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CAPÍTULO 1. NATURALEZA EVOLUTIVA DE LA APREHENSIÓN DEL

1.3. Elementos constitutivos de la imputabilidad/inimputabilidad

1.3.4. Esfera valorativa

been reported where dietary Ca was associated with an increased excretion of faecal fat and

fatty acids regardless of the form (either dairy products, Ca-fortified food products or Ca

supplements) in which the Ca was consumed (see Table 2-2). However, there are a few

studies which did not report an increase in faecal fat excretion with increased dietary Ca

(Ditscheid et al. 2005; Boon et al. 2007; Hjerpsted et al. 2011). A common factor in the latter

studies where no effect of high dietary Ca intake on faecal fat excretion was reported was

that the diets used contained greater amounts of protein than those used in the studies where

a “Ca effect” was observed (Boon et al. 2007; Hjerpsted et al. 2011). For example, Jacobsen et

al. (2005) reported conflicting effects for high dietary Ca concentrations on faecal fat

excretion within the same study when the protein intake differed for the subject groups. The

latter research group compared three different diets (see Table 2-2) varying in Ca (500 vs

1800 mg d-1) and in protein content (15 vs 23 E% protein) and observed that when the dietary

Ca content was increased and the protein content remained the same (15 E% protein) faecal

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54

excretion was observed when the increased dietary Ca intake was accompanied by an

increase in the protein content of the diet (23 E% protein). Similarly, a Ca-intervention study

in subjects with a daily protein intake of 20% of energy showed no differences in faecal fat

excretion for varying concentrations of dietary Ca (400 vs 1200 vs 2500 mg Ca d-1) (Boon et al.

2007). High protein diets have been reported to improve Ca absorption (Kerstetter et al.

1998), thereby likely leaving less Ca available for fatty acid complexation. In another study,

Jacobsen et al. (2005) reported a 66% higher urinary Ca excretion for the high-Ca/high-protein

diet compared to the high-Ca/normal-protein diet which suggested greater absorption of Ca

from the intestinal tract for the high protein diet, which would have resulted in less Ca being

available for Ca-fatty acid soap formation. A diet high in protein has been suggested to

improve Ca absorption (Kerstetter et al. 1998) by keeping Ca in solution and preventing

precipitation with interfering components such as phosphates (Scholz-Ahrens and

Schrezenmeir 2000; Camara-Martos and Amaro-Lopez 2002). As mentioned in section 2.3.2.1,

phosphopeptides derived from casein bind ionized Ca to the phosphate group of serine

residues, this would render Ca less available for the formation of Ca-fatty acid soaps and

explain the lower excretion of fatty acids in the presence of dairy protein.

2.5.2.

Fatty acid soap formation with divalent cations other than Ca

Ca is a divalent cation, and therefore when ionised has the ability to bind two free fatty acid

molecules forming a Ca-fatty acid complex that is suspected to be insoluble at the pH of

intestinal fluids and therefore passes through the gut unabsorbed (Gacs and Barltrop 1977).

However, Ca is not the only divalent cation present in the gastrointestinal tract. Nutritionally

relevant minerals such as Mg, Zn, Fe and Cu are also present in the gastrointestinal tract and

Chapter Two: Literature Review 55

fatty acids. Moreover, considering that dietary fat can impair Ca absorption most likely via

soap formation, the absorption of other divalent minerals may be similarly affected. Despite

the obvious similarity between Ca and other divalent minerals little work has been

conducted examining the ability of divalent cations other than Ca to form soaps. A few

studies investigating the effect of dietary fat on Ca retention have included observations

about faecal Mg excretion and digestibility. Of the limited information available, Kaup et al.

(1990) reported that, feeding rats diets containing 20 g 100 g-1 of butter fat resulted in a 20%

lower apparent faecal magnesium digestibility and greater faecal Mg excretion when

compared to rats receiving a diet containing 5 g 100 g-1 of butter fat suggesting that Mg-fatty

acid soaps may be the mechanism by which Mg absorption was altered. Moreover, and as is

the case for Ca, Mg absorption appears to be influenced to a greater degree by the presence

of saturated fatty acids as opposed to unsaturated fatty acids. The latter is evidenced by a

study where rats receiving a diet containing 25 g 100 g-1 triolein absorbed up to 55% of the

ingested Mg (apparent Mg digestibility was comparable to the fat-free control diet) whereas

for diets containing 25 g 100 g-1 diet of tripalmitin or tristearin only, 30% of the ingested Mg

was absorbed (Tadayyon and Lutwak 1969b). However, not all published studies have

reported an effect of dietary fat consumption on increased faecal Mg excretion or reduced

