NZ206542A

NZ206542A - Enzyme-containing liquid detergent compositions

Info

Publication number: NZ206542A
Application number: NZ206542A
Authority: NZ
Inventors: M C Crossin
Original assignee: Colgate Palmolive Co
Priority date: 1982-12-13
Filing date: 1983-12-09
Publication date: 1985-12-13
Also published as: DK159501B; DE3344097C2; NO159288C; DK159501C; CA1208578A; IT1172373B; PT77817A; GR79133B; AU2229183A; ZM8783A1; MX161813A; GB2131826B; NO159288B; SE460726B; BR8306834A; FR2537597B1; ZA839050B; SG40189G; PT77817B; FI834555A

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number £06542 206542 Priority Date(s): K<5. Complete Specification Filed: Class: OJ.Q3/.3Z& Publication Date: '.t?.. PKm.. P.O. Joumsl. No*. f.f'. NO DRAWINGS Patents Fonn No. 5 Number PATENTS ACT 1953 Dated COMPLETE SPECIFICATION LIQUID DETERGENT COMPOSITION WITH MIXED ENZYME FORMULATION KWe COLGATE-PAUOLIVE COMPANY, a corporation organised under the laws of the State of Delaware, United States of America of 300 Park Avenue, New York., New York 10022, United States of America do hereby declare the invention for which i/we pray that a Patent may be granted to Klt/us. and the method by which it is to be performed, to be particularly described in and by the following statement: 1 206542 This Invention relates, in general, to enzyme-containing liquid detergent compositions which are suitable for laundering or pre-soak formulations. More particularly, the invention relates to detergent compositions containing mixtures of protease and amylase enzymes In defined proportions which provide particularly effective soil and stain removal during laundering. The formulation of enzyme-containing liquid detergent compositions has been the focus of much attention in the prior art. The desirability of incorporating enzymes into detergent compositions is primarily due to the effectiveness of proteolytic and amylolytic enzymes In decomposing proteinaceous and starchy materials found on soiled fabrics, thereby facilitating the removal of stains such as, gravy stains, blood stains, chocolate stains and the like during laundering. However, enzymatic materials suitable for laundry compositions, particularly proteolytic enzymes, are-relatively expensive. Indeed, they generally are the most expensive ingredients in a typical commercial liquid detergent composition, even when present in relatively minor amounts. Moreover, an excess of enzymes is generally required in the detergent formulation. Because of the known instability of enzymes in aqueous compositions an excess of enzymes is generally added to the formulation to compensate for the expected loss of enzyme activity during prolonged periods of storage. Consequently, the expense associated with the use of enzymeB in liquid detergent , compositions has heretofore been a significant deterrent to their widespread commercial use -2- 206542 Detergent compositions containing mixtures of enzymes, e.g. proteases and amylases, have been broadly described in the prior art. Thus, for example, U.S. Patent 3,630,930 describes a granular detergent composition containing from about 0.5 to 20$ of enzyme carrier granules, the enzyme granules being comprised of from about 0.001 to 10$ of mixtures of protease and amylase enzymes in a weight ratio of protease to amylase of 50:1 to 1:5. British Patent specification 1,240,058 discloses a granular detergent composition containing a mixture of protease and amylase enzymes in a weight ratio of protease to amylase of 30:1 to 3:1, the weight percent of amylase In the composition being from 0.0003 to 3$. In U.S. Patent 3,931,034 to Inamorato et al there is disclosed a granular detergent composition containing a mixture of alkaline protease ando^-amylase enzymes in a ratio of activity varying from 100,000 to 400,000 Novo amylase units of amylase per Anson unit of protease. Accordingly, while the use of enzyme mixtures in granular deter gent compositions is generally disclosed in the patent literature, the mixtures themselves are, in most instances, so broadly described as to encompass, for example, mixtures wherein the percent protease may vary within 5 orders of magnitude (British Patent 1240058), or the percent amylase may vary within 5 orders of magnitude (U.S. Patent 3,630,930), thereby encouraging the belief that within such broad ranges, the greater.the amount-of enzyme used, the more effective the resultant stain removal. In addition, the aforementioned patents, in common with the Inamorato et al patent are strictly related to granular compositions and hence provide no teaching regarding the use of enzyme mixtures in liquid compositions 3977 SUMMARY OT' THE INVENTION The present invention provides an enzyme-containing liquid detergent composition comprising: (a) from about 5 to about 75$, hy weight, of one or more detergent surface active agents selected from the group consisting of anionic, nonionic, cationic, ampholytic and zwitterionic detergent compounds; (b) from about 25 to 85$ water; and (c) an enzyme mixture consisting essentially of an alkaline protease enzyme and an °£amylase enzyme in relative proportions such that the ratio of the enzyme activities is from about 4,000 to about " 80,000 Novo amylase units of e<-amylase per Anson unit of protease, said protease being present in an amount to provide from about 0.25 to about 2.5 Anson units per 100 grams of detergent composition. In accordance with the process of the invention, laundering of stained and/or soiled materials is effected by contacting such materials with an aqueous solution of the above-defined liquid detergent composition. The present invention is predicated on the discovery that the amount of alkaline protease enzyme which is ordinarily necessary for the removal of proteinaceous stain can be significantly.reduced by the combination therewith of an amount of amylase enzyme in accordance with the Invention so as to provide a detergent composition having equivalent or improved stain removal capabilities but at a considerably reduced formulation expense. Unlike the disclosures in the art which recommend mixing proteases and amylases over broad ranges in detergent compositions, the enzyme mixtures herein described are characterized by a synergistic interaction of -4- 206542. protease and amylase, and encompass only those mixtures having a « narrowly defined ratio of activities. The activities of the alkaline protease and cxs-amylase enzymes are expressed in Anson units of protease, and Novo amylase units of amylase, respectively. These are units commonly used in the art to describe the activity, under common conditions, of enzyme formulations containing protease or amylase enzymes. In a preferred embodiment of the invention the enzyme mixtures contain relative amounts of alkaline protease and cf-amylase to provide from about 10,000 to 50,000 Novo amylase units of o^-amylase per Anson unit of protease, an activity ratio of from about 15,000 to 40,000 being even more preferred and a ratio of from about 30,000 to 40,000 being especially desirable. The amount of the enzyme mixture present In the liquid detergent composition will, of course, depend to some extent on the amount of the composition which is to be added to the wash solution. For"detergent compositions intended for use at concentrations of about 0.15$ in the wash solution of an automatic home laundry machine, a suitable amount of mixture will provide from about 0.25 to about 2.5 Anson units of protease per hundred grams of detergent composition, a ratio of from about 0.5 to 2.0 being preferred, and about 1.5 Anson units/ 100 grams of composition being a particularly preferred protease concentration. DETAILED DESCRIPTION OF THE INVENTION The activity of the alkaline protease enzyme is, as noted above, measured in terms of Anson units. The Aneon hemoglobin method for the measurement of Anson