FLAVOUR ADDITIVES

Abstract

The present invention relates to the use of one or more nucleotides selected from the group consisting of GMP (guanosine monophosphate) and IMP (inosine monophosphate), a first amino acid selected from the group consisting of alanine, histidine and glycine or a mixture thereof; a second amino acid selected from the group consisting of proline and threonine or a mixture thereof; and furaneol for increasing the palatability of a foodstuff to a companion animal. The invention also relates to a pet food-stuff or supplement comprising one or more nucleotides selected from the group consisting of GMP and IMP, a first amino acid selected from the group consisting of alanine, histidine and glycine or a mixture thereof; a second amino acid selected from the group consisting of proline and threonine or a mixture thereof; and furaneol, and also to a method of increasing the palatability of a food-stuff to a companion animal.

Claims

1-18. (canceled)

19. Water for cats that has increased palatability and comprises: (a) one or more nucleotides selected from the group consisting of guanosine monophosphate (GMP), inosine monophosphate (IMP), and a combination thereof that is present at a concentration between about 0.5 mM and about 10 mM; (b) a first free amino acid that activates a cat umami receptor that is present at a concentration between about 20 mM to about 50 mM; (c) a second free amino acid that does not activate the cat umami receptor that is present at a concentration between about 20 mM to about 50 mM; and (d) furaneol that is present at a concentration between about 1 ppm and about 10 ppm; wherein the first free amino acid is histidine and the second free amino acid is proline.

20. The water of claim 19, wherein the one or more nucleotides are a combination of GMP and IMP.

21. The water of claim 20, wherein the combination of GMP and IMP is present at a concentration between about 2 mM to about 3 mM.

22. The water of claim 19, wherein the one or more nucleotides is GMP.

23. The water of claim 22, wherein the GMP is present at a concentration between about 2 mM to about 3 mM.

24. The water of claim 19, wherein the first free amino acid is present at a concentration between about 20 mM to about 30 mM, and the second free amino acid is present at a concentration between about 20 mM to about 30 mM.

25. The water of claim 24, wherein the first free amino acid is present at a concentration of 25 mM, and the second free amino acid is present at a concentration of 25 mM.

26. The water of claim 19, wherein the furaneol is present at a concentration between about 2 ppm and about 5 ppm.

27. Water for cats that has increased palatability and comprises: (a) one or more nucleotides selected from the group consisting of guanosine monophosphate (GMP), inosine monophosphate (IMP), and a combination thereof that is present at a concentration between about 0.5 mM and about 10 mM; (b) a first free amino acid that activates a cat umami receptor that is present at a concentration between about 20 mM to about 50 mM; (c) a second free amino acid that does not activate the cat umami receptor that is present at a concentration between about 20 mM to about 50 mM; and (d) furaneol that is present at a concentration between about 1 ppm and about 10 ppm; wherein the first free amino acid is histidine and the second free amino acid is threonine.

28. The water of claim 27, wherein the one or more nucleotides are a combination of GMP and IMP.

29. The water of claim 28, wherein the combination of GMP and IMP is present at a concentration between about 2 mM to about 3 mM.

30. The water of claim 27, wherein the one or more nucleotides is GMP.

31. The water of claim 30, wherein the GMP is present at a concentration between about 2 mM to about 3 mM.

32. The water of claim 27, wherein the first free amino acid is present at a concentration between about 20 mM to about 30 mM, and the second free amino acid is present at a concentration between about 20 mM to about 30 mM.

33. The water of claim 32, wherein the first free amino acid is present at a concentration of 25 mM, and the second free amino acid is present at a concentration of 25 mM.

