METHOD OF HYDROLYZING RICE PROTEIN CONCENTRATE WITH

PROTEASE ENZYMES

RELATED APPLICATION

This application claims benefit of priority to provisional application serial number 60/563,841 filed April 20, 2004, which is incorporated by reference to the same extent as though fully replicated herein.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention is broadly concerned with hydrolyzed rice protein and uses thereof, particularly in connection with cosmetic products such as shampoos, shampoo conditioners, hair styling gels, hair conditioners, hair reparative, hair tonics, hair fixatives, hair mousses, bath and shower gels, liquid soaps, moisturizing sprays, makeup, pressed powder formulations, lip products, bath additives, sanitizing wipes, hand sanitizers, premoistened towelettes, skin lotions and creams, shaving creams, sunscreens, and anti-wrinkle and color cosmetics. More particularly, it is concerned with hydrolyzed rice protein which is preferably in the form of a mixture of amino acids, peptides and/or protein fractions derived from the hydrolysis of naturally occurring rice protein; such mixtures, when used in cosmetic products, provide enhanced properties.

2. Description of the Prior Art

Current marketing trends in personal care products have created increased interest and demand for natural ingredients due to influence from animal rights activists, consumer advocates, lobbyists, and the BSE (mad cow disease) scare. Consumers are also becoming environmentally conscious and are looking for more healthy lifestyles, which include more natural products in terms of foods, cosmetics, and packaging. Thus, many manufacturers and marketers of cosmetic products are seeking to include hydrolyzed proteins of plant or vegetable origin in their products and avoid the use of animal proteins.

A closer look at the ingredient listing in most product packages reveals the use

of hydrolyzed grain and oilseed proteins from wheat, corn, oat, and soy among others, in hair and skin care products. Animal-based ingredients such as hydrolyzed collagen and keratin are becoming rare. Hydrolyzed grain and oilseed proteins provide benefits to hair by improving the body, manageability, shine, dry combing and moisture retention. In particular, the quateraized derivatives of hydrolyzed plant or vegetable proteins have excellent conditioning properties and provide greater ease of wet combing. In skin care formulations, hydrolyzed grain and oil seed proteins promote effective moisturizing and dermal conditioning, combat chapping and irritation, and resist pollutant and irritant damage. Another potential source of cosmetic protein from grain is rice, which is the staple food in the densely populated countries of Asia, especially Bangladesh, Burma, Cambodia, China, Indonesia, Korea, Laos, Vietnam, Thailand, Sri Lanka, and the Philippines. Rice provides as much as 75% of the daily caloric intake of people in these Asian countries. The major components of rice are starch and protein representing about 90% of the grain. Rice starch exists as compound granules, and the individual granule has an angular or polyhedral shape. The dimension of rice starch is around 3-5 microns in diameter. In general, rice has 7-9% protein content. The protein component of rice consists of four fractions: glutelin, prolamin, globulin, and albumin. The major amino acids in rice proteins are glutamic acid, aspartic acid, arginine, and leucine.

SUMMARY OF THE INVENTION

The present invention is directed to a new form of rice protein, namely hydrolyzed rice protein and derivatives thereof, as well as uses of such protein products in cosmetic formulations. The preferred hydrolyzed rice protein and derivatives thereof in accordance with the invention comprise a mixture of amino acids, peptides and/or protein fractions derived from the hydrolysis of the naturally occurring rice protein. Such hydrolysis is preferably carried out enzymatically, but if desired acid hydrolysis can also be employed.

In more detail, the hydrolyzed rice protein is typically in the form of a mixture having an amino acid, peptide and/or protein fragment molecular weight range of from about 75-5,000 with an average molecular weight of from about 1,500-2,500.

During processing, it is often desirable to membrane filter the hydrolysis products in order to segregate the peptides and/or protein fragments to achieve different molecular weight profiles. In one such preferred method, a hydrolyzed rice protein product of relatively high molecular weight is provided, with a molecular weight range for the respective peptides and/or fragments therein of from about 1,000-5,000 and with an average molecular weight of from about 3,000-4,000. Similarly, a lower molecular weight fraction (sometimes referred to as a rice amino acid fraction) is produced wherein the respective amino acids and peptides exhibit a molecular weight range of from about 75-1,000, with an average molecular weight of from about 100- 300.