unit activity is a procedure well known in the art for determining the activity of proteolytic 206542 enzymes, and is set forth in the "Journal of General Physiology", volume 22, pages 79-89 (1939), such disclosure being incorporated herein by reference. The modified Anson hemoglobin method may also be used for measuring the proteolytic activity, such modified method being described in the article "Alkali-Resistant Enzyme for Detergents", S. R. Green, Soap and Chemical Specialities, pages 86, 88, 90, 9^ and 133, May 1968, which disclosure Is incorporated herein by reference. In principle, the method employs the alkaline protease enzyme to digest a denatured hemoglobin substrate at standard conditions in a buffered aqueous medium at the selected pH, and the amount of digested material is determined by a color test with phenol reagent. The activity of the ®<-amylase enzyme is, as previously noted, measured in terms of Novo amylase units. The standard procedure for the measurement of such Novo units is a modification of the SKB method (Sandstedt, Kneen & Blish, Cereal Chemistry 16, 712, (1939)) without addition of beta-amylase. In this procedure 20 ml of a buffered starch solution (prepared by the method described below) are measured in a test tube (diameter 24 mm, length 190 mm) and placed in a wafrer thermostat at a temperature of 37°C. After a few minutes pre-warming, 10 ml of the amylase solution to be tested (or v ml amylase solution + (10-v) ml water) is added. The contents of the tube are thoroughly mixed and at the same time a stopwatch is started. At appropriate time intervals 1 ml of the reaction mixture is added to 5 ml of a dilute iodine solution (prepared by the method described below), shaken and transferred to a comparison tube, and the color is compared with the standard color. If the color endpoint is reached in less than 10 minutes, a more dilute amylase solution or a smaller volume of amylase solution is used. As colorimeter the Hellige Comparator 607 is used with the glass c*{-amylase standard. (cf. Redfern Methods for determination of amylase, Cereal Chemistry 24, 259, (1947)). The of-amylase -6- 206542 activity of the sample may be calculated by using the following form-< ula: A = 1^30 x V where t x a x v A = ^amylase activity in Novo amylase units per gram ; t - time to reach the color endpoint (minutes) a = weight of sample in grams V = volume to which the sample is diluted (ml) v = volume of amylase solution used (ml) The factor "l430" is not strictly constant but depends to some degree upon the starch quality used. For exact determinations, the value of the factor should be calculated by means of a commercially available standard amylase preparation with known activity. The "dilute iodine solution" mentioned above is prepared by dissolving 1 ml of "stock Iodine solution" and 20 g of potassium iodide in sufficient water to make 500 ml; the "stock iodine solution" is prepared by dissolving 11 g of iodine crystals and 22 g of potassium iodide in sufficient water to make 500 ml. The "buffered starch solution" mentioned above is prepared as follows: 10 g soluble starch (e.g. Merck, Amylum solubile, Soluble Starch, Erg. B.6) calculated as dry matter are made into a slurry with some water. The slurry is added to about 200 ml of boiling water. "When the starch is completely dissolved, the solution is cooled, transferred to a 1 liter volumetric flask and made up to the mark with water. The starch solution (made by dissolving 9.36 g NaCl, 69.00 g KHgPO^, 4.80 g NagHPOj^, 2 H2O in sufficient water to make 1 liter). Finally the solution is saturated with toluene. The pH of the finished buffered starch solution should be 5.7. The starch solution must be as freshly prepared as possible but can be stored in the refrigerator for not more than 2k hours. Distilled water is used In all cases. 206542. The"enzyme activity of proteolytic and amylolytic enzyme preparations is ordinarily determined, as a practical mattery without conducting the above-described assay procedures. For the majority of commercially available liquid enzyme preparations containing protease or amylase enzymes, the enzyme activity is provided by the manufacturer and is expressed in Anson units, or Novo amylase units (or units directly proportional thereto). Alternatively, the activity of a given enzyme preparation can be readily determined analytically by a procedure wherein the enzyme reactivity with a protein or starch substrate, as the case may be, is measured at standard conditions and then compared with the reactivity of reference enzyme preparations of known activity. In such analytical procedure, enzyme reactivity may be conveniently expressed as the optical density of a test solution containing the enzyme'preparation and the protein or starch substrate when measured at standard conditions, the higher the optical density, the greater the activity. The suitable alkaline proteolytic enzymes include the various commercial liquid enzyme preparations which have been adapted for use,-in detergent compositions, enzyme preparations in powdered form being also useful although, as a general rule, less convenient for incorporation into the present liquid detergent compositions. Thus, suitable liquid enzyme preparations include "Alcalase" and "Esperase" sold by Novo Industries, Copenhagen, Denmark, and "Maxa-tase" and "AZ-Protease" sold by Gist-Brocades, Delft, The Netherlands. "Alcalase" is particularly preferred for the present compositions. Among the suitable<y-amylase liquid enzyme preparations are those sold by Novo Industries and Gist-Brocades under the tradenames "Termamyl" and "Maxamyl", respectively. An organic solvent is preferably used in combination with water to serve as the solvent for the liquid detergent composition. The preferred organic solvent is a lower alkanol of 1 to 4 carbon atoms 206542 having from 1 to 3 hydroxy groups, preferably 1 or 2. The lower alkanol is most preferably ethanol or a mixture of ethanol and isopropanol, with lower monoalcohols such as propanol and butanol, and lower polyols of 2 to 3 carbon atoms such as ethylene glycol and propylene glycol being useful albeit less preferred. The use of primary, secondary and tertiary butanol or n-propanol as the lower alkanol is generally restricted to mixtures of same with ethanol, ethanol being preferably at least 80 to 90$ of such mixtures. It is highly preferred to use ethanol as the sole alkanol and organic solvent. In mixtures of ethanol and isopro-panol it is preferred that ethanol be the major component, ethanol being usually from 60 to 90$ of the mixture, preferably about 75$ (i.e., in a 3:1 ratio). Of course, other mixtures of the various alkanols may be used, such as ethanol and propylene glycol, and in such mixtures it is also preferred that ethanol be the major component. 206542 The compositions of the present invention contain one or more surface active agents selected from the group of anionic, nonionic, cationic, ampholytic and zwitterionic detergent compounds. The synthetic organic detergents employed in the practice of the invention may be any of a wide variety of such compounds which are well known and are described at length in the text Surface Active Agents, Vol. II, by Schwartz, Perry and Berch, published In 1958 by Interscience Publishers, the relevant disclosures of which are hereby incorporated by reference. The nonionic detergents are usually poly-lower alkoxylated lipophiles wherein the desired hydrophile-lipophile balance Is obtained from addition of a hydrophilic poly-lower alkoxy group to a lipophilic moiety. For the present compositions the nonionic detergent employed is preferably a poly-lower alkoxylated higher alkanol wherein the alkanol is of 10 to 18 carbon atoms and wherein the number of moles of lower alkylene oxide (of 2 or 3 carbon atoms) is from 3 to 12. Of such materials it is preferred to employ those wherein the higher alkanol is a higher fatty alcohol of 11 or 12 to 15 carbon atoms and which contain from 5 to 8 or 5 to 9 lower alkoxy groups per mole. Preferably, the lower alkoxy is ethoxy but In some Instances It may be desirably mixed with propoxy, the latter, if present, usually being a minor (less than 50$) constituent. Exemplary of such compounds are those wherein the alkanol is of 12 to 15 carbon atoms and which contain about 7 ethylene oxide groups per mole, e.g., Neodo^' 25-7 and Neodol 23-6.5, which products are made by Shell Chemical Company, Inc. The former Is a condensation product of a mixture of higher fatty alcohols averaging about 12 to 15 carbon atoms, with about 7 moles of ethylene oxide and the latter is a corresponding mixture wherein the carbon atom content of the higher fatty alcohol is 12 to 13 and the number of ethylene oxide groups per mole averages about 6.5. The higher alcohols are primary alkknols. 