blood-cleansing ability of sodium chloride solutions

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1 Csmet. Sci., 49, (March/April 1998) Evaluatin f the effect f surfactants n the bld-cleansing ability f sdium chlride slutins MARTIN E. CHANDLER, JUDY BATEMAN, and THOMAS G. WOOD, Research and Develpment Department, C. B. Fleet C. Inc., 4615 Murray Place, Lynchburg, VA Received fr publicatin March 23, Synpsis This study investigated the effect f three different surfactants n the bld-cleansing ability f sdium chlride slutins. Three surfactants, sdium lauryl sulfate (SLS), cetyl pyridinium chlride (CPC1), and decyl glucside, and three sdium chlride slutins, 0.9% sdium chlride, 2.0% sdium chlride, and a sdium chlride-baseduche slutin, were used. The cleansing was based n the percent bld remval frm a clth. In all three sdium chlride slutins, the surfactant interacts, altering the bld-cleansing ability f the sdium chlride slutins. SLS, CPC1, and decyl glucside lwered the bld cleansing ability f the sdium chlride slutins, althugh the decyl glucside can be used effectively, in lwer cncentratins, with sdium chlride slutins. INTRODUCTION Physilgical saline is used as a bilgical cleanser and irrigant because its istnic nature makes it cmpatible with bdy fluids and tissues; therefre, red bld cells can be suspended in it withut being lysed (1). Surfactants are als effective cleansing agents fr bth bilgical and nn-bilgical substances because they separate particles frm surfaces and allw them t be washed away by the slutin (2). Theretically, if sdium chlride slutins and the ideal surfactant culd be cmbined in the prper prprtin, a pwerful all-purpse cleanser culd be prduced. Researc has shwn that salt inhibits a surfactant's cleansing ability, but there is little infrmatin n hw surfactants affect the bilgical cleansing f sdium chlride slutins (3). By studying the influence f several surfactants n sdium chlride slutins, the interactin between the tw substances when used t remve bld and ther bilgical materials can be bserved. If prperly cntrlled, a slutin culd use bth sdium chlride and surfactant prvide a bilgical and nn-bilgical cleansing. The ultimate gal f this research is t demnstrate hw surfactants affect the cleansing abilities f sdium chlride slutins. Sdium lauryl sulfate, CH3(CH2) CH2OSO3Na, in aqueuslutin, carries its surfaceactive prperties as anins; therefre it is referred t as an aninic surfactant. It is a ll

2 102 JOURNAL OF COSMETIC SCIENCE cmmn ingredient in detergents and tthpaste (4). SLS is knwn t be incmpatible in acids (ph < 2.5), where the SLS is in a nn-inized frm, which is highly inactive in slutin. Cetyl pyridinium chlride, a quaternary ammnium cmpund, is a catinic surfactant, with the fllwing frmula: /NCH2(CHz) 14CH3CL CPC1 is nt a very effective detergent, althugh it is ppularly used as a preservative (5). The third surfactant, decyl glucside, is a nninic surfactanthat des nt dissciate int ins when in slutin. Decyl glucside is an alkyl plyglycside, a surfactant cnsisting f numerus alkyl chains lengths. Decyl glucside is a hydrcarbn with an average chain length f 12 carbn atms (6) and is an effective cleanser because it cntains plar glucse grups, with a typical mlecule cntaining an average f tw plar glucse grups. By prviding pwerful cleansing ability with little r n irritatin t the skin and mucus membranes, alkyl plyglycsides have becme an exciting alternative in surfactant technlgy. The three sdium chlride slutins were tested against surfactants that representhree f the main classes f surfactants. Each f the surfactants was tested at varius cncen- tratins arund the critical micelle cncentratin. EXPERIMENTAL MATERIALS The SLS was btained frm the Stepan Cmpany, Nrthfield, Illinis; CPC1 frm Penta Manufacturing, Fairfield, New Jersey; and decyl glucside (Plantaren 2000 ) frm Henkel Crpratin, Hbken, New Jersey. The duche slutin with sdium chlride was Summer's Eve Extra Cleansing Vinegar and Water, supplied by C. B. Fleet C. Inc., Lynchburg, Virginia. All materials were used withut further purificatin. CLEANSING PROCEDURE All bld samples were cllected in Vacutainer tubes with EDTA as an anticagulant. The plasma was remved, and the cells were rinsed and adjusted t a 40% hematcrit with the 0.9% sdium chlride and stred at 4øC. Three-inch-diameter Veratec Flexiln (70% rayn/30% plyester) clth pads were cut int eight pie-shaped pieces alng the diagnal. A 50- fi aliqut f bld was expelled nt the tip f the clth using an Eppendrf pipettr. After ne minute, the clth was picked up with frceps and placed int an 80-ml beaker cntaining 8 ml f the slutin under study, and placed n a Fisher clinical rtatr at 100 rpm. After swirling fr 15 secnds, the rtatr was stpped and the clth remved. Excess liquid was drained by gently tuching the clth alng the inside wall f the beaker. Ptassium cyanide reagent was added t the beaker as necessary. The reagent bund t the hemglbin, changing the smtic pressure arund the cell, which caused the red