Although the hydrolyzed rice protein and derivatives thereof can be produced as a dry powder, this is sometimes difficult and normally the product is in the form of an aqueous dispersion containing rice protein or derivatives at a level of from about 18-35% by weight, more preferably from about 23-27% by weight. As used herein, "derivatives" of hydrolyzed rice protein refers to changes in the structure of the individual amino acids, peptides and/or protein fragments produced by amino acid addition, deletion, replacement, substitution and/or modifications; mutants produced by recombinant and/or DNA shuffling; quaternized species; and all other chemically synthesized/modified forms of the individual amino acids, peptides and/or protein fragments which retain at least in part some activity of the initial hydrolyzed amino acids, peptides and/or protein fragments. One particularly preferred class of hydrolyzed rice protein derivatives is the lipid derivatives, especially those synthesized using C12-C22 fatty acids. The term "hydrolyzed rice protein" is intended to embrace and cover not only the amino acids, peptides and/or protein fractions derived from the hydrolysis of naturally occurring rice protein but also all "derivatives" as herein defined.

The rice products of the invention can be used to good effect in a variety of cosmetic formulations which include at least one ingredient selected from the group consisting of humectants, emollients, conditioners, thickeners, moisturizing agents, opacifiers, pearl agents, buffering agents, slip agents, feel agents, anti-static agents, acidifiers, preservatives, film formers, plasticizers, setting agents and suspending agents (usually, each of the foregoing ingredients when used is present at a level of

from about 0.05-10% by weight). An amount of hydrolyzed rice protein or derivative thereof is incorporated into this type of cosmetic formulation, usually at a level of from about 1-10% by weight, more preferably from about 3-8% by weight. Inasmuch as the preferred rice protein products are in the form of liquid dispersions, it is a simple matter to add the rice protein product to the cosmetic formulations during preparation thereof. Generally, the formulations of the invention comprise from about 10-95% by weight water, more preferably from about 20-75% by weight water.

Among the cosmetic products which can benefit from incorporation of the rice products of the invention are those selected from the group consisting of shampoos, shampoo conditioners, hair styling gels, hair conditioners, hair reparatives, hair tonics, hair fixatives, hair mousses, bath and shower gels, liquid soaps, moisturizing sprays, makeup, pressed powder formulations, lip products, bath additives, sanitizing wipes, hand sanitizers, premoistened towelettes, skin lotions and creams, shaving creams, sunscreens and anti-wrinkle and color cosmetics. In products of these types, the shampoos and shampoo conditioners further comprise at least about 6% by weight detergent; the hairstyling gels further comprise a gel-forming polymer system; the hair conditioners further comprise at least about 0.3% by weight cationic hair conditioner; the hair reparatives further comprise at least about 2% by weight cationic hair conditioner; the bath and shower gels further comprise at least about 25% by weight surfactant; the skin lotions and creams further comprise at least about 2% by weight of a cream former; the sunscreens further comprise a sunblocking agent; and the shaving creams further comprise at least about 10% by weight detergent and have a basic pH.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic flow diagram illustrating the steps involved in the preferred process for the production of hydrolyzed rice protein.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

The following examples set forth preferred procedures for the production of hydrolyzed rice protein, and use thereof in cosmetic products. It is to be understood, however, that these examples are provided by way of illustration and nothing therein

should be taken as a limitation upon the overall scope of the invention.

EXAMPLE 1

This example describes a preferred procedure for the preparation of hydrolyzed rice protein, using rice protein concentrate as a starting material. The concentrate was obtained by methods known in the art. The following Table sets forth the amino acid profile of the protein fraction of the concentrate.