206542 Other examples of such detergents include Tergitol® 15-S-7 and Tergitol 15-S-9, both of which are linear secondary alcohol ethoxy- lates made by Union Carbide Corporation, The former is a'mixed ethoxylation product of an 11 to 15 carbon atom linear secondary alkanol with seven moles of ethylene oxide and the latter is a similar product but with nine moles of ethylene oxide being reacted. Also useful in the present compositions are higher molecular weight nonionics, such as Neodol 45-11, which are similar ethylene oxide condensation products of higher fatty alcohols, with the higher fatty alcohol being of 14 to 15 carbon atoms and the number of ethylene oxide groups per mole being about 11. Such products are also made by Shell Chemical Company. Other useful nonionics are represented by Plurafac B-26 (BASF Chemical Company), the reaction product of a higher linear alcohol and a mixture of ethylene and propylene oxides. In the preferred poly-lower alkoxylated higher alkanols, the best balance of hydrophlllc and lipophilic moieties are obtained when the number of lower alkoxies are from about 40$ to 100$ of the number of carbon atoms in the higher alcohol, preferably 40 to 60$ thereof. The nonionic detergent is preferably comprised of at least 50$ of the preferred ethoxylated alkanols. Higher molecular weight alkanols and various other normally solid nonionic detergent compounds-and surfactants may contribute to gelation of the liquid detergent composition and consequently, are normally omitted or limited in quantity in the present compositions, although minor proportions thereof may be employed for their cleaning properties, etc. With respect to both preferred and less preferred nonionic detergents, the alkyl groups present therein are preferably linear although a minor degree of slight branching may be tolerated, such as at a carbon next to or two carbons removed from the terminal carbon of the straight chain and away from the ethoxy phain 206542 with the proviso that such branched alkyl is no more than three carbons in length. Normally the proportion of carbon atoms in such a branced configuration will be minor, rarely exceeding 20$ of the total carbon atom content of the alkyl. Similarly, although linear alkyls which are terminally joined to the ethylene oxide chains are highly preferred and are considered to result in the optimum combination of detergency, blodegradabillty and non-gelling characteristics, medial or secondary joinder to the ethylene oxide in the chain may occur. In such instance, it is usually in only a minor proportion of such alkyls, generally lesB than 20$ but aB is in the case of the aforementioned Tergitols, may be greater. • Also, when propylene oxide is present in the lower alkylene oxide chain, it will usually be less than 20$ thereof and preferably less than 10$ thereof. With the nonionic detergent, which is the major synthetic organic detergent in the present liquid detergent compositions, there may be employed an anionic detergent. The most preferred anionic detergent compounds are the higher (10 to 18 or 20 carbon atoms) alkyl benzene sulfonate salts wherein the alkyl group preferably contains 10 to 15 carbon atoms, most preferably being a straight chain alkyl radical of 12 or 13 carbon atoms. Preferably, such an alkyl benzene sulfonate has a high content of 3- (or higher) phenyl isomers and a correspondingly low content (usually well below 50$) of 2- (or lower) phenyl isomers; in other words, the benzene ring is preferably attached in .large part at the 3, 4, 5, 6 or 7 position of the alkyl group and the content of isomers in which the benzene ring Is attached at the 1 or 2 position is correspondingly low. Typical alkyl benzene sulfonate surface active agents are described in U.S. Patent 3,320,174. Of course, more highly branched alkyl benzene sulfonates may also be employed but usually are not preferred, due to their lack of biodegradability. 1 2 I' 9S4 iv: fftoevfl© ( 206542 Other anionic detergents which are useful are the olefin sulfonate salts. Generally, these contain long chain alkenyl sulfonates or long chain hydroxyalkane sulfonates (with the OH being on the carbon atom which is not directly attached to the .carbon atom bearing the -SO^H group). The olefin sulfonate detergent usually comprises a mixture of such types of compounds in varying amounts, often together with long chain diBulfonates. or sulfate-sulfonates. Such olefin sulfonates are described in many patents, such as U.S. Patent Nos. 2,06l,6l8; 3,409,637; 3,332,880; 3,420,875; 3,428,654; 3,506,580; and British Patent No. 1,129,158. The number of carbon atoms in the olefin sulfonate is usually within the range of 10 to 25, more commonly 10 to 18 or 20, e.g., a mixture principally of C12, and 0jg, having an average of about 14 carbon atoms, or a mixture principally of C-j_4, C^g and 0^3, having an average of about 16 carbon atoms. Another class of useful anionic detergents is that of the higher paraffin sulfonates. These may be primary paraffin sulfonates made by reacting long chain alpha-olefins and bisulfites, e.g., sodium bisulfite, or paraffin sulfonates having the sulfonate groups distributed along the paraffin chain, such as the products made by reacting a long chain paraffin with sulfur'dioxide and. oxygen under ultraviolet light, followed by neutralization with sodium hydroxide or other suitable base (as in U.S. Patents 2,503,280 2,507,088; 3,260,741; and 3,372,188), The paraffin sulfonates preferably contain from 13 to 17 carbon atoms and will normally be the monosulfonate but if desired, may be di-, tri- or higher sulfonates. Typically, the di- and polysulfonates will be employed In admixture with a corresponding monosulfonate,', for example, as a mixture of mono- and disulfonates containing up to about 30$ of the disulfonate. The hydrocarbon substituent thereof will preferably be linear but If desired, branched chain paraffin i NXPATSMF OFPItB r — -re; '?ss . - 13 - i RECEIVED 206542- sulfonates can be employed, although they are not as good with respect to biodegradability. Other suitable anionic detergents are sulfated ethoxylated is a fatty alkyl of from 10 to 18 or 20 carbon atoms, m is from 2 to 6 or 8 (preferably having a value from about 1/5 to 1/2 the number of carbon atoms in R) and M is a solubllizing salt-forming cation, such as an alkali metal, ammonium, lower alkyl-amino or lower alkanolamino, or a higher alkyl benzene sulfonate wherein the higher alkyl Is of 10 to 15 carbon atoms. As is the case with the preferred nonionic detergent, it will be preferred for the alkyl in the anionic alkoxylate detergent to be a mixture of different chain lengths, such as 11, 12, 13, l4 and 15 carbon atom chains or 12 and 13 carbon atom chains, rather than all of one chain length. Ethylene oxide is the preferred lower alkylene oxide of the anionic alkoxylate detergent, as