WO2005096837A2

WO2005096837A2 - A ready-to-use therapeutic food composition or nutritional supplement & a method of making the same

Info

Publication number: WO2005096837A2
Application number: PCT/GB2005/001342
Authority: WO
Inventors: Stephen Collins; Christiani Jeyakumar Henry
Original assignee: Valid International Ltd
Priority date: 2004-04-07
Filing date: 2005-04-06
Publication date: 2005-10-20
Also published as: WO2005096837A3; GB2412834A; GB0407931D0

Abstract

One embodiment of the present invention provides a ready-to-use therapeutic food or nutritional supplement that comprises a mixture of at least one ground, roasted cereal, and at least one ground, roasted legume or cooked leguminous protein extract, said at least one cereal and said at least one legume or leguminous protein extract being selected to be mutually complementary so as to provide a PDCAAS of at 5 least 60. Another embodiment relates to a method of making such a ready-to-use food or nutritional supplement, which comprises mixing at least one ground, roasted cereal with at least one ground, roasted legume or leguminous protein extract.

Description

A ready-to use therapeutic food composition or nutritional supplement & a method of making the same

The present invention relates to a ready-to-use therapeutic food composition or nutritional supplement and to a method of making such a composition. It is estimated that approximately one billion people around the World currently suffer from malnutrition of varying degrees of acuity; about 1-2% of children are severely malnourished; and about 40% of the World's population in total is chronically malnourished. Accordingly there is a great need for ready-to-use therapeutic food compositions ('RUTFs') that are capable of treating such malnourishment by providing the daily energy, protein and micronutrient (minerals, vitamins, and the like) requirements of human beings. An ideal RUTF should have a good nutritional quality, and good stability; it should be palatable, and have a consistency suitable for feeding in children and adults who may find it difficult to masticate food; preferably it should require little or no additional processing prior to feeding. In particular, an RUTF should be protein-rich, providing the essential amino acids that the body is incapable of synthesising itself at a sufficiently high rate. A joint FAO/WHO/UNU Expert Consultation in 1985¹ prescribed an amino acid reference pattern based on the essential amino acid requirements of a pre-school age child. In 1989 a joint FAO/WHO Expert Consultation on Protein Quality Evaluation² concluded that protein quality could be assessed adequately by expressing the content of the first limiting essential amino acid of a test protein as a percentage of the content of the same amino acid in the reference pattern. This percentage may be corrected for the true fecal digestibility of the test protein. It has now been recognized by both FAO and WHO that, in the absence of animal/human studies to evaluate protein quality, this scoring method, known as the protein digestibility-corrected amino acid score (PDCAAS), is the best alternative, and it has been adopted as the preferred method for the measurement of the protein value in human nutrition.

¹ FAO/WHO UNU Expert Consultation (1985) Energy and Protein Requirements. Technical Report Series 724. World Health Organisation, Geneva.

² FAO/WHO Expert Consultation (1990) Protein Quality Evaluation. Food and Agricultural Organisation of the United Nations, FAO Food and Nutrition Paper 51, Rome PDCAAS(%) = — x E x 100

where T = mg of Umiting amino acid in lg of test protein, R = mg of same amino acid in lg of reference protein, and F is the true fecal digestibility (%). The 'limiting amino acid' is the amino acid in a given protein showing the greatest percentage deficit in comparison with the amino acid pattern judged optimal for human growth and metabolism.

An RUTF should also be energy-rich, providing at least 4.0kcal/g, and should desirably contain the necessary micronutrients for supporting recovery and growth, including calcium, iron, and zinc, where appropriate.

It is also important for an RUTF to have a maximal protein, energy and nutrient density. Children aged one to three years can only eat about 200ml to 300 ml of solid food at one time, and if a child is only eating three small meals a day, then each one must have a sufficiently high concentration of energy and nutrients to meet the child's requirements over twenty-four hours.

A typical RUTF comprises a staple as the main ingredient, preferably a cereal, a protein supplement, and an energy supplement such as fat, oil or sugar to increase the energy concentration of the mix.

Such food compositions can also be used for other vulnerable individuals such, for example, as post-operative patients, and cancer patients suffering from cachexia, as well as for supplementing the diets of military personnel, sportsmen and women, and others needing or desiring a compact, energy-rich, protein-rich food supplement.

US 6346284 Bl discloses an oxidation-stable complete food or nutritional supplement having a moisture content of less than 10 % wt. and comprising a mixture of food-grade products that is coated with at least one lipid-rich substance derived from oleaginous seeds, and enriched with vitamins, soluble or insoluble mineral salts, enzymes or mixtures thereof. In one embodiment the coating substance contains a paste derived from peanuts, and the food-grade products include sl immed milk, powdered yogurt or whey as a protein supplement. This food is available commercially under the trade name "Plumpy'nut®".

However, a great many malnourished people live in developing countries where milk and other animal-derived products are expensive. Accordingly there is a need for a therapeutic food composition that does not include such products. Furthermore, peanuts are well known to suffer from a number of disadvantages. Specifically, peanuts are known to be susceptible to mycotoxins, particularly aflatoxin. They are also common allergens, and have a high phytate:zinc ratio, leading to poor mineral absorption. Accordingly there is a need to provide an RUTF that is peanut-free. In recent years, the advantages of community-based treatment of malnutrition have been recognised. It would be advantageous for local communities to use locally available food products in therapeutic food compositions since such food products are generally more available and less expensive than imported produce, and are more likely to be tolerated by the digestive systems of the local population. Moreover, the use of local produce could be expected to stimulate the local economy.

It is therefore an object of the present invention to provide a novel therapeutic food composition or food supplement that can be made from locally available food products.

Another object of the present invention is to provide such a therapeutic food composition or food supplement that is free or substantially free of peanuts, and it is yet another object of the present invention to provide such a therapeutic food composition or food supplement that is free or substantially free of animal-derived products, including dairy products such as milk and milk powder.

Yet another object of the present invention is to provide such a therapeutic food composition or food supplement that is protein-rich and comprises a staple cereal product. Yet another object of the present invention is to provide such a therapeutic food composition or food supplement that has a maximal PDCAAS score.

Yet another object of the present invention is to provide such a therapeutic food composition or food supplement that has good stability.

