STUDIES ON PERMEABILITY OF MEMBRANES : VII. CONDUCTIVITY OF ELECTROLYTES WITHIN THE MEMBRANE.

Two methods of measuring the electrical conductivity of the dried collodion membrane in contact with an electrolyte solution are described and the results of such measurements with different electrolytes in different ranges of concentration recorded. Some of the difficulties encountered in making these measurements are outlined. Of special interest was the fact that each membrane with each electrolyte showed a maximum level of resistance at a certain point in the dilution scale, a level which was not surpassed by further dilution. It is believed that this level was fixed by the collodion itself rather than by the contiguous electrolyte solution. Its existence limited the results available for reasonable interpretation. In relatively concentrated solutions the conductivity was shown to be approximately proportional to the concentration. With different electrolytes in the same concentration it was shown that the conductivities varied much more than in simple solutions without a membrane and that they fell in the order HCl > KCl > NaCl > LiCl. A method was described whereby the electrolyte content of a membrane in contact with different chloride solutions could be determined. It was shown that a membrane saturated with either 0.5 N HCl or 0.5 N KCl had practically the same total electrolyte content whereas the same membrane in contact with 0.5 N LiCl contained only half the quantity. These results were used in interpreting the conductivity data, the evidence presented strongly suggesting that two factors are operative in causing the widely divergent conductivities recorded with different electrolytes. The first factor depended on the quantity of electrolyte which can enter the membrane pores, a quantity dependent on the size of the pores and the volume of the larger of the two hydrated ions of the electrolyte. This factor was the chief one in determining the difference in conductivity between KCl and LiCl. The second factor was concerned with differences in the mobility of the various cations within the membrane brought about by friction between the moving ions and the pore walls. With KCl and HCl the quantity of electrolytes entering the membrane was in each case the same, being determined by the size of the larger Cl^- ion. The widely different conductivity values were explained as due to the changes in the mobility of the two cations within the membrane pores.     

THE ELIMINATION OF DISCREPANCIES BETWEEN OBSERVED AND CALCULATED P.D. OF PROTEIN SOLUTIONS NEAR THE ISOELECTRIC POINT WITH THE AID OF BUFFER SOLUTIONS

1. It had been noticed in the previous experiments on the influence of the hydrogen ion concentration on the P.D. between protein solutions inside a collodion bag and aqueous solutions free from protein that the agreement between the observed values and the values calculated on the basis of Donnan''s theory was not satisfactory near the isoelectric point of the protein solution. It was suspected that this was due to the uncertainty in the measurements of the pH of the outside aqueous solution near the isoelectric point. This turned out to be correct, since it is shown in this paper that the discrepancy disappears when both the inside and outside solutions contain a buffer salt. 2. This removes the last discrepancy between the observed P.D. and the P. D. calculated on the basis of Donnan''s theory of P.D. between membrane equilibria, so that we can state that the P.D. between protein solutions inside collodion bags and outside aqueous solutions free from protein can be calculated from differences in the hydrogen ion concentration on the opposite sides of the membrane, in agreement with Donnan''s formula.     

CATAPHORETIC CHARGES OF COLLODION PARTICLES AND ANOMALOUS OSMOSIS THROUGH COLLODION MEMBRANES FREE FROM PROTEIN

1. It had been shown in previous papers that when a salt solution is separated from pure water by a collodion membrane, water diffuses through the membrane as if it were positively charged and as if it were attracted by the anion of the salt in solution and repelled by the cation with a force increasing with the valency. In this paper, measurements of the P.D. across the membrane (E) are given, showing that when an electrical effect is added to the purely osmotic effect of the salt solution in the transport of water from the side of pure water to the solution, the latter possesses a considerable negative charge which increases with increasing valency of the anion of the salt and diminishes with increasing valency of the cation. It is also shown that a similar valency effect exists in the diffusion potentials between salt solutions and pure water without the interposition of a membrane. 2. This makes it probable that the driving force for the electrical transport of water from the side of pure water into solution is primarily a diffusion potential. 3. It is shown that the hydrogen ion concentration of the solution affects the transport curves and the diffusion potentials in a similar way. 4. It is shown, however, that the diffusion potential without interposition of the membrane differs in a definite sense from the P.D. across the membrane and that therefore the P.D. across the membrane (E) is a modified diffusion potential. 5. Measurements of the P.D. between collodion particles and aqueous solutions (ε) were made by the method of cataphoresis, which prove that water in contact with collodion particles free from protein practically always assumes a positive charge (except in the presence of salts with trivalent and probably tetravalent cations of a sufficiently high concentration). 6. It is shown that an electrical transport of water from the side of water into the solution is always superposed upon the osmotic transport when the sign of charge of the solution in the potential across the membrane (E) is opposite to that of the water in the P.D. between collodion particle and water (ε); supporting the theoretical deductions made by Bartell. 7. It is shown that the product of the P.D. across the membrane (E) into the cataphoretic P.D. between collodion particles and aqueous solution (ε) accounts in general semiquantitatively for that part of the transport of water into the solution which is due to the electrical forces responsible for anomalous osmosis.     

