ELECTROLYTE SOLUTIONS CONTAINING POLYANIONIC MATERIALS

Abstract: The use of non-diffusible organic polyionic materials in aqueous electrolyte solutions wherein the electrolyte types and concentrations are suitable for use in treating living animal cells (including mammals and man). In these solutions, the Na:Cl ratio is normalizable without the use of unphysiological or demonstrably pathological amounts or ratios of small diffusible anions. Effective plasma expanders and plasma substitutes are provided. Near equilibrium couples may be incorporated if desired.

Description: ELECTROLYTE SOLUTIONS CONTAINING POLYANIONIC MATERIALS DESCRIPTION This invention lies in the field of electrolyte fluids and processes for their preparation and use. fluids designed to contact mannalian cells all have as a general characteristic an osmotic pressure above about 260 millios oles/liter (mOs/L) . The most common fluid given in 5% dextrose. in H 2 0. The second most common fluid given is normal saline (0.9N or 0.095% NaCl) . It has been known (Black DAK, Lancet L:305-312,1954) that giving to adult humans much over 500 ml of normal saline per day leads to hyperchloremic acidosis since normal plasma contains about 136-145 mEq/L (milliequivalents per liter) plasma Na and about 100-106 mEq/L plasma of Cl for an average Na:Cl milliequivalent ratio of about 1:36. There has been a long standing interest in creating an artificial plasma which dates back to 1883 with the origin of S. Ringer's solutions (J Physiol : 29-42,1883) which are still in use today. It is now recognized that plasma is an "unmakable" solution since the law of electrical neutrality requires that the number of positive ions (cations) equal the number of negative ions (anions) . Plasma itself has an exceedingly complex composition. As any practicing clinician knows, since plasma contains 25-28 mEq/L of HC0^~, the number of measurable + ( ) cations in plasma is greater than the number of measurable ( ) anions (mainly Cl~, HCO,~, and small amounts of Pi —1 * 8) in plasma by about 10-17 mEq/L plasma. The difference between cations is called ' the "anion gap". Efforts to cure the anion gap have been attempted since about the time of Ringer. A similar gap exists in each of extracellular and intracellular fluids. The anio gap is now known ' to be caused mainly by the presence of polyanionic proteins, especially albumin which in man and mammals generally is a protein of about 68^000 M. . (molecular weight) and which has about 20 negative (anion) charges/mole at the physiological pH of blood which is about 7.35 to 7.45 (see Tanford C.J Am Che Soc 42; 441-451, 1950) . Since the normal albumin concentration in, for example, mammalian blood is about 0.65mM/L (millimoles per liter), about 13 mEq/L of the plasma anion gap is due to this source. Although albumin is found in all mammaliam plasma, its chemical structure differs from species to species. If albumin from an animal such as the cow is intravenously intro¬ duced into man, an allergenic response promptly results. Therefore, only albumin specific to a species may be used repeatedly in a therapeutic situation. Although (suitably purified) albumin from one human can be so introduced into another without an allergenic response, human albumin is costly, a potential source of infection with viral agents, such as hepatitis or AIDS, and is difficult to obtain in quantity at the present time, as for therapeutic purposes. Thus, electrolyte solutions for therapeutic use which do not require use of a material such as albumin are still useful. The history of electrolyte solutions including dialysis media can be brefly related: 1) Sodium chloride. The earliest solutions used in medical therapy (Latta T.Lancet i_: 274-277, 1832) contained sodium cations (Na ) and chloride anions (Cl ) . Today, normal saline, which is isomolar NaCl (0.9-0.95%) is still given to patients intravenously. The problems with such solution are that it does not regulate pH and it induces hyperchloremic acidosis when given at much over 1 L/70kg man/day. 2) Ringer's. The second attempt to make a fluid which was not immediately lethal for contacting human cells was designed by S. Ringer in the 1880' s and is still in use today. This fluid composition is essentially 130-145 mEq/L Na +" , 2-4 mEq/L K , 3.00 mEq/L Ca 2+ , 100-134 - -1 8 mEq/L Cl , 7-14 mEq/L i , and optionally up to 30-45 mEq/dl-lactate or acetate, when a normal Na:Cl ratio is to be obtained. Ringer's lactate is now known to cause profound difficulties with the cellular redox state (see equation 4) and has poor buffering capacity. This led in the 1920's and 30's to development of a series of physiologi- cally compatible fluids designed by the great names of modern biology: Warburg, Locke, and Tyrode, and culminated in 1932 with the development of Krebs-Henseleit solution. 3) Krebs-Henseleit. (Krebs HA, Henseleit KA. Hoppe-Seyler's Z_ Physiol Chem 210 : 33-66, 1932) Krebs- Henseleit solution has a composition as given in Table 2 herein, the essential advance being that it normalizes the HC0 3 "* /C0 2 ratio, thus achieving adequate pH control. The value of this fluid is attested to by the fact that from it, or its partner, the Krebs Phosphate-Ringers (Krebs HA.Hoppe-Seyler's Z_ Physiol Chem 217 : 193 1933) , have evolved all the first modern renal dialysis fluids, and many special fluids for tissue perfusion or incubation. The problem with Krebs-Henseleit solution, aside from too high Ca 7 (See Burritt MF, Pierides AM, Offord KP Mayo Clm Proc 55; 606-613, 1980) and Mg 2 by factors of about 2, and S04 2 by even more, is that the remaining anion gap was made up by Krebs with Cl " . Thus again, normal Cl " is 100-106 mEq/L in plasma, but Krebs-Henseleit