Yet another object of the present invention is to provide such a therapeutic food composition or food supplement that has an acceptable palatability, and preferably a consistency that is suitable for children.

A different object of the present invention is to provide a method of making a therapeutic food composition or food supplement which is inexpensive and easy to carry out, especially in developing countries. Yet another object of the present invention is to provide a method of making a therapeutic food composition or food supplement which is free or substantially free of peanuts and animal-derived products and utilises locally available food products. In accordance with one aspect of the present invention therefore there is provided a ready-to-use therapeutic food or nutritional supplement comprising a mixture of at least one ground, roasted cereal, and at least one ground, roasted legume or cooked leguminous protein extract, said at least one cereal and said at least one legume or leguminous protein extract being selected to be mutually complementary so as to provide a PDCAAS of at least 60.

Suitably the cereal and the legume or leguminous protein extract may be selected so as to give a maximal PDCAAS as defined by FAO/WHO, 1989. Preferably the PDCAAS of the food or supplement is at least 65, and more preferably at least 70. In some embodiments the RUTF or supplement according to the present invention may comprise a simple mix of one cereal and one legume or leguminous protein extract. However, in more complex embodiments, three or more cereals and legumes may combined to optimise the PDCAAS of the composition.

Said at least one cereal may comprise any cereal known to those skilled in the art. This has the advantage that it is generally possible to use a cereal product that is grown locally to the site of production of the RUTF or supplement.

In some embodiments, said at least one cereal may be selected from oats, maize, quinoa, barley, wheat, rye, triticale, sorghum, millet, teff, and rice. All cereals form a good basis for a young child's diet. They contain 7-14% wt. protein, up to 75% wt. carbohydrates and, apart from oats, quinoa and maize, negligible amounts of fat. Suitably, the RUTF or supplement in accordance with the present invention may be free or substantially free of any oleaginous seed or grain, i.e. any cereal or legume comprising more than about 20% wt. fat, such, for example, as peanuts, almonds, and pistachio nuts. Although oat is seldom grown in the tropics, oats or oat products are available in many markets and the protein value is superior to that of all other cereals. For this reason, a non-milk based, oat-substituted RUTF may an appropriate alternative for use in young children's diets. Likewise, although rice shows a low protein value (7% wt.), next to oats it is the best in protein quality; rice is easily absorbed owing to its low cellulose content. Overall, dry whole-cereal grain is also a valuable source of tMamine, riboflavin and niacin; it also contains calcium and iron. However, variation in cereals' nutrient content is not uncommon. For instance, maize differs in its pattern of amino acids and its lower niacin availability. However, these factors are less important in a well balanced ready-to-use food. Said at least one legume or leguminous protein extract may comprise any legume known to those skilled in the art. This has the advantage that it is generally possible to use a legume or legumes that are grown close to the place where the RUTF or supplement is made, provided that the at least one legume and at least one cereal complement one another to give the desired PDCAAS. In some embodiments, said legume may be selected from chick peas, soybeans, lentils, split peas, haricot beans, mung (green) beans, fava bean, pigeon pea, and kidney beans.

Many different legumes are eaten in traditional diets, and most have similar nutrient values. Their chief asset for use in infant food is that they are not only inexpensive but also contain a high amount of protein. The protein content of dried legumes varies between 20% wt. and 30 % wt. and, although it is of only moderate protein quality, it is valuable in supplementing other food proteins, notably cereals. Legumes are useful sources of mia in, riboflavin and niacin as well as folate, calcium and iron. However, it should be noted that nearly all legumes contain some form of natural toxin, which should be removed by careful preparation or processing. Most toxins are either soluble in water or are removed by normal cooking procedures. Most legumes contain little fat, with the exception of soybeans.

Said cereal and legume should be roasted since the use of raw non-roasted commodities could lead to the presence of potentially high levels of anti-nutritional factors and phytates. Roasting also improves flavour development, and alter the textural and physico-chemical properties of the cereal or legume. Preferably the cereal and legume are dry roasted to avoid the risk of rancidity associated with roasting in oil. In a village context the roasting step may be carried out using hot sand. Accordingly it is difficult to give a time-temperature coefficient for the roasting, but preferably roasting is carried on until substantially all anti-nutritional factors have been destroyed or removed.

Said at least one roasted cereal and said at least one roasted legume are ground such that they are included in the RUTF or supplement in particulate form. In some embodiments the cereal or legume may be milled to form a flour. Suitably the ground cereal and or the ground legume may have a particle size of less than 200μ, preferably less than lOOμ. For example, the ground cereal and legume may have a particle size in the range 50μ to lOOμ such that the RUTF or supplement is not 'gritty', thereby improving the acceptability of the composition, especially to young children. In some embodiments the ground, roasted cereal and legume components are then intimately mixed. By ''leguminous protein extract" is meant a product of leguminous origin in which the proteins have been concentrated. Said proteins may have been concentrated by an adequate process to contain at least 50% crude protein, as related to the dry matter, and which may be restructured or textured. A leguminous protein extract may be used in the RUTF or supplement according to the present invention instead of, or in addition to, one or more ground, roasted legumes to supplement the protein content of the at least one cereal by complementation. A protein extract made from any suitable legume may be used, including those mentioned above, but in some embodiments said RUTF or supplement may comprise soy bean extract which is widely available and inexpensive. A suitable soy bean extract is commercially available from Degussa Texturant Systems Ltd. under the trade name 'Soyamin 90'.

In some embodiments, the RUTF or nutritional supplement according to the present invention may further comprise at least one ground, roasted oilseed in addition to said at least one ground, roasted cereal and said at least one ground, roasted legume or leguminous protein extract. Advantageously, the oilseed may be selected to enhance the taste, flavour, or nutritional qualities of the RUTF or supplement. For example, the oilseed may be selected to increase the micronutrient content of the RUTF or supplement.

Preferably said at least one oilseed may comprise sesame seed. The use of sesame seeds in the formulation may be advantageous for a number of reasons. Sesame seeds provide acceptable taste and flavour. They are also a good source of polyunsaturated fatty acids. Further, they are a significant source of calcium as compared to many other oilseeds. (See Table 1 below). This is significant since, as mentioned above, the RUTF or supplement according to the present invention preferably excludes the use of milk powder, which is a rich source of calcium.