INFLUENCE OF THE CONCENTRATION OF ELECTROLYTES ON THE ELECTRIFICATION AND THE RATE OF DIFFUSION OF WATER THROUGH COLLODION MEMBRANES

1. When a watery solution is separated from pure water by a collodion membrane, the initial rate of diffusion of water into the solution is influenced in an entirely different way by solutions of electrolytes and of non-electrolytes. Solutions of non-electrolytes, e.g. sugars, influence the initial rate of diffusion of water through the membrane approximately in direct proportion to their concentration, and this. influence begins to show itself under the conditions of our experiments when the concentration of the sugar solution is above M/64 or M/32. We call this effect of the concentration of the solute on the initial rate of diffusion of water into the solution the gas pressure effect. 2. Solutions of electrolytes show the gas pressure effect upon the initial rate of diffusion also, but it commences at a somewhat higher concentration than M/64; namely, at M/16 or more (according to the nature of the electrolyte). 3. Solutions of electrolytes of a lower concentration than M/16 or M/8 have a specific influence on the initial rate of diffusion of water through a collodion membrane from pure solvent into solution which is not found in the case of the solutions of non-electrolytes and which is due to the fact that the particles of water diffuse in this case through the membrane in an electrified condition, the sign of the charge depending upon the nature of the electrolyte in solution, according to two rules given in a preceding paper. 4. In these lower concentrations the curves representing the influence of the concentration of the electrolyte on the initial rate of diffusion of water into the solution rise at first steeply with an increase in the concentration, until a maximum is reached at a concentration of M/256 or above. A further increase in concentration causes a drop-in the curve and this drop increases with a further increase of concentration until that concentration of the solute is reached in which the gas pressure effect begins to prevail; i.e., above M/16. Within a range of concentrations between M/256 and M/16 or more (according to the nature of the electrolyte) we notice the reverse of what we should expect on the basis of van''t Hoff''s law; namely, that the attraction of a solution of an electrolyte for water diminishes with an increase in concentration. 5. We wish to make no definite assumption concerning the origin of the electrification of water and concerning the mechanism whereby ions influence the rate of diffusion of water particles through collodion membranes from pure solvent to solution. It will facilitate, however, the presentation of our results if it be permitted to present them in terms of attraction and repulsion of the charged particles of water by the ions. With this reservation we may say that in the lowest concentrations attraction of the electrified water particles by the ions with the opposite charge prevails over the repulsion of the electrified water particles by the ions with the same sign of charge as that of the water; while beyond a certain critical concentration the repelling action of the ion with the same sign of charge as that of the water particles upon the latter increases more rapidly with increasing concentration of the solute than the attractive action of the ion with the opposite charge. 6. It is shown that negative osmosis, i.e. the diminution of the volume of the solution of acids and of alkalies when separated by collodion membranes from pure water, occurs in the same range of concentrations in which the drop in the curves of neutral salts occurs, and that it is due to the same cause; namely, the repulsion of the electrified particles of water by the ion with the same sign of charge as that of the water. This conclusion is supported by the fact that negative osmosis becomes pronounced when the ion with the same sign of charge as that of the electrified particles of water carries more than one charge.     

ON THE CAUSE OF THE INFLUENCE OF IONS ON THE RATE OF DIFFUSION OF WATER THROUGH COLLODION MEMBRANES. I

1. In three previous publications it had been shown that electrolytes influence the rate of diffusion of pure water through a collodion membrane into a solution in three different ways, which can be understood on the assumption of an electrification of the water or the watery phase at the boundary of the membrane; namely, (a) While the watery phase in contact with collodion is generally positively electrified, it happens that, when the membrane has received a treatment with a protein, the presence of hydrogen ions and of simple cations with a valency of three or above (beyond a certain concentration) causes the watery phase of the double layer at the boundary of membrane and solution to be negatively charged. (b) When pure water is separated from a solution by a collodion membrane, the initial rate of diffusion of water into a solution is accelerated by the ion with the opposite sign of charge and retarded by the ion with the same sign of charge as that of the water, both effects increasing with the valency of the ion and a second constitutional quantity of the ion which is still to be defined. (c) The relative influence of the oppositely charged ions, mentioned in (b), is not the same for all concentrations of electrolytes. For lower concentrations the influence of that ion usually prevails which has the opposite sign of charge from that of the watery phase of the double layer; while in higher concentrations the influence of that ion begins to prevail which has the same sign of charge as that of the watery phase of the double layer. For a number of solutions the turning point lies at a molecular concentration of about M/256 or M/512. In concentrations of M/8 or above the influence of the electrical charges of ions mentioned in (b) or (c) seems to become less noticeable or to disappear entirely. 2. It is shown in this paper that in electrical endosmose through a collodion membrane the influence of electrolytes on the rate of transport of liquids is the same as in free osmosis. Since the influence of electrolytes on the rate of transport in electrical endosmose must be ascribed to their influence on the quantity of electrical charge on the unit area of the membrane, we must conclude that the same explanation holds for the influence of electrolytes on the rate of transport of water into a solution through a collodion membrane in the case of free osmosis. 3. We may, therefore, conclude, that when pure water is separated from a solution of an electrolyte by a collodion membrane, the rate of diffusion of water into the solution by free osmosis is accelerated by the ion with the opposite sign of charge as that of the watery phase of the double layer, because this ion increases the quantity of charge on the unit area on the solution side of the membrane; and that the rate of diffusion of water is retarded by the ion with the same sign of charge as that of the watery phase for the reason that this ion diminishes the charge on the solution side of the membrane. When, therefore, the ions of an electrolyte raise the charge on the unit area of the membrane on the solution side above that on the side of pure water, a flow of the oppositely charged liquid must occur through the interstices of the membrane from the side of the water to the side of the solution (positive osmosis). When, however, the ions of an electrolyte lower the charge on the unit area of the solution side of the membrane below that on the pure water side of the membrane, liquid will diffuse from the solution into the pure water (negative osmosis). 4. We must, furthermore, conclude that in lower concentrations of many electrolytes the density of electrification of the double layer increases with an increase in concentration, while in higher concentrations of the same electrolytes it decreases with an increase in concentration. The turning point lies for a number of electrolytes at a molecular concentration of about M/512 or M/256. This explains why in lower concentrations of electrolytes the rate of diffusion of water through a collodion membrane from pure water into solution rises at first rapidly with an increase in concentration while beyond a certain concentration (which in a number of electrolytes is M/512 or M/256) the rate of diffusion of water diminishes with a further increase in concentration.     

