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Journal of the Science of Food and Agriculture

J Sci Food Agric 83:330337 (online: 2003) DOI: 10.1002/jsfa.1316

Plant sterols in vegetables, fruits and berries


Vieno Piironen,1* Jari Toivo,1 Riitta Puupponen-Pimia2 and Anna-Maija Lampi1
1 2

Department of Applied Chemistry and Microbiology, PO Box 27 (Latokartanonkaari 11), FIN-00014 University of Helsinki, Finland VTT Biotechnology, PO Box 1500, FIN-02044 VTT, Finland

Abstract: The plant sterol contents of the most important vegetables, fruits and berries available in Finland were determined by capillary gas chromatography. The sample preparation procedure included both acid and alkaline hydrolysis to liberate sterols from their conjugates. The plant sterol contents of fresh vegetables ranged from 51 to 370 mg kg1 fresh weight (fw) in samples obtained from retail sale. The highest contents (>300 mg kg1) were measured in broccoli, Brussels sprouts, cauliower and dill and the lowest (51 mg kg1) in potato. The range of sterol contents on a dry weight (dw) basis was high, 2464100 mg kg1 dw. Considerable variation was also observed when individual samples of some vegetables were compared. Sitosterol was the main sterol (proportion 4386%) in all vegetables except cucumber and spinach, in which D7-sterols dominated. The total sterol contents were in the range 116228 mg kg1 fw in all fresh fruits except avocado, which contained more sterols, 752 mg kg1 fw. In fresh berries the corresponding range was 60279 m kg1 fw. The wild berries lingonberry and blueberry were better plant sterol sources than the cultivated berries blackcurrant, redcurrant and strawberry. In fruits and berries the proportion of sitosterol ranged from 6193% total sterols. On the basis of the results, the contribution of vegetables, fruits and berries to the total average daily plant sterol intake was estimated to be ca 60 mg. # 2003 Society of Chemical Industry

Keywords: plant sterols; phytosterols; vegetables; fruits; berries

INTRODUCTION

Plant sterols have been shown to decrease serum total and LDL cholesterol when sterol-enriched foods providing 1.52 g of plant sterols per day are consumed.14 Furthermore, epidemiological and experimental studies suggest that dietary plant sterols may offer protection against several types of cancer.5 These results have aroused general interest in the signicance of plant sterols in foods. However, little is known about the effects of plant sterol intake from non-enriched foods. Plant sterols were suggested as being at least partially responsible for the differences in plasma cholesterol levels and synthesis observed when 16 normolipidemic subjects were given experimental diets including either corn or olive oil.6 Agren et al 7 showed that both serum total and LDL cholesterol levels of patients with rheumatoid arthritis were signicantly lowered by a vegan diet containing on average 732 mg of sitosterol and 164 mg of campesterol per day, compared with an omnivorous diet. In addition, Ellegard et al 8 concluded that the effect of the current dietary recommendation to reduce saturated fat and increase dietary bre may be partly explained by the plant sterol content of the diet. In a casecontrol study by De Stefani et al 9 the total plant

sterol intake was strongly inversely related to the incidence of stomach cancer; the relationship remained after controlling the effects of antioxidants such as vitamin C. On the other hand, a higher dietary intake of plant sterols was not associated with a lower risk of colon and rectal cancer in a prospective epidemiological study (the Netherlands Cohort Study on Diet and Cancer).10 Vegetable oils and cereals are generally known to be the best natural sources of dietary plant sterols.11 However, as Normen et al 10 concluded, high consumption of fruits and vegetables, which seldom have a sterol concentration above 200300 mg kg1 fresh weight, also contributes substantially to the total sterol intake. In their study the contribution of fruits and vegetables to the total plant sterol intake was calculated to be on average 21% total sterols. The total plant sterol intake from vegetables, fruits and berries is composed of a large number of individual sterols.12 These can be divided into subgroups based on the structure of the ring system (ie the number and location of double bonds and methylation at the C-4 position) and side chain (alkylation, double bonds). The most common sterols in vegetables, fruits and berries are 4-desmethylsterols such as sitosterol,