3 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS 103 bld cells t lyse. This methd was used t determine the amunt f hemglbin released int the slutin by the lysed bld cells. The percent transmittance (%T) at 550 nanmeters was determined fr each f the hemglbin slutins using a Perkin- Elmer Lambda 3 UV/VIS spectrphtmeter with a tungsten light surce. Standard curves were prepared fr each surfactant cncentratin, as well as fr each new bld sample. These standard curves were based n bld dilutins and their respective percent transmittances. All f the standard curves were essentially linear, with crrelatin cefficients clse t 1. By using standard curve values, the percent transmittance was cnverted t milligrams f hemglbin per milliliter f slutin. The percentage f bld remved was calculated by dividing the mg/ml f hemglbin by 6.25, which was the theretical amunt f hemglbin delivered in the 50-pl aliqut f bld, and multiplying by 100. All repetitins in an individual series were averaged, with the averages fr each sample based n at least ten trials. DISCUSSION Figures 1 thrugh 9 plt the % bld remval vs the cncentratin f surfactant in the sdium chlride slutins. These curves prvide infrmatin that can be used t explain the chemical interactin between the sdium chlride in slutin and the surfactants. SODIUM LAURYL SULFATE The ph f the 0.9% NaCI/SLS and 2.0% NaCI/SLS slutins (Figures 1, 2) remained between 6.0 and 7.0. The duche slutins (Figures 3, 6, and 9) maintained a ph f 3.32 t 3.38 due t the acetic acid (vinegar) in slutin. In acidic slutins, the SLS is unable t inize t a great degree, restricting its surface-active capabilities. The surface-active anins f SLS, which are hydrphbic alkyl chains with a negative in attached t ne end, frm micelles that attract the plar water mlecules less than the small, highly plar Na + and C1-. Bld cells are affected by the water arund them and, depending n the nature f the slutin, the cells can lyse r crenate. The SLS micelies interact and cmpete with the bld cell membrane and prevent the water frm maintaining the pressure utside f the bld cell. Theretically, the nn-plar prtins f the surfactant mlecules are attracted t the cell membrane by van der Waals frces, with the phsphlipid prtin entering the cell membrane. This wuld decrease the cell's permeability t water. As the water pressure utside the bld cell decreases, the pressure inside the cell causes the bld cell t lyse and release its cntents int slutin. Once in slutin, the bilgical cntents f the cell are denatured, and SLS micelles are then capable f remving this denatured material frm the clth. The cleansing drps ff as the SLS cncentratin increases t 0.08% (Figure 1), and the cleansing then increases fr the next tw successive SLS cncentratins. SLS alne in aleinized water is able t cleanse the denatured cellular material frm the clth, but at the lwer SLS cncentratins n lysing ccurs. At these lwer cncentratins f SLS, the sdium chlride is respnsible fr the bld remval. As the cncentratin f SLS increases, mre surface-active artins enter the slutin. This