TABLE 1

The hydrolysis method is schematically illustrated in FIG. 1, and was carried out as follows. First, 750 gallons of soft water was heated to 140 degree F and placed in a reaction tank. 1,000 pounds of the above-described rice protein concentrate was added to the reaction tank with agitation to completely disperse the meal. At this

point, 22 pounds of 50% NaOH solution was added to the reaction tank for 1 hour to solubilize the proteins and create the alkaline dispersion 10 of FIG. 1.

In the next step, the protein slurry was treated with 15 pounds of protease enzyme (step 12), using a commercially available enzyme (Alcalase, Novo) and hydrolyzed for 2 hours with agitation. During this hydrolysis, the pH was maintained between 7.5-8.0 using 50% NaOH. A second dose of the Alcalase protease enzyme (15 pounds) was then added to the slurry followed by agitation for 2 hours. During this period, there was no pH adjustment.

In the next step 14, the pH of the slurry was adjusted to 6.5 using lactic acid, whereupon a second protease treatment (step 16) was carried out. This treatment included addition of 10 pounds of a second protease enzyme (Flavorzyme, Novo) followed by agitation for 4 hours. Thereupon, 3 pounds of Dual Protease Enzyme (Enzyme Development Corp.) was dispersed in 1 gallon of tap water and added to the slurry followed by the addition of 10 pounds of Neutrase (Novo). The slurry was then agitated for an additional 2 hours to continue the hydrolysis process.

In step 18, lactic acid was added to the slurry to lower the pH to 4.5, followed by the addition of 8 pounds sodium metabisulfite with agitation for 10 minutes. Next, in step 20, the slurry was heated to 160 degree F to deactivate all protease enzymes. The slurry containing hydrolyzed rice proteins was then processed in a rotary vacuum filter (step 22) to remove insolubles, and the filtrate was clarified by passage through a packed-house filter unit (step 24).

The clarified filtrate from step 24 was then fed to a nanofiltration membrane system, in order to generate a permeate and a retentate having different molecular weight profiles (step 26) after the retentate reached about 31% by weight solids using a refractometer. The membrane system was selected so that the lower molecular weight permeate proteins were generally below a molecular weight of 1,000, whereas the retentate proteins had a molecular weight of above about 1,000.

The retentate fraction was first treated by the addition of preservatives (step 28), namely 1% by weight Germaben II and 0.3% by weight Dowicil 200. The retentate was then chilled to 34-35 degree F and allowed to stand for 16-24 hours

(step 30). The chilled retentate was then cold-filtered in a packed-house filter unit to remove haziness (step 32), and additional preservatives were added (15% extra

Germaben II and Dowicil 200).

The resulting retentate was aged 1-2 weeks (step 34) filtered (step 36) and packed in aqueous liquid form in 5-gallon jugs or 55-gallon drums (step 38) as hydrolyzed rice protein 39. This liquid contained about 10% by weight protein. The permeate fraction was conventionally evaporated (step 40) to achieve a solids content of about 34% by weight, using a refractometer. Preservatives were then added (1% Germaben II and 0.3% Dowicil 200) followed by mixing for 1 hour (step 42). The retentate was then subjected to the same aging, filtration and packaging steps 34, 36 and 38 as the retentate, to produce aqueous liquid rice amino acid product 44. This liquid product contained about 4.5% by weight protein.

A molecular weight analysis of the hydrolyzed rice protein (from retenate) and amino acid products (from permeate) revealed that the higher molecular weight protein product was a mixture of peptides and/or protein fragments with a range of molecular weights of from about 1,000-5,000 and an average molecular weight of 3,500; the amino acid product was a mixture of amino acids and peptides with a range of molecular weights of from about 75-1,000, and average molecular weight of 200. In particular, the rice amino acid product was analyzed to contain (wb) 0.28% aspaitic acid, 0.13% threonine, 0.15% serine, 0.38% glutamic acid, 0.09% proline, 0.17% glycine, 0.17% alanine, 0.03% cysteine, 0.14% valine, 0.04% methionine, 0.09% isoleucine, 0.17% leucine, <0.01 % tyrosine, 0.13% phenylalanine, 0.02% histidine, 0.08% lysine, 0.11% arginine, and <0.01% tryptophan.