* Correspondence to: Vieno Piironen, Department of Applied Chemistry and Microbiology, PO Box 27 (Latokartanonkaari 11), FIN-00014 University of Helsinki, Finland E-mail: vieno.piironen@helsinki. Current address: National Technology Agency of Finland, PO Box 69, FIN-00101 Helsinki, Finland Contract/grant sponsor: Academy of Finland Contract/grant sponsor: National Technology Agency of Finland (Received 11 July 2002; revised version received 27 August 2002; accepted 23 October 2002)

# 2003 Society of Chemical Industry. J Sci Food Agric 00225142/2003/$30.00

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campesterol, stigmasterol and avenasterols. Monomethyl and dimethyl sterols serve mainly as precursors of the end-products.12 Plant sterols occur both as free sterols and as bound conjugates, ie fatty acid esters (mainly C16 and C18 fatty acids), esters of phenolic acids, glycosides (most commonly with b-D-glucose) and acylated glycosides (esteried at the 6-hydroxy group of the sugar moiety).13 The plant sterol content in a given plant may vary depending on many factors, such as genetic background, growing conditions, tissue maturity and postharvest changes.11 In plants, sterols have both a structural function and a metabolic role.1316 They are integral membrane components which serve to regulate the uidity and permeability of membranes, affecting various membrane functions such as simple diffusion, carriermediated diffusion and active transport across the membrane, as well as modulating the activities of membrane-associated proteins including enzymes, receptors and signal transduction components. In addition, they are precursors of other bioactive steroids. They are biogenic precursors of so-called brassinosteroids, a special class of growth substances, and substrates for the synthesis of numerous secondary plant metabolites. Although the level of general interest in plant-based foods as natural plant sterol sources has increased, very few plant sterol compositional studies producing data suitable for food composition databases or tables have been published. Validated food composition data are needed in order to be able to investigate further the effects of natural plant sterol levels. In these compositional studies, both validation of the analytical method for each food group to be studied and representative sampling are crucial. Recent studies have focused mainly on individual vegetables or on the role of sterols in plants. The most comprehensive recent study was by Normen et al,17 in which vegetable and fruit samples (34 items) obtained from two shops were analysed for plant sterols; no berries were included in the sampling. Earlier, Weihrauch and Gardner18 collected data published up to 1977, and Jonker et al 19 analyzed plant sterols in a few vegetables. The German food composition tables20 and the USDA food composition database21 also include some data on plant sterols. However, only total plant sterol contents20,21 or sitosterol, campesterol and total sterol contents20 are given and the source of the information is unspecied. This study was undertaken in order to produce plant sterol composition data which could be used to update the Finnish national food composition database maintained by the Public Health Institute (Helsinki, Finland). Earlier, data on cereal products were published.22 In this study, samples of the most important vegetables, fruits and berries available in Finland were analysed for plant sterols using a validated gas chromatographic method. The sample preparation method included both acid and alkaline hydrolysis to liberate all bound steryl conjugates.
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MATERIALS AND METHODS Sampling

The most important vegetables, fruits and berries available in Finland were selected for analysis. The sampling also included selected vegetable and fruit products, almonds, peanuts and several other items (see Tables 15). Most samples (see Tables 1, 4 and 5) were purchased from retail stores (most of the samples in Tables 1, 4 and 5) or marketplaces (berries, domestic apples) in 1996 and 1997 as described by Koivu et al. 23 The sampling was supplemented in 2001. The 10 retail stores, representing the major food chains in Finland, and four marketplaces were located in the Helsinki area. Most vegetables were of domestic origin and were purchased at the peak of their season, and all fruits except apple were imported. Eight to 10 subsamples of 0.21 kg were taken from each food item. In general, one pooled sample was prepared representing each item, and the samples were determined as consumed (edible parts were analysed). Each subsample was diced if needed, and identical amounts of each subsample were incorporated in the pool.23 The pooled samples were kept at 18 C until being freeze-dried for analysis. The freeze-dried samples were stored at 18 C and homogenised prior to extraction. In addition, samples of some vegetables were taken from other sources in 19972000 in order to provide information about variation in sterol contents (see Tables 2 and 3). Different cabbages were obtained directly from a food company (Lannen Ltd, Sakyla, Finland). These samples (ca 2 kg) represented one industrial lot used for the production of frozen vegetable products (see Table 2). Furthermore, samples of pea (see Table 3) taken to represent three cultivars and from two or three growing years were obtained from the same company. All pea samples were grown at the same experimental farm in Finland. In addition, samples (24 kg) of different domestic onions (grown in the same area in Finland) were obtained from a wholesale store (see Table 2). All these samples were freeze-dried, homogenised with a household homogenizer (Bamix, Type M133, Esge AG, Mettlen, Switzerland) and thereafter stored at 18 C.
Sample preparation for sterol analysis