contains 127.8 mEq/L. Krebs realized this deficiency and attempted (Krebs HA. Biochem Biophys Acta 4_: 249-269.1950) to remedy this problem with the cration of Krebs Serum substitute (see Table 2) . Because he failed to understand from a theoretical point of view how such a problem could be solved, he picked anions.on the basis of 0 2 consumption measurement in tissue slices. The anions picked to make up the 13 mEq/L anion gap were glutamate -, fumarate2-, and pyruvate , which are inappropriate in living cells (although not in tissue slices where cut surfaces of cells are exposed) because glutamate and fumarate cannot readily penetrate the cell membrane. .1 This was really how matters stood until the development of widesprad renal dialysis in the 1960's. The pioneering of these life saving techniques largely by Scribner in Seattle, Scribner in Washington, and Merrill 5 and his group at Harvard led to the need for a cheap, convenient fluid. Krebs-Henseleit solution (with only very slight variations, see Table 2) was used by the Harvard group in open baths where the necessary C0 2 was lost to the atmosphere with a resultant rise in pH (see 10 equation 1) and the conversion of HCO, to CO, 2 salts. This simple problem was inconvenient to the physicians in charge and led them, well-meaning but misguidedly, to seek a more convenient substitute for Krebs-Henseleit solution. 4) Gilman-Mudge-Scribner and the Substitution 15 of Acetate for HCO-, in High-Volume Fluids. Alfred Gilman of Columbia University was, in effect, the dean of American pharmacologists, and he and his students in the middle 1940*s reasoned that acetate is ultimately metabolized to C0 2 , and since it readily penetrated cell 20 walls, it could be used as an alternative source of HCO, " (Mudge GH, Manning JA, Gilman A. Proc Soc Exptl Biol Med" 71 :136-138,1949) . While all of this is true, it ignores the profound upsets in mineral and energy metabolism which, at the time, no one recognized, but which are now 25 clear and which have led to the present invention, since it is now absolutely clear that acetate containing fluids causes profound toxicity which can easily be overcome and therefore can ' no longer be tolerated in iew of the new art described here. 3.0 About 80% of all current hemodialysis fluids in the U.K. use 35mM/l acetate " in combination with 130-150 mM Na + , 1-1.75/mmole Ca 2+ , 0-lmM/L Mg 2+ , and 100/mM/L Cl " (See Parsons FM, Stewart WK In: Drukker W, Parsons FM, Maher JF,. eds. Replacement of Renal Function by dialysis. 35 2nd Edition, 1984, Martinus Nijhoff: Hingham, pp 148-170). -5- Minor alterations in commercial fluids involve the use ofd,l-lactate (35-50mM/L0 in place of acetate, but this alternative from commercial sources is almost as un¬ satisfactory as acetate and is submitted to be no longer tolerable for patient care. Prior art illustrative of electrolyte solutions are provided in Tables I, II, and III herewith. Prior art electrolyte solutions which incorporated albumin are illustrated in Table III herewith. In the prior art solutions, even those containing albumin, the soldium cation to chloride anion milliequivalent ratio was never normalized or made to fall in a range associated with normal animal cells in a manner which would not induce measureable taxic effects in the cells so contacted (or with the particular animal fluid which was to be mimicked by a particular fluid, for example, human blood plasma compared to Kresbs-Henseleit solution) . So far as now known, the only organic polyanionic substance heretofore employed in aqueous electrolyte solutions has been albumin, "hemocel" or gelatin and no such electrolyte solution is believed to have either a Nai'Cl milliequivalent ratio in the physiologically normal range or an electrolyte composition comparable to normal mammalian (or human) blood plasma. Furthermore, so far as is now known, no albumin containing such electrolyte solution has ever previously been employed in in vivo mammalian therapy (e.g., parenterally, intravenously, or otherwise administered) . In even the field of plasma expanders (which is regarded as an application for aqueous electrolyte solutions suitable for contacting living animal cells) , it has heretofore been thought by those skilled in the art (see, for example, Mudge on "Agents Affecting Volume and Composition of Body Fluids", pp. 848-884 of Goodman & Gilman*s The Pharacologic Basis of Therapeutics published in 1980 by Mac illan, New York, that an ideal plasma expander should be, among other properties, pharmacologically inert. While many substances have been invesl igated as plasma expanders, organic polyanionic Table I Prior Art Fluids to Which Nacroaolecules Have Been Added (1) (2) (3) (4) (5) (6) Units Norul 5≠ Normal Ringer's Lactated Acetated Range Plasma Dextrose Saline Injectable Ringer's Ringer's of ■moles N.E.J.N. in 0.9. Peritoneal 283, 1285 Uater Dialysis L fluid 1970 (Cowercial) Na 136 - 145 154 147 130 140 131-141.5 K 3.5 - 5.0 4 4 10 0-4 Ca 2.1 -2.6 2.5 1.5 2.5 1.75-2.0 free £Ca2+] £ 1.06 ) Ng 0.75 - 1.25 2.5 1.5 1.5 0.25-0.75 free ttg2+] £ 0.53 3 " Σ £q Cations 142.7-153.2 154 156 137 158 135-151 l 100- 106 154 156 109 103 96-106 HC03 26-28 EPi ' 1 - 1.45 ∞4 0.32 - 0.94 L - lactate 0.6 - 1.8 28(d, l) β(d, l) 3S-45(d, l) pyruvate Lact/pyr D-fl-OHbutyrate •cetoacetate fl-HB/ acac acetate 47 45 Other ΣrnEq anions 128.7-139.4. 154 156 137 158 135-151 Na/Cl 1.28 - 1.45 1.00 0.94 1.19 1.36 1.36-1.33 Glucose 3.9 - 5.6 278 83-236 or others ∞2 0.99 - 1.39 pH 7.35-7.45 55.5-6.5 S5.5-6.5 S6.0-6.5 =6.0-*.5 S6.0-6.5 S6.0-6.5 I mOs 285 - 295 278 310 309 272 312 347-535 Use: Hydration Nal Multiple IV Fluid Electro- Peritoneal t Nutrition Replace- Blood lyte Dialysis ent Products Replace- Administration ment 1) Facts and Comparisons Lippincott, St Louis, 1981 2) Facts and Comparisons Lippincott, St Louis, 1981 3> Facts and Comparisons pp 35d-53, Oct '81- Aug '83, JB Lippincott, St Louis. Ringer S. I Physio 4: 29-42 1883. 