Table 1: Calcium levels in nuts and seeds

Food Calcium content (mg/lOOg)

Peanuts 60

Pecan nuts 61

Pine nuts 11

Pistachio nuts 61

Sesame seeds 670 Sunflower seeds 110

Source: (McCance, Widdowson's 1991³)

In one embodiment, the RUTF or nutritional supplement according to the present invention may comprise roasted rice, roasted sesame seeds and soya bean extract. Preferably, the RUTF or supplement may comprise about 18 to 22% wt. roasted rice flour, 6 to 10% wt. soy bean extract and 27 to 31% wt. roasted sesame seed paste. The RUTF may further comprise one or more oils, fats or sugars as an energy supplement, together with appropriate vitamins or minerals to make a total of 100% wt.

In another embodiment of the present invention, the RUTF or supplement according to the invention may comprise roasted barley as a cereal component, roasted sesame seeds as an oilseed, and a soy bean extract. Preferably, the RUTF or supplement comprises 13 to 17% wt. roasted barley flour, 25 to 29% wt. roasted sesame seed paste, and 7 to 11% wt. soya bean extract, with the balance comprising one or more energy supplements and vitamin-mineral premixes.

In yet another embodiment of the invention, the RUTF or supplement may comprise roasted maize as a cereal component, roasted sesame seeds as an oilseed, and roasted chick peas as a legume component. Preferably, the RUTF or supplement may comprise about 31 to 35% wt. roasted maize flour, 11 to 15% wt. sesame seed paste, and 23 to 27% wt. roasted chick pea flour, with the balance again being made up from energy supplements such as oil, fats or sugars, and vitam -mineral premixes. Such combinations of cereals, legumes and oil seeds have been found to be particularly advantageous having regard to their amino acid complementation, product stability, availability as raw materials, and their organoleptic properties.

In accordance with the present invention, the RUTF or nutritional supplement may suitably further comprise an external energy supplement. Said energy supplement may comprise one or more of oil, fat, and sugar. Any suitable energy-rich oils or fats known to those skilled in the art may be used, but preferably said oils and fats are selected from vegetable oils and fats.

³ McCance and Widdowson's (1991). The composition of foods (fifth edition), the Royal Society of Chemistry and MAFF. In some embodiments, said RUTF or supplement may comprise a vegetable oil selected from sunflower oil, safflower oil, corn oil, palm oil, soy bean oil, or any other vegetable oil that is available in the vicinity of the location where the RUTF or supplement in accordance with the present invention is to be made. Suitably the vegetable oil may also be selected on the basis of its nutritional qualities. For example, sunflower oil may be used in order to meet (n-3) and (n-6) fatty acids requirements. It is usually specified that at least 3 to 10% of total energy should be provided by (n-6) fatty acids, and 0.3 to 2.5% by (n-3) fatty acids.

Preferably, said vegetable oil has a free fatty acid content of less than about 0.5% wt. to avoid rancidity, and more preferably less than about 0.1% wt. Further the vegetable oil should have a peroxide value of less than about 20 mEq. of O2/kg of fat, more preferably less than about lOmEq. In some embodiments, the vegetable oil may have a peroxide value of less than 5mEq.

Said oil or fat should be mixed thoroughly with the cereal/legume mix and may form a paste which may advantageously be easy for severely malnourished people, particularly children, to eat.

The RUTF or supplement in accordance with the present invention may comprise more than about 10% wt. water, and preferably less than about 25% wt. water. Typically the RUTF or supplement comprises about 11-15% wt. water, e.g. 12% wt., 13% wt. or 14% wt. The RUTF or supplement may thus form a water-in-oil emulsion. However, the majority of the water is bound water, and suitably the water activity of the RUTF or supplement is less than about 0.5, preferably less than about 0.4. In some embodiments the

RUTF or supplement in accordance with the present invention may have a water activity in the range of about 0.2 to 0.3. Said sugar may comprise any suitable mono- or disaccharide, but sucrose is especially preferred since it is generally inexpensive and has a good water-holding capacity.

In some embodiments said energy supplement may comprise a vegetable oil, such as sunflower oil, and sugar. Suitably the RUTF or nutritional supplement according to the present invention may further comprise one or more supplementary micronutrients such, for example, as vitamins, soluble and insoluble mineral salts, and mixtures thereof. It may also include enzymes. As the product is not in aqueous solution, it is of no consequence whether the mineral salts used are soluble or insoluble in water. Insoluble mineral salts have the advantage of being less expensive and insipid.

Preferably the minerals include one or more of calcium, iron and zinc. Zinc is especially important to support growth. However, severely malnourished individuals cannot tolerate iron, and so in some embodiments the present invention provides an RUTF that is free or substantially free of iron. Examples of mineral salts which may be mentioned are calcium carbonate, zinc oxide, copper sulfate, potassium chloride, magnesium oxide, and iron salts. Minerals may be included in the composition by means of a suitable mineral pre- mix that is admixed with the other ingredients.

WHO, 1999⁴ recommends the following composition for a mineral mix:

Table 1

Mineral Parts

Potassium chloride 89.5

Tripotassium citrate 32.4

Magnesium chloride(MgCl2.6H2θ) 30.5

Zinc acetate 3.3

Copper sulfate 0.56

Sodium selenate^a lOxlO^"3

Potassium iodide^a 5xl0-³ aIf it is not possible to weigh very small amounts accurately, this substance may be omitted.

Vitamins may also be included in the RUTF or supplement, for example in the form of a pre-mix that is admixed with the other components. Said vitamins may be provided to meet or supplement the daily needs of a person taking the RUTF or supplement. In some embodiments, said vitamins may comprise one or more of Nitamin A,

⁴ Management of severe malnutrition: a manual for physicians and other senior health workers, 1999. World Health Organisation, Geneva. Vitamin D, Vitamin E, Vitamin C, Vitamin Bl, Vitamin B2, Vitamin B6, Vitamin B12, Vitamin B9, Vitamin PP, Vitamin B5, Vitamin B8, and Vitamin K.