STUDIES ON PERMEABILITY OF MEMBRANES : I. INTRODUCTION AND THE DIFFUSION OF IONS ACROSS THE DRIED COLLODION MEMBRANE.

The theoretical aspects of the problem of sieve-like membranes are developed. The method of preparing the dried collodion membrane is described, and the method of defining the property of a particular membrane is given. It consists of the measurement of the Co P, that is the P.D. between an 0.1 and an 0.01 M KCl solution separated by the membrane. Co P is in the best dried membranes 50 to 53 millvolts, the theoretically possible maximum value being 55 millivolts. Diffusion experiments have been carried out with several arrangements, one of which is, for example, the diffusion of 0.1 M KNO₃ against 0.1 M NaCl across the membrane. The amount of K⁺ diffusing after a certain period was in membranes with a sufficiently high Co P (about 50 millivolts or more) on the average ten times as much as the amount of diffused Cl^-. In membranes with a lower Co P the ratio was much smaller, down almost to the proportion of 1:1 which holds for the mobility of these two ions in a free aqueous solution. When higher concentrations were used, e.g. 0.5 M solution, the difference of the rate of diffusion for K⁺ and Cl^- was much smaller even in the best membranes, corresponding to the fact that the P.D. of two KCl solutions whose concentrations are 10:1 is much smaller in higher ranges of concentration than in lower ones. These observations are confirmed by experiments arranged in other ways. It has been shown that, in general, the diffusion of an anion is much slower than the one of a cation across the dried collodion membrane. The ratio of the two diffusion coefficients would be expected to be calculable in connection with the potential difference of such a membrane when interposed between these solutions. The next problem is to show in how far this can be confirmed quantitatively.     

THE COLLOIDAL BEHAVIOR OF PROTEINS

1. It is well known that neutral salts depress the osmotic pressure, swelling, and viscosity of protein-acid salts. Measurements of the P.D. between gelatin chloride solutions contained in a collodion bag and an outside aqueous solution show that the salt depresses the P.D. in the same proportion as it depresses the osmotic pressure of the gelatin chloride solution. 2. Measurements of the hydrogen ion concentration inside the gelatin chloride solution and in the outside aqueous solution show that the difference in pH of the two solutions allows us to calculate the P.D. quantitatively on the basis of the Nernst formula See PDF for Equation if we assume that the P.D. is due to a difference in the hydrogen ion concentration on the two sides of the membrane. 3. This difference in pH inside minus pH outside solution seems to be the consequence of the Donnan membrane equilibrium, which only supposes that one of the ions in solution cannot diffuse through the membrane. It is immaterial for this equilibrium whether the non-diffusible ion is a crystalloid or a colloid. 4. When acid is added to isoelectric gelatin the osmotic pressure rises at first with increasing hydrogen ion concentration, reaches a maximum at pH 3.5, and then falls again with further fall of the pH. It is shown that the P.D. of the gelatin chloride solution shows the same variation with the pH (except that it reaches its maximum at pH of about 3.9) and that the P.D. can be calculated from the difference of pH inside minus pH outside on the basis of Nernst''s formula. 5. It was found in preceding papers that the osmotic pressure of gelatin sulfate solutions is only about one-half of that of gelatin chloride or gelatin phosphate solutions of the same pH and the same concentration of originally isoelectric gelatin; and that the osmotic pressure of gelatin oxalate solutions is almost but not quite the same as that of the gelatin chloride solutions of the same pH and concentration of originally isoelectric gelatin. It was found that the curves for the values for P.D. of these four gelatin salts are parallel to the curves of their osmotic pressure and that the values for pH inside minus pH outside multiplied by 58 give approximately the millivolts of these P.D. In this preliminary note only the influence of the concentration of the hydrogen ions on the P.D. has been taken into consideration. In the fuller paper, which is to follow, the possible influence of the concentration of the anions on this quantity will have to be discussed.     