4 104 JOURNAL OF COSMETIC SCIENCE loo E I I I I % SLS I O NaC1 Slutin/SLS DI Water DI Water/SLS Figure % NaC1/SLS slutins. increases the amunt f micelies within the slutin, thereby reducing the slutin's ability t remve the bld. At higher cncentratins f SLS, the bld cells are being lysed by the SLS, and sdium chlride is nt effective in remving the lysed cellular material. With an abundance f material depsited n the clth, the SLS is respnsible fr the cleansing ccurring in the slutin. The same trend can be seen in the 2.0% NaC1/SLS slutin (Figure 2). The cleansing remains steady thrughuthe slutins, and des nt drp ff as much as in the 0.9% NaCI. With mre Na + and CI- in slutin, mre SLS is required t create micelles because the sdium chlride restricts micelle frmatin. The sdium chlride can sus- pend the cells and distribute them int the slutin, and the SLS lyses these cells and remves the denatured cellular material. Althugh the micelies can lyse the cells at lw cncentratins f SLS, there are nt enugh micelies t adequately cleanse the denatured

5 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS [ I % SLS NaCI Slutin/SLS 0 DI Water DI Water/SLS Figure 2. 2% NaC1/SLS slutins. cellular material depsited n the clth. Overall, the cleansing decreases slightly because in lwer cncentratins f SLS, the micelles are able t lyse the cells efficiently, but they are nt as efficient in cleansing the abundance f denatured cellular material depsited n the clths. In the duche slutin (ph = ), the SLS is incapable f sufficient inizatin (Figure 3). Micelie frmatin is nt present; therefre, the surfactant actin f SLS is greatly restricted and cleansing is at a minimum. The bld cells are lysed in the slutin, but the denatured cellular material depsited n the clths cannt be cleansed by the sdium chlride. Therefre, the nninized SLS interferes with the salt slutin's ability t remve the bld frm the clth. In deinized water (darkened square, Figure 3) the SLS lysed the bld cells and remved

6 106 JOURNAL OF COSMETIC SCIENCE , 70 _ O % SLS O Duche/SLS DI Water DI Water/SLS DI Water (ph=4)/sls Figure 3. Sdium chlride-baseduche/sls slutins. 88% f the material, but in deinized water, with its ph adjusted t apprximately 4 with HCI (darkened triangle, Figure 3), the cleansing drpped t 76%. The ability f SLS t cleanse by surfactant actin relies n its ability t frm micelies, but in acidic slutin, SLS is nt inized t any great extent, and s n micelies are frmed. If micelles are nt frmed, then the surfactant activity f SLS is reduced, resulting in drastically reduced bld cleansing. CETYL PYRIDINIUM CHLORIDE CPCI, as a catinic surfactant, is nt recgnized as a particularly pwerful surfactant;

7 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS 107 therefre, the cleansing assciated with the CPC1/sdium chlride slutins can be attributed mainly t the sdium chlride (7, Figure 5). In deinized water alne (darkened square, Figure 4), a CPC1 slutin (0.01%) cleansed 57% f the bld. Althugh the surface-active catin is able t lyse the cells in a similar fashin as SLS, the surfactant activity f CPC1 is nt as adept at cleansing the lysed cell material. As the cncentratin f CPC1 increases in the sdium chlride slutins (Figure 5), the cleansing decreases. In these situatins the CPC1 tyses the cells befre they can be carried away frm the clth by the sdium chlride, and nce the denatured cellular material is depsited n the clth, the CPC1 is less effective in cleansing. Accrding t the data, the majrity f the cleansing in these slutins is dne by the sdium chlride where it lifts the bld cells ff the clth and they are distributed within the slutin. The cells begin t lyse as the cncentratin f CPC1 increases, and the sdium chlride cannt lift the cellular material away frm the clth. "øø t 9O 8O 7O 6O 5O 4O 3O % CpCl NaC1 Slutin/CPCl DI Water DI Water/CPC1 Figure % NaC1/CPC1 slutins.