4) Facts and Comparispns pp 35d S3, Oct '81- Aug '83, JB Lippincott, St Louis. HartmannAF. M&dffcsoc 03$.1349-1354.1934. 5) Facts and Comparisons pp 35d 53, Oct 1 81- Aug '83, JB Lippincott, St Louti. Fox α, Hinfield JM, Slobody LB, Smindler CM, Uttimer JK. J _m Med Assoc 148: 827-833, 1952. 6> F"*s and Comparisons pp 35d-53, Oct '81- Aug '83, JB Lippincott, St Louis. Table II Fluids to Which Complex Macromolecules Are Added (8) (9) (10) (11) (12) (13) Units Normal Hyper- Tyrode's Krebs Brighaa Scribner's Range Plasma tonic Henseleit Dialysis Dialysis of mmoles N.E.J.M. NaCl Acetate 283, 1285 (Resusitation) Hemodialysis L fluid 1970 Na 136 - 145 1200 150.1 143 140 135 130-145 K 3.5-5.0 5.9 5.9 4 1.5 0-5 Ca . 2.1 -2.6 1.8 2.5 1.25 1.25 1.25-2.0 free ECa2*3 £ 1.063 Hg 0.75 - 1.25 0.45 1.2 0.5 0.5 0-1 free EMg2+3 £ 0.533 EmEq Cations 142.7-153.21200 160.5 156.3 147.5 140 138-147 Cl 100 - 106 1200 147.48 127.8 120.7 • 105 92-111 HCO3 26-28 11. 25 26.8 I Pi 1 - 1.45 1.22 1.18 SO* 0.32 - 0.94 1.18 L - lactate 0.6- 1.8 pyruvate Lact/pyr D-β-OHbutyrate acetoacetate fl-HB/ acac acetate 35 35-45 Other Σ Eq anions 128.7-139.4 1200 161.6 157.3 147.5 140 138-147 Na/Cl 1.28-1.45 1.00 0.96 1.12 1.16 1.29 1.29-1.31 Glucose 3.9 - 5.6 5.6 10 or others C02 0.99 - 1.39 - 1.24 1.24 H 7.35-7.45 85.5-6.5 7.1 7.4 7.4 »5m3~T}»5 85.5-6.5 £ mOs 285,.- 295 2400 318.8 308 304.8 278.2 258-309 Use: Hemorrhage Perfusion General Hemo¬ Hemo¬ Hemo¬ 1 Shock dialysis dialysis dialysis 8) fiasco IT. Pontieri V. Rocha M, Silva E. Lopes OU. Am J Physiol 239: H664-673, 1 Hypertonic Ringer's Lacate has also been advocated in treatment of hemorrhage. See Nerlich M, Gunther R, De ling RH. Circ Shock 10 :179-188, 1983. Both »re inadequate. 9) Tyrode NJ. Arch int Pharmacodyn 20: 205, 1910. 10) Krebs HA, Henseleit KA. Hoppe-Seyler's Z Physiol Chem 210: 33-66, 1932. 11) Murphy UP, Swan RC, Halter C, Heller JM, Merrill JP. J Lab Clin Med 40: 436-445, 1952. 12 ) Mion CM, Hegstrom RM, Boen ST, Scribner BH. Trans Am Soc Artif Intern Organs 10: 110-113, 1964. The use of acetate in physiological fluids mas first proposed by: Nudge GH, Manning JA, Gilnun A. Proc Soc Exptl Biol Med 71ι 136-138, 1949. 13 ) Parsons FM, Stewart HK. In: Replacement of Renal Function bχ Dialysis (Drukker U, Parsons FM, Maher JF, eds) 2nd Edition, Martinus Nijhoff, Hingha, pp 148-170. Table HI. 'Prior Art' Fluids To Uhich Macromolecules Have Been Added (15) (16) (17) (IB) (19) (20) Units Normal Krebs Schimassek Krebs Heoatocyte Bahl an Fulgraff Plasma Liver Per- Liver Kidney Incubation Kidney Kidney moles N.E.J.M. fusion mith Perfusion Perfusion Perfusion Perfusion 283, 1285 Bovine serum L fluid 1970 Albumin and Red Cells Na 136 - 145153 151.54 148 153 147 143 K 3.5 - 5.0 5.9 5.9 5.9 5.9 4.9 4.74 .Ca 2.1 -2.6 2.5 l.β 2.5 2.5 2.56 1.25 free £Ca2 +3 £ 1.06 3 Ng 0.75 - 1.25 1.2 0.49 1.2 1.2 1.2 0.59 free EMg2+3 £ 0.53 3 EmEq Cations 142.7-153.2 166.3 162.02 161.3 166.3 159.4 151.15 Cl 100 - 106 127.8 147.48 127.6 127.8 127 113.04 HCO3 26 - 28 25 11.9 25 25 24.5 25 I Pi 1 - 1.45 1.18 1.22 1.18 1.18 1.18 1.18 SO4 0.32-0.94 1.18 - 1.2 1.2 1.18 1.18 L - lactate 0.6 - 1.8 (10 Na-1 Lac) 1.33 5 Na 1-Lac 9.09 2.75<d,l) 3.5(?d,l» pyruvate 0.09 0.91 0.25 0.25 Lact/pyr 14.8 10 10 7 or 14 D-θ-OHbutyrate acetoacetate fl-HB/ acac acetate 5.0 Other . ΣnEq anions 128.7-139.4 167.0 162.81 162.3 167.0 159.1 151.31 Na/Cl 1.28-1.45 1.12 1.03 1.16 1.20 1.20 1.26 (1.20) 61ucose 3.9 - 5.6 5.45 6.2 or others 6.7 urea 6.7 urea ∞2 0.99-1.39 1.25 1.24 1.24 1.24 1.24 1.24 00 PH 7.35-7.45 7.4 7.1 7.4 7.4 7.4 7.4 I wDsm 285 - 295 328 321 318 328 327 307.9 Albumin (g≠) 3.5-5 3.9 2.5 5 2.5 5.5 0.05 • Artificial perfusion fluid generally add 1.5 to 8 g≠ albumin, dialyzed against a medium listed in Table I; that is Krebs-Henseleit (10), Krebs-Ringer Phosphate (11), Tyrode's (9), Locke's (8), or Krebs-Henseleit with a lowered Ca 2+ to the 1 *. range, particularly in heart perfusion. They may or may not contain red cells. Krebs-Henseleit is known to contain about twice the amount of ionized C2 + as serum. 15) Hems R, Ross BD, Berry MN, Krebs HA. Biochem J 101, 284, 1966; Krebs Henseleit (10) with 3.9 g≠ bovine albumin. 16) Schmassek H. Biochem Z 336, 460, 1963. Essentially Tyrode's ( 9 ) with added lactate and pyruvate. 17) Nishiitsutsuji-Uwo JM, Ross BD, Krebs HA. Biochem J 103, 852-862, 1967; Krebs-Henseleit (10) with 5g≠ albumin, dry. IB) Crow KE, Cornell NU, Veech RL. Biochem J 172, 29-36, 1978, Krebs-Henseleit (10) with 2.5 g≠ dialysed albumin plus 1-lactate plus pyruvate. 19 ) Bahlman J. et al. fa J Phvsiol 212, 771967; Krebs Henseleit (10) with lactate and pyruvate and 5.5 g≠ bovine albumin 20 ) Fulgraff et al. Arch int Pharmacodvn 172, 49, 1972; Krebs-Henseleit (10) with 1/2 Hg and Ca plus lactate and pyruvate plus 5 mt, acetate, plus 0.05 g≠ albumin plus 2 g≠ hemocel. -9- substances (whether natural or synthetic in origin) do not appear to have previously been considered. Dextran (see Mudge, reference cited) appears to be regarded as the best known artificial plasma expander, yet dextvan is a branched polysaccharide of about 200,000 glucose units with a mole¬ cular weight of about 40 million and which has no anionic charge. In my copending U.S. patent applications U.S. S.N. 623510, U.S. S.N. 623102 and U.S. S.N. 623443, identi- fied by attorney's docket numbers P-83,2198, P-83,2213, P83,1655 and P-85,1402, P-85,1403 P-85,1404 provide aqueous electrolyte fluids which are also useful for contacting living animal cells, but