Plant sterols were extracted and puried from vegetable, fruit and berry samples according to the protocol presented earlier for cereal samples using similar reagents and solvents.22 All samples were analysed in triplicate. At the beginning of the determination an internal standard, either cholesterol (99%; Aldrich Chemical Co, Milwaukee, WI, USA) or dihydrocholesterol ($95%; Sigma Chemical Co, St Louis, MO, USA), was added to the homogenised freeze-dried sample (0.5 g). With fresh samples the sample size was 1.0 g. The samples were subjected to acid hydrolysis with HCl in ethanol at 8085 C for 60 min and
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alkaline hydrolysis with KOH at the same temperature for 30 min in order to liberate sterols from conjugates. However, for cucumber and spinach samples, only alkaline hydrolysis was used, because they contain D7-sterols which are prone to isomerisation or decomposition under acidic conditions. Thereafter 12 ml of water was added, the unsaponiable lipids were extracted with 20 ml of cyclohexane, and the extracts were puried using either C18 cartridge (Varian MegaBond Elut, 1.0 g; Harbor City, CA, USA) or SiOH cartridge (Varian Bond Elut, 0.5 g) solid phase extraction.22 The evaporated sterol fraction residue was dissolved in 500 ml of dichloromethane, and an aliquot of 100 ml was evaporated to dryness and silylated. The samples were silylated with BSTFA/TMCS (99:1) in pyridine.22 The TMS ether derivatives were dissolved in 200 ml of hexane before analysis by gas chromatography.
Gas chromatographic (GC) analysis of sterols

vegetable samples was 98.2 6.6% (N = 5). Furthermore, the GC separation and quantication were monitored daily by injecting a standard sterol mixture containing cholesterol, dihydrocholesterol and stigmasterol. The detection limit of the GC analysis calculated per 0.5 ml injection was 0.75 ng. Thus the lowest sterol concentration given when the normal sample size, sample preparation and GC analysis procedures were applied to a sample with an average moisture content was 1.6 mg kg1 fresh weight.
Moisture determination

The moisture contents of the fresh products were determined gravimetrically by heating the samples (1 5 g) at 103 2 C overnight. Residual moisture in the freeze-dried samples was measured either gravimetrically or by the Karl Fischer titrimetric method using a Mettler DL18 titrator (Mettler Instruments, Greifensee, Switzerland) according to the manufacturers instructions.

GC separation of sterol TMS ethers was performed using an RTX-5w/Integra fused silica capillary column (5% diphenyl/95% dimethyl polysiloxane, 60 m 0.32 mm id, 0.1 mm lm with 10 m Integra-Guard column; Restek Corp, Bellefonte, PA, USA) with oncolumn injection and ame ionisation detection as described by Piironen et al. 22 The injection volume was 0.5 ml. Quantication was based on an internal standard method as reported earlier.22 A blank test without any internal standard was run for each material to ascertain that there were no interfering peaks at the same retention time. When cholesterol was used as an internal standard, trace amounts of cholesterol found in the samples were taken into account when calculating the results. Identication of sterols was based on their retention times relative to those of commercially available sterols (sitosterol 95%, campesterol 98%, stigmasterol 95% and sitostanol 96.7%; Sigma Chemical Co), mass spectral analysis (GC/MS) and comparison with literature data.2426 In the GC/MS analyses, sterol TMS ethers were separated in a column similar to that described above, and running conditions were the same as those described by Mattila et al. 27
Quality control of plant sterol analyses

