d, and the range of mean intakes in previous studies was

0.048 mg/d in the Finnish study (Bobe et al., 2008) to 8.08 mg/d in China (Cao et al., 2010).

Matairesinol belongs to the group of lignans. Its dietary intake has been reported from

countries including Denmark (Tetens et al., 2013), Finland (Tetens et al., 2013; Valsta et al.,

2003), Italy (Tetens et al., 2013), the Netherlands (Boker et al., 2002), Sweden (Tetens et al.,

2013), native and immigrant populations in the UK (Tetens et al., 2013; Ward et al., 2010), and

the USA (Bandera et al., 2011). The methods applied were food frequency questionnaires

(Bandera et al., 2011; Boker et al., 2002), the 7 day mixed survey technique (Tetens et al.,

2013), food records (Tetens et al., 2013; Ward et al., 2010) and 24 h dietary recalls (Tetens et

al., 2013; Valsta et al., 2003). The matairesinol intake from PFS was also low (the average

intake estimate in the present study was 0.00004 mg/d) compared with the mean dietary

intake (ranging from 0.006 mg/d (Bandera et al., 2011) to 0.09 mg/d (Nurmi, Mursu, Penalvo,

Poulsen and Voutilainen, 2010). Similarly, the summed dietary intakes of lignans were much

higher than the total lignan intake from PFS. In most of the previous studies, the lignan group

has consisted of the sum of matairesinol, secoisolariciresinol, lariciresinol and pinoresinol

(Bandera et al., 2011; McCann et al., 2010; Milder et al., 2005; Nurmi et al., 2010; Pellegrini et

al., 2010; Suzuki et al., 2008; Tetens et al., 2013; Zamora-Ros et al., 2012), sometimes only of

matairesinol and secoisolariciresinol (Kuhnle et al., 2011; Valsta et al., 2003; Ward et al., 2010),

and in one study of the sum of enterolactone and enterodiol (Hanna et al., 2010). Intakes have

been reported from Australia (Hanna et al., 2010), Denmark (Tetens et al., 2013; Zamora-Ros

et al., 2012), Finland (Nurmi et al., 2010; Tetens et al., 2013; Valsta et al., 2003), France

(Zamora-Ros et al., 2012), Germany (Zamora-Ros et al., 2012), Greece (Zamora-Ros et al.,

2012), Italy (Pellegrini et al., 2010; Tetens et al., 2013; Zamora-Ros et al., 2012), the

Netherlands (Milder et al., 2005; Zamora-Ros et al., 2012), Norway (Zamora-Ros et al., 2012),

Spain (Zamora-Ros et al., 2012) , Sweden (Tetens et al., 2013; Zamora-Ros et al., 2012), the UK

(Tetens et al., 2013) and the USA (Bandera et al., 2011). In the present study, the average

15
intake estimate from PFS was 0.000263 mg/d, and the mean dietary intake in previous studies

was from 0.244 mg/d (McCann et al., 2010) to 2.71 mg/d (Hanna et al., 2010).

To the best of our knowledge, intake estimates in human populations for trans-anethole, p-

coumaric acid and rosmarinic acid have not previously been published. Trans-anethole is used

in the food industry as a flavouring agent. Trans-anethole is hepatotoxic in rats, and the

World Health Organization has set an acceptable daily intake (ADI ) value of 0-2 mg/ kg body

weight for its use as a food (Joint FAO/WHO expert committee on food additives, 2000). For a

60 kg adult, this would mean an intake of 120 mg/d. The median intake of the maximum intake

scenario for trans-anethole from fennel-containing PFS was 21.4 mg/g, and the highest

individual intake of the maximum intake scenario was 87.0 mg/d.