these electrolyte fluids do not, in contrast to the fluids of the present invention, require the use of organic polyionic substances. For present disclosure purposes, the entire disclosure and contents of these copending applications is incorp- ated herein by reference. Definitions used in such copending applications, for example, are incorporated herein.to by reference. This invention relates to new and improved aqueous electrolyte solutions suitable for contacting living animal cells and to processes for making and using the same. The solutions (and processes of use) of this invention are employable in, for example: 1. intravenous electrolyte and fluid thereapy for mammals (including man) ; 2. dialysis fluids (both hemo and peritoneal) r - 3. parenteral nutrition (when administered in combination with nutrients) ; 4. perfusion media (with or without nutrients) ; 5. incubation media (with or without nutrients) ; 6. tissue culture (usually used with nutrients) ,* 7. plasma expanders and substitutes; and the like. -10- Aqueous electrolyte solutions of this invention comprise: (a) at lease one inorganic cation, (b) at least one inorganic anion, and (c) at least one non diffusible polyionic material. In any given solution, the total positive charges equal the total negative charges. The concentration of any one of such polyionic material is always less than about 10,000 millimoles per liter and the minimum concentration of any one such ionic material is always at least about 0.1 millimoles per liter, and similarly for each of said inorganic cation(s) and said inorganic anion (s). The charge Z asso-iated with a molecule of any given such polyionic material can vary from greater than zero to infinity. The molarity (number of moles per liter) of such polyionic material multiplied by Z produces a charge value indicating the total anionic equivalents of a given polyionic material in a given solution. The polyionic material can be anionic, cationic, or even a mixture of such. One class of the elctrolyte solutions of this invention incorporates at least one organic polyanionic substance of predeterminable and variable anionic charge. Such a substance is employed to fill, in whole or in part, the anion gap of a given aqueous electrolyte solution, and thus normalizes the Na:Cl milliequivalent ratio of the resulting solution. Another class of the electrolyte solutions of ' this invention incorporates at least one organic polycat- ionic substance of predeterminable and variable cationic charge. Such a substance is employed to fill, in whole or in part, a cation gap existing in a given aqueous electrolyte solution existing between, for example, inorganic cations and anions in an aqueous electrolyte solution. -11- For purposes of contacting living cells (e.g. physiological purposes) , an organic polyanionic substance useful in the practice of this invention is characterized by having an anionic charge valve which is equal to (molar concentration) times (total anionic charge per molecule) measured at a pH value to be associated with the particular electrolyte solution involved. Broadly, the pH range of a physiologic solution of this invention can range from about 5 to 9, but physio- logical pH values in the range from about 6. ' 8 to 7.6 are more preferred, and such pH values in the range from about 7.2 to 7.6 are presently most preferred for in vivo usage and desired physiologic pH. The anionic charge value for an organic polyanionic substance used in such practice of this invention should range from about -2 to -1000 net milliequivalents/millimole, and preferably from about -13 to -15 milliequivalents/millimole, and most preferably, about -14 milliequivalents/millimole (at a pH of about 7.4). In general the organic polyionic substances useful in the practice of this invention are characterized by being substantially non diffusible through a semiper- meable membrane, such as a semipermeable membrane of the type employed in hemodialysis and substantially impermeant to the mammalian cell membrane by means other than endocy- tosis. Particularly when a given organic polyanionic substance is to be used in a physiologic solution that will be employed in mammalian in vivo application (s) , it is, of course, preferred that such be: (1) non-antigentic; (2) non-pyretic; (3) metabolizable if taken up b ^ the reticulo- endothelial system. -12- It is more preferred that the metabolizable products, produced during enzymatic breakdown of a given organic polyanionic substance constitute safe entry points into the cellular metabolic processes. For present purposes, the polyionic non diffusible materials used are classifiable into three distinct groups or classes, as follows: Class I comprises of naturally occurring proteinaceous substances such as those which occur in blood and blood products; Class II comprised of polyanionic synthetic polymeric substances such. as. those which contain pendant acidic groups such as carboxyl, sulf te, sulfonyl, and the like; and Class III comprised of polycationic synthetic polymeric substances such as those which contain pendant basic groups, such as amines and the like. Examples of Class I substances include albumin, sodium albuminate, washed red cells, and the like. Examples of Class II substances include carboxymethyl starch, carboxyethyl starch, poly-d-betahydroxybutyrate, carboxy¬ methyl cellulose, cation exchange resins, and the like. Examples of Class III substances include anion exhange resins, and the like. As used herein, the term ion exhange resin refers to resins that are insoluble solid acids or bases which have the property of exchanging ions from solutions. "Cation exchange resins" contain fixed electronegative (anionic) charges which interact