RESULTS AND DISCUSSION

In this study the total plant sterol contents of vegetable, fruit and berry samples were analysed using a sample preparation procedure enabling the determination of total sterol contents, ie both free sterols and sterols bound to conjugates (esters and glycosides) were measured. The main focus was on producing data which could be utilised for updating food composition databases. Therefore plant sterols occurring in amounts signicant as dietary components are reported separately (see Tables 1, 2, 4 and 5). The values for stanols thus include both sitostanol and campestanol, the values for avenasterols include both D5- and D7-avenasterol, and the values for other sterols include minor desmethyl sterols, monomethyl sterols and dimethyl sterols identied by mass spectral data as sterols.
Vegetables

The overall validation of the sample preparation and the GC method was reported by Toivo et al. 28 They showed that the recoveries of cholesterol, cholesteryl palmitate and glycosidic sterols added to whole wheat our (N = 6) were 93.8%, 100.1% and 91.2% respectively. In our routine work, one of the two inhouse reference samples, freeze-dried pea and whole grain wheat our, was analysed on each day of analysis to monitor the analytical consistency. The coefcients of variation for sitosterol and campesterol were 7.1 and 7.6% in the pea sample (N = 49) and 7.1 and 7.6% in the whole grain wheat our sample (N = 25) respectively. The recovery of stigmasterol added to various
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In fresh vegetables the total plant sterol contents were from low to moderate, ranging from 51 (potato) to 370 (Brussels sprouts) mg kg1 fresh weight (fw), when retail samples were compared (Table 1). The highest values, >300 mg kg1, were measured in broccoli, Brussels sprouts, cauliower and dill. The contents were < 100 mg kg1 in cucumber, potted lettuce, onion, potato and tomato, 100200 mg kg1 in carrot, Chinese cabbage, leek, red beet, swede and white cabbage, and 200300 mg kg1 in parsley, pea and sweet pepper. The range of sterol contents of the analysed vegetables was surprisingly high, 246 4100 mg kg1, when the plant sterol contents were calculated on a dry weight (dw) basis. The contents were > 4000 mg kg1 in cauliower, 3000 4000 mg kg1 in broccoli and 20003000 mg kg1 in Brussels sprouts, Chinese cabbage, cucumber, dill, potted lettuce and sweet pepper. In most other samples the contents were in the range 1000
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Plant sterols in vegetables, fruits and berries

2000 mg kg1. The lowest values, <1000 mg kg1, were found in leek, onion and potato. In the frozen vegetable products the contents varied in the range 10331677 mg kg1 dw, spinach being the richest source of sterols. The differences in sterol contents may be largely explained by the anatomical structure of the tissue, ie the proportion of membrane-rich tissues such as meristematic tissue and chlorenchyma. Vascular tissue and starch-rich tissue, on the other hand, contain fewer membranes. For example, ower heads of cauliower and broccoli are mainly composed of meristematic tissue, which thus may explain their higher sterol contents. It is interesting to note that the highest contents were on a similar level to those of many vegetable oils.11 For example, the total plant sterol contents of rened soybean oil are in the range 22103280 mg kg1. In whole grain wheat, rye, barley and oats the contents were found to be 466985 mg kg1 fw.22 Some differences were observed when the pooled samples representing vegetables on the retail market (Table 1) were compared with the individual samples obtained directly from industry (Table 2). The

