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The effect of salatrim, a low-calorie modified triacylglycerol, on appetite and energy intake^1,2,3

¹ From the Department of Human Nutrition, Centre for Advanced Food Studies, Faculty of Life Sciences (LBS, HTC, MTA, CB, and AA), the Department of Biomedical Sciences, The Panum Institute (JJH), and the Department of Clinical Biochemistry, Rigshospitalet (JFR), University of Copenhagen, Copenhagen, Denmark

² Supported by Danisco A/S and The Danish Council for Strategic Research for Food and Health.

³ Reprints not available. Address correspondence to LB Sørensen, Department of Human Nutrition, Centre for Advanced Food Studies, Faculty of Life Sciences, University of Copenhagen, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark. E-mail: lbs{at}life.ku.dk.

	ABSTRACT

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

 
Background:Salatrim is modified triacylglycerol that is rich in short-chain fatty acids and stearic acid. It is used as a lower-calorie fat replacer. In addition, it has been hypothesized that salatrim's reduced absorption in the small intestine may lead to greater amounts of fat in the gastrointestinal tract, which may decrease appetite and energy intake through the release of appetite-regulating gastrointestinal hormones.

Objective:We aimed to compare the effects of salatrim and traditional fat on appetite, ad libitum energy intake, and gastrointestinal hormones.

Design:Twenty-two healthy, young, normal-weight men participated in a randomized, double-blind, crossover study. Test meals were a traditional fat meal and a salatrim meal with a mixture of traditional fat and salatrim. Visual analogue scales were used to record appetite and well-being every 30 min, and blood was sampled frequently. An ad libitum lunch was served 4.5 h after the test meal.

Results:The salatrim meal increased fullness (P = 0.04) and decreased hunger (P = 0.06) significantly more than did the traditional fat meal. The traditional fat meal increased well-being (P = 0.02). There was no significant difference in ad libitum energy intake or overall energy intake between the 2 test days. No significant differences in blood glucose, insulin, triacylglycerol, ghrelin, cholecystokinin, glucagon-like peptide-1, or peptide YY concentrations were found. A significantly (P = 0.01) smaller increase in free fatty acids was observed after the salatrim meal than after the traditional fat meal.

Conclusions:Salatrim had a modestly more suppressive effect on appetite than did a traditional fat. Gastrointestinal hormones did not seem to be involved.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

 
Obesity is a major health problem worldwide, and it is a risk factor for several chronic disorders (1-3). Even small changes in energy intake (EI) that lead to a positive energy balance can result in weight gain over time (4). Thus, slight modifications in food intake, such as the inclusion of foods that affect energy balance, may prevent weight gain and even facilitate weight loss. Replacing traditional dietary fat with low-calorie fat, such as modified triacylglycerols, could be a way to reduce EI. An example is the structured triacylglycerol salatrim (5). The salatrim family of triacylglycerols contains a mixture of short-chain and long-chain fatty acids (FAs); the short-chain FAs are acetate, propionate, and butyrate, and the long-chain FAs predominantly are stearic acid (6). The different salatrim types differ in the content of short-chain FAs; some contain all 3 FAs, others only 2 FAs, and some just 1 FA. Because the high amounts of short-chain FAs supply fewer kilojoules per gram than do high amounts of long-chain FAs, the caloric value of salatrim is lower than that of traditional fat (7). In addition, because of the high proportion of stearic acid, which is poorly absorbed, the caloric availability of salatrim is assumed to be lower than that of traditional fat (7, 8). Thus, compared with the 38 kJ/g (9 kcal/g) in traditional fat, salatrim is estimated to deliver 21 kJ/g (5 kcal/g), according to US regulation (5) or 25 kJ/g (6 kcal/g) according to European Union regulation (9). The exact caloric value is dependent on the composition of the specific type of salatrim (7).