34. The water of claim 27, wherein the furaneol is present at a concentration between about 2 ppm and about 5 ppm.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0050] The invention will now be described in reference to the following Figures and Examples in which:

[0051] FIG. 1 shows the results of a difference test of a composition comprising 25 mM histidine+25 mM proline+2.5 mM Ajitide+4 ppm furaneol with a composition comprising 50 mM histidine+2.5 mM Ajitide+4 ppm furaneol;

[0052] FIG. 2 shows the results of a difference test of a composition comprising 25 mM histidine+25 mM proline+2.5 mM Ajitide+4 ppm furaneol with a composition comprising 50 mM proline+2.5 mM Ajitide+4 ppm furaneol;

[0053] FIG. 3 shows the results of a difference test of a composition comprising 25 mM histidine+25 mM threonine+2.5 mM Ajitide+4 ppm furaneol with a composition comprising 50 mM threonine+2.5 mM Ajitide+4 ppm furaneol;

[0054] FIG. 4 shows the results of a difference test of a composition comprising 25 mM histidine+25 mM proline+2.5 mM GMP+4 ppm furaneol with a composition comprising 25 mM histidine+25 mM proline+2.5 mM GMP;

[0055] FIG. 5 shows the results of a difference test of a composition comprising 75 mM alanine+25 mM proline+2.5 mM Ajitide+4 ppm furaneol with a composition comprising 25 mM histidine+2.5 mM Ajitide+4 ppm furaneol;

[0056] FIG. 6 shows the resulting dose response curves of each first amino acid of the invention that were screened in vitro for their ability to activate the T1R1/T1R3 receptor in the presence of 0.2 mM IMP. The corresponding EC.sub.50 values are shown in the table.

[0057] FIG. 7 shows the resulting dose response curves of each second amino acid of the invention that were screened in vitro for their ability to activate the T1R1/T1R3 receptor in the presence of 0.2 mM IMP.

[0058] FIG. 8 shows the dose response curves of nucleotides of the invention that were screened in vitro for their ability to activate the T1R1/T1R3 receptor in the presence of 20 mM alanine. The corresponding EC.sub.50 values are shown in the table.

[0059] FIG. 9 shows the predicted structure of the T1R1/T1R3 umami receptor.

[0060] FIG. 10 shows a schematic of the predicted structure of the umami receptor; and

[0061] FIG. 11 shows a sequence alignment of the human, feline, canine, mouse and rat umami receptors.

EXAMPLES

[0062] All amino acids used in the examples are of the L-form. Ajitide is a 50:50 mixture of GMP:IMP.

Example 1

[0063] Cats were allowed access to water containing 25 mM histidine+25 mM proline+2.5 mM Ajitide+4 ppm furaneol and to water containing 50 mM histidine+2.5 mM Ajitide+4 ppm furaneol The methodology used a 2-bottle choice test with 24 cats (the final number of cats for each test can vary due to data being discarded by spillage, etc.). Cats were housed individually during trial periods and had free access to water available between testing periods. The test involved a choice test between the tastant/mixture at a given concentration dissolved in deionised water versus deionised water only or another tastant/mixture. Control was made for positional bias (e.g. A/B exposure 1 and B/A exposure 2) and evaporation loss. The testing time was 36 hours (i.e. 18 hours per day, allowing a two-day crossover). Following two consecutive days of each testing, cats had two consecutive days of rest. Cats were offered a dry diet as a single meal at the start of the test period for one hour, calculated to meet the individual requirements for each cat.

[0064] The results are shown in the table below, and in FIG. 1.

TABLE-US-00002 ANOVA Table for Fixed Effects Degrees of Freedom Factor Numerator Denominator F-value P-value Product Difference 1 24 0.54 0.4715

TABLE-US-00003 Table of Mean Product Difference, Standard Error & 95% Confidence Intervals Standard 95% Confidence Interval Product Difference Mean Error Lower Upper His + Pro + Aji + Fur − 7.38 10.09 −13.45 28.22 2xHis + Aji + Fur

[0065] Intake of the combination of histidine+proline+Ajitide+furaneol, was on average 7.38 g more that the intake of histidine+Ajitide+furaneol alone.

Example 2

[0066] Cats were allowed access to water containing 25 mM histidine+25 mM proline+2.5 mM Ajitide+4 ppm furaneol and to water containing 50 mM proline+2.5 mM Ajitide+4 ppm furaneol

[0067] The results are shown in the table below, and in FIG. 2.