difference was about 30% in the sterol contents of white cabbage and broccoli when the contents were expressed on a fresh weight basis. In cauliower the range of total sterol contents was 274411 mg kg1 fw. Furthermore, the total sterol contents of different onion samples (Table 2) varied considerably, from 59 to 192 mg kg1 fw (7331451 mg kg1 dw). In the case of pea (Table 3) the total sterol contents of the three cultivars grown in two or three years varied from 214 to 270, from 190 to 249 and from 179 to 258 mg kg1 fw respectively when samples of approximately the same maturity level were compared. Comparison of peas of varying maturity levels measured as tenderometer values (ca 100200) indicated that higher sterol contents are found at higher maturity levels. Previously, maturity level, water stress during the growing period, pre-storage treatment and storage were shown to cause considerable variation in the sterol contents and compositions of vegetables.2932. One additional factor which may cause variation in the published results arises from differences in the separation of edible portions of vegetables. Earlier we showed that the ower heads of cauliower and

Table 1. Plant sterol contents in vegetables (mg kg1 fw)

Sample Broccoli (Brassica oleracea var italica Plenck) Brussels sprouts (Brassica oleracea var gemmifera DC) Carrot (Daucus carota ssp sativus (Hoffm) Schubler & G Martens) Cauliower (Brassica oleracea var botrytis L) Chinese cabbage (Brassica pekinensis (Lour) Rupr) Cucumber (Cucumis sativus L) Dill (Anethum graveolens L) Leek (Allium porrum L) Onion, yellow (Allium cepa L) Parsley (Petroselinum crispum (Miller) Nyman ex AW Hill) Pea (Pisum sativum L) Potato (Solanum tuberosum L) Potted lettuce (Lactuca sativa var crispa L) Red beet (Beta vulgaris var conditiva Alef) Swede (Brassica napus L) Sweet pepper, red (Capsicum annuum L) Tomato (Lycopersicon esculentum Miller) White cabbage (Brassica oleracea var capitata L) Bean, frozen Spinach, frozen Sweet corn, frozen

Total Total plant Dry Other plant sterols matter D5- D7(g kg1) Brassicasterol Campesterol Stigmasterol Sitosterol avenasterols Stanols sterols sterols (mg kg1 dw) 108 155 113 30 20 67 1 71 2 20 0 80 30 27 0 285 5 277 5 104 4 20 80 2 0a 367 6 9 0 370 6 2 0 153 4 3408 56 2388 41 1361 36

76 47 38 131 224 154 149 222 210 35 167 104 85 64 99 110 61 276

20 20 30

72 2 22 0 16 2 13 1 60 12 1 36 2 60 60 27 1 42 1 50 31 1 81 20 54 2

16 1 20 133 4 12 0 115 2 12 0 30 24 0 57 3 20 16 0 36 3 15 1

216 7 103 1 155 2 166 6 70 2 136 3 212 3 32 3 37 1 91 3 103 2 164 3 33 1 114 4 54 3 152 4

20 20 10 1 20 20 20 40 40 15 2

3 0 310 10 4100 127 a 130 0 2790 10 2051 33 2480 71 865 34 603 20 1943 43 1337 21 246 20 2434 43 1023 41 1265 27 2583 44 1167 34 1500 43 1124 56 1677 18 1033 24

76 1 78 1 6 1 12 1 325 9 14 0a 194 8 4 0a 93 3 18 2 5 0 288 6 5 1 22 1 297 5 14 1a 51 4 5 0 11 0 85 2 50 80 20 12 1 171 7 132 3 6 0 220 4 9 1a 74 2 148 4

12 1 14 1 124 6 92 1 102 1 32 3 17 1 285 7

, trace (<2 mg kg1); , not detected. a In addition, minor amount of cholesterol.