In the present study, we tested the hypothesis that both salatrim's lower caloric value and its reduced absorption in the small intestine will result in a lower EI than is seen with traditional fat. A lower absorption of salatrim leaves undigested FAs in the middle and lower intestine, which may generate greater feelings of satiety and decrease EI because of appetite-regulating gastrointestinal hormones such as cholecystokinin, glucagon-like peptide-1 (GLP-1), and peptide YY (PYY). Thus, the objective of the present study was to compare the effects of salatrim and traditional fat on appetite sensations, ad libitum EI, and gastrointestinal hormones.

	SUBJECTS AND METHODS

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

 
Study design and subjects
The subjects were tested on 2 different occasions in a double-blind, randomized, crossover design. On 2 test days, a breakfast meal containing either traditional fat only or a mixture of salatrim and traditional fat was served; 4.5 h later, an ad libitum lunch was served. The test days were separated by a 2–4-wk wash-out period. Twenty-two healthy, normal-weight men who were nonsmokers and not elite athletes and who had no history of obesity, diabetes, or liver disease were included in the study.

All subjects provided written informed consent before the start of the study. Ethical approval was obtained from the Ethics Committee of Copenhagen and Frederiksberg (KF 01 275625), and the study was performed in accordance with the Helsinki II declaration.

Test meals
The breakfast test meal
Two test meals that were rich in fat were prepared for breakfast. Both test meals consisted of a roll, raspberry jam, and water for drinking. In the salatrim roll, both salatrim (Benefat C-IP; Danisco A/S, Brabrand, Denmark; material no. 1211962, batch no. 4010259499) and margarine (AMA; Dragsbæk A/S, Thisted, Denmark) were incorporated. In the roll with traditional fat, only margarine (AMA; Dragsbæk A/S) was incorporated (Table 1). The Benefat and the margarine were softened gently before being mixed with the other ingredients of the rolls.

The ad libitum lunch
The ad libitum lunch was a homogenous mixed cold dish consisting of pasta, minced ham, grated carrots, green peas, and sour cream dressing (15% of energy from protein, 30% from fat, and 55% from carbohydrate; 616 kJ/100 g).

Experimental protocol
Before each test day, the subjects followed a standardized weight-maintenance diet (22% of energy from fat, 18% from protein, and 60% from carbohydrate; 2.0 g/MJ fiber). To ensure equally filled glycogen stores, the subjects were not allowed to engage in strenuous physical activity for 1 d before the test day (11).

On the test day, the subjects arrived at the department at 0800 after having fasted for 12 h. They had been instructed to use a nonstrenuous means of transportation. Each subject's body weight was measured to the nearest 0.1 kg by using a digital scale (SECA model 707; Seca, Copenhagen, Denmark). After a 15-min rest, a catheter (Venflonpro 393206; Becton-Dickinson, Helsingborg, Sweden) was inserted in an antecubital arm vein; after another 15-min rest, a fasting blood sample was taken at 0830 (time = –15 min). The breakfast test meal was served at 0845 (time = 0 min) and was consumed within 15 min. The time spent consuming the test meal was recorded and reproduced on the second test day. Appetite ratings were made on 100-mm visual analogue scales (VASs) with text expressing the most negative and the most positive rating anchored at each end. The VASs were used to assess hunger, satiety, fullness, prospective food consumption, desires for special foods, well-being, and palatability of the test meal (ie, taste, smell, visual appeal, aftertaste, and overall palatability) (12). Appetite sensations were recorded every half-hour, and postprandial blood samples were taken 15, 45, 75, 135, 195, and 255 min after meal initiation. Palatability ratings were assessed immediately after consumption of the test meal. At 1330 (time = 285 min), lunch was served, and the subjects were instructed to eat ad libitum until comfortably satisfied (Figure 1). The amount of food eaten was measured by weighing.

View larger version (4K): [in this window] [in a new window]   FIGURE 1.. Protocol of the test day. Visual analogue scales (VAS) were used to register palatability of the test meal, satiety, hunger, fullness, prospective food consumption, well-being, thirst, and the desire to eat something sweet, salty, savory, or fatty.