DONNAN EQUILIBRIUM AND THE PHYSICAL PROPERTIES OF PROTEINS : I. MEMBRANE POTENTIALS.

1. It is shown that a neutral salt depresses the potential difference which exists at the point of equilibrium between a gelatin chloride solution contained in a collodion bag and an outside aqueous solution (without gelatin). The depressing effect of a neutral salt on the P.D. is similar to the depression of the osmotic pressure of the gelatin chloride solution by the same salt. 2. It is shown that this depression of the P.D. by the salt can be calculated with a fair degree of accuracy on the basis of Nernst''s logarithmic formula on the assumption that the P.D. which exists at the point of equilibrium is due to the difference of the hydrogen ion concentration on the opposite sides of the membrane. 3. Since this difference of hydrogen ion concentration on both sides of the membrane is due to Donnan''s membrane equilibrium this latter equilibrium must be the cause of the P.D. 4. A definite P.D. exists also between a solid block of gelatin chloride and the surrounding aqueous solution at the point of equilibrium and this P.D. is depressed in a similar way as the swelling of the gelatin chloride by the addition of neutral salts. It is shown that the P.D. can be calculated from the difference in the hydrogen ion concentration inside and outside the block of gelatin at equilibrium. 5. The influence of the hydrogen ion concentration on the P.D. of a gelatin chloride solution is similar to that of the hydrogen ion concentration on the osmotic pressure, swelling, and viscosity of gelatin solutions, and the same is true for the influence of the valency of the anion with which the gelatin is in combination. It is shown that in all these cases the P.D. which exists at equilibrium can be calculated with a fair degree of accuracy from the difference of the pH inside and outside the gelatin solution on the basis of Nernst''s logarithmic formula by assuming that the difference in the concentration of hydrogen ions on both sides of the membrane determines the P.D. 6. The P.D. which exists at the boundary of a gelatin chloride solution and water at the point of equilibrium can also be calculated with a fair degree of accuracy by Nernst''s logarithmic formula from the value pCl outside minus pCl inside. This proves that the equation x² = y ( y + z) is the correct expression for the Donnan membrane equilibrium when solutions of protein-acid salts with monovalent anion are separated by a collodion membrane from water. In this equation x is the concentration of the H ion (and the monovalent anion) in the water, y the concentration of the H ion and the monovalent anion of the free acid in the gelatin solution, and z the concentration of the anion in combination with the protein. 7. The similarity between the variation of P.D. and the variation of the osmotic pressure, swelling, and viscosity of gelatin, and the fact that the Donnan equilibrium determines the variation in P.D. raise the question whether or not the variations of the osmotic pressure, swelling, and viscosity are also determined by the Donnan equilibrium.     

THE INFLUENCE OF ELECTROLYTES ON THE CATAPHORETIC CHARGE OF COLLOIDAL PARTICLES AND THE STABILITY OF THEIR SUSPENSIONS : I. EXPERIMENTS WITH COLLODION PARTICLES.

1. When collodion particles suspended in water move in an electric field they are, as a rule, negatively charged. The maximal cataphoretic P.D. between collodion particles and water is about 70 millivolts. This is only slightly more than the cataphoretic P.D. found by McTaggart to exist between gas bubbles and water (55 millivolts). Since in the latter case the P.D. is entirely due to forces inherent in the water itself, resulting possibly in an excess of OH ions in the layer of water in contact and moving with the gas bubble, it is assumed that the negative charge of the collodion particles is also chiefly due to the same cause; the collodion particles being apparently only responsible for the slight difference in maximal P.D. of water-gas and water-collodion surfaces. 2. The cataphoretic charge of collodion particles seems to be a minimum in pure water, increasing as a rule with the addition of electrolytes, especially if the cation of the electrolyte is monovalent, until a maximal P.D. is reached. A further increase in the concentration of the electrolyte depresses the P.D. again. There is little difference in the action of HCl, NaOH, and NaCl or LiCl or KCl. 3. The increase in P.D. between collodion particles and water upon the addition of electrolyte is the more rapid the higher the valency of the anion. This suggests that this increase of negative charge of the collodion particle is due to the anions of the electrolyte gathering in excess in the layer of water nearest to the collodion particles, while the adjoining aqueous layer has an excess of cations. 4. In the case of chlorides and at a pH of about 5.0 the maximal P.D. between collodion particles and water is about 70 millivolts, when the cation of the electrolyte present is monovalent (H, Li, Na, K); when the cation of the electrolyte is bivalent (Mg, Ca), the maximal P.D. is about 35 to 40 millivolts; and when the cation is trivalent (La) the maximal P.D. is lower, probably little more than 20 millivolts. 5. A reversal in the sign of charge of the collodion particles could be brought about by LaCl₃ but not by acid. 6. These results on the influence of electrolytes on the cataphoretic P.D. between collodion particles and water are also of significance for the theory of electrical endosmose and anomalous osmosis through collodion membranes; since the cataphoretic P.D. is probably identical with the P.D. between water and collodion inside the pores of a collodion membrane through which the water diffuses. 7. The cataphoretic P.D. between collodion particles and water determines the stability of suspensions of collodion particles in water, since rapid precipitation occurs when this P.D. falls below a critical value of about 16 millivolts, regardless of the nature of the electrolyte by which the P.D. is depressed. No peptization effect of plurivalent anions was noticed.     