it is with the nonionic detergent, and the proportion thereof in the polyethoxylated higher alkanol sulfate is preferably 2 to 5 moles of ethylene oxide groups present per mole of anionic detergent, with three moles being most preferred, especially when the higher alkanol is of 11 or 12 to 15 carbon atoms. To maintain the desired hydrophile-lipo-phile balance, when the.carbon atom content of the alkyl chain is in the lower portion of the 10 to 18 carbon atom range, the ethylene oxide content of the detergent may be reduced to about two moles per mole whereas when the higher alkanol is of l6 to 18 carbon atoms, in the higher part of the range, the number of ethylene oxide groups may be increased to 4 or 5 and in some cases to as high as 8 or 9. Similarly, the salt-forming cation may be altered to obtain the best solubility. It may be any suitably solubllizing metal or radical but will most frequently be alkali metal, e.g., higher fatty alcohols of the formula RO^gH^O^SO^M, wherein R 1 4 2065*2. sodium, or ammonium. If lower alkylamine or alkanolatnine groups are utilized the alkyls and alkanols will usually contain from 1 to 4 carbon atoms and the amines and alkanolamlnes may be mono-, di- and tri-substituted, a.s in monoethanolamine, diisopropanol-amine and trimethylamine. The poly-lower alkoxy higher alkanol sulfates maybe employed in place of"or in combination with other preferred anionic detergents such as the higher alkyl benzene sulfonates to supplement the nonionic detergent in the present liquid detergent compositions. A preferred polyethoxylated alcohol sulfate detergent Is available from Shell Chemical Company and is marketed as Neodol 25-3S. Examples of the higher alcohol polyethenoxy sulfates which may be employed In the liquid detergent compositions of the invention include: mixed C_„ normal- or primary alkyl tri- Xd~ Xz> ethenoxy sulfate, sodium salt; myristyl triethenoxy sulfate, potassium salt; n-decyl dlethenoxy sulfate, diethanolamine salt; lauryl dlethenoxy sulfate, ammonium salt; palmityl tetraethenoxy sulfate, sodium salt; mixed C^ ^ normal primary alkyl mixed trl- and tetra-ethenoxy sulfate, sodium salt; stearyl penta-ethenoxy sulfate, trimethylamine salt; and mixed C^0 normal primary alkyl triethenoxy sulfate, potassium salt. Other useful anionic detergents include the higher acyl sarcosinates, e.g., sodium N-lauroyl sarcosinate; higher fatty alcohol sulfates^ such as sodium lauryl sulfate and sodium tallow alcohol sulfate; sulfated oils; sulfates of mono- or diglycerides of higher fatty acids, e.g., stearic monoglyceride monosulfate; although, of these, the sodium higher alcohol sulfates have been found to be inferior to the polyethoxylated sulfates in detergency; aromatic poly(lower alkenoxy) ether sulfates, such as the sulfates of the condensation products of ethylene oxide and nonyl phenol 206.5^2. (usually having 1 to 20 oxyethylene groups per molecule, preferably 2 to 12); polyethoxy higher alcohol sulfates and alkyl phenol polyethoxy sulfates having a lower alkoxy (of 1 to 4 carbon atoms, e.g., methoxy) substituent on a carbon close to that carrying the sulfate group, such as monomethyl ether monosulfate of a long chain vicinal glycol, e.g., mixture of vicinal alkane diols of l6 to 20 carbon atoms in a straight chain; acyl esters of isethionic acid, e.g., oleyl isethlonates; acyl N-methyl taurides, e.g., potassium N-methyl lauroyl or oleyl taurides; higher alkyl phenyl polyethoxy sulfonates; higher alkyl phenyl disulfonates, e.g., pentadecyl phenyl disulfonate; and higher fatty acid soaps, e.g., mixed coconut oil and tallow soaps in a 1:4 ratio. Among the aforementioned types of anionic detergents, the sulfates and sulfonates are generally preferred but the corresponding organic phosphates and phosphonates may also be employed when their contents of phosphorus are not objectionable. Generally, the water soluble anionic synthetic organic detergent, (including soaps), as was previously indicated, are salts of alkali metal cations, such as potassium, lithium, and especially sodium, although salts of ammonium and substituted ammonium cations, such as those previously described, e.g., triethanolajmine, triisopropylamine, may be used too. In the above exemplifications of water-soluble anionic detergent it should be considered that the sodium, potassium, ammonium and alkanolammonium salts are individually recited for each detergent. Ampholytic detergents may be employed In the present compo- • sitions in minor proportions In replacement of the anionic detergent or a part thereof or in replacement of part of the nonionic detergent. Ampholytic detergents include the higher fatty carboxy-lates, phosphates, sulfates or sulfonates which contain a cationic substituent such as an amino group, which may be quaterhized, e.g., with a lower alkyl group, or chain extended at the amino group by 206542- condensation with a lower alkylene oxide, e.g., ethylene oxide. Generally the compositions containing such ampholytic or catlonic detergents will not be as effective and may have a greater tendency to gel or thicken on standing. Therefore they are often avoided. However, if such properties are unobjectionable, minor proportions of ampholytlcs such as Miranol C2M, sold by Mlranol Chemical Company, or Deriphat 151, a sodium N-coco betaamine propionate, sold by General Mills, Inc., may be utilized. A catlonic detergent that may sometimes be useful is distearyl dimethyl ammonium chloride (it has fabric softening activity) and the higher fatty amine oxides, such as bis(2-hydroxyethyl) octadecyl amine oxide. - . The viscosity control agent utilized to maintain the desired viscosity of the liquid detergent composition, prevent gelation at low temperatures and allow a reduction in lower alkanol solvent content is preferably a water soluble formate. Sodium formate is preferred but alkali metal formates may be utilized, e.g., potassium formate and various other water soluble formates, Including formic acid, which may be added to the liquid detergent composition wherein it dissolves, ionizes and/or reacts to produce essentially the same type of liquid detergent as results from the addition of the alkali metal formate In salt form. Other formates that .may be employed are those of water soluble cations, such as previously described as salt-forming cations for the anionic detergents. Although it is preferred to employ the formate viscosity control agent, it has been found that various salts of dibasic acids can also be successfully used, among which the best appears to be disodium adipate, referred to herein as sodium adipate. Other salts of dibasic acids of the formula (CH2)n(C00H)2 where n is 1 to 6, may also be employed 20654Z and in some instances the salts of mono-unsaturated acids of the same chain lengths and configurations may be used. However, it is highly preferred to utilize the saturated aliphatic straight chain terminally carboxylated compounds. It is more preferable to employ those wherein n Is 3 to 5, most preferably 4, and wherein the acid Is fully neutralized, but the acid salts may also be used. Among the dibasic acids that may be employed, either as the mono- or disalts, are malonic, succinic, glutaric, adlpic and pimelic acids. An unsaturated dibasic acid, maleic acid, can also be used, at least in part. The acids may be employed without prior neutralization or may be used as their salts, such as disodium malonate, monopotasslum succinate, di-triethanolamine glutarate, disodium adipate and monosodium pimelate. To assist in solubllizing the detergents and optical bright-eners which may be present in the liquid detergent compositions, a small proportion of alkaline material or a mixture of such materials is often included in the present formulations. Suitable alkaline materials include mono-, di- and trialkanolamlnes, alkyl amines, ammonium