Assays
Blood samples were drawn into test tubes. For glucose analyses, they were drawn into tubes containing EDTA or fluoride; for analyses of free FA (FFA), cholecystokinin, GLP-1, PYY, and ghrelin, they were drawn into tubes containing EDTA; and for insulin and triacylglycerol analyses, they were drawn into tubes with no additives. The tubes (except those for insulin and triacylglycerol analyses) were kept on ice. Plasma glucose was analyzed by using standard enzymatic methods (Roche Diagnostic Systems GmbH, Mannheim, Germany). Serum insulin was measured by using a chemiluminescent immunometric assay (Diagnostic Products Corporation, Los Angeles, CA). Serum triacylglycerol was analyzed by using the Test-Combination Triacylglycerols GPO-PAP-method (Roche Diagnostic Systems GmbH, Basel, Switzerland). Plasma FFA concentrations were analyzed by using the ACS-ACOD-MEHA method in the Wako NEFA C test kit (Wako Chemicals GmbH, Neuss, Germany). GLP-1 was measured against standards of synthetic GLP-1(7–36) by using antiserum code no. 89390, which is specific for the amidated C-terminus of GLP-1 of intestinal origin (13). The radioimmunoassay of PYY in plasma was performed by using antiserum code no. 8412-2II (14). This reacts equally with PYY1-36 and PYY3-36. Synthetic human PYY (Peninsula Laboratories, St Helens, United Kingdom) was used for standards. We obtained ¹²⁵I-PYY1-36 (code no. IM259) from Amersham Biosciences (Little Chalfont, United Kingdom). Plasma ghrelin concentrations were measured by using a radioimmunoassay performed with the use of a kit (catalog no. GHRA-88HK; Linco Research, St Charles, MO) for active (octanoylated) ghrelin (15). Plasma cholecystokinin concentrations were measured with an accurate radioimmunoassay by using antiserum code no. 92128, which is specific for cholecystokinin peptides, and which does not bind any homologous gastrins (16). Quality controls were within accepted limits for all assays.

Statistical analyses
Baseline values were compared by using a paired t test. The effect of meal type on VAS scores was tested by using a repeated-measures analysis of covariance (ANCOVA) with the mixed procedure in SAS software (version 9.1; SAS Institute, Cary NC). Baseline values were included as covariates (Figure 2 and Figure 3). Because VAS scores are confined to the interval from 0 to 100, the scores were transformed (if X_i denotes the scores, then Y_i = arcsin Y_i = arcsin).

View larger version (18K): [in this window] [in a new window]   FIGURE 2.. Mean (±SEM) appetite scores (hunger, satiety, fullness, and prospective food consumption) during the 2 test days. n = 20. Visual analogue scales equal to 100 mm correspond to "I cannot eat another bite" (satiety), "I have never been hungrier" (hunger), "I am totally full" (fullness), and "I can eat a lot" (prospective food consumption). The breakfast test meal was served at 0 min. There were no significant differences between baseline values (paired t test). Repeated-measures ANCOVA was used to test differences between postprandial appetite scores. Hunger, P = 0.06; satiety, P = NS; fullness, P = 0.04; prospective food consumption, P = NS.

View larger version (12K): [in this window] [in a new window]   FIGURE 3.. Mean (±SEM) well-being scores during the 2 test days. n = 20. Visual analogue scales equal to 100 mm correspond to "I feel really good." The breakfast test meal was served at 0 min. There were no significant differences between baseline values (paired t test). Repeated-measures ANCOVA was used to test differences between postprandial scores (P = 0.02).