CONTRIBUTION TO THE THEORY OF PERMEABILITY OF MEMBRANES FOR ELECTROLYTES

On page 39, Vol. viii, No. 2, September 18, 1925, multiply the right-hand side of formula (2) by the factor See PDF for Equation. On page 44, immediately after formula (1) the text should be continued as follows: Let us suppose a membrane to be separated by two solutions of KCl of different concentrations K₁ and K₂ and these concentrations and the corresponding concentrations of K⁺ within the membrane, which are in equilibrium with the outside solutions, to be so high that the H⁺ ions may be neglected. When a small electric current flows across the system, practically the K⁺ ions alone are transferred and that in a reversible manner. Therefore the total P.D. is practically See PDF for Equation This P.D. is composed of two P.D.''s at the boundaries and the diffusion potential within the membrane. Suppose the immobility of the anions is not absolute but only relative as compared with the mobility of the cations, KCl would gradually penetrate into the membrane to equal concentration with the outside solution on either side and no boundary potential would be established. In this case the diffusion P.D. within the membrane is the only P.D., amounting to See PDF for Equation but, V being practically = 0, it would result that See PDF for Equation So the definitive result is the same as in the former case. Now cancel the printed text as far as page 48, line 13 from the top of the page, but retain Fig. 1. On page 50, line 19 from the top of the page, cancel the sentence beginning with the word But and ending with the words of the chain.     

STUDIES ON PERMEABILITY OF MEMBRANES : VIII. THE BEHAVIOR OF THE DRIED COLLODION MEMBRANE TOWARD BIVALENT CATIONS.

A study of the behavior of the dried collodion membrane toward the bivalent calcium ion showed that: 1. There is almost no potential difference established across a membrane separating two calcium chloride solutions of 0.1 and 0.01 N concentrations. 2. The transfer numbers of chlorine and calcium, as measured in electrical transfer experiments, are both close to 0.5. 3. A sample of membrane in equilibrium with a solution of calcium chloride has an extremely high electrical resistance, greater than is observed with solutions of the chlorides of any of the monovalent cations. 4. The total electrolyte content of a membrane in equilibrium with a solution of calcium chloride was only 20 per cent of that observed when the solution was lithium chloride and 10 per cent of that found when the solution was potassium chloride. In explaining these various results it is supposed that (1), (2) and (3) are all the result of (4), that is, of the inability of the calcium ion to penetrate any but the largest of the membrane pores. As the total quantity of electrolyte able to penetrate the membrane is very small the electrical conductivity must also be very small. Moreover, the few larger pores that are large enough to transport the hydrated calcium ion are too large to exert any appreciable effect in decreasing the mobility of the anion. Thus the membrane has no effect in modifying the potentials established across concentration chains with CaCl₂ and the transfer numbers determined experimentally are what one would expect if no membrane were present.     

PROTOPLASMIC POTENTIALS IN HALICYSTIS : III. THE EFFECTS OF AMMONIA

The nature and origin of the large "protoplasmic" potential in Halicystis must be studied by altering conditions, not only in external solutions, but in the sap and the protoplasm itself. Such interior alteration caused by the penetration of ammonia is described. Concentrations of NH₄Cl in the sea water were varied from 0.00001 M to above 0.01 M. At pH 8.1 there is little effect below 0.0005 M NH₄Cl. At about 0.001 M a sudden reversal of the potential difference across the protoplasm occurs, from about 68 mv. outside positive to 30 to 40 mv. outside negative. At this threshold value the time curve is characteristically S-shaped, with a slow beginning, a rapid reversal, and then an irregularly wavering negative value. There are characteristic cusps at the first application of the NH₄Cl, also immediately after the reversal. The application of higher NH₄Cl concentrations causes a more rapid reversal, and also a somewhat higher negative value. Conversely the reduction of NH₄Cl concentrations causes recovery of the normal positive potential, but the threshold for recovery is at a lower concentration than for the original reversal. A temporary overshooting or increase of the positive potential usually occurs on recovery. The reversals may be repeated many times on the same cell without injury. The plot of P.D. against the log of ammonium ion concentration is not the straight line characteristic of ionic concentration effects, but has a break of 100 mv. or more at the threshold value. Further evidence that the potential is not greatly influenced by ammonium ions is obtained by altering the pH of the sea water. At pH 5, no reversal occurs with 0.1 M NH₄Cl, while at pH 10.3, the NH₄Cl threshold is 0.0001 M or less. This indicates that the reversal is due to undissociated ammonia. The penetration of NH₃ into the cells increases both the internal ammonia and the pH. The actual concentration of ammonium salt in the sap is again shown to have little effect on the _P.D. The pH is therefore the governing factor. But assuming that NH₃ enters the cells until it is in equilibrium between sap and sea water, no sudden break of pH should occur, pH being instead directly proportional to log NH₃ for any constant (NH₄) concentration. Experimentally, a linear relation is found between the pH of the sap and the log NH₃ in sea water. The sudden change of P.D. must therefore be ascribed to some system in the cell upon which the pH change operates. The pH value of the sap at the NH₃ threshold is between 6.0 and 6.5 which corresponds well with the pH value found to cause reversal of P.D. by direct perfusion of solutions in the vacuole.     