The intake of p-coumaric acid from fennel-containing PFS was low compared with the supply

from common foods. According to previous analyses, the average intake of p-coumaric acid

from PFS (0.0068 mg/d) would be supplied from less than 1 g to 17 g of tomato, depending on

the variety (Barros et al., 2012) or 0.5 g of lettuce (Bayram et al., 2012). We did not find

concentration information for rosmarinic acid in other foods than herbs.

In conclusion, the intakes of daidzein, genistein, total isoflavones, kaempferol, quercetin,

apigenin, luteolin, matairesinol and lignans from fennel-containing PFS were low in comparison

to what is known of their dietary supply in different populations. The intake of estragole, a

potentially harmful compound, from PFS was usually moderate when compared with intake

from other dietary sources. However, the individual range of intake was quite large, and a

heavy consumption of fennel-containing PFS may lead to a comparably high intake of

estragole. Similarly, the intake of trans-anethole from fennel-containing PFS did not exceed the

ADI value in the present study even in the upper range of the maximum intake scenario, but

PFS sources should be taken into account when assessing the total exposure. To the best of

our knowledge, this study has provided the first intake estimates for trans-anethole, p-

coumaric acid and rosmarinic acid in human populations.

16
Acknowledgements

We wish to thank all the respondents of the PFS Consumer Survey. We are grateful to all the

partners of the PlantLIBRA Project, who made this study possible. Special thanks to colleagues

who have contributed to the PlantLIBRA PFS Consumer Survey and the ePlantLIBRA Database

(see front page for members of the Working Groups); thanks to the PlantLIBRA partners who

added the fennel composition data to the database: Qi Chang and Rui-Le Pan (Institute of

Medicinal Plant Development, China), Lucia Steenkamp (Council for Scientific and Industrial

Research, South Africa) and Jackie Hughes (Institute of Food Research, UK); thanks also to

Viktoria Knaze, Raimon Mil-Villarroel and Liliana Vargas-Murga (Fundacin para la

Investigacin Nutricional, Barcelona Science Park, University of Barcelona, Barcelona, Spain);

The research leading to these results has received funding from the European Communitys

Seventh Framework Programme (FP7/20072013) under grant agreement n 245199. It was

carried out within the PlantLIBRA project (website: www. plantlibra.eu). This article does not

necessarily reflect the Commission views or its future policy on this area.

17
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Figure 1. Calculation of the consumed amounts of plant food supplements

24
Table 1. Characteristics of Finnish plant food supplement (PFS) consumers

Consumers of fennel- Total consumers (n=401)

containing PFS (n=21)
Sex
Male 6 (28.6%) 193 (48.1%)
Female 15 (71.4%) 208 (51.9%)
Mean age, years 45.5 48.3
Level of educationa
Lowest 1 (4.8%) 47 (11.7%)
Medium 11 (52.4%) 237 (59.1%)
Highest 9 (42.9%) 117 (29.2%)
BMI
Underweight 0 (0.0%) 9 (2.2%)
Normalweight 11 (52.4%) 188 (46.9%)
Overweight 7 (33.3%) 147 (36.7%)
Obese 3 (14.3%) 57 (14.2%)
Perceived health status
Good or very good 15 (71.4%) 306 (76.3%)
Neither good nor bad 5 (23.8%) 77 (19.2%)
Bad or very bad 1 (4.8%) 18 (4.5%)
Regular medication during
the last 12 mo
Yes 12 (57.1%) 194 (48.4%)
No 9 (42.9%) 207 (51.6%)
Consumption of other
dietary supplements
Yes 19 (90.5%) 305 (76.1%)
No 2 (9.5%) 84 (20.9%)
Not sure 0 (0.0%) 12 (3.0%)
Use of complementary or
alternative health care
Yes 12 (57.1%) 218 (54.4%
No 9 (42.9%) 183 (45.6%)
Special diet
Yes 7 (33.3%) 117 (29.2%)
No 14 (66.7%) 284 (70.8%)
Smoking status
Never smoker 10 (47.6%) 182 (45.4%)
Current smoker 4 (19.0%) 90 (22.4%)
Ex-smoker 7 (33.3%) 129 (32.2%)
a
=Classification according to the Finnish education system: Lowest = Primary or lower
secondary education, Medium = Upper secondary level or lowest level tertiary education,
Highest = Lower-degree or higher-degree level tertiary education (Statistics Finland)

25
Table 2: Characteristics of the plant food supplements that contain fennel (Foeniculum vulgare) consumed by 21 consumers among the sample of 401
Finnish PFS consumers of the PlantLIBRA PFS Consumer Survey.