Figure 4

Figure 5

Figure 6

Figure 7

View larger version (23K): [in this window] [in a new window]   FIGURE 4.. Mean (±SEM) and individual values for energy intake at the ad libitum lunch and total energy intake. n = 20 for both. We used 21 kJ salatrim/g in the calculation. Paired t tests were used to test differences in the effect of meal type on ad libitum and total energy intakes. None of the differences were significant.

View larger version (14K): [in this window] [in a new window]   FIGURE 5.. Mean (±SEM) triacylglycerol and free fatty acid concentrations during the 2 test days. n = 20. The breakfast test meal was served at 0 min. There were no significant differences between baseline values (paired t test). Repeated-measures ANCOVA was used to test differences between postprandial concentrations. Data for triacylglycerol concentrations were log transformed before statistical analyses. Triacylglycerol, P = NS; free fatty acids, P = 0.01.

View larger version (13K): [in this window] [in a new window]   FIGURE 6.. Mean (±SEM) glucose and insulin concentrations during the 2 test days. n = 20. The breakfast test meal was served at 0 min. There were no significant differences between baseline values (paired t test) or between postprandial concentrations (repeated-measures ANCOVA).

View larger version (12K): [in this window] [in a new window]   FIGURE 7.. Mean (±SEM) glucagon-like peptide-1 (GLP-1) concentrations during the 2 test days. n = 20. The breakfast test meal was served at 0 min. There were no significant differences between baseline values (paired t test) or postprandial scores (repeated-measures ANCOVA).

	RESULTS

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

Table 2

There was no significant difference between the ad libitum EI after the salatrim meal (3414 kJ) and after the traditional fat meal (3331 kJ) (P = 0.63). The overall numerical difference in EI (energy from the test meal and the ad libitum meal) between the salatrim meal and the traditional fat meal was –256 kJ (95% CI: –611, 98; P = 0.15) when 21 kJ salatrim/g was used in the calculation and –172 kJ (95% CI: –527, 182; P = 0.32) when 25 kJ salatrim/g was used (Figure 4).

No significant differences were found in glucose and insulin concentrations between the 2 test days, although a trend for a time x meal interaction for glucose was observed (P = 0.07): glucose concentrations were higher at time 45 min after the traditional fat meal than after the salatrim meal (Figure 5). There were no significant differences in triacylglycerol concentrations, although postprandial concentrations were numerically lower after the salatrim meal than after the traditional fat meal (Figure 6). There was a significantly smaller increase in FFAs after the salatrim meal than after the traditional fat meal (P = 0.01) (Figure 6). No differences in ghrelin, cholecystokinin, GLP-1, or PYY concentrations were observed between the 2 test days (Figure 7).

The traditional fat meal increased well-being (P = 0.02) more than did the salatrim meal (Figure 3). There was a decrease in well-being at time 15 min, right after consumption of the salatrim meal, which was mainly due to the very low scores of 2 subjects. Fourteen of the 20 subjects did not report any gastrointestinal side effects 24 h after consumption of the breakfast test meals on the 2 test days, and no differences in subjective gastrointestinal side effects were reported by the rest of the subjects. There were no significant differences in the palatability ratings between the salatrim meal and the traditional fat meal (P > 0.30 for all variables).

	DISCUSSION

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

 
In the present study, a meal with a mixture of salatrim (Benefat C-IP) and traditional fat had a modestly greater effect on subjective appetite sensations than did a meal containing only traditional fat. The ad libitum and total EIs did not differ significantly on the 2 test days. Thus, the hypothesis that the ingestion of salatrim will result in less appetite and a lower subsequent EI than will the ingestion of traditional fat was only partly confirmed.