CHEMICAL ANTAGONISM OF IONS : III. EFFECT OF SALT MIXTURES ON GELATIN ACTIVITY.

1.25 per cent gelatin solutions containing enough NaOH to bring them to pH 7.367 (or KOH to pH 7.203) were made up with various concentrations of NaCl, KCl and MgCl₂, alone and in mixtures, up to molar ionic strength. The effects of these salts on the pH were observed. MgCl₂ and NaCl alone lower the pH of the Na gelatinate or the K gelatinate, in all amounts of these salts. KCl first lowers the pH (up to 0.01 M K⁺), then raises the pH. Mixtures of NaCl and KCl (up to 0.09 M of the salt whose concentration is varied) raise the pH; then (up to 0.125 M Na⁺ or K⁺) lower the pH; and finally (above 0.125 M) behave like KCl alone. Mixtures of MgCl₂ and NaCl raise the pH up to 0.10 M Na⁺, and lower it up to 0.15 M Na⁺ regardless of the amount of MgCl ₂. Higher concentrations of NaCl have little effect, but the pH in this range of NaCl concentration is lowered with increase of MgCl₂. Mixtures of MgCl₂ and KCl behave as above described (for MgCl₂ and NaCl) and the addition of NaCl plus KCl to gelatin containing MgCl₂ produces essentially the same effect as the addition of either alone, except that the first two breaks in this curve come at 0.07 M and 0.08 M [Na⁺ + K⁺] and there is a third break at 0.12 M. In this pH range the free groups of the dicarboxylic acids and of lysine are essentially all ionized and the prearginine and histidine groups are essentially all non-ionized. The arginine group is about 84 per cent ionized. Hence we are studying a solution with two ionic species in equilibrium, one with the arginine group ionized, and one with it non-ionized. It is shown that the effect of each salt alone depends upon the effect of the cation on the activity of these two species due to combination. The anomalous effects of cation mixtures may be qualitatively accounted for if one or both of these species fail to combine with the cations in a mixture in proportion to the relative combination in solutions of each cation alone. Special precautions were taken to ensure accuracy in the pH measurements. The mother solutions gave identical readings to 0.001 pH and the readings with salts were discarded when not reproducible to 0.003 pH. All doubtful data were discarded.     

THE COMBINATION OF SALTS AND PROTEINS. I

1. It has been found that the ratios of the total concentrations of Ca, Mg, K, Zn, inside and outside of gelatin particles do not agree with the ratios calculated according to Donnan''s theory from the hydrogen ion activity ratios. 2. E.M.F. measurements of Zn and Cl electrode potentials in such a system show, however, that the ion activity ratios are correct, so that the discrepancy must be due to a decrease in the ion concentration by the formation of complex ions with the protein. 3. This has been confirmed in the case of Zn by Zn potential measurements in ZnCl₂ solutions containing gelatin. It has been found that in 10 per cent gelatin containing 0.01 M ZnCl₂ about 60 per cent of the Zn⁺⁺ is combined with the gelatin. 4. If the activity ratios are correctly expressed by Donnan''s equation, then the amount of any ion combined with a protein can be determined without E.M.F. measurements by determining its distribution in a proper system. If the activity ratio of the hydrogen ion and the activity of the other ion in the aqueous solution are known, then the activity and hence the concentration of the ion in the protein solution can be calculated. The difference between this and the total molar concentration of the ion in the protein represents the amount combined with the protein. 5. It has been shown that in the case of Zn the values obtained in this way agree quite closely with those determined by direct E.M.F. measurements. 6. The combination with Zn is rapidly and completely reversible and hence is probably not a surface effect. 7. Since the protein combines more with Zn than with Cl, the addition of ZnCl₂ to isoelectric gelatin should give rise to an unequal ion distribution and hence to an increase in swelling, osmotic pressure, and viscosity. This has been found to be the case.     

STUDIES ON THE PERMEABILITY OF MEMBRANES : III. ELECTRIC TRANSFER EXPERIMENTS WITH THE DRIED COLLODION MEMBRANE.

The transfer numbers of the ions of electrolytes in the dried collodion membrane, as determined in a previous paper indirectly from the E.M.F. of concentration chains, can also be determined directly by electrical transfer experiments. It is shown that the difficulties involved in such experiments can be overcome. The transfer numbers obtained by the two methods are in satisfactory agreement. The experimental results obtained in the transfer experiments furnish an additional argument in favor of maintaining the theory that the electromotive effects observed in varying concentrations of different electrolytes with the dried collodion membrane may be referred to differences in the mobilities of the anions and cations within the membrane. As was shown by the method of the previous paper, the transfer number depends largely on concentration. There are some minor discrepancies between the values of the transfer numbers obtained by the two methods which, as yet, cannot be completely explained.     