It will be appreciated that the two products developed using this process comprise peptides and/or protein fragments derived from naturally occurring rice protein, and that the reference to "hydrolyzed rice protein" and "rice amino acid" is a convenience, merely referring to the fact that the respective mixtures have different molecular weight profiles. In both cases, however, the products are "hydrolyzed rice protein" and this term is used herein to refer to both of these products and for that matter any product containing amino acids, peptides and/or protein fragments derived from naturally occurring rice protein via hydrolysis.

EXAMPLE 2

In this example, a rice amino acid product is produced by acid hydrolysis of

rice protein concentrate. In the process, 450 gallons of warm (160 degree F) water is transferred to a homomixer tank, followed by the addition of 2 gallons of concentrated HCl (36%). The mixture is stirred and 950 pounds of rice protein concentrate is added, with continued stirring for 30 minutes. The acidified slurry is then transferred to a glass-lined reactor and 448 gallons of concentrated HCl (36%) is added to the reactor. The temperature of the mixture within the reactor is raised to 212-220 degree F using a heat exchanger, followed by mixing for 24 hours. At the end of the 24 hour mixing period, the hydrolyzate is cooled to 120-140 degree F, and 50% NaOH solution is added to adjust the pH to 6.0-7.0. The neutralized hydrolyzate is then clarified using a rotary vacuum filter to remove solid particulates, and the clarified hydrolyzate is concentrated to the desired solids level (20-30%) in an evaporator.

Preservatives (1% Germaben and 0.3% Dowicil 200) are added. The product is then chilled at 34-35 degree F for 16-24 hours. After chilling, the product is filtered using a packed-house filter unit to remove haziness. The product is then aged for 1-2 weeks, and a final filtration is carried out using a packed-house filter unit. The product is then packaged in suitable containers such as 5 gallon jugs or 55 gallon drums.

EXAMPLE 3

In this example, quaternized derivatives of hydrolyzed rice protein or rice amino acid are prepared. In the first step, 100 parts of the rice product (either hydrolyzed rice protein or hydrolyzed rice amino acid) are added to a reaction tank. The pH of the mixture is adjusted to 9.1-9.3 using 50% NaOH.

Seventeen parts by weight of 3-chloro-2-hydroxypropyl-N, N,N-dimethyldo- decylammonium chloride (Quab 342, 40% active) and 6 parts by weight of tap water are mixed in a separate container, followed by 1.7 parts by weight of 50% NaOH with agitation. The reaction is allowed to proceed for 10 minutes, giving a final pH typically between 10-11 (a small amount of additional 50% NaOH was added if the pH fell below 10). Alternatives to the Quab 342 product are 3-chloro-2- hydroxypropyl-N,N,N-dimethyloctadecylammonium chloride (Quab 426) or 3- chloro-2-hydroxypropyl-N,N,N-trimethylammonium chloride (Quat 188) to produce other derivatives. The solution in the separate container is then added to the pH-

adjusted rice mixture, followed by agitation for 16-24 hours. The pH of the solution is then adjusted 4.4-4.6 using lactic acid (88%) solution. The product is then filtered using a packed-house filter unit to remove particulates, followed by packaging.

Chemically, the finished product is laurdimonium hydroxypropyl hydrolyzed rice protein (or laurdimonium hydroxypropyl rice amino acid). If Quab 426 is used, the product is steardimonium hydroxypropyl hydrolyzed rice protein (or steardimonium hydroxypropyl rice amino acid). If Quat 188 is used, the product is called hydroxypropyl trimonium hydrolyzed rice protein (or hydroxypropyl trimonium rice amino acid).

EXAMPLE 4

Hydrolyzed rice starch is produced by enzyme hydrolysis. 88.5% by weight (based on total weight of mixture) 750 degree C water is transferred into a homomixer tank followed by the addition of 0.5% by weight α-amylase. The mixture is stirred for at least 1.5 hours, and 10.0% by weight of rice starch is added during stirring. At the completion of hydrolysis, the enzyme in the slurry is deactivated by increasing the temperature of the slurry to 850 degree C for at least 1 hour. 96% by weight (based on total solids) hydrolyzed rice protein is added, together with 1% by weight (total solids basis) of a preservative comprising phenoxyethanol, isopropylparaben, isobutylparaben and butylparaben (sold by ISP as LiquiPar). The slurry is then chilled and filtered and adjusted to solids of 15% to 35%.