Specification Dose form Amount of fennel Number of consumers Average amount of Average consumption
in product consumption/da days/year
Water extract of fennel, Liquid 1.36 mg/ml 1 1.7 ml 90
fennel oil
Fennel seed Liquid 9.6 mg/ml 6 23 ml 27
Fennel Liquid 8.3 mg/ml 4 11 ml 60
Fennel oil Tablet 3 mg/tablet 4 2.25 tablets 169
Water extract of fennel Liquid 1.75 mg/ml 5 30 ml 51
seeds
Ground fennel seed Tablet 14.1 mg/tablet 2 14 tablets 5
a
During the reported consumption period

26
Table 3: Intake of bioactive components from plant food supplements that contain fennel (Foeniculum vulgare ) among the 21 Finnish consumers.

Bioactive component Daily intake during consumption period (median, range) Daily intake during previous 12 months (median, range)
Minimum estimate Average estimate Maximum Minimum Average estimate Maximum
estimate estimate estimate
Estragole (mg) 0.01 (0.00-0.95) 0.20 (0.01-7.95) 0.78 (0.03-32.26) 0.00 (0.00-0.15) 0.02 (0.00-1.27) 0.06 (0.01-5.14)
Rosmarinic acid (mg) 0.04 (0.00-2.59) 0.09 (0.00-5.61) 0.15 (0.00-9.41) 0.00 (0.00-0.51) 0.01 (0.00-1.38) 0.01(0.00-2.32)
Trans-anethole (mg) 0.41 (0.03-33.41) 1.15 (0.05-63.07) 21.44 (0.07-86,98) 0.03 (0.01-5.32) 0.12 (0.01-10.04) 0.17 (0.01-13.85)
p-Coumaric acid (mg) - 0.0068 (0.00-0.38) - - 0.0007 (0.00-0.06) -
Daidzein (g) - - 0.0007 (0.0002- - - 0.0001 (0.00005-
0.288) 0.046)
Genistein (g) - - 0.0007 (0.0002- - - 0.0001 (0.00005-
0.288) 0.046)
Kaempferol (mg) - 0.0034 (0.00-0.19) - - 0.0004 (0.00-0.03) -
Apigenin (g) - - 0.049 (0.00-2.02) - - 0.004 (0.00-0.32)
Luteolin (g) - 0.0525 (0.00-2.88) - - 0.0055 (0.00-0.46) -
Quercetin (mg) - 0.0246 (0.00-1.35) - - 0.0026 (0.00-0.21) -
Matairesinol (g) - 0.0066 (0.00-0.36) - - 0.0007 (0.00-0.06) -
Total isoflavones (g) - - 0.0007 (0.0002- - - 0.0007 (0.0002-
0.288) 0.288)
Mixture of lignans - 0.0412 (0.00-2.26) - - 0.0043 (0.00-0.36) -
(g)

27
 Number/
amount
usually
taken

Average
Consumption consumption
frequency per during
day consumption
period
Average
consumption
during last
12 months

Proportion of
consumption
period:
Consumption
days per
year/365

28
 Intake of estragole from fennel-containing plant food supplements (PFS) was moderate
Intake of trans-anethole from fennel-containing PFS was lower than ADI value
Intakes of beneficial compounds from PFS were low compared with dietary intake
First intake estimates of p-coumaric and rosmarinic acids are presented

29