WHO, 1999⁵ recommends the following composition for a vitamin mix:

Table 2

Vitamin Amount per litre of liquid diet

Water-soluble:

Tlnamine (vitamin Bl) 0.7mg Riboflavin (vitamin B2) 2.0mg Nicotinic acid lOmg Pyridoxine (vitamin B6) 0.7mg Cyanocobalamin (vitamin B12) lmg Folic acid 0.35mg

Ascorbic acid (vitamin C) lOOmg Pantothenic acid (vitamin B5) 3mg Biotin O.lmg

Fat-soluble:

Retinol (vitamin A) 1.5mg Calciferol (vitamin D) 30mg α-Tocopherol (vitamin E) 22mg Vitamin K 40mg

According to another aspect of the present invention, said RUTF or nutritional supplement may comprise:

10 to 40 % wt. ground, roasted cereal;

5 to 35 % wt. ground, roasted legume or leguminous protein concentrate;

0 to 30 % wt. ground, dry roasted oilseed;

10 to 30 % wt. oil;

10 to 30 % wt. sugar;

' Ibid. 0 to 5 % wt. micronutrients; wherein the total of said ingredients is 100 % wt.

In some embodiments, said RUTF or supplement may comprise 20-30% wt. of ground, roasted legume. Alternatively, said RUTF or supplement may comprise 5-10% wt., preferably 8-9% wt., leguminous protein extract, e.g. soybean extract.

Where a protein extract is used, 25-30% wt. oilseed may also be included, preferably about 26-29% wt., but where the RUTF or supplement comprises a ground, roasted legume, then a lower proportion of oilseed may be used, such for example as 10- 15% wt., typically 13-14% wt.

Similarly, where the RUTF or supplement comprises a protein extract, 15-25% wt. vegetable oil may also be included, preferably about 19-24% wt., but where the RUTF or supplement comprises a ground, roasted legume, then a lower proportion of vegetable oil may be used, such for example as 10-15% wt., typically 13-14% wt. Where the RUTF or supplement comprises a protein extract, 20-25% wt. sugar may also be included, preferably about 22-24% wt., but where the RUTF or supplement comprises a ground, roasted legume, then a lower proportion of sugar may be used, such for example as 12-18% wt., preferably 13-16% wt.

Said RUTF or supplement may comprise 1-2% wt., preferably 1.5-1.7% wt., of a mineral premix.

The RUTF or supplement in accordance with the present invention may have an energy content of greater than 5.0kcal/g, and a protein content of more than 12% wt. The RUTF or supplement may also have a carbohydrate content of about 40% wt. or more, and a fat content of 25% wt. or more. According to a different aspect of the present invention there is provided a method of making an RUTF or nutritional supplement in accordance with the present invention, which method comprises mixing at least one ground, roasted cereal with at least one ground, roasted legume or leguminous protein extract.

Preferably the cereal-legume mix is then mixed with an oil or fat, which oil or fat may carry sugar or a micronutrient premix.

The cereal-legume mix may further comprise at least one ground, roasted oilseed, which may typically be provided in the form a paste. The equipment should be clean and properly dried before starting production, since the presence of water in the RUTF or supplement would significantly increase the water activity, leading to possible microbial spoilage. A uniform product mixture is desired to deliver the therapeutic function of the RUTF by ensuring that substantially the same amount of nutrients is present in every portion that is eaten by a subject. Thus, sufficient mixing is important to guarantee such an outcome. For this reason, mixing should preferably be done mechanically.

The finished RUTF or supplement should then be transferred into suitable clean, dry containers, and stored away from heat and light. Advantageously the RUTF or supplement according to the present invention does not require any special storage or refrigeration, and may have a shelf-life of at least six months.

In yet another aspect of the present invention, the RUTF or nutritional supplement may comprise an inoculum of at least one probiotic microorganism.

By "probiotic" is meant a live microbial supplement which beneficially affects the host by improving its intestinal microbial balance. Various acid-resistant, intestinal species of lactobacilli and bifidobacteria are well known to those skilled in the art for use as probiotics, e.g. Lactococcus raffinolactis 32-77, Pediacoccu pentoseceus 5-33:3, Lactobacillus paracesei subsp. paracesei iP,and Lactobacillus plantarum 2362. These are commercially available from Synbiotics AB, Sweden, under the trade name "Synbiotic 2000 forte"™.

It has been found that the inclusion of a probiotic culture in an RUTF or nutritional supplement according to the present invention may serve to stabilise the lining of the large intestine in a malnourished individual, as well as in other groups suffering from disruption of the gut lining such for example as HTV/AIDS sufferers, and post-operative and cancer- cachexia patients. Desirably, the probiotic bacteria should remain inactive before consumption of the RUTF or supplement, and become active once the RUTF or supplement has been ingested.

In accordance with yet another aspect of the present invention therefore there is provided an RUTF or nutritional supplement comprising one or more food-grade products having a water activity of no more than 0.4, and an inoculum of at least one probiotic microorganism.

Advantageously, said RUTF or supplement may further comprise an effective amount of a prebiotic supplement. Known prebiotic substances are derived from insoluble and dietary fibres, resistant starches, carbohydrates and oligosaccharides . In some embodiments said prebiotic supplement may comprise one or more of beta-glucan, pectin, inulin, allicin, and fructo oligo saccharide.

In yet another aspect of the present invention there is provided a method of therapy or prophylaxis of diseases of the intestinal lining which comprises administering to a person in need thereof an effective therapeutic or prophylactic amount of an RUTF according to the present invention. Said diseases may include ailments of the gut associated with malnourishment, HIV/AIDS, cancer-cachexia, and post-operative conditions. In yet another aspect of the present invention there is provided the use of an

RUTF according to the present invention in the manufacture of a medicament for the treatment or prophylaxis of diseases of the intestinal lining.

Following is a description by way of example only with reference to the accompanying drawings of embodiments of the present invention. In the drawings:

FIG. 1 shows schematically a method of making of making a first RUTF or nutritional supplement in accordance with the present invention; and

FIG 2 shows schematically a method of making a second RUTF or nutritional supplement according to the present invention.

Example 1: Method of making an RUTF or nutritional supplement

In accordance with a first embodiment of the present invention, an RUTF or nutritional supplement is manufactured generally by roasting and milling cereal-legume- oilseed mixtures. This represents a simple and easily operable process in most developing countries. With reference to FIG. 1, the required quantities of one or more cereals 10, one or more legumes 12, and optionally one of more oilseeds 14 (see below) are first weighed out in predetermined proportions.