with mobile counterions (cations) having the opposite, or positive, charge. "Anion exchange resins" have fixed electro positive charges and exchange negatively charged anions. Ion exchange resins are three-dimensional macromolecules or insoluble polyelectrolytes having fixed charges distributed uniformly throughout the structure. -13- Although especially for physiologic in vivo solutions it is presently preferred that the polyanionic material used in the practice of this invention be in a form such that it forms a true solution (presently most preferred) , or true stable disperision in water such that individual units of the material in the aqueous medium are below about 0.01 micron in average particle size, those skilled in the art will appreciate that microscopic particle sizes may be associated with a given polyanionic material when iii vitro and other non biochemical appli¬ cations are contemplated. For example, in the case of using a cation-exchange resin in a dialysis fluid the resin may be in the form of uniform spheres of styrene- divinyl benzene (S-DVB) copolymers having diameters ranging from about 0.3 to 1.0mm. In physiologic electrolyte solutions of this invention, especially those intended for in vivo appli¬ cations indicated, sodium cations and chloride anions are incorporated. The ratio of sodium cation milliequivalents per liter to the chloride anion milliequivalents per liter is within the range found in normal mammalian blood plasma. The specified milliequivalent ratio of sodium to chloride in normal mammalian blood and in mormal animal extracellular fluid and intracellular fluid based on available information is believed to be in the range from about 1.24:1 to 1.47:1. In the case of anormal human adult, this range is now believed to extend (based on published information) from about 1.24:1 to 1.47:1, and preferably from about 1.33:1 to 1.42:1 and most preferably from about 1.36:1 to 1.42:1. These ratios are employed in solutions used in the practices of this invention. However, solutions employed in the practice of this invention can contain a Na:Cl milliequivalent ratio which is somewhat broader than the range considered normal, such as a ratio which is in the range from about 1.24:1 to 1.6:1 depending upon the mammal" (or patient), i condit¬ ion, the purposes of the physician or clinician, and the conditions of treatment. -14- By the term "milliequivalent ratio" as sometimes used herein, is meant the ratio of milliequivalents per liter of one substance to milliequivalent per liter of another substance in an aqueous medium. Characteristically, a physiologic solution of this invention contains from about 1 to 2400 millimoles per liter of sodium cations, and more preferably from about 120 to 165 millimoles per liter of sodium cations, and more preferably from about 129 to 163.5 mM/1 and most preferably from about 136 to 145 mM/1. In addition, a solution contains sufficient chloride anions to produce the milliequivalent ratio of sodium cations to chloride anions as above defined. In general, the maximum amount of organic polyanionic material which is used in a physiologic electrolyte solution of this invention is such as to supply an entire anion gap selected for that solution and thereby achieve a particular Na:Cl milliequivalent ratio sought for such solution. The minimum amount of organic polyanionic material which is used in a physiologic electrolyte solution of this invention is such as to make up only a part of the desired anion gap and achieve the desired Na:Cl ratio when more than only the organic polyanionic material is sued to help make up such anion gap desired and achieve such desired Na:Cl ratio. As a practical matter, no inherent biochemical minimum effective amount for organic polyanionic material which must be present in a solution of this invention is known or believed to exist. Preferably, however, the minimum total quantity of organic polyanioniς material used in any given solution is at least such as to produce a charge value in the range from about 10 to 18 mole charges per liter. The preferred anionic material contains carboxyl groups with a charge density of about 10-30 charges/70000 MW. Greater density or strength such as SO. 2- groups may cuase bleeding abnormalities and can denature protein. -15- Preferred but optional additives which may be used in combination with organic polyanionic material to close the anion gap and" achieve the Na:Cl ratio for a given physiologic solution are described herein below. Since, as described above, electrolyte solutions of the prior art containing albumin did not contain a normal milliequivalent ratio of Na:Cl, those compositions of the invention which contain albumin and which have such a normalized Na:Cl ratio are submitted to be novel and to constitute a patentable advance in this art since normalization of the Na:Cl ratio avoids acidosis. Particularly since no prior art electrolyte physiologic solutions are known which contain Class II polyanionic materials as described above, such solutions which contain a normalized Na:Cl ratio are likewise sub¬ mitted to be novel and to constitute a patentable advance in this art. In addition, however, electrolyte solutions which contain- such Class II polyanionic materials and which contain a Na:Cl ratio within the somewhat broader range above characterized are believed to be novel and useful and never previously known to the art since extremes of Na:Cl ratio in combination with Class II polyanionic materials offer new patentably useful fields of use and applications under both iii vivo and in vitro use conditions not heretofore possible. Optionally, in addition to sodium, a solution of this invention can conatin one or more of the