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Table 2. Plant sterol contents in some individual vegetable samples (mg kg1 fw)

Sample Cabbages Broccoli Cauliower, 1 Cauliower, 2 Cauliower, 3 White cabbage Onions Yellow onion, 1 Yellow onion, 2 Yellow onion, 3 Red onion

Total Dry Total plant matter D5- D7Other plant sterols 1 (g kg ) Brassicasterol Campesterol Stigmasterol Sitosterol avenasterols Stanols sterols sterols (mg kg1 dw) 108 78 67 72 77 57 131 125 132 90 14 0 20 4 88 1 91 1 57 2 68 2 19 0 30 14 0 50 24 5 60 33 0 12 1 16 0 52 15 1 90 20 4 381 8 285 17 196 8 205 1 86 1 46 2 81 4 76 6 109 21 20 21 30 30 20 20 20 30 16 0 51 91 3 0a 23 1a 3 0a 35 5a 494 9 411 19 274 11 301 2 105 2 59 5 137 4 92 8 192 31 4575 80 5274 249 4092 169 4180 28 1367 20 1029 81 1043 30 733 64 1451 238

, trace (<2 mg kg1); , not detected; 13, individual samples. a In addition, minor amount of cholesterol.

broccoli were better sources of plant sterols than less membrane-rich parts of these vegetables.33 Their contents were 4180 and 4210 mg kg1 dw, whereas the contents of the other parts were 12101730 and 2960 mg kg1 respectively. On the other hand, the peels of several vegetables had about twofold higher sterol contents than the edible portions. All these factors leading to variation in the sterol contents set demands for the sampling and sample description in food compositional studies. The number of subsamples should always be considered in order to ensure reliability of the results in food compositional studies. In all vegetables except spinach and cucumber, sitosterol was the main plant sterol. Its proportion varied from 43 to 86% of the total sterols (TS). Campesterol was the second most important sterol in most vegetables. However, in some vegetables the proportion of stigmasterol was the second highest, up to a maximum of ca 40% (dill, parsley). The stanol

contents ranged from 0 (not detected) to 11%, being 2% TS in most samples; those of the other sterols ranged from 0 (not detected) to 27%. According to previous studies, D7-sterols dominated in cucumber and spinach.34 The main sterols in cucumber and spinach were spinasterol and dihydrospinasterol, contributing 34% and 33% of the total sterols in cucumber and 62% and 27% in spinach respectively. In general, the distribution of sterols among species in the family Cucurbitaceae has been found to constitute one of the most complex patterns known in the plant kingdom. The analysed total plant sterol contents of vegetables were generally similar to those published by Normen et al,17 who also used both acid and alkaline hydrolysis in sample preparation. In their study, samples were obtained from two shops. However, some differences were also noticed; for example, their value for leek was 81 mg kg1 fw (our value 194 mg kg1) and for Chinese cabbage 85 mg kg1 fw (in this

Table 3. Plant sterol contents in three cultivars of pea grown in three years

Pea variety (maturity level a) Bikini (105) Avola (100) Avola (115) Avola (160) Avola (196) Bikini (100) Bikini (103) Bikini (150) Bikini (185) Snake (103) Avola (119) Snake (101) Avola (118) Snake (119)
a

Year 1997 1998 1998 1998 1998 1998 1998 1998 1998 1998 1999 1999 2000 2000

Dry matter (g kg1) 196 176 209 239 272 210 194 230 266 226 224 214 207 213

Total plant sterols (mg kg1 fw) 233 15 233 8 249 10 353 21 502 14 270 13 214 3 426 8 474 11 258 6 216 6 216 12 190 8 179 3

Total plant sterols (mg kg1 dw) 1187 76 1326 44 1192 48 1477 87 1845 51 1286 62 1102 16 1851 35 1781 41 1141 27 966 28 1008 54 918 39 839 14

Tenderometer value.

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study 130 mg kg1). Earlier, Jonker et al 19 published data on few vegetables, utilising a sample preparation method including acid hydrolysis. Their results for cauliower and potato are comparable with our results. On the other hand, the results collected by Weihrauch and Gardner18 from earlier studies and those of the German food composition table20 are generally somewhat lower, which may be due mainly to differences in analytical methods.

contents of oil in mature fruits (820 g kg1) were lower than those in immature fruits (1162 g kg1).
Berries