THE INFLUENCE OF THE CHEMICAL NATURE OF SOLID PARTICLES ON THEIR CATAPHORETIC P.D. IN AQUEOUS SOLUTIONS

1. The effect of eight salts, NaCl, Na₂SO₄, Na₄Fe(CN)₆, CaCl₂, LaCl₃, ThCl₄, and basic and acid fuchsin on the cataphoretic P.D. between solid particles and aqueous solutions was measured near the point of neutrality of water (pH 5.8). It was found that without the addition of electrolyte the cataphoretic P.D. between particles and water is very minute near the point of neutrality (pH 5.8), often less than 10 millivolts, if care is taken that the solutions are free from impurities. Particles which in the absence of salts have a positive charge in water near the point of neutrality (pH 5.8) are termed positive colloids and particles which have a negative charge under these conditions are termed negative colloids. 2. If care is taken that the addition of the salt does not change the hydrogen ion concentration of the solution (which in these experiments was generally pH 5.8) it can be said in general, that as long as the concentration of salts is not too high, the anions of the salt have the tendency to make the particles more negative (or less positive) and that cations have the opposite effect; and that both effects increase with the increasing valency of the ions. As soon as a maximal P.D. is reached, which varies for each salt and for each type of particles, a further addition of salt depresses the P.D. again. Aside from this general tendency the effects of salts on the P.D. are typically different for positive and negative colloids. 3. Negative colloids (collodion, mastic, Acheson''s graphite, gold, and metal proteinates) are rendered more negative by low concentrations of salts with monovalent cation (e.g. Na) the higher the valency of the anion, though the difference in the maximal P.D. is slight for the monovalent Cl and the tetravalent Fe(CN)₆ ions. Low concentrations of CaCl₂ also make negative colloids more negative but the maximal P.D. is less than for NaCl; even LaCl₃ increases the P.D. of negative particles slightly in low concentrations. ThCl₄ and basic fuchsin, however, seem to make the negative particles positive even in very low concentrations. 4. Positive colloids (ferric hydroxide, calcium oxalate, casein chloride—the latter at pH 4.0) are practically not affected by NaCl, are rendered slightly negative by high concentrations of Na₂SO₄, and are rendered more negative by Na₄Fe(CN)₆ and acid dyes. Low concentrations of CaCl₂ and LaCl₃ increase the positive charge of the particles until a maximum is reached after which the addition of more salt depresses the P.D. again. 5. It is shown that alkalies (NaOH) act on the cataphoretic P.D. of both negative and positive particles as Na₄Fe(CN)₆ does at the point of neutrality. 6. Low concentrations of HCl raise the cataphoretic P.D. of particles of collodion, mastic, graphite, and gold until a maximum is reached, after which the P.D. is depressed by a further increase in the concentration of the acid. No reversal in the sign of charge of the particle occurs in the case of collodion, while if a reversal occurs in the case of mastic, gold, and graphite, the P.D. is never more than a few millivolts. When HCl changes the chemical nature of the colloid, e.g. when HCl is added to particles of amphoteric electrolytes like sodium gelatinate, a marked reversal will occur, on account of the transformation of the metal proteinate into a protein-acid salt. 7. A real reversal in the sign of charge of positive particles occurs, however, at neutrality if Na₄Fe(CN)₆ or an acid dye is added; and in the case of negative colloids when low concentrations of basic dyes or minute traces of ThCl₄ are added. 8. Flocculation of the suspensions by salts occurs when the cataphoretic P.D. reaches a critical value which is about 14 millivolts for particles of graphite, gold, or mastic or denatured egg albumin; while for collodion particles it was about 16 millivolts. A critical P.D. of about 15 millivolts was also observed by Northrop and De Kruif for the flocculation of certain bacteria.     

THE STRUCTURE OF THE COLLODION MEMBRANE AND ITS ELECTRICAL BEHAVIOR : XI. THE PREPARATION AND PROPERTIES OF "MEGAPERMSELECTIVE" COLLODION MEMBRANES COMBINING EXTREME IONIC SELECTIVITY WITH HIGH PERMEABILITY

1. The electronegative membranes described in the literature which show a high degree of ionic selectivity (permitting cations to pass and restricting the anions) have serious shortcomings: their absolute permeability is extremely low, much too small for convenient experimentation; their ionic selectivity in most cases is not as perfect as would be desirable, and is moreover adversely affected by prolonged contact with electrolyte solutions. 2. A method has been worked out to prepare membranes substantially free from these defects. Porous collodion membranes were cast on the outside of rotating tubes and then oxidized with 1 M NaOH. By allowing the oxidized porous membranes to dry in air on the tubes membranes of desirable properties are obtained. These membranes are smooth, have a well defined shape, and allow considerable handling without breaking. 3. This new type membrane when tested for ionic selectivity by the measurement of the "characteristic concentration potential," consistently gives potentials of 54 to 55 mv., the maximum thermodynamically possible value (at 25°C.) being 55.1 mv. This high degree of ionic selectivity is not lost on prolonged contact with water, and is only very slowly affected by electrolyte solutions. 4. The absolute permeability of the new type membranes can be varied over a very wide range by changing the time of oxidation. Under optimum conditions membranes can be obtained with a resistance in 0.1 N KCl solution of only 0.5 ohms per 50 cm.² membrane area. The absolute rate of cation exchange through these membranes between solutions of different uni-univalent electrolytes is very high, in one case, e.g. 0.9 m.eq. cations per 4 hours, the anion leak being 0.02 m.eq. Thus, the absolute permeability of the new type membranes is two to four orders of magnitude greater than the permeability of the dried collodion membranes and the oxidized ("activated") dried collodion membranes used heretofore. Because of the characteristic properties of the new type membranes the term "megapermselective" (or "permselective") collodion membranes is proposed for them.     