8 108 JOURNAL OF COSMETIC SCIENCE The same effect can be seen in the duche slutin (Figure 6). The CPC1 is able t inize in the acidic slutin and is capable f lysing the bld cells, but the majrity f bld cleansing is carried ut by the sdium chlride in this slutin. Part f the decreased cleansing with increased cncentratins f CPCI culd be the catinic substantivity t the fabric. DECYL GLUCOSIDE The decyl glucside/sdium chlride results differ frm the ther slutins because the cncentratins had t be higher than thse f SLS r CPC1. The decyl glucside wuld nt disslve in the sdium chlride slutins belw 0.09% because at this cncentratin and belw, the micelies are unable t frm and the nninic decyl glucside mlecules are suspended in slutin. At lwer cncentratins, the slutins appear cludy because loo % CPCl O NaCI Slutin/CPCl DI Water DI Water/CPCl Figure 5. 2% NaC1/CPC1 slutins.

9 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS J I I I % SLS l:)cuf L-"P 0 DI Wfdnr m DIW l Figure 6. Sdium chlride-baseduche/cpc1 slutins. the surfactant is salted ut f slutin. Decyl glucside is a nninic surfactanthat des nt inize in the slutin, but like SLS and CPC1, it is able t cmpete with the bld cells fr the water mlecules nce micelies have frmed. The micelles frmed by decyl glucside are able t attract water because f the plar glucse grups n the mlecules. Decyl glucside in deinized water (darkened square, Figure 7) is able t cleanse 76% f the bld, an amunt almst identical t that f the 74% cleansed by aleinized water alne. In the lwer cncentratins (belw 0.09%), the cleansing accmplished by these slutins is by the sdium chlride in slutin. The cleansing decreases ver the next tw cncentratins (0.11% and 0.15%) because the decyl glucside is able t lyse the cells befre the sdium chlride can remve them frm the clth, and the denatured cellular material is then nly partially cleansed (Figures 7, 8). As the decyl glucside increases, the cleansing rises ver the next three cncentratins (0.175%, 0.20%, and 0.30%)

10 110 JOURNAL OF COSMETIC SCIENCE m 0 I I I I I % Dec' Glucslde O DI Wmlim' Figure % NaC1/decyl glucside slutins. (Figures 7, 8). At the lwest and highest cncentratins, the decyl glucside des nt inhibit the bld cleansing, but in fact, it seems that it and sdium chlride are an effective team in cleansing the bld. In the duche slutin (Figure 9), the cleansing des decrease slightly, but the reductin is nt as drastic as seen in the ther sdium chlride slutins. Since decyl glucside is a nninic surfactant, the ph f the acidic duche slutin des nt affect its cleansing ability. CONCLUSIONS Overall, the bld cleansing ability f the surfactants is inferir t that f the sdium chlride slutins. Sdium chlride remves the bld by lifting the entire cell frm the

11 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS 111 loo 8 70 E 60 5O 4O 3O % Decyl Glucside 0, NaC1 Slutin/Decyl Glucside DI Water [] DI Water/Dccyl Glucside Figure % NaC1/decyl glucside slutins. surface f the clth. The surfactants lyse the bld cells, denature the cellular material, and remve the material by carrying it away with their micelies. The tw cleansing methds vary greatly, and the percentage bld remval can fluctuate as the surfactant cncentratin is increased within a slutin (8, 9). At lw cncentratins f the surfactant, the majrity f the cleansing is dne by the sdium chlride. As the surfactant cncentratin rises, the cleansing is affected by the interactin between the surfactant and sdium chlride in slutin. At this pint, there is a transitin between the cleansing by sdium chlride and that by the surfactant. When ne lks at the figures, ne ntes that the transitin is dented by a general decrease in cleansing and a large standard deviatin. Cleansing by sdium chlride is quite gentle, but the cleansing by the surfactant invlves lysing the cells and remving