Cereals form a basic component of an RUTF or nutritional supplement in view of their good protein, carbohydrate, mineral and vitamin contents. However, most cereals generally lack one or more essential amino acids, and thus have a low PDCAAS score as defined above. Examples of amino acid deficiencies in various commodities used are shown in Table 3: Table 3: Amino acid deficiencies in various products used in weaning-food preparation

Amino acid

Main deficiency Good source of

Cereal grains

Rice (White) Lysine-Tryptophan Methionine

Wheat flour Lysine

Rice polishing Methionine Lysine

Oilseeds

Soybean Methionine Lysine

Sesame Lysine Methionine

Peanut Lysine-Methionine

Food legumes

All kinds Methionine Lysine

In accordance with the present invention, said one or more cereals are combined with one or more legumes, which cereals and legumes are selected such that their amino acid profiles complement one another as described above to produce an improved combined PDCAAS score of at least 60-62, preferably at least 65-67, and more preferably at least 70 or more, e.g. 71, 72, 73 or 74. In some embodiments, the cereals and legumes may also be combined with at least one oilseed to enhance further the amino acid and/or mineral content of the final RUTF or supplement. Examples of specific cereal-legume- oilseed mixtures are given below.

The weighed portions of cereals, legumes and oilseeds are then individually roasted to remove anti-nutritional factors, and to improve their organoleptic and physico-chemical properties; dry roasting is especially preferred. Each of the roasted products is then individually ground or milled to form a flour. Said flours may have a particle size in the range 50μ to lOOμ such that the final product is not 'gritty' on the tongue. In some embodiments said oilseed may be provided in the form of a paste. Instead of, or in addition to, a roasted and milled legume, a leguminous protein extract may be used. The roasted cereals, legumes, and optional oilseeds are then premixed in a clean, dry bowl 20 for at least 5 to 10 minutes. At this stage a predetermined proportion of a mineral and/or vitamin premix 24 may be added and intimately mixed with the other flours. A suitable mineral premix is available commercially from Nutriset, France, under the trade name "CMV". Said premix 24 may comprise any suitable mixture of vitamins or soluble or insoluble mineral salts.

A suitable premix 24 as recommended by WHO is recited above. Another suitable premix comprises:

Product Amount (in grams per 600 grams)

Vitamin A 8.68 Vitamin D 5.04 Vitamin E 31.50 Vitamin C 45.53 Vitamin Bl 0.47 Vitamin B2 1.42 Vitamin B6 0.47 Vitamin B 12 0.42 Vitamin B9 0.17 Vitamin PP 4.17 Vitamin B5 2.44 Vitamin B8 0.05 Vitamin K 0.33 Zinc oxide 9.30 Calcium carbonate 93.06 Copper sulfate 3.24 Potassium chloride 393.71 Total 600

Yet another suitable premix comprises: Product Amount (in grams per 4000 grams)

Vitamin A 6.12 Vitamin D 2.80 Vitamin E 13.68 Vitamin C 59.74 Vitamin Bl 1.20 Vitamin B2 1.50 Vitamin B6 0.67 Vitamin B12 1.61 Vitamin B9 0.31 Vitamin PP 9.29 Vitamin B5 7.14 Zinc oxide 12.63 Iron sulfate 10.26 Calcium carbonate 1244.54 Copper sulfate 1.18 Potassium chloride 1205.45 Magnesium oxide 185.18 Dipotassium phosphate 1185.31 Tricalcium phosphate 51.31 Total 4000

In a separate clean, dry bowl 22, a predetermined amount of sugar 16 is premixed with a predetermined amount of vegetable oil 18, again for at least 5 to 10 minutes. Preferably sucrose (icing sugar) is used as the sugar.

The premixed cereals, legumes, optional oilseeds, and minerals and vitamins are then mixed thoroughly with the sugar-oil mixture using a mechanical mixing device 30 to form a viscous paste. As mentioned above, it is important to form a uniform product mixture, and accordingly at this stage the ingredients should be mixed for at least 15 minutes to ensure the same amount of nutrients in every portion of the product.

The product is then transferred into clean, dry pots, preferably using a plastic or metal spatula. Lids are then fitted tightly onto the pots, and the pots are labelled with a use- by date. Ideally the pots should then be stored prior to use away from heat and light, but the RUTF or supplement according to the invention does not require special storage conditions or refrigeration.

Examples 2 - 14

In some embodiments the RUTF or supplement according to the present invention may comprise a non-milk based, nutritionally complete, cereal-legume based food that is scientifically formulated to treat severe malnutrition, or to be used as a supplementary food for vulnerable children. It has all the micronutrients and macronutrients necessary for growth. It is appealing in taste and has good product stability.

Thirteen cereal-legume mixtures as shown in Table 4 below were manufactured according to the process of Example 1 above. The foods were prepared from roasted or processed ingredients with the total exclusion of water, and were expected to have a low dietary bulk, low bacterial contamination, and to be substantially ready-to-use. The commodities chosen had the most appropriate energy density and high biological value of protein. Moreover, the proposed foods advantageously had a soft consistency, making them easy to swallow, which is desirable for infant feeding for example. The mixtures were then evaluated on the basis of the following criteria:

(a) Amino acid complementation for maximal protein quality (b) Product stability

(c) Ease of raw materials available in developing countries

(d) Taste/texture and flavour of final product

Table 4: Examples of RUTF formulations

Food Macronutrients Energy/lOOg

Protein Carbohydrate Fat

Weight (%) (kcal)

2 Sesame seeds (Sesamum spp.), and maize (Zea mays) 11.9 44.8 26.5 465

3 Brown chickpeas (Cicer aretinum), sesame seeds, and barley (Hordeum vulgare) 12.4 44.8 29.6 495

4 Sesame seeds, rice (Oryza spp.), chickpeas, and maize 12.3 49.4 21 436

5 Barley, sesame seeds, and Soyamin-90 ® (90% protein) 14.1 40 39 567

6 Sesame seeds, Soyamin-70 ® (70% protein), and wheat (Triticum spp.) 13 44.3 29.6 496

7 Soyamin-70, and rice 12.7 40.9 34.1 521

8 Sesame seeds, Soyamin-70 ®, and barley 11.6 39.3 37.6 542

9 Soybean (Glycine max ), lentils (Lens culinaris) and wheat 13.9 41.5 33.1 520

10 Sesame seeds, rice, and Soyamin-90 ® 13.8 43 36 551

11 Sesame seeds, maize, and Soyamin-70 ® 12.2 36.4 38.5 541

12 Split peas (Pisum sativum), rice, and Soyamin-90 ® 13.2 44.2 30.3 502

13 Maize, sesame seeds, and chickpeas 13.4 50.6 28.6 513

14 Soybean, oat (Avena sativa), and wheat 13.1 40.8 33.6 518

Plumpy'nut® (comparative example) 14.5 43 33.5 530

*AU cereals and legumes were roasted. Appropriate proportion of sugar, oil and vitamins and minerals were added.