hydroxides. Of these, the preferred materials are the alkanolamines, preferably the trialkanolamlnes and of these, especially triethanolamine. The pH of the final liquid detergent composition containing such a basic material will usually be neutral or slightly basic. Satisfactory pH ranges are from 7 to 10, preferably from about 7.5 to 9, and most preferably from about 7.5 to 8.5. The optical fluorescent brighteners or whiteners employed in the liquid detergent compositions are important constituents of modern detergent compositions which give washed laundry and materials a bright appearance so that the laundry is not only clean but also appears clean. Although it Is possible to utilize a single brlghtener for a specific Intended purpose in the present 206542 liquid detergent it is generally desirable to employ mixtures of brighteners which will have good brightening effects on cotton, nylons, polyesters and blends of such materials and which are also bleach stable, A good description of such types of optical brighteners is given In the artlfcle "The Requirements of Present Day Detergent Fluorescent Whitening Agents" by A. E. Siegrist, J, Am. Oil Chemists Soc., January 1978 (Vol. 55). That article and U.S. Patent 3,812,041, issued May 21, 1974, both of which are hereby incorporated by reference for their relevant disclosures, contain detailed descriptions of a wide variety of suitable optical brighteners. Among the brighteners that are useful in the present liquid detergent compositions are: Calcofluor 5BM (American Cyanamid); Calcofluor White ALF (American Cyanamid); J30F A-2001 (Ciba); CDW (Hilton-Davis); Phorwite RKH, Phorwite BBH and Phorwite BHC (Verona); CSL, powder, acid (American Cyanamid); FB 766 (Verona); Blancophor PD (GAF);'UNPA (Geigy); Tinopal RBS 200 (Geigy). The acid or "nonionic" forms of the brighteners tend to be solubilized by alcohols of the present formulas, while the salts tend to be water soluble. Adjuvants may be present in the liquid detergent composition to give it additional properties, either functional or aesthetic. Included among the useful adjuvants are soil suspending or anti-redoposition agents, such as polyvinyl alcohol, sodium carboxy-methyl cellulose, hydroxypropylmethyl cellulose; thickeners, e.g., gums, alginates, agar agar; foam improvers, e.g., lauric myristic diethanolamide; foam destroyers, e.g., silicones; bactericides., e.g., tribromosalicylanillde, hexachlorophene; dyes; pigments (water dispersible); preservatives; ultraviolet absorbers; fabric softeners; opacifying agents, e.g., polystyrene suspensions; and perfumes. Of course, such materials will be selected for the 20654& properti^s desired in the finished product and to be compatible with the other constituents thereof. Other adjuvants that may be employed are dihydric or trihydric lower alcohols which, in addition to being solvents and reducing the flash point of the product, can act as anti-freezing constituents and may improve compatibilities of the solvent system with particular product components. Among these compounds the most preferred group, includes the lower polyols of 2 to 3 carbon atoms, e.g., ethylene glycol, propylene glycol and glycerol, but the lower alkyl (C-^-C^) etheric derivatives of such compounds, known as Cello-solves^ may also be employed. The proportions of such substitutes for the lower alkanols will be limited, normally being held to no more than 20$ of the total alcohol content of the liquid detergent. Another category of useful additives are hydrotropes which serve to enhance the solubility in aqueous solution of components which otherwise have limited solubility in water. Useful hydro-tropes include the alkali metal, ammonium and ethanolamine salts of the following acids; (1) aryl sulfonic acids, such as benzene sulfonic acid and C^-C^ alkyl-substituted benzene sulfonic acids, e.g., toluene sulfonic acid and xylene sulfonic acid; and (2) C5-C6 alkyl sulfuric acids, such as hexyl sulfuric acid. The proportions of the various components of the present liquid detergent compositions are important for the manufacture of a uniform product of desirable viscosity and acceptable heavy duty laundering action which does not gel at low temperatures or upon standing in an open container at room temperature. To promote solubility of the fluorescent brighteners and other constituents in the detergent composition and to make a clear, homogeneous and readily pourable liquid product, from 10 to 60$ of the total liquid detergent concentrate should be nonionic detergent. Preferably, especially when an anionic detergent is present 20654-2 in the liquid product, the proportion of the nonionic detergent is from 20 to 40$ and more preferably it is 30 to 40$, with the best formula known at the present time including about 32$. The proportion of anionic detergent will usually be in the range of 3 to 15$, preferably 4 to 12$ and most preferably 6 to 10$ with the best formula known at the present including about 7$ thereof. The ratio of total nonionic detergent to anionic detergent will normally be from 15:1 to 1:1, with 8:1 to 2:1 being preferred and 5:1 to 3:1 being most preferred. The lower alkanol in the liquid detergent compositions will generally be present in a sufficient proportion to aid in dissolving and/or stabilizing the various constituents in the final product. The proportions of lower alkanol used will normally be from about 3 to 15$, preferably 4 to 12$, more preferably 4 to 8$ and at the present time most preferably about 5$. The viscosity control agent utilized or a mixture of such agents will normally be from about 0.5$ to 5$ of the final liquid detergent composition, preferably about 0.5 to 3$, and most preferably about 1$. The percentage of water, the main solvent in the present compositions, will usually be from about 25 to 85$, preferably 35 to 65$ and most preferably from about 40 to 55$, hy weight, of the liquid composition. In the most preferred formulations there will be about 45 to 50$ water. The content of the alkaline additive, such as triethanolamine, In the liquid composition is usually from about 0.1 to 5$ of the composition and preferably 1 to 3$, by weight, thereof. The total proportion of optical brlghtener is normally from about 0.05 to 1.5$, preferably about O.l to 1$ and most preferably about 0.2 to 0.5$. 206542 The liquid detergent compositions of the present invention, can be made by simple manufacturing techniques. In a typical manufacturing method the optical brighteners are slurried in the monohydric alcohol, after which water is added to the slurry together with a small amount of a base, such as triethanolamine, .which helps to partially dissolve the previously suspended material. Addition of the anionic detergent compound usually results in the remainder of the brightener dissolving to make a clear solution. The viscosity control agent is then added as the acid, acidic salt or completely neutralized salt, preferably the sodium or potassium salt, and agitation is continued until the solution becomes clarified, which may normally take about 5 to 10 minutes. At this point the principal detergent, the non-Ionic, is added along with a minor amount of acid for purposes of pH regulation, the pH being generally adjusted to a value at which the proteolytic enzyme used is most stable. This is followed by agitation of the solution and the addition of adjuvants, such as, perfume and dye which give the product its final desired properties. The protease and oframylase enzyme preparations are then added to th.: solution and mixed therewith as the final step in producing the product liquid detergent composition. If desired the viscosity control additive may be incorporated earlier in the procedure. The above operations may be effected at room temperature, although suitable temperatures within the range of 20° to 