TABLE-US-00004 ANOVA Table for Fixed Effects Degrees of Freedom Factor Numerator Denominator F-value P-value Product Difference 1 23 5.50 0.0280

TABLE-US-00005 Table of Mean Product Difference, Standard Error & 95% Confidence Intervals Standard 95% Confidence Interval Product Difference Mean Error Lower Upper His + Pro + Aji + Fur − 25.74 10.98 3.03 48.45 2xPro + Aji + Fur

[0068] Intake of the combination of histidine+proline+Ajitide+furaneol, was on average 25.74 g more that the intake of 2xproline+Ajitide+furaneol, which is a significant difference and shows a clear preference for the four component combination.

[0069] The results of Examples 1 and 2 together show that a combination of histidine+proline +Ajitide+furaneol is preferable to either of the three component combinations tested.

Example 3

[0070] A difference test was carried out as described in Example 1 to compare a composition containing 25 mM histidine+25 mM threonine+2.5 mM Ajitide+4ppm furaneol with a composition containing 50 mM threonine+2.5 mM Ajitide+4ppm furaneol.

[0071] The results are shown in the table below and in FIG. 3.

TABLE-US-00006 ANOVA Table for Fixed Effects Degrees of Freedom Factor Numerator Denominator F-value P-value Product Difference 1 24 17.12 0.0004

TABLE-US-00007 Table of Mean Product Difference, Standard Error & 95% Confidence Intervals Standard 95% Confidence Interval Product Difference Mean Error Lower Upper His + Thr + Aji + Fur − 27.85 6.73 13.96 41.74 2xThr + Aji + Fur

[0072] It can be seen that the intake of the combination of histidine+threonine+Ajitide+furaneol was, on average, 27.85 g more than the intake of 2xthreonine+Ajitide+furaneol which is a significant difference.

Example 4

[0073] Cats were allowed access to water containing 25 mM histidine+25 mM proline+2.5 mM GMP+4 ppm furaneol and to water containing 25 mM histidine+25 mM proline+2.5 mM GMP.

[0074] The results are shown in the table below and in FIG. 4.

TABLE-US-00008 ANOVA Table for Fixed Effects Degrees of Freedom Factor Numerator Denominator F-value P-value Product Difference 1 24 60.93 0.0000

TABLE-US-00009 Table of Mean Product Difference, Standard Error & 95% Confidence Intervals Standard 95% Confidence Interval Product Difference Mean Error Lower Upper His + Thr + Aji + Fur − 39.62 5.08 29.15 50.10 2xThr + Aji + Fur

[0075] A P-value of 0.0000 indicates a value of less than 0.0001.

[0076] It can be seen that the intake of histidine+proline+GMP+furaneol was, on average, 39.62 g more than the intake of histidine+proline+GMP, which is a significant difference.

Example 5

[0077] Cats were allowed access to water containing 25 mM histidine+2.5 mM Ajitide+4 ppm furaneol and to water containing 75 mM alanine+25 mM proline+2.5 mM Ajitide+furaneol.

[0078] The results are shown in the table below and in FIG. 5.

TABLE-US-00010 ANOVA Table for Fixed Effects Degree of Freedom Factor Numerator Denominator F-value P-value Product Difference 1 24 17.59 0.0003

TABLE-US-00011 Table of Mean Product Difference, Standard Error & 95% Confidence Intervals Standard 95% confidence Interval Product Difference Mean Error Lower Upper His + Thr + Aji + Fur − −45.34 10.81 −67.64 −23.03 2xThr + Aji + Fur

[0079] It can be seen that the intake of alanine+proline+Ajitide+furaneol was, on average, 45.34 g more than the intake histidine+Ajitide+furaneol, which is a significant difference.

Example 6

[0080] In vitro screening was carried out in order to establish which first amino acids bind and activate the umami receptor. Results are shown in FIG. 6.

Example 7

[0081] In vitro screening was carried out in order to establish which second amino acids bind and activate the umami receptor. Results are shown in FIG. 7.

Example 8

[0082] In vitro screening was carried out in order to establish which nucleotides bind and activate the umami receptor. Results are shown in FIG. 8.