Of the above formulations, three were deemed to be particularly advantageous in terms of the different criteria mentioned above and were developed further, using the complementary effect of combining oilseeds with grains, legumes or soy protein concentrate - "Soyamin 90®" (Degussa Texturant Systems Ltd.) - along with appropriate vitamins and minerals.

RUTF-1: "Rice-Sesame" based (Example 10)

Ingredients:, Roasted rice flour, roasted sesame seeds paste, Soyamin 90, sunflower oil, icing sugar, vitamin and mineral premix (Nutriset).

Quantities (%)

• Roasted rice flour 20%

• Soyamin 90 8%

• Roasted sesame seeds paste 29%

• Sunflower oil 19.4%

• Icing sugar 22%

• Premix 1.6%

Total 100.0%

RUTF-2: "Barley-Sesame" based (Example 5)

Ingredients: Roasted pearl barley flour, roasted sesame seeds paste, Soyamin 90, sunflower oil, icing sugar and vitamin, mineral premix (Nutriset):

Quantities (%)

• Roasted pearl barley flour 15%

• Roasted sesame seeds paste 27%

• Soyamin 90 9%

• Sunflower oil 24%

• Icing sugar 23.4%

• Premix 1.6%

• Total 100.0%

RUTF-3: "Maize-Sesame" based (Example 13)

Ingredients: Roasted sesame seeds paste, roasted maize flour, roasted chickpeas flour, sunflower oil, icing sugar and vitamin/mineral premix (Nutriset).

Quantities (%)

• Roasted maize flour 33.4%

• Roasted sesame seeds paste 13%

• Roasted chickpeas flour 25%

• Sunflower oil 12%

• Icing sugar 15%

• Premix 1.6%

Total 100.0% Analysis

The following quantitative analyses were performed on the RUTF-1, RUTF-2 and RUTF-3 formulations. The analytical and bacteriological tests and the protein digestibility corrected amino acid score results are presented accordingly. Protein Fat

Carbohydrate Moisture Ash • Water activity

Minerals Peroxide value Free fatty acids Microbial • Protein digestibility-corrected amino acid score (PDCAAS)

Gross nutritional composition of RUTF samples

The RUTF gross nutritional composition is presented in Table 5. For comparative purposes, the Nutriset - "Plumpy'nut" ® - nutritional information is also included. Protein and fat are reported to contribute 11% and 57% in energy content, respectively. Likewise, total energy and moisture are reported to be 530 kcal/lOOg and <5%, respectively (Nutriset, 2002⁶).

From Table 5, it is evident that the "Rice-Sesame" based RUTF-1, the "Barley- Sesame" based RUTF-2, and the "Maize - Sesame" based RUTF-3 are close to "Plumpy'nut" ® in their chemical composition. Using the basic formulation outlined above, it would be easy to alter the amount of macronutrient and/or micronutrient of the products by varying the oilseed: cereal: legume combinations and/or mineral/vitamin mixtures.

' Nutriset, (2002), Production "Plumpy'nut" ® locally, draft report. Table 5: RUTF-1, RUTF-2, RUTF-3 and Nutriset "Plumpy'nut" © nutritional composition per lOOg and percentage contribution to energy

Nutrients RUTF-1 Energy RUTF-2 Energy RUTF-3 Energy Plumpy'nut ® (**) Energy

% % % 100g %

Energy (1) kcal 551 567 512 530

Energy kjoules 2307 10 2373 10 2142 11 2218 11

Protein g 13.8 31 14.1 28 13.4 39 14.5 32

Carbohydrate (*) g 43 59 39.9 62 50.2 50 43 57

Fat g 36 39 28.6 33.5

Ash g 4.3 3.9 4.9 4

Moisture g 2.9 3.1 2.9 <5

(*) Carbohydrate is by difference assuming protein to be nitrogen (N) times 6.25

(_L) The energy has been calculated using Atwater factors

(**) Protein and fat are reported to contribute 11% and 57% in energy input. Total energy is reported to be 530 kcal/lOOg and moisture <5%

Mineral analysis

Table 6: Mineral analysis for RUTF products

RUTF-1 RUTF-2 RUTF-3 Plumpy'nut ® mg/kg

Cu 2.1 2.7 1.8 1.7

Zn 10.9 15.1 10.2 13

Ca 338.1 315.7 209.8 310

Na 256.5 250.5 189.9 <290

Mg 118.4 115.5 119.1 86

Fe 5.6 5.4 4.4 12.45

The mineral content, as evident from Table 6, for the three formulations compares very favourably with "Plumpy'nut" ® for all micronutrients except iron. This could be easily rectified by increasing the level of iron added to the premix.

Water activity (a_w)

Table 7: Water activity (a_w) in three RUTF samples

RUTF samples Water activity

Rice-Sesame

(RUTF- 1) 0.290

Barley-Sesame

(RUTF- 2) 0.279

Maize - Sesame

(RUTF- 3) 0.260

Plumpy'nut ® 0.241

The a_w for RUTF-1, RUTF-2 and RUTF-3 is shown in Table 7. The interaction between water activity and microbial proliferation in food is illustrated in Table 8. Table 8: Interaction between aw and microbial proliferation in some foods

Water activity Foods Microorganisms

0.98 Fresh meats, fish Most food spoilage and vegetables, milk food-borne pathogenic organisms grow

0.85 - 0.60 Flour, cereals, nuts No pathogenic bacteria grow

0.60 Confectionery noodles, Microorganisms do not dried milk multiply but can remain viable for long periods

0.30- 0.20 Biscuits, instant No viable microbial Coffee growth

Source: Fellows, 2000⁷

From Table 8 it can be deduced that the a_w of RUTF 1, RUTF-2 and RUTF-3 is at an acceptable level. All three formulations are therefore likely to maintain microbial safety.