According to the hypothesis tested in the present study, salatrim should reduce the appetite because of less absorption of fat in the small intestine, which would lead to a greater release of appetite-regulating hormones in the gastrointestinal tract. However, there were no differences in concentrations of cholecystokinin, GLP-1, or PYY during the 2 test days. This finding suggests that there was no difference in fat absorption from the gastrointestinal tract. Because fecal fat excretion and composition were not measured in the present study, it is not known whether there actually was a difference in fat absorption due to differences in stearic acid content in the test meals. Finley et al (7, 8) investigated the absorption of stearic acid from salatrim in human subjects. Thirty-six subjects received a maintenance diet [either 1800 kcal/d (7.5 MJ/d) or 2500 kcal/d (10.5 MJ/d)]. For 4 d, the subjects received 45 g/d of either salatrim (23CA) (short-chain FAs acetate and propionate) or coconut oil or 60 g/d of either salatrim (23CA) or coconut oil in a parallel design. The investigators reported that the absorption of stearic acid in the subjects receiving 1800 and 2500 kcal salatrim/d (26 and 34 g stearic acid/d, respectively) was 72.4% and 63.5%, respectively (7). In a recent study, Tuomasjukka et al (17) investigated stearic acid absorption from 30 g of a different type of salatrim (Benefat B-R) (short-chain FA acetate) in a single meal (hamburger) in a double-blind crossover study. Consumption of the salatrim meal with 16.7 g stearic acid resulted in an apparent absorption of 87% stearic acid from salatrim. The 2 studies mentioned differ in design and in the type of salatrim used, which could explain some of the contrast in the results. The present study is most similar to the study by Tuomasjukka et al because it is a single-meal study, although the type of salatrim is not the same; among other things, the composition of the short-chain FAs is different. However, it is not known whether this difference in short-chain FA composition influenced the absorption of stearic absorption. In comparing the 2 single-meal studies, it could be expected that stearic acid excretion would be greater in the present study because the proportion of natural fat was lower than that in the study by Tuomasjukka et al, and this lower proportion of natural fat has been shown in rat studies to result in greater fecal excretion of stearic acid (7, 18, 19).

The salatrim meal was expected to result in a smaller postprandial increase in serum triacylglycerol than would the traditional fat meal both because of the high content of short-chain FAs in salatrim and because of the higher content of stearic acid in salatrim, which would lead to poor digestibility. However, there was no significant difference in postprandial triacylglycerol response between the 2 meals. Three studies have investigated the effect of salatrim on triacylglycerol concentrations: a 12-wk crossover study (20), the study by Finley et al (8), and a single-meal study (21). In the long-term study (20), there were no differences in triacylglycerol between the salatrim diet and other test fat diets. In the study by Finley et al (8), no differences in triacylglycerol were observed, although fecal stearic acid excretion was greater in the salatrim group. In the single-meal study conducted by Sanders et al (21), a meal consisting of a muffin with 30 g salatrim (23SO) as test fat and a milkshake with 20 g sunflower oil resulted in a significantly smaller increase in triacylglycerol than did 2 meals with either high-oleate sunflower oil or cocoa butter as test fat. The excretion of fat was not measured in the study of Sanders et al. It is possible that the reason that those investigators observed a difference in postprandial triacylglycerol response, whereas we did not, is that the test meal in the 2 studies consisted of 50 and 40 g fat, respectively, and 40 g fat may be too low a dose when measuring postprandial lipemia. In addition, the study of Sanders et al had 35 subjects, whereas the present study had 20. The numerical difference in postprandial triacylglycerol may indicate that the lack of statistical significance could be due to insufficient study power.

A significant difference in postprandial FFA profiles was observed in the present study. FFAs were measured for only 4 h after the test meal was served. Therefore, it is not clear whether the smaller increase in concentration of FFAs with salatrim would have been followed by an increase later on, as was seen in the study by Tholstrup et al (22), who investigated the effects of different FAs on the postprandial lipid profile. Thus, it is not known whether the result reflects slower or less-efficient stearic acid absorption after the salatrim meal than after the traditional meal.