The parameters of this process may be varied in several respects. Examples include (weight bases as stated above): starting water - 70 - 90% protease enzyme - 0.2 - 1.0% rice starch - 5% - 15% hydrolysis time - 1.5 - 18 hours hydrolyzed rice protein (mix) - 1.0% - 99% hydrolyzed rice starch (mix) - 1.0% - 99%

EXAMPLE 5 The following Tables reflect products produced using the rice protein products of the type described above.

Table 2

Ultra Shine Hair Reparative Shampoo

Aqua Pro® II QRt

010328 DM-B

QS = Quality Standard pH = 5.75±0.5

Viscosity = 8,000 ±15% (RVDV Spindle # 3, 10 rpm @ 25 degree C)

Procedure:

1. In a suitable primary tank meter out required amount of distilled water, in order.

2. Add remaining ingredients with good mixing.

3. Add Part B and C with good mixing.

Table 3

Moisturizing Hand & Body Lotion

Aqua Pro® II QRt

010622 DM-B

QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure:

1. In a suitable primary tank meter out required amount of water add ingredients of Phase A in order and heat to 75 degree C.

2. In a secondary tank weigh out all the ingredients in Phase B in order heat to 75 degree C.

3. Add Phase B to Phase A at 75 degree C with good mixing.

4. Add Part D in order at 35 degree C.

Table 4

Refreshing Facial Mask

Aqua Pro® QRt

030102 DM-D

QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-F, 10 rpm 25 degree C

Procedure:

1. Mix Part A in a primary tank and heat to 75 degree C. 2. In a secondary tank mix all ingredients of Part B and heat to 75 degree C.

3. Add Part B to A and with good mixing add Part C to AB.

4. Cool batch to 35 degree C and add Part D with good mixing.

5. Add Part E to batch at 25 degree C.

Apply on clean face and neck, avoiding eye and lip areas. Relax for 10-15 minutes and rinse well with warm water. For external use only discontinue use if sensitivity is experienced.

Table 5

Gentle Conditioner

Aqua Pro® II QRt

030113 DM-C

1. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben QS = Quality Standard pH = 3.5±0.5

Viscosity = 12,000 ±10% (RVDV Spindle # 4, 10 rpm @ 25 degree C) Procedure: 1. In a suitable primary tank meter out required amount of distilled water.

2. Add ingredients in Part-A in order and heat to 75 degree C when all in solution cool batch to 35 degree C.

3. Add Part B and Part-C in order.

4. Adjust pH 3.0-4.0 with 25% citric acid. 5. Add fragrance and preservative.

Table 6

Control Fine Line Lotion

Aqua Pro® II QRt

030522 DM-C

1. Hydroxypropyltrimoπium Hydrolyzed Rice Protein

QS = Quality Standard Procedure:

1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A and heat to 75 degree C. 3. In a secondary tank, weigh out all the ingredients in Part B in order and heat to

75 degree C.

4. Add Part B to Part A at 55 degree C.

5. Add Premix Part C at 40 degree C.

6. Add Part E at 30 degree C.

Table 7

Hair Color Protect and Reparative Shampoo

Aqua Pro® H QRt

030630 DM-E

1 Hydroxypropyltrimonium Hydrolyzed Rice Protein

2 Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH = 5.75±0.5

Viscosity = 8,000 ±15% (RVDV Spindle # 3, 10 rpm @ 25 degree C) Procedure:

In a suitable primary tank meter out required amount of distilled water, in order add remaining ingredients with good mixing. Add Part-B and C with good mixing. Adjust pH to 5.5 with 25% citric acid. Adjust viscosity with 10% NaCl.