following additional metallic cations in the respective quantity as indicated below: Quantity range Cation (millimoles per liter) Component broad preferred potassium 0 - 90 0 - 5 calcium 0 - 60 0 - 2.5 magnesium 0 - 15 0 - 1 -16- Optionally a solution of this invention can have additionally incorporated (dissolved) therein from 0 to about 550 millimoles per liter of at least one substantially nonionic (including zwitterionic) osmotically active sub- stance (which is preferably metabolizable) . Examples of usable such nonionic substances include glucose, glycerol, fructose, sorbitol, and the like. Glucose is presently most preferred. A physiologic electrolyte solution used in the practice of this invention is further characterized by generally having: (1) sufficient total substances in the absence of any nonionics dissolved therein to produce an osmolarity ranging from about 260 to 5000 milliosmoles (mOs) and preferably from about 265 to 550 mOs and most preferably from about 28.0 to 320 in mOs; (2) the relationship between total ionic sub¬ stances is such that the pH ranges from about 5 to 9, and preferably from about 6.9 to 8.6; and most preferably from about 7.35 to 7.55; (3) the charges of all cations equal the charges of. all anions; and (4) the minimum total concentration of all such near equilibrium couple (s) present is at least about 0.1 millimoles per liter, and preferably is at least about 0.5 mM/1, and more preferably about 2mM/l, while the maximum concentration thereof is preferably not more than about 465 mM/L and more preferably is nqt more than about 61 mM/1 and most preferably is not more than about 50 mM/1. (5) Non-physiological electrolyte solutions of this invention may be made in like manner and may be used to prepare ion exhange resins, active enzyme solutions and the like. -17- Various additional objects, aims, and purposes, features, advantages, applications, variations, and the like will be apparent to those skilled in the art from the teachings of the present specification taken with the claims. Optional Anion Gap Components In addition to the organic polyanionic materials in the anion gap of a given electrolyte solution of the present invention one may include in the anion gap at least one near equilibrium couple selected from the group consisting of (1) bicarbonate and carbon dioxide, (2) 1-lactate " and pyruvate , and (3) d-betahydroxybutyrate and acetoacetate . Thus, one first optional class of solutions of this invention contains in addition to organic polyanionic material an inorganic class of anions comprised of chloride and bicarbonate. These solutions have a physiological pH which is broadly in the range from about 6.9 to 8.6, and more preferably in the range from about 7.35 to 7.45, and most preferably is about 7.4 (for human use) . Dissolved carbon dioxide must also be present in these solutions with bicarbonate anions. When administered, these solu¬ tions not only tend to maintain a mammal's normal blood (and plasma) ratio of sodium to chloride, but also tend to set (regulate) the mammal's normal blood (plasma) pH at a normalized value. Another second optional class .(preferred) of such solutions characteristically contains a class of carboxylate anionic couple pairs comprised of at least one of (a) a mixture of D-betahydroxybutyrate anions and acetoacetate anions, and (b) a mixture of both (a) and (b) . These solutions have a physiological pH which is as above defined in connection with such first optional class of solutions. When administered, these solutions not only tend to maintain the mammal's redox state within a normal range, but also tend to maintain that mammal's phosphorylation potential witt .n a normal range. -18- Another (third) class (more preferred) of such solutions characteristically not only contains both chloride anions, and bicarbonate/carbon dioxide mixture, as in such (first) class of solutions, but also utilizes (contains) such class of carboxylate anionic couples, as in such (second) class of solutions. When administered, these solutions achieve the above idicated effects obtained from the use of such first optional class of solutions and the above indicated effects obtained from the use of such second optional class of solutions. The total quantity, or sum (sigma) , bicarbonate anions and carbon dioxide present in a solution of this invention ranges from 0 to about 465 millimoles per liter of solution, and preferably 0 to 55 millimoles/L. The ratio of bicarbonate milliequivalents per liter to dissolv¬ ed carbon dioxide milliequivalents per liter in a solution of this invention can range from about 0.1/1 to 55/0.1 and preferably 11/1 to 24/1. More preferably, such total ranges from about 10 to " 45 mM/1 and such ratio ranges from about 18:1 to 26:1, and still more preferably, such total ranges from about 23 to 35 mM/1 while such ratio ranges from about 19:1 to 21:1. A ratio of 19.95 for [HC0 3 " ]/ [CO-3 gives a pH 7.4 which is presently particularly preferred. The total quantity, or sum (sigma) of 1-lactate anions and pyruvate anions present in a solution of this invention ranges from 0 to about 465 (preferably 0 to 55) millimoles per liter of solution. The ratio of 1-lactate anion milliequivalents per liter to pyruvate anion illi- equivalents per liter in a solution of this invention can range from about 20:1 to 1:1. Preferably, such total quantity ranges from about 0.5 to 10 mM/1 and such ratio ranges from 3:1 to 15:1, and more preferably such total quantity ranges from about 2 to 8 mM/1 while such ratio ranges from about 5:1 to 12:1. -19- The total quantity, or sum (sigma) of d-beta¬ hydroxybutyrate anions and acetoacetate anions present in a solution of this invention ranges from about 0 to