Although the postprandial triacylglycerol response indicates a difference in fat absorption between the 2 groups in the present study, no differences in concentrations of cholecystokinin, GLP-1, or PYY were observed during the 2 test days. Nevertheless, a significant difference in some of the subjective appetite variables was observed. No studies investigating the effect of salatrim on appetite sensations and EI have been published. The World Health Organization Food Additives Series (23) cited unpublished observations of increased satiation after a salatrim meal in a single-meal study including supplementation with either salatrim (23SO) or cocoa butter. In that study, 4 men and 4 women received a single breakfast meal supplemented with 30 g of either salatrim (23SO) or cocoa butter in a crossover design. Two to 4 h after the salatrim meal, the women experienced discomfort, such as feeling bloated, cramped, or nauseated, whereas the men did not experience any discomfort but felt satiated and did not feel like eating for the rest of the day. The difference by sex in the occurrence of side effects was probably due to differences in body weight, an effect that was also seen in one of the studies by Finley et al (8). In the single-meal study conducted by Tuomasjukka et al, 30 g salatrim in a meal with other types of fat did not cause gastrointestinal problems in male subjects (17). In the single-meal study by Sanders et al (21), possible side effects were not investigated. In the other studies in which salatrim was investigated, salatrim was incorporated in cookies, muffins, bonbons, or ice cream, and the salatrim load was spread over the whole day. The general picture is that salatrim in doses of >30 g/d can cause gastrointestinal upsets and headaches, especially in women, and that mixing the salatrim fat with natural fat seems to reduce side effects (8, 20). This was confirmed in a pilot study carried out at the Department of Human Nutrition in which different salatrim test meals were tested (C Bitz, A Astrup, unpublished observations, 2005). In that study, consumption of test meals in which salatrim was the only type of fat led to nausea and gastrointestinal problems 2 h after consumption.

In the present study, subjects experienced a difference in well-being between the 2 test days, and it could be speculated that the differences in the subjects' appetite on the 2 test days may simply reflect this difference in well-being. The question is how to explain the difference in well-being. In general, the well-being reported on both test days was relatively high, and the difference between the 2 days was only modest when the initial level was considered. Except for a small decrease in well-being right after the salatrim test meal, the well-being scores reported are characterized by being at the same level as the initial well-being scores, whereas the well-being after the traditional fat test meal increased immediately after consumption of the test meal and stayed at this higher level for the rest of the test period. These results suggest that the difference in well-being was not due to side effects caused by salatrim, because the course of the salatrim test day in the present study does not resemble the course of the test day described in the unpublished study cited by the World Health Organization or observed in the unpublished pilot study.

The decrease in well-being after the salatrim meal cannot be explained by a difference in how the salatrim meal was perceived by the subjects, because there were no significant differences in the palatability ratings of the salatrim meal and the traditional fat meal. There were no differences in subjective gastrointestinal side effects 24 h after the consumption of the test meals on the 2 test days, which supports the notion that the salatrim meal in the present study did not cause gastrointestinal problems.

In conclusion, in the present study, the salatrim meal had a modestly more suppressive effect on appetite than did the traditional fat meal. Gastrointestinal hormones do not seem to be involved.

	ACKNOWLEDGMENTS

 
We thank Yvonne Fatum, Charlotte Kostecki, Søren Andreasen, and the rest of the technical staffs at the Department of Human Nutrition, the Department of Medical Physiology, and the Department of Clinical Biochemistry for expert assistance, and we gratefully acknowledge the participation of all the persons involved in the study.

The authors' responsibilities were as follows—CB and AA: designed the study; HTC and MTA: conducted data collection and data entry and participated in writing the first draft of the manuscript; JJH: analyzed GLP-1, PYY, and ghrelin; JFR: analyzed CCK; LBS: the statistical analysis; AA and LBS: interpreted data; LBS: wrote the initial draft of the manuscript; and all authors: reading and critiquing of the manuscript. None of the authors had a personal or financial conflict of interest.

	REFERENCES

TOP ABSTRACT INTRODUCTION SUBJECTS AND METHODS RESULTS DISCUSSION REFERENCES

 

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