50°C. may be employed, as desired, with the proviso that when volatile materials, such as perfume., are added, the temperature should be low enough so as to avoid objectionable losses. The product obtained will usually have a pH within the range of 7 to 10, and a density within the range of from 0.9 to 1.1, preferably from 0.95 to 1.05. The viscosity of the final product at 24°C. will be in the range of 60 to 150 centipolses, preferably from about 80 to 140 centipolses, and most preferably from about 115 to 135 centipolses, according to measurements that are made with a Brookfield vlscosimeter at room temperature. - 22 - 2065*2 The present liquid compositions are efficient and easy to use. Compared to heavy duty laundry detergent powders, much smaller volumes of the present liquids are employed to obtain comparable cleaning of soiled laundry. For example, using a typical preferred formulation of this invention, only about 60 grams or 1/4 cup of liquid Is needed for a full tub of wash in a top-loading automatic washing machine in which the water volume is 15 to 18 gallons (55 to 75 liters); and even less (about 1/2) is needed for front-loading machines. Thus, the concentration of the liquid detergent composition in the wash water is on the order of about 0.1$. Usually, the proportion of the liquid composition in the wash solution will range from about 0.05 to 0.3$, preferably from 0.08 to 0.2$ and most preferably from about 0.1 to 0.15$. The proportions of the various constituents of the liquid composition may vary accordingly. Equivalent results can be obtained by using greater proportions of a more dilute formulation but the greater quantity needed will require additional packaging and will generally be less convenient for consumer use. Also, more highly diluted products will be more apt to freeze in cold weather, and may be more subject to hydrolysis and chemical changes on storage. 23 3 0 AUG 1984 Example 1 20^542 A liquid detergent composition (containing no enzymes) designated as composition "A" was prepared at room temperature by mixing the following components in the Btated proportions: Component Weight Percent Ethoxylated C22~C15 32.0 primary alcohol (7 moles EO/mole alcohol) Sodium dodecyl benzene 7-0 sulfonate \ Triethanolamine 2.8 Ethanol 5.0 Sodium formate 1.0 h2S04 (cone.) 0.7 Optical brighteners^) 0.27 Dye(2) 0.01 Perfume 0.35 Water balance (1) A mixture of Phorwite RKH and Phorwite BHC brighteners manufactured by Verona. (2) Polar Brilhant Blue (PBB) manufactured by Ciba-Geigy. 206542 Enzyme-containing liquid detergent compositions B-U were formulated by adding various amounts of protease and alpha-amylase enzymes to the above-described composition A. The enzyme concentration In each of the detergent compositions is shown In Table 1, expressed in terms of percent of enzyme formulation based on the weight of the composition. The protease enzyme employed was a liquid enzyme.formulation sold under the name "Alcalase" by Novo Industries of Copenhagen, Denmark having a concentration of 2.5 Anson units per gram of enzyme preparation. The alpha-amylase enzyme employed was a liquid ^nzyme formulation sold under the name "Termamyl" by Novo'Industries having a concentration of 120,000 Novo amylase units per gram of liquid enzyme preparation. Test Procedure A total of 6 cotton swatches, 3 stained with beef liver blood and 3 stained with grass, were placed In each of 4 buckets of a Tergotometer vessel manufactured by U.S. Testing Company. A series of laundering tests were conducted using a different liquid detergent composition from compositions A-U in each bucket of the Tergotometer under the following test conditions: • Liquid detergent concentration 0.09$ Agitation 100 rpm Agitation time 10 minutes Water temperature 120°F Water hardness About 150 ppm as calcium carbonat At the end of the wash, the test swatches were rinsed in tap water and then dried. The percent stain removal was measured by taking a reflectance reading for each stained test swatch prior to and after the washing using a Gardner XL-20 Colorimeter,. and the percent stain removal ($ S.R.) was calculated as follows; 206542- fo S.R, = (Rd after washing) - (Rd before Washing) (Rd before staining) - (Rd before washing) wherein "Rd before washing" represents the Rd value after staining. The values of percent stain removal calculated for each of the three test swatches having a common stain were averaged for each liquid detergent composition tested. The results are shown In Table 1 which sets forthvthe percent S.R. for each of the liquid detergent compositions tested (compositions A-U) and the enzyme concentration of such detergent compositions. £3:sv»- 26 20654Z Table 1 Comparative Stain Removal with Enzyme-Containing Detergent Compositions Composition Weight $/-j\ Protease*1 } Weight $ oc-Amylasel^; $ Stain Removal Beef liver blood Grass A 0.0 0.0 42.0 31.5 B 0.2 0.0 50.8 37.6 C 0.4 0.0 52.8 36.7 D 0.6 0.0 53.9 36.8 E 0.8 0.0 55.5 36.1 P 0.0 0.2 44.2 37.6 ' G 0.2 0.2 55.3 42.4 H 0.4 0.2 58.4 44.8 X 0.6 0.2 60.6 44.5 .J 0.8 0.2 60.5 44.1 K 0.0 0.4 47.1 38.3 L 0.2 0.4 56.2 43.4 M 0.4 0.4 58.6 45.0 N 0.6 0.4 63.3 44.8 0 0.8 0.4 63.2 45.0 P 0.0 0.6 47.7 37.6 Q 0.2 0.6 55.4 41.3 R 0.4 0.6 57.5 42.1 S " 0.6 0.6 60.9 43.7 T 0.8 0.6 62.0 44.5 U 0.0 0.8 47.7 36.5 (1) The proteolytic activity of Alcalase is 2.5 Anson urtlts per gram. Thus, a concentration of, for example, 0.2$ of Alcalase in the liquid detergent composition corresponds to a protease enzyme activity of 0.5 Anson units (0.2 x 2.5) per 100 grams of detergent composition. (2) The araylolytlc activity of Termamyl is 120,000 Novo amylase units per gram. Thus, a concentration of 0.2$ Termamyl in the liquid detergent composition corresponds to an ©^.-amylase enzyme activity of 24,000 Novo amylase units (0,2 x 120,000) per 100 grams of detergent composition. 206542. As indicated in Table 1, the percent stain removal (S.R.) achieved with composition A represents the S.R. achieved in the absence of enzymes in the detergent composition. Referring to the S.R. data for the beef liver blood stain, a comparison of the S.R. achieved with compositions P, K, P and U, all of which contain amylase enzyme, but no protease and are therefore not in accordance with the invention, shows that composition P containing 0.6 wt. $ Amylase provided the maximum improvement in S.R. (47.7$) achievable with amylase enzyme, i.e., an increase of about 5.7$ relative to the S.R. value of 42.0$ for the enzyme-free composition A, Similarly, a comparison of the S.R. achieved with compositions B, C, D and E, all of which contain protease enzyme, but no amylase, shows that composition E'containing 0.8$ protease provided the maximum improvement in S.R. (55.5$) with protease enzyme, i.e., an increase of about 13-5$ relative to the 42$ S.R. achieved with enzyme-free composition A. The synergistic interaction of protease and amylase enzymes for the removal of proteinaceous stains is evident from Table 1. Thus, for example, composition G containing 0.2 wt. $ protease and 0.2 wt. $ amylase enzymes (corresponding to an araylase/protease enzyme activity ratio of 24,000 Novo amylase units per 0.5 Anson units) provided nearly the same improvement in S.R. (relative to enzyme-free composition A) as was achieved with 0.8 wt. $ protease in composition E. Prom an economic standpoint, the use of composition G containing a mixture of enzymes in accordance with the invention clearly represents a substantial reduction in the requirement for the relatively expensive protease enzyme, as compared to composition E. The highest percentage of stain removal was achieved with composition N. Specifically, the combination of 0.6 wt. $ protease and 0.4 wt. $ amylase in composition N provided a greater than 21$ increase In the percent S.R. for the blood stain relative to enzyme- j 4SKP3984 „ </div>