Table 8

Liquid Make-UP

Aqua Pro® QRt

031119 DM-D

1. Hydroxypropyltrimonium Hydrolyzed Rice Protein QS = Quality Standard Procedure:

1. Heat Part A to 75 degree C. Press parts E through colloid mill with some propylene glycol from Part C and recirculate until pigments are evenly dispersed.

2. Pre-mix Part C under mixer and add at 65 degree C on cooling.

3. Premix Part B to 75 degree C and add to main batch.

4. Sprinkle Part F to main batch at 65 degree C on cooling.

5. Pre-mix Part D and add at 65 degree C on cooling. 6. At 45 C add Part G and H to main batch on cooling.

Table 9

Hair Gel

Aqua Pro® II RO 021128 DM-C

1. Hydrolyzed Rice Protein and Hydrolyzed Rice Starch QS = Quality Standard pH = 5.00+0.5

Viscosity = RVDV T-B, 5 rpm @ 25 degree C)

Procedure:

1. Premix Carbomer in 25% water.

2. Add Part B to Part- A with good mixing.

3. Add Part C in order.

Table 10

Facial Mask

Aqua Pro® π RO

030102 DM-F

1. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben QS = Quality Standard pH = 6.0+0.5

Viscosity = RVDV Spindle T-F, 10 rpm @ 25 degree C

Procedure: 1. Mix Part A in a primary tank and heat to 75 degree C.

2. In a secondary tank mix all ingredients of Part B and heat to 75 degree C.

3. Add Part B to A and with good mixing add Part C to AB.

4. Cool batch to 35 degree C and add Part D with good mixing.

5. Add Part E to batch at 25 degree C. Apply on clean face and neck, avoiding eye and lip areas. Relax for 10-15 minutes and rinse well with warm water. For external use only discontinue use if sensitivity is experienced.

Table 11

Gentle Conditioner

Aqua Pro® π RO

030113 DM-E

1. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben QS = Quality Standard pH = 3.5±0.5

Viscosity = 12,000 ±10% (RVDV Spindle # 4, 10 rpm @ 25 degree C)

Procedure:

1. In a subtle primary tank meter out required amount of distilled water.

2. Add ingredients in Part A in order and heat to 75 degree C when all in solution cool batch to 35 degree C.

3. Add Part B and Part C in order.

4. Adjust pH 3.0-4.0 with 25% citric acid.

5. Add fragrance and preservative.

Table 12

Hair Reparative Shampoo Aqua Pro® RO 030501 DM-B

1. Steardimonium Hydroxypropyl Hydrolyzed Rice Protein

2. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH - 5.75±0.5

Viscosity = 8,000 ± 15% (RVDV Spindle # 3, 10 rpm @ 25 degree C) Procedure:

1. In a suitable primary tank meter out required amount of distilled water.

2. Add ingredients of Part A in order.

3. Add Part B and C with good mixing.

4. Adjust pH with 25% Citric Acid. 5. Adjust Viscosity with 25% NaCI.

Table 13

Diminishing Fine Line and Wrinkle Cream Aqua Pro® RO 030522 DM-B

1. Hydrolyzed Rice Protein and Hydrolyzed Rice Starch QS = Quality Standard pH = 6.0+0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure: 1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A and heat to 75 degree C.

3. In a secondary tank, weigh out all the ingredients in Part B in order and heat to 75 degree C.

4. Add Part B to Part A. 5. Add Part C and Part D at 35 degree C.

6. Add Part E at 30 degree C.

Table 14

Film Forming and Hair Color Protective Shampoo

Aqua Pro® II RO

030630 DM-H

1. Hydrolyzed Rice Protein and Hydrolyzed Rice Starch

2. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH = 5.75+0.5

Viscosity = 8,000 ± 15% (RVDV Spindle # 3, 10 rpm @ 25°C)

Procedure:

In a suitable primary tank meter out required amount of distilled water, in order add remaining ingredients with good mixing. Add Part B and C with good mixing. Adjust pH to 5.5 with 25% citric acid. Adjust viscosity with 10% NaCI.