Fruits

The total plant sterol contents ranged between 116 (banana) and 228 (orange) mg kg1 fw in all fresh fruits except avocado, which contained signicantly more sterols, 752 mg kg1 (Table 4). On a dry weight basis the contents ranged between 471 and 2929 mg kg1. The main component was again sitosterol (7286%), but considerable differences were observed in the sterol compositions. Plum and banana contained only a few sterols in signicant amounts, whereas the proportion of other sterols was highest (12%) in grapes, which also contained signicant amounts of stanols. The total sterol contents are generally in agreement with the values published by Normen et al 17 and Jonker et al,19 although in the case of individual fruits some differences were noticed. For example, our value for kiwi fruit (182 mg kg1 fw) is higher than that of Normen et al 17 (91 mg kg1). Weihrauch and Gardner18 and the German food composition table20 reported lower total sterol contents for several fruits. Only a few recent studies on fruits as sources of plant sterols have been published. The recent interest has been mainly in changes in sterol contents during maturation. In Chardonnay grape berry skins35 and peach fruits,36 sterol contents declined continuously during maturation. An increase occurred at peak maturity in grapes and could be related to the phenomenon of over-maturation.35 Similarly, Lozano et al 37 showed that the plant sterol

The plant sterol contents in fresh berries ranged from 60 (red currant) to 279 (lingonberry) mg kg1 fw, being signicantly lower in the cultivated berries blackcurrant, redcurrant and strawberry than in the wild berries lingonberry and blueberry (Table 5). Cultivated raspberry was richer in sterols than the other cultivated berries. As in the case of vegetables and fruits, a high variation in the sterol contents was observed when these were calculated on a dry weight basis. The plant sterol contents ranged from 372 to 2160 mg kg1, being considerably lower in blackcurrant and redcurrant and somewhat lower in strawberry than in the other berries. Interestingly, differences between berries could not be explained by the seed size, the proportion of seeds in the berries or the oil content of the berries. For example, lingonberry and blueberry were comparable to raspberry as sterol sources, although the proportion of seeds in these berries is considerably lower than that in raspberry (14 and 29 versus 101 g kg1).38 In addition, their oil contents are lower (41 and 85 g kg1 fresh berries) than that of raspberry (223 g kg1).38 The main sterol in berries was sitosterol; its proportion was 6193%, being lowest in lingonberry. The proportion of other sterols was considerable, especially in lingonberry and strawberry, in which it exceeded 20% of the total sterols. The proportion of stanols was also signicant in lingonberry (12%) and blueberry (6.5%). Previously, interest in berries as plant sterol sources has been very limited. In earlier studies, strawberry was found to contain 120 mg sterols kg1 fw.18 The plant sterol content of Bulgarian berry seed oils differed considerably, being 1.2 g kg1 in chokeberry seed oil, 1.4 g kg1 in blackcurrant seed oil and only 0.4 g kg1 in rosehip seed oil.39 In rosehip seed the total oil content was the lowest, only one-third of that

Table 4. Plant sterol contents in fruits (mg kg1 fw)

Sample Apple (Malus domestica Borkh) Avocado (Persea americana Miller) Banana (Musa paradisiaca L) Grape, green (Vitis spp) Kiwi fruit (Actinidia deliciosa (A Chew) C S Liang & A R Ferguson) Orange (Citrus sinensis (L) Osbeck) Orange juice Plum, red (Prunus domestica L)

Total Total plant Dry Other plant sterols matter D5- D71 (g kg ) Brassicasterol Campesterol Stigmasterol Sitosterol avenasterols Stanols sterols sterols (mg kg1 dw) 122 257 246 144 140 90 41 2 13 1 14 0 50 31 13 1 20 23 0 157 4 618 5 84 8 143 3 137 1 70 39 2 8 1 183 7 5 1 46 29 752 25 6 0 116 9 15 1 23 1 200 4 4 1 12 1 182 2 1499 57 2929 97 471 38 1390 29 1302 13

123 98 142

61

34 1 10 1 11 0

90 50 70

170 4 69 6 106 3

40

20

12 1a 228 6 4 1a 90 8 130 2

1854 52 921 83 913 15

, trace (<2 mg kg1); , not detected. a In addition, minor amount of cholesterol.