Microbial analysis

Table 9: Microbial analysis for RUTF products

Microorganisms RUTF-1 RUTF-2 RUTF-3

TVC at 37oC cfu/g <10 <10 <10

Salmonella spp in 25g N.D. N.D. N.D.

Coliforms cfu/g <10 <10 <10

Coagulase-positive <10 <10 <10

Staphilococcus cfu/g

Mould cfu/g <10 <10 <10

Yeast cfu/g <10 <10 <10

* N.D. not detected

⁷ Fellows, P.J. (2000), Food processing technology, principles and practice (2nd edition), Woodhead, Cambridge. As illustrated in Table 9, the results are within microbial specification for this type of foods (Shapton & Shapton, 1998^s).

Free fatty acids (FFA) Oils may be noticeably rancid when the FFA ranges from 0.5% to 1.5%. The FFA for the oil used to prepare RUTF-1, RUTF-2 and RUTF-3 was measured as 0.07 %. Thus, the FFA level is less than the range for rancidity.

Peroxide value The peroxide value for the oil used to prepare RUTF-1, RUTF-2 and RUTF-3 was measured as 4.32 mEq. of 02 kg fat. It has been reported that fresh oils usually show a value less than lOmEq/kg, and rancidity may be detected when the peroxide value is between 20 mEq. and 40 mEq. (Pearson, 1976⁹). The peroxide value of the oil used to prepare RUTF-1 and RUTF-2 indicates that the oil is not rancid.

Protein digestibility-corrected amino acid score in RUTF formulation (PDCAAS)

An important feature is the protein quality of the three formulations based on the PDCAAS. The RUTF protein digestibility - corrected amino acid score (PDCAAS) - is presented in Table 10.

Shapton D. A and Shapton N. F. (1998) Principles and Practice for the Safe Processing of Foods, Woodhead , England, Cambridge. Table 10: Protein Digestibility-Corrected Amino Acid Score in RUTF samples.

RUTF samples PDCAAS (%)

Rice-Sesame

(RUTF- 1) 71.2

Barley-Sesame

(RUTF- 2) 74.3

Maize - Sesame

(RUTF- 3) 70

Plumpy'nut ® 83

It is evident from the foregoing that the "Barley-Sesame" (RUTF-2) formulation had the best protein quality attributes.

Example 15: Method of making an RUTF or nutritional supplement with probiotic microorganisms

In some embodiments it may be desirable as described above to include in an RUTF or nutritional supplement according to the present invention an inoculum of probiotic bacteria such, for example, as lactobaciUi or bifidobacteria. With reference to

FIG.2, such an RUTF or supplement may be made in a similar manner to Example 1 above, with the addition of the probiotic bacteria 32 at the main mixing stage. At this time, or earlier, an amount of prebiotic material 33 such, for example, as pectin or inulin may also be added. In some embodiments, said prebiotic material 33 may be added to the cereal-legume-oilseeds premix.

Example 16: RUTF with probiotic culture

An RUTF according to the present invention was made as described in Example 15 above with the following ingredients: Quantities (%_.

Roasted maize flour 32.4%

Roasted sesame seeds paste 12%

Roasted chickpeas flour 23%

Sunflower oil 11%

Icing sugar 15%

Premix 1.6%

Inulin beta-glucan, and pectin 5%

"Synbiotic 2000 forte"™* (-10¹¹ bacteria)

Total 100.0%

* A mixture of e.g. Lactococcus raffinolactis 32-77, Pediacoccus pentoseceus 5- 33:3, Lactobacillus paracesei subsp. paracesei 19, Lactobacillus plantarum 2362, from Synbiotics AB, Sweden, under the trade name "Synbiotic 2000 forte"

The RUTF formulations as hereinbefore described are low cost, ready-to-use therapeutic weaning foods that are free of milk powder and peanuts. No-animal based ingredients are used in the formulations. Said RUTF formulations have been developed on the basis of the complementary effect of combining two or more ingredients, mainly oilseeds, grains and legumes, with appropriate vitamin and minerals. The RUTFs do not contain any trace of nuts (peanut) and/or nut products, mainly to minimize mycotoxin contamination. With increasing concerns about the use of peanuts in infant food (owing to allergenic and mycotoxin contamination), the products according to the present invention have considerable global promise. The gross compositions of RUTF-1, RUTF-2 and RUTF-3 are comparable with prior art RUTF formulations, and the protein quality and the mineral contents of those formulations compare favorably with such prior art compositions. The RUTF products according to the invention have an acceptable level of protein quality and energy density and are suitable for feeding young children and vulnerable groups.

Claims

1. A ready-to-use therapeutic food or nutritional supplement comprising a mixture of at least one ground, roasted cereal, and at least one ground, roasted legume or cooked leguminous protein extract, said at least one cereal and said at least one legume or leguminous protein extract being selected to be mutually complementary so as to provide a PDCAAS of at 5 least 60.

2. A food or nutritional supplement as claimed in claim 1, wherein the PDCAAS of the food or supplement is at least 65, and more preferably at least 70.

3. A food or nutritional supplement as claimed in claim 1 or claim 2, wherein said at least one cereal is selected from oats, maize, quinoa, barley, wheat, rye, triticale, sorghum, millet, teff, and rice.

4. A food or nutritional supplement as claimed in any preceding claim, which is free or substantially free of any oleaginous seed or grain, i.e. any cereal or legume comprising more than about 20% wt. fat, such, for example, as peanuts, almonds, and pistachio nuts.

5. A food or nutritional supplement as claimed in any preceding claim, wherein said legume is selected from chick peas, soybeans, lentils, split peas, haricot beans, mung (green) beans, fava bean, pigeon pea, and kidney beans.

6. A food or nutritional supplement as claimed in any preceding claim, wherein said cereal and legume are roasted.

7. A food or nutritional supplement as claimed in claim 6, wherein said at least one roasted cereal and said at least one roasted legume are ground such that they are included in the ready-to-use food or supplement in particulate form.

8. A food or nutritional supplement as claimed in claim 6 or claim 7, wherein the cereal or legume are milled to form a flour.