Mg absorption. For example, comparing fat-free diets to diets supplemented with oleic or

palmitic acid and with varying Ca concentrations did not show decreased Mg retention in

chicks when fat was present (Atteh and Leeson 1984). Moreover, a study in rats investigating

the effect of the positional distribution of stearic and oleic acid in the triacylglycerol molecule

did not show an influence of the structured triglycerides on faecal excretion and the apparent

digestibility of Mg (Brink et al. 1995). In general, the results from some of the latter studies

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56

acids and therefore that Mg-fatty acid soap formation is possible. However, Mg-fatty acid

soap formation does not seem to be as profound as Ca-fatty acid soap formation, as studies

showing an increase in faecal Ca excretion did not always show an effect on Mg excretion

(Atteh and Leeson 1984). Studies investigating the effect of dietary fat on divalent trace

mineral absorption (eg Zn, Fe, Cu) are inconclusive (Lukaski et al. 1986; Johnson et al. 1992;

Wapnir and Sia 1996). It has been suggested that an increase in dietary fat increases Fe

absorption in rats, regardless of the fat source (Johnson et al. 1987). However, some studies

suggest a decrease in the absorption of Zn and Fe in the presence of polyunsaturated fatty

acids (Lukaski et al. 1986; Wapnir and Lee 1990). For example, a study performed with

endurance athletes showed increased faecal Zn and Fe excretion in the presence of a diet rich

in polyunsaturated fatty acids compared to a diet high in carbohydrates or saturated

medium chain fatty acids (Lukaski et al. 1986). Moreover, in an in vivo study using intestinal

perfused rats, arachidonic acid (C20:4) inhibited zinc removal from the intestinal lumen.

However, the saturated long chain fatty acid, palmitic acid, had the opposite effect and

stimulated zinc absorption. Another study in rats showed improved Fe absorption in the

presence of saturated fatty acids (Johnson et al. 1992). However, not all studies have showed

such an effect of saturated fatty acids on trace element (Fe and Cu) absorption (Johnson et al.

1987; Wapnir and Sia 1996). For example, Cu absorption had been reported to decrease

significantly in the presence of free palmitic acid and stearic acid when investigated in rats

using jejunal perfusion (Wapnir and Sia 1996). The inconclusive results and the limited

research conducted regarding divalent cations other than Ca and their ability for fatty acid

Chapter Two: Literature Review 57

2.6.

Concluding Comments

An increased dietary Ca intake has been repeatedly reported to increase faecal fat excretion

in animals and humans. It has been hypothesised that this Ca-induced increase in faecal fat

output leads to a decreased uptake of energy which might further result in a reduction of

body weight and body fat. The mechanism by which dietary Ca increases faecal fat output

has been suggested to be the formation of Ca-fatty acid complexes, so called Ca-fatty acid

soaps, which are insoluble in the intestinal environment and evade absorption in the small

intestine.

Although the formation of insoluble Ca-fatty acid soaps is believed to be the cause of the

reduced fat absorption, few researchers have attempted to isolate Ca-fatty acid soaps from

faecal samples. The currently available methods described in the literature to extract Ca-fatty

acid soaps from faecal material appear to poorly recover soaps. However, to determine what

impact Ca-fatty acid soap formation has on faecal fat excretion, it is necessary to be able to

determine the amount of fat excreted in the form of fatty acid-soaps.

The formation of Ca-fatty acid soaps is believed to not only alter the absorption of fat but has

also been shown to reduce the absorption of Ca from the gastrointestinal tract in studies

conducted in animals and human infants. If the formation of Ca-fatty acid soaps could

potentially impair Ca absorption, other divalent cations may also be able to complex with

fatty acids leading to decreased absorption of essential minerals from the intestinal tract.

Further research is necessary to definitively address the hypothesis that Ca-fatty acid soap

formation is the mechanism for increased faecal fatty acid excretion in the presence of dietary

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58

this has not been done well. Moreover, understanding where in the gastrointestinal tract

soaps are formed would provide important knowledge about the behaviour of dietary Ca

Chapter Three: Assay Development for Ca-Fatty Acid Soap Determination 59

CHAPTER THREE:

Development of an Assay to Determine