about 465 (preferably 0 to 55) millimoles per liter of solution. The ratio of d-betahydroxybutyrate anion milliequivalents per liter to acetoacetate milliequivalents per liter in a solution of this invention can range from about 6:1 to 0.5:1. Preferably, such total ranges from about 1 to lOmM/1 and such ratio ranges from about 4:1 to 1:1, and more preferably such total ranges from about 2 to 8 mM/1 while such ratio ranges from about 3:1 to 1.5:1. . One of the three near equilibrium couples optionally employed in the practice of this invention (the bicarbonate~/carbon dioxide couple) tends, as used in this invention, to regulate the concentration of hydrogen ions in blood (plasma) and in the treated mammal's cells, and each one of such couples tends to normalize the redox state of each of the three pyridine mucleotide couples. The phosphorylation potential also tends to be normalized. Also, each such near equilibrium couple when used as herein described constitutes a safe entry point into the metabolic system of a mammal. Further, each such near equilibrium couple, if utilized in this invention, exhibits a distribution between intracellular fluid and extracellular fluid such that the ratio of the con-entrations in, respectively, intracellular fluid to extracellular fluid ranges from about 1:1 to 1.5:1 in most all mammalian cells. One skilled in the art may wish to include with a polyanionate an aqueous electrolyte solution of this inven¬ tion containing a total of not less than about 0.05 to 0.T mM/1 of at least one. of such three near equilibrium couples and not more than about 465 mM/1 thereof and preferably from 0 to 55 millimoles/liter. -20- The prefered cationic salt form of the poly- anionate for purposes of plasma expansion is the sodium form, however the other cations listed hereinbelow may be used. For the purposes of plasma expansion pure (Na ) Polyanionatez- may be given as far superior to the un¬ charged materials currently used for the purpose of plasma expansion. We prefer however to administer the material in a solution with the Na:Cl ratio in the range from 1.24 to 1.6. Electrolyte Solutions Electrolyte solutions of this invention which are presently preferred for physiologic usage are characterized as being comprised of water which has dissolved therein each of the following components in the amounts indicated: -21- Table V Quantity Component range mM/L preferred broad Total Cations 125- ■ 160 1 - 2400 (1) sodium 125- ■ 160 1 - 2400 (2) potassium 0 - 6 0 - 90 (3) cal .ci ■um2+ 0 - 1.5 0 - 60 2+ (4) magnesium 0 - 1 0 - 15 Total Anions 125 - 160 1 - 2400 (5) chloride " 100 - 130 0 - 2000 (6) bicarbonate 0 - 55 0 - 465 (7) inorganic phosphate 2" 0 - 5 0 - 22 (8) 1-lactate + pyruvate 0 - 55 0 - 465 (9) d-B Hydroxybutyrate " acetoacetate 0 — 55 0 _ 465 (10) non diffusible polyanionate in mEq/L 25 - 160 0. 2 - 2400 Total Nonionics 0 - 550 0 - 575 (11) carbon dioxide 0 - 10 0 — 25 (12) osmotically active substance 0 - 550 0 - 575 the interrelationships between components being such that: -mEq ratio of [bicarbonate]/[CO-] 1/1 - 55/0.1 -mEq ratio of [1-lactate " ]/[pyruvate r ] 20/1 - 1/1 -mEq ratio of [d-hydroxybutyrate~]/[acetoacetate-] 6/1 - 0.5/ -mEq ratio of [Na ]/[Cl~] 1.24 - 1.6 (1) 0 - infinity total of (6) , (8), (9), (10) 0.2 - 2400 -milliosmolarity 260 - 5000 -pH 5 - 9 (1) preferred -22- Preparation of polyanionic material Any convenient preparative procedure may be used, for example: Suitable Class II polyanionics are prepared as follows: Albumin is taken in water (about 1.5 mM/1) and dialyzed for 48 hours against deionized water a 4 degrees C. It is treated with NaOH to a pH of 7.4 at which pH it has become about sodium 20 albummate --20 Carboxymethyl starch prepared in sizes comparably to those available for Dextran (see Mudge GH. In: Gilman AG, Goodman LS, Gilman A. eds. The Pharmacological Basis of Therapeutics, 6th edition, Macmillan, New York, 1980, 859-862.) has a pK , of about 4 and can be prepared in the same manner as albumin. Likewise carboxymethyl dextrans are similarly prepared. Carboxymethylation can be con¬ veniently accomplished, for example, by titration with chloroacetate. Any carboxy-lower alkyl groups may be used, but methyl is presently preferred. Each of these products has a Z[cone ] term of about -14mEq/l for use as a plasma substitute or plasma expander, for example, in the treatment of burns and war wounds, or as a counter polyanion in hemodialysis or peritoneal dialysis to cancel the need for adding toxic amounts of small organic anions such as d,1-lactate, acetate or chloride to such fluids to obtain a Na:Cl ratio of from 1.24 to 1.6. Plasma Substitutes and Plasma Extenders , Electrolyte solutions of this invention containing non-diffusible polyanionics are useful as plasma substitutes and plasma extenders, and, in such useage, the presently most preferred non-diffusible polyanionics are serum albumin and sodium albuminate, although other Class II polyanionics may be employed, such as carboxymethyl starch, poly gamma glutamate, and the like, preferably in the solidum form for human use. -23- Serum albumin is normally present in plasma at about 0.65 mM/1 and at pH 7.4 has -20 charges per molecule for a Z[albumin concentration] term of about 13 to 14 mEq/L. From the relationship: [C]RT this osmotic 20- pressure of serum albumin (which is a 20 albuminate ) is about 264 mmHg. (Eqn 6.) In contrast, the osmotic pressure for dextran (molecular weight 68,500 with no charge) is about 12.6. mmHg on the basis of Equation 6 above. For dextran to be equivalent to sodium albuminate in its osmotic effect, one would have to make the plasma contain about 13 mM/1 dextran. or about 90% solution. Such a solution would be so viscous