9. A food or nutritional supplement as claimed in any of claims 6-8, wherem me ground cereal and or the ground legume have a particle size of less than 200 μm, preferably less than lOOμm, more preferably in the range 50μm to lOOμm.

10. A food or nutritional supplement as claimed in any preceding claim, which comprises soy bean extract.

11. A food or nutritional supplement as claimed in any preceding claim, further comprising at least one ground, roasted oilseed such as sesame seed.

12. A food or nutritional supplement as claimed in any preceding claim, which comprises roasted rice, roasted sesame seeds and soy bean extract.

13. A food or nutritional supplement as claimed in claim 12, which comprises about 18 to 22% wt. roasted rice flour, 6 to 10% wt. soy bean extract and 27 to 31% wt. roasted sesame seed paste.

14. A food or nutritional supplement as claimed in any of claims 1-11, which comprises roasted barley as a cereal component, roasted sesame seeds as an oilseed, and a soy bean extract.

15. A food or nutritional supplement as claimed in claim 14, which comprises 13 to 17% wt. roasted barley flour, 25 to 29% wt. roasted sesame seed paste, and 7 to 11% wt. soya bean extract, with the balance comprising one or more energy supplements and vitamin-mineral premixes.

16. A food or nutritional supplement as claimed in any of claims 1-11, which comprises roasted maize as a cereal component, roasted sesame seeds as an oilseed, and roasted chick peas as a legume component.

17. A food or nutritional supplement as claimed in claim 16, which comprises about 31 to 35% wt. roasted maize flour, 11 to 15% wt. sesame seed paste, and 23 to 27% wt. roasted chick pea flour, with the balance being made up from energy supplements such as oil, fats or sugars, and vitamin-mineral premixes.

18. A food or nutritional supplement as claimed in any preceding claim, which further comprises an external energy supplement such as one or more of fat, sugar, and oil, such as vegetable oil selected from sunflower oil, safflower oil, corn oil, palm oil, soy bean oil.

19. A food or nutritional supplement as claimed in claim 18, which comprises a vegetable oil having a free fatty acid content of less than about 0.5% wt. and more preferably less than about 0.1% wt.

20. A food or nutritional supplement as claimed in claim 18 or claim 19, which comprises a vegetable oil having a peroxide value of less than about 20 mEq. of O₂/kg of fat, more preferably less than about lOmEq, even more preferably less than 5mEq.

21. A food or nutritional supplement as claimed in any preceding claim, which comprises more than about 10% wt. water, and preferably less than about 25% wt. water.

22. A food or nutritional supplement as claimed in claim 21, wherein the majority of the water is bound water, and the water activity of the food or supplement is less than about 0.5, preferably less than about 0.4, more preferably in the range of about 0.2 to about 0.3.

23. A food or nutritional supplement as claimed in any preceding claim, further comprising one or more supplementary micronutrients such, for example, as vitamins, soluble and insoluble mineral salts, and mixtures thereof, and/or one or more enzymes.

24. A food or nutritional supplement as claimed in claim 23, wherein said one or more mineral salts include one or more of calcium, iron and zinc, such as calcium carbonate, zinc oxide, copper sulfate, potassium chloride, magnesium oxide, and iron salts.

25. A food or nutritional supplement as claimed in any preceding claim, comprising 10 to 40 % wt. ground, roasted cereal; 5 to 35 % wt. ground, roasted legume or leguminous protein concentrate; 0 to 30 % wt. ground, dry roasted oilseed; 10 to 30 % wt. oil; 10 to 30 % wt. sugar; 0 to 5 % wt. micronutrients; wherein the total of said ingredients is 100 % wt.

26. A food or nutritional supplement as claimed in any preceding claim, which comprises 20-30% wt. of ground, roasted legume.

27. A food or nutritional supplement as claimed in claim 26, which comprises 10- 15%wt, preferably 13-14% wt, of oilseed; and or 10-15% wt., preferably 13-14% wt, of vegetable oil; and or 12-18% wt., preferably 13-16% wt, of sugar.

28. A food or nutritional supplement as claimed in any of claims 1-25, which comprises 5-10% wt., preferably 8-9% 5 wt., leguminous protein extract, e.g. soybean extract.

29. A food or nutritional supplement as claimed in claim 28, which comprises 25- 30%wt, preferably 26-29% wt, of oilseed; and/or 5-25% wt., preferably about 19-24% wt., of vegetable oil; and or 20-25%) wt., preferably about 22-24% wt., of sugar.

30. A food or nutritional supplement as claimed in any preceding claim, which comprises 1-2% wt., preferably 1.5-1.7% wt., of a mineral premix.

31. A food or nutritional supplement as claimed in any preceding claim, which has an energy content of greater than 5.0kcal/g, and a protein content of more than 12% wt, and/or a carbohydrate content of about 40% wt. or more, and/or a fat content of 25% wt. or more.

32. A food or nutritional supplement as claimed in any preceding claim, further comprising an inoculum of at least one probiotic microorganism.

33. A ready-to-use food or nutritional supplement comprising one or more food-grade products having a water activity of no more than 0.4, and an inoculum of at least one probiotic microorganism.

34. A food or nutritional supplement as claimed in any preceding claim, which further comprises an effective amount of a prebiotic supplement, which may be selected from insoluble and dietary fibres, resistant starches, carbohydrates and oligosaccharides, such as beta-glucan, pectin, inulin, allicin, and fructo oligo saccharide.

35. A method of therapy or prophylaxis of a disease of the intestinal lining which comprises administering to a person in need thereof an effective therapeutic or prophylactic amount of a food or nutritional supplement according to any preceding claim.

36. A method as claimed in claim 35, wherein said disease is selected from ailments of the gut associated with malnourishment, HIV/AIDS, cancer-cachexia, and postoperative conditions.

37. Use of a food or nutritional supplement according to any of claims 1-34 in the manufacture of a medicament for the treatment or prophylaxis of diseases of the intestinal lining.

38. A method of making a ready-to-use food or nutritional supplement in accordance with any of claims 1-34, which method comprises mixing at least one ground, roasted cereal with at least one ground, roasted legume or leguminous protein extract.

39. A method as claimed in claim 38, wherein the cereal-legume mix is mixed with an oil or fat, which oil or fat may carry sugar or a micronutrient premix.