Table 15

Sunscreen Moisturizing Lotion Aqua Pro® RO 031015 DM-B

1. Hydrolyzed Rice Protein and Hydrolyzed Rice Starch QS = Quality Standard pH = 6.0+0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure: 1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A and heat to 75 degree C.

3. In a secondary tank, weigh out all the ingredients in Part B in order and heat to 75 degree C.

4. Add Part B to Part A. 5. Add Part C in order at 30 degree C.

Table 16

Facial Mask

Aqua Pro® QRL

030102 DM-C

QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-F, 10 rpm @ 25 degree C

Procedure:

1. Heat Part A to 75 degree C. 2. Pre-heat Part B to 75 degree C.

3. Add to Part B to Part A.

4. Add Part C in order to Part A + B.

5. Add Part D to Part A B C with good mixing.

6. Cool batch to 35 degree C. 7. Add Part E with good mixing.

Apply on clean face and neck, avoiding eye and lip areas. Relax for 10-15 minutes and rinse well with warm water. For external use only discontinue use if sensitivity is experienced.

Table 17

Gentle Conditioner

Aqua Pro® H QRL

030113 DM-D

1. Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben QS = Quality Standard pH = 3.5±0.5

Viscosity = 12,000 ±10% (RVDV Spindle # 4, 10 rpm @ 25 degree C)

Procedure:

1. In a suitable primary tank meter out required amount of distilled water.

2. Add ingredients in Part A in order and heat to 75 degree C when al in solution cool batch to 35 degree C.

3. Add Part B and Part C in order.

4. Adjust pH 3.0-4.0 with 25% citric acid.

5. Add fragrance and preservative.

Table 18

Moisturizing Cream

Aqua Pro® QRL

030320 DM-B

1. Laurdimonium Hydroxypropyl Hydrolyzed Rice Protein QS = Quality Standard pH = 6.0+0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure:

1. Heat Part A to 75 degree C.

2. Heat Part B to 75 degree C.

3. Add Part B to Part A with good mixing at 65 degree C.

4. Add Part C and cool to 35 degree C.

5. Add Part D in order.

Table 19

Reduce Fine Line Lotion

Aqua Pro® II QRL

030522 DM-D

1. Laurdimonium Hydroxypropyl Hydrolyzed Rice Protein

QS = Quality Standard pH = 6.0+0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure: 1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A and heat to 75 degree C.

3. In a secondary primary tank, weight out all the ingredients in Part B in order and heat to 75 degree C.

4. Add Part B to Part A at 55 degree C. 5. Add Premix Part C at 40 degree C.

6. Add Part E at 30 degree C.

Table 20

Hair Strengthening and Reparative Shampoo

Aqua Pro® QRL

040409 DM-A

1 Laurdimonium Hydroxypropyl Hydrolyzed Rice Protein

QS = Quality Standard pH = 5.75±0.5

Viscosity = 8,000 +15% (RVDV Spindle # 3, 10 rpm @ 25 degree C)

Procedure:

1. In a suitable primary tank meter out required amount of distilled water heat to 75 degree C.

2. Add ingredients in Part A with good mixing cool at 35 degree C.

3. Add ingredients of Part B and C in order with good mixing.

4. Adjust pH 5.5±0.5.

Table 21

Hair Color Protect and Conditioning Shampoo

Aqua Pro® II QRL

030630 DM-F

1 Laurdimonium Hydroxypropyl Hydrolyzed Rice Protein

2 Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH = 5.75+0.5

Viscosity = 7,500 ±15% (RVDV Spindle # 3, 10 rpm @ 25 degree C) Procedure:

In a suitable primary tank meter out required amount of distilled water, in order add remaining ingredients with good mixing. Add Part B and C with good mixing. Adjust pH to 5.5 with 25% citric acid. Adjust viscosity with 10% NaCI.