THE ACTIVATION OF STARFISH EGGS BY ACIDS : II. THE ACTION OF SUBSTITUTED BENZOIC ACIDS AND OF BENZOIC AND SALICYLIC ACIDS AS INFLUENCED BY THEIR SALTS.

1. Comparison of the rates of activation of unfertilized starfish eggs in pure solutions of a variety of parthenogenetically effective organic acids (fatty acids, carbonic acid, benzoic and salicylic acids, chloro- and nitrobenzoic acids) shows that solutions which activate the eggs at the same rate, although widely different in molecular concentration, tend to be closely similar in C_H. The dissociation constants of these acids range from 3.2 x 10^–7 to 1.32 x 10^–3. 2. In the case of each of the fourteen acids showing parthenogenetic action the rate of activation (within the favorable range of concentration) proved nearly proportional to the concentration of acid. The estimated C_H of solutions exhibiting an optimum action with exposures of 10 minutes (at 20°) lay typically between 1.1 x 10^–4 M and 2.1 x 10^–4 M (pH = 3.7–3.96), and in most cases between 1.6 x 10^–4 M and 2.1 x 10^–4 M (pH = 3.7–3.8). Formic acid (C_H = 4.2 x 10^–4 M) and o-chlorobenzoic acid (C_H = 3.5 x 10^–4 M) are exceptions; o-nitrobenzoic acid is ineffective, apparently because of slow penetration. 3. Activation is not dependent on the penetration of H ions into the egg from without, as is shown by the effects following the addition of its Na salt to the solution of the activating acid (acetic, benzoic, salicylic). The rate of activation is increased by such addition, to a degree indicating that the parthenogenetically effective component of the external solution is the undissociated free acid. Apparently the undissociated molecules alone penetrate the egg freely. It is assumed that, having penetrated, they dissociate in the interior of the egg, furnishing there the H ions which effect activation. 4. Attention is drawn to certain parallels between the physiological conditions controlling activation in the starfish egg and in the vertebrate respiratory center.     

CHANGES OF APPARENT IONIC MOBILITIES IN PROTOPLASM : I. EFFECTS OF GUAIACOL ON VALONIA

In normal cells of Valonia the order of the apparent mobilities of the ions in the non-aqueous protoplasmic surface is K > Cl > Na. After treatment with 0.01 M guaiacol (which does not injure the cell) the order becomes Na > Cl > K. As it does not seem probable that such a reversal could occur with simple ions we may assume provisionally that in the protoplasmic surface we have to do with charged complexes of the type (KX _I)⁺, (KX _II)⁺, where X _I and X _II are elements or radicals, or with chemical compounds formed in the protoplasm. When 0.01 M guaiacol is added to sea water or to 0.6 M NaCl (both at pH 6.4, where the concentration of the guaiacol ion is negligible) the P.D. of the cell changes (after a short latent period) from about 10 mv. negative to about 28 mv. positive and then slowly returns approximately to its original value (Fig. 1, p. 14). This appears to depend chiefly on changes in the apparent mobilities of organic ions in the protoplasm. The protoplasmic surface is capable of so much change that it does not seem probable that it is a monomolecular layer. It does not behave like a collodion nor a protein film since the apparent mobility of Na⁺ can increase while that of K⁺ is decreasing under the influence of guaiacol.     

THE CHEMISTRY OF HUMAN SKIN : IV. THE ELECTROKINETIC EFFECT OF VARIOUS IONS UPON SUSPENDED PARTICLES OF STRATUM CORNEUM

1. The electrophoretic activity of particles of human skin in distilled water and different concentrations of salt solutions has been studied. The electrokinetic potential and the charge density were determined and comparisons made with results obtained by electroendosmosis. 2. The electrokinetic potential is ultimately decreased if sufficient salt is added. The order of inhibition is Al > Ca > Ba > K > Na. 3. The lyotrophic series Li > Na > K > Rb and Cl > I > Br express respectively the comparative effect of the monovalent cations and anions upon the electrokinetic potential of the skin. 4. Since both electrophoresis and electroendosmosis are dependent upon the electrokinetic potential, it follows from the results obtained, that the greatest rate of flow through the human skin under the force of an applied electrical current would be given by concentrations of neutral salts between 0.0001 M and 0.0002 M.