12 112 JOURNAL OF COSMETIC SCIENCE loo i ( ' 3O % Decyl Glucside SE Duche/Decyl Glucside DI Water DI Water/Decyl Glucside Figure 9. Sdium chlride-baseduche/decyl glucside slutins. the denatured cell material. At the transitin pint, there is lysed cell material in slutin as well as whle bld cells. Therefre, the amunt f cleansing can vary greatly frm trial t trial, resulting in increased standard deviatins. Once the cncentratin f the surfactant is increased, the majrity f the cleansing is dne by the surfactant, and the standard deviatins decrease (10-14). The SLS and decyl glucside are pwerful surfactants, but CPC1 is nt an effective cleansing agent. It is capable f lysing the cells, but it nly remves a prtin f the denatured cellular material. SLS and decyl glucside are pwerful surfactants that are capable f cleansing lysed bld cells. When cmparing SLS and decyl glucside, ne bserves that SLS is a little better at cleansing in deinized water but that decyl glucside is far superir in the acidic duche slutin. Als, decyl glucside is mre likely t be used because it is less irritating (6).

13 SURFACTANTS AND BLOOD CLEANSING BY SLS SOLUTIONS 113 Surfactants inhibit the cleansing f sdium chlride slutins. Inic interactins, van der Waals frces, and the ability t lyse bld cells all influence the slutins' ability t cleanse bld frm the surface f the clth. The results indicate that surfactants can inhibit the cleansing f bld by sdium chlride slutins by lysing the cells and allwing the denatured cellular material t depsit nt the surface f the clth. Once such material is n the surface f the clth, the sdium chlride slutins cannt remve it. ACKNOWLEDGMENTS We wuld like t thank C. B. Fleet C., Inc., fr supprting the research, and Dinner Bell Meat Prducts and Packers fr prviding bvine bld. REFERENCES (1) A. N. Martin, J. Swarbrick, and A. Cammarata, Physical Pharmacy, 2nd ed., 1969, pp (2) M. M. Rieger, Surfactants and Csmetics (Marcel Dekker, New Yrk, 1985), pp (3) L. Afr, The use f quaternary ammnium cmpunds as preservatives in plivirus vaccine. Preliminary reprt, State Bacteril. Lab., Stckhlm. Acta Pathl. Micrbil, Scand., 61(1), (1964). (4) The Merck Index, 12th ed. (Merck Research Labratries, Divisin f Merck & C., Inc., Whitehuse Statin, NJ), pp , (5) I. Chen and A. Libackyj, Cacervating sap-electrlyte slutins. I. Charge prperties. II. Generalized treatment f cacervating sap-electrlyte systems. J. Cllid Sci., 19(6), (1964). (6) K. Hill, W. yn Rybinski, and G. Stll, Alkyl Plyglycsides (Weinheim, New Yrk, 1997). (7) M. M. Rieger, Surfactant Encyclpedia, 2nd ed. (Allured Publishing Crp., Carl Stream, IL, 1996). (8) J. Falbe, Surfactants in Cnsumer Prducts (Heidelberg, New Yrk, 1987). (9) Th. F/3rster, H. Hensen, R. Hffman, and B. Salka, Using alkyl plyglycsides in persnal-care prducts, Csmet. Tilerr., 110 (April 1995). (10) V. L. Sneyink and D. Jenkins, Water Chemist (Jhn Wiley & Sns, New Yrk, 1980), pp (11) J. S. Frutn and S. Simmnds, General BichemiJtry, 2nd ed. (Jhn Wiley & Sns, New Yrk, 1958). (12) Y. Marcus, In Slvatin (Jhn Wiley & Sns, New Yrk, 1985), pp (13) J. Burgess, Ins in Slutin (Jhn Wiley & Sns, New Yrk, 1988), p. 15. (14) D. Myers, Surfaces, Interfaces, and Cllids (VCH Publishers, New Yrk, 1991), pp

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