Table 22

Make-Up Concealer

Aqua Pro® QRs

011009 DM-C

1 Steardimonium Hydroxypropyl Hydrolyzed Rice Protein QS = Quality Standard Procedure:

1. Heat Part A to 75 degree C.

2. Heat Part B to 75 degree C. 3. Add Part B to A with high shear mixing.

4. Add Part C in order with good mixing.

5. Add Part D at 35 degree C.

Table 23

Facial Mask

Aqua Pro® QRs

030102 DM-E

QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-F, 10 rpm @ 25 degree C

Procedure:

1. Mix Part A in a primary tank and heat to 75 degree C. 2. In a secondary tank mix all ingredients of Part B and heat to 75 degree C.

3. Add Part B to A and with good mixing add Part C to AB.

4. Cool batch to 35 degree C and add Part D with good mixing.

5. Add Part E to batch at 25 degree C.

Apply on clean face and neck, avoiding eye and lip areas. Relax for 10-15 minutes and rinse well with warm water. For external use only discontinue use if sensitivity is experienced.

Table 24

Moisturizing Lotion for Sensitive Skin Aqua Pro® QRs 030113 DM-B2

1 Steradimonium Hydrolxypropyl Hydrolyzed Rice Protein QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure:

1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A in order and heat to 75 degree C.

3. Pre-mix ingredients in Part B to 75 degree C and add to Part A.

4. When all in solution cool batch to 35 degree C.

5. Add Part C in order.

Table 25

Reduce Fine Line

Aqua Pro® QRs

030522 DM-D

1 Steardimonium Hydroxypropyl Hydrolyzed Rice Protein QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure: 1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A and heat to 75 degree C.

3. In a secondary tank, weigh out all the ingredients in Part B in order and heat to 75 degree C.

4. Add Part B to Part A at 55 degree C. 5. Add Part C at 40 degree C.

6. Add Part D at 30 degree C.

Table 26

Hair Color Protect and Voluminizing Shampoo

Aqua Pro® II QRs

030630 DM-G

1 Steardimonium Hydroxypropyl Hydrolyzed Rice Protein

2 Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH = 5.75±0.5

Viscosity = 8,000 ±15% (RVDV Spindle #3, 10 rpm @ 25 degree C) Procedure:

In a suitable primary tank meter out required amount of distilled water, in order add remaining ingredients with good mixing. Add Part B and C with good mixing. Adjust pH to 5.5 with 25% citric acid. Adjust viscosity with 10% NaCl.

Table 27

Reparative and Conditioning Shampoo

Aqua Pro® QRs

040409 DM-B

1 Steardimonium Hydrolxypropyl Hydrolyzed Rice Protein

2 Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS = Quality Standard pH = 5.75±0.5

Viscosity = 8,000 ±15% (RVDV Spindle # 3, 10 rpm @ 25 degree C) Procedure:

1. In a suitable primary tank meter out required amount of distilled water mix ingredients in Part A in order when all in solution.

2. Add ingredients of Part B .

3. Add Part C to main batch. 4. Add Part D in order.

5. Adjust pH to 5.5 ±0.50.

6. Adjust desire viscosity with 25% NaCl.

Table 28

Eye Serum

Aqua Pro® QRs

031020 DM-D

1 Steardimonium Hydroxypropyl Hydrolyzed Rice Protein QS = Quality Standard pH = 6.0±0.5

Viscosity = RVDV Spindle T-B, 5 rpm @ 25 degree C

Procedure:

1. In a suitable primary tank, meter out required amount of distilled water.

2. Add ingredients of Part A in order.

3. Add Part B and mix.

4. Add Part C in order.

Table 29

Hair Reparative Conditioner

Aqua Pro® II QRs

040225 DM-B

1 Propylene Glycol (and) Diazolidinyl Urea (and) Methylparaben (and) Propylparaben

QS - Quality Standard pH = 3.5±0.5

Viscosity = 12,000 ±10% (RVDV Spindle # 4, 10 rpm 25 degree C)

Procedure:

1. In a suitable primary tank meter out required amount of distilled water.

2. Add ingredients in Part A in order and heat to 75 degree C.

3. Cool batch to 35 degree C and add Part B and Part C in order.

4. Adjust pH to 3.0-4.0 with 25% citric acid.

5. Add Part C fragrance and preservative as desire.