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Table 5. Plant sterol contents in berries and some miscellaneous samples (mg kg1 fw)

Sample Berries Blackcurrant (Ribes nigrum L) Blueberry (Vaccinium myrtillus L) Lingonberry (Vaccinium vitis-idaea L) Raspberry (Rubus idaeus L) Redcurrant (Ribes rubrum L) Strawberry (Fragaria ananassa Duch) Others Alfalfa, seed (Medicago sativa L) Alfalfa, shoots Almond (Prunus dulcis (Miller) D A Webb) Peanut (Arachis hypogaea L)

Dry matter D5- D7(g kg1) Brassicasterol Campesterol Stigmasterol Sitosterol avenasterols Stanols 198 122 156 143 162 91 50 10 1 60 90 20 20 21 81 1 222 11 169 3 233 6 56 2 73 3 20 10 0 31 17 0 35 1 20

Other sterols 13 2 67 1 20 0 22 2

Total plant sterols

Total plant sterols (mg kg1 dw)

88 1 446 7 264 13 2163 109 279 3 274 6 60 2 100 1 1774 9 1918 42 372 13 1100 8

921 59 956 940

48 1 80 33 1 162 3

416 13 82 2 60 120 0

593 39 57 7 1175 21 716 3

144 73 15 4 61 0 75 3

224 12 533 27 1959 97 2127 106 10 0 20 1 14 2 27 0 199 10 3377 165 89 6 1384 27 1448 28 89 1 1176 6 1251 6

, trace (<2 mg kg1); , not detected.

in the other two berries. Recently, sea buckthorn berries were shown to contain 340520 mg sterols kg1, whereas the corresponding values in seeds and fresh pulp/peel were 12001800 and 240 400 mg kg1 respectively.40
Other samples

According to previous data, almonds and peanuts contain considerable amounts of sterols, 1348 and 1176 mg kg1 fw respectively. Previously published values for different nuts and almonds range between 220 and 2200 mg kg1.1821 We also selected seeds and shoots of alfalfa as sample material, because the shoots are consumed in signicant amounts, especially by vegetarians. The total sterol content of shoots on a fresh weight basis (199 mg kg1) was within the range of vegetables, and the content on a dry weight basis (3377 mg kg1) was among the highest measured in vegetables (Table 5).

CONCLUSION

(National Public Health Institute dietary survey for Finnish adults) 1997 survey data, the plant sterol intake of the Finnish adult population was estimated to be 305 and 237 mg day1 for men and women respectively (Valsta L et al, in preparation). Cereal products and margarines and oils were the major contributors. However, the contribution of vegetables, fruits and berries was also shown to be signicant; they supplied about 200 mg g1 of the total plant sterol intake for men and 250 mg g1 for women. Furthermore, based on food consumption gures of the Food Balance Sheet42 and Vegetable Balance Sheet,43 the average daily plant sterol intake from vegetables, fruits and berries was estimated to be ca 60 mg. This is an approximate estimate based on total food consumption gures, which do not take into account food waste or variation in food consumption between individuals or population groups. In the vegetable group the greatest amount of plant sterols was estimated to be supplied by potato, although its sterol content is low. The other main individual contributors were various cabbages and carrot.

The results of this study were successfully used to update the Finnish national food composition database maintained by the National Public Health Institute (Valsta L et al, in preparation). In this study, mainly fresh vegetables, fruits and berries were analysed. However, both our own study41 and that of Normen et al 17 showed that cooking of vegetables does not cause signicant losses of plant sterols. Therefore the data can also be used to calculate plant sterol intakes from cooked foods. The database thus enables estimation of plant sterol intakes and can be utilised in epidemiological studies for investigating the signicance of plant sterols on the basis of their natural dietary levels. Using the database and the FINDIET
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ACKNOWLEDGEMENTS

This study was nancially supported by the Academy of Finland, the National Technology Agency and several Finnish food companies. The authors thank L Laukkanen, R Ronkainen, M Toivo, P Hakala, R-L Hovi, A Majanen and K Risunen for their technical assistance.

REFERENCES
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