Claims

<div class="application article clearfix printTableText" id="claims"> 206542 free composition A. Thus, the 63.3$ S.R. achieved with composition N Is significantly higher than the maximum S.R. that could be achieved with detergent compositions containing protease enzyme in the absence of c*-Amylase. The amylase/protease enzyme activity ratio of such composition N is 48,000 Novo amylase units per 1.5 Anson units. The synergistic interaction of protease and alpha-amylase enzymes is likewise evident in the S.R. data for the grass stain. Thus, for example, composition H containing 0.2$ amylase and 0.4$ protease enzymes provided a substantially higher S.R. than could be achieved with detergent compositions containing either protease or amylase as individual enzymes • in the composition. WHAT WE CLAIM IS; 206542-

1. An enzyme-containing liquid detergent composition comprising; (a) from substantially 5 to substantially 75%, by weight, of one or more detergent surface active agents selected from the group consisting of anionic, nonionic, cationic, ampholytic and zwitterionic detergent compounds; (b) from substantially 25 to 85%, by weight, water; and (c) an enzyme mixture consisting essentially of an alkaline protease enzyme and an «• -amylase enzyme in relative proportions such that the ratio of the respective enzyme activities in said mixture is from substantially 4,000 to substantially 80,000 Novo amylase units of ot-amylase per Anson unit of protease, said protease being present in an amount to provide from substantially 0.25 to substantially 2.5 Anson units per 100 grams of detergent composition.

2. A detergent composition according to claim 1 wherein the ratio of the enzyme activities in said enzyme mixture is from substantially 15,000 to substantially 40,000 Novo amylase units of oi-amylase per Anson unit of protease, and said protease is present in an amount to provide from substantially 0.5 to substantially 2.0 Anson units per 100 grams of detergent composition.

3. A detergent composition according to claim 2 wherein said ratio of enzyme activities is from substantially 30,000 to a Li Lid njlpatentqffbce :antially 40,000 Novo amylase units of ot -amylase per ".r'.p- Anso i unit of protease. -8 OCT 1985

4. A detergent composition according to claim 1 wherein said detergent surface active agents consist essentially of from substantially 20% to substantially 40%, by weight, of a water-soluble C2-C3 alkoxylated C10-C13 alkanol nonionic - 30 - 206542. detergent and from substantially 4 to 12%, by weight, of a water-soluble salt of a C10-C15 alkyl benzene sulfonate anionic detergent; and which contains, in addition, from substantially 3 to 15%, by weight, of a lower alkanol selected from the group consisting of a lower monoalcohol having 1 to 4 carbon atoms, a lower polyol having 2 to 3 carbon atoms and mixtures thereof.

5. A detergent composition according to claim 1 which contains, in addition, from substantially 0.5 to 5%, by weight, of a viscosity control agent selected from the group consisting of water-soluble formate salts and dibasic acids of the formula (CH2)n<COOH)2 wherein n is 1 to 6.

6. An enzyme-containing liquid detergent composition consisting essentially of (a) <b) (c) ItZrPATENT OFFICE - G OCT ]?85 R'-C^tVSO from substantially 20 to substantially 40%, by weight, of a nonionic detergent compound consisting essentially of a water-soluble C2-C3 alkoxylated C10-Ci8 alkanol; from substantially 4 to substantially 12%, by weight, of an anionic detergent compound consisting essentially of a water-soluble salt of a Cio~Ci5 alkyl benzene sulfonate; from substantially 3 to 15%, by weight, of a lower alkanol selected from the group consisting of a lower monoalcohol having 1 to 4 carbon atoms, a lower polyol having 2 to 3 carbon atoms and mixtures thereof; from substantially 0.5 to 5%, by weight, of a viscosity control agent consisting essentially of a water-soluble formate salt; (e) from substantially 35 to 65%, by weight, water; and (f) an enzyme mixture consisting essentially of an alkaline protease enzyme and an 06-amylase enzyme in relative proportions such that the ratio of the - 31 - -W- ZObi respective enzyme activities in said mixture is from substantially 4,000 to substantially 80,000 Novo amylase units of ot-amylase per Anson unit of protease, said protease being present in an amount to provide from substantially 0.25 to substantially 2.5 Anson units per 100 grams of detergent composition.

7. A detergent composition according to claim 6 which, in addition, contains from 0.1 to 5%, by weight, of an alkanol-amine.

8. A detergent composition according to claim 6 wherein the ratio of enzyme activities in said enzyme mixture is from substantially 15,000 to substantially 40,000 Novo amylase units of ^-amylase per Anson unit of protease, and said protease is present in an amount to provide from substantially 0.5 to substantially 2.0 Anson units per 100 grams of detergent composition.

9. A detergent composition according to claim 8 wherein said ratio of enzyme activities is from substantially 30,000 to substantially 40,000 Novo amylase units of «-amylase per Anson unit of protease.

10. A detergent composition according to claim 6 wherein said nonionic detergent compound is a polyethoxylated C12-C15 alkanol having from 3 to 12 ethylene oxide groups per mole; said anionic detergent is a C12 or c13 alkyl benzene sulfonate; said lower alkanol is ethanol or a mixture of MJ^PATEWTOFPItf^ Lno1 ant^ isopropanol; and said viscosity control agent is -8 OCT 1985 sod RECEIVED um formate.

11. A method of laundering comprising contacting the stained and/or soiled fabrics to be laundered with an enzyme-containing liquid detergent composition comprising: - 32 - • 206543 (a) from substantially 5 to substantially 75%, by weight, of one or more detergent surface active agents selected from the group consisting of anionic, nonionic, cationic, ampholytic and zwitterionic detergent compounds; (b) from substantially 25 to 85%, by weight, water; and (c) an enzyme mixture consisting essentially of an alkaline protease enzyme and an »£-amylase enzyme in relative proportions such that the ratio of the respective enzyme activities in said mixture is from substantially 4,000 to substantially 80,000 Novo amylase units of pt-amylase per Anson unit of protease, said protease being present in an amount to provide from substantially 0.25 to substantially 2.5 Anson units per 100 grams of detergent composition.

12. A method according to claim 11 wherein the ratio of enzyme activities in said enzyme mixture is from substantially 15,000 to substantially 40,000 Novo amylase units of ^-amylase per Anson unit of protease, and said protease is present in an amount to provide from substantially 0.5 to substantially 2.0 Anson units per 100 grams of detergent composition.

13. A method according to claim 11 wherein said liquid detergent composition consists essentially of M? pATeMTncgiffil^" from substantially 20 to substantially 40%, by weight, of a nonionic detergent compound consisting essentially of a water-soluble C2-C3 alkoxylated C]_o~Ci8 alkanol; -8 OCT 1985 RECEIVED (b) from substantially 4 to substantially 12%, by weight, of an anionic detergent compound consisting essentially of a water-soluble salt of a C10-C15 alkyl - 33 - 20654a benzene sulfonate; (c) from substantially 3 to 15%, by weight, of a lower alkanol selected from the group consisting of a lower monoalcohol having 1 to 4 carbon atoms, a lower polyol having 2 to 3 carbon atoms and mixtures thereof; (d) from substantially 0.5 to 5%, by weight, of a viscosity control agent consisting essentially of a water-soluble formate salt; (e) from substantially 35 to 65%, by weight, water; and (f) an enzyme mixture consisting essentially of an alkaline protease enzyme and an od-amylase enzyme in relative.proportions such that the ratio of the respective enzyme activities in said mixture is from substantially 4,000 to substantially 8O7-OOO Novo amylase units of ot-amylase per Anson unit of protease, said protease being present in an amount to provide from substantially 0.25 to substantially 2.5 Anson units per 100 grams of detergent composition. NX. PATENT < WEST-WALKEF1, McCABE -80C7J985 per: ATTORiui: 1 o run i HE APPLICANT RECSIVIQ - 34 - </div>