as to be fatal in some circumstances. It is concluded that artificial plasma expanders must have net fixed negative charges at pH 7.2 to 7.4 and preferably be given in the sodium form to achieve the desired effect of expansion of the intravascular volume. It is further concluded that the currently described and widely used "ideal plasma expanders" (See Mudge in Goodman and Gilman 1980, cited above) are essentially worthless for the purposes for which they ar.e currently used in medical practice. Examples of non-diffusible polyanionic materials useful in plasma substitutes and plasma extenders include albumin, sodium albuminate, carboxymethyl starch, any carboxy-lower alkyl starch, carboxymethyl cellulose, carboxymethyl dextrose, gamma polyglutamate, red cells, polyacrylamides, and polysulfonates. The sodium forms of these polyanionates are particularly preferred for the purpose of plasma volume expansion. Because of the higher charge density of the polysulfonates, this class is usable, but not preferred; - - Utilization of Polyanionics Polyanions of the type described have a particular application in the treatment of war wounds and burns, where stable preparations of effective plasma expanders are required. The use of these types of material in their Na-polyanionate form would be extremely effective and with lower toxicity than the current practice of using uncharged dextrans (See Williams TG, Riley TRD, Moody RA. Brit Med J 2J3ι6: 775-777, 1983; Williams TG, Brit Med J 296: 790-792, 1983). They may be used as a less toxic alter¬ native of hemorrhage (See Velasco IT et al Am J Physiol 239: H664-673, 1980, or to large volumes of crystaloid solutions such as Ringer's Lactate (See Nerlich M, Gunther. R, Demling RH. Circ Shock 10: 179-188, 1983). Insoluble, non-diffusible polyanionics have a variety of uses, in filling the anion gap. They can be used in tissue culture medium or on the surface of tissue culture plates to create a suitable culture medium that carries a sodium (or cation) carbox late such that the proper cationic composition can be obtained with a normal Na:Cl ratio. Such polyanionic materials can also be used in dialysis fluids to obtain a desired Na:Cl ratio and to overcome the anion gap as an alternative to small anions, such as shown in my copending case, P-83,2198 and P-85,1402. Their large molecular size is such that they do not pass through the semipermeable membranes employed in hemodialysis, A preferred class of solid insoluble non- diffusible polyanionics ' comprises -arboxy (lower alkyl) polymers. Examples include carboxylated styrene-divinyl- benzene ion resins and acrylic ion exchange resins such as those available from such companies as Dow Chemical Company ("Dowex resins") and Rohm and Haas Company. The sulfonated resins may be used as anions, in, for example hemodialysis or on tissue culture plates to obtain the desired Na:Cl ratio, but should not be used intravascularly, where they may interfere with the clotting mechanism of blood (such as occurs with heparin) . -25- Use of Polycationic Materials Non-diffusible polycationic materials can be used in accord with this invention to treat alkalosis and thus increase the solidum to chloride ratio without giving ammonia, which is undesirable because excessive ammonia administration: (1) causes what can be an excessive urea synthesis; (2) causes what can be major upsets in the cellular redox state and the cellular phosphorylation potential; and (3) can induce hepatic coma and death. Determination of the Charge Value Z [polyion 2 r ~j As follows from Eqn. 2, below, the number Z x [concentration of polyion] where Z is the absolute value of the net charge, - or +/mole on polyionic substance, and [ ] indicates the molar concentration. From such values, the value Z [polyion ] can be determined by placing the polyion in a dialysis sac through which it is impermeant and dialyzing it at equal pressure against an excess volume of diffusible salt solution, for example, NaCl, at any pH of interest. The ratios of [Na ] and [Cl ] in the dialyzing solution and inside the dialysis sac containing the polyion are related to the term Z [polyion ] according to Eqn 2 following: [Na + ]o/[Na + ]i = [Cl " ]i/[Cl " ]o = [Cl " ]o/£[Cl " ]i + Z [polyion 2 ]} The present invention is further illustrated by reference to the following examples. Those skilled in the art will appreciate that other and further embodiments are obvious and.within the spirit and scope of this invention from the teachings of these present examples taken with the accompanying specification. Examples 1-2 Illustrative electrolyte solutions of this invention are provided. Example 1 is a modified Krebs- Henseleit solution wherein the Na:Cl ratio is normalized with sodium albuminate. Example 2 is artificial plasma expander which is an alternative to a dextran containing expander. It has 20 times the osmotic pressure of dextran. Table Examples of New Polyanionate Solutions for Contacting living Cells, Either I.V., Through Dialysis Membranes, or In Vitro . Units Normal ( 1 ) ( 2 ) Plasma Krebs' Veech's mmoles W.E.J.M. Albuminate Polyanionate 283, 1285 I fluid 1970 Na + 136 - 145 138 142 K+ 3.5 - 5.0 4.0 4.5 ΣCa 2.1 -2.6 2.25 1.1 free.[Ca2+l I 1.06 ] ΣMg 0.75 - 1.25 1.00 0.56 ° free [Hg2+] I 0.53 J Σ y-Eq Cations 142.7-153.2 148.5 149.73 Cl" 100 - 106 103 102 ICO3- 26-28 29 29 Σ Pi 1 - 1.45 1.2 1.1- St_,2- 0.32-0.94 - - L- lactate' 0.6- 1.8 pyruvate" Lact/pyr D-fl-OHbutyrate" acetoacetate" β-HB/ acac acetate' Other 0.73 M 0.70 H Albuminate Carboxymethyl-Starch (-14.6 nEq) (-16.7 mEq) Σ mEq anions 128.7-139.4 148.8 149.73 Na + /Cr 1.28 - 1.45 1.34 1.39 Glucose 3.9 - 5.6 _ - or others PH 7.35 - 7.45 7.4 7.4 Σ tisa 285 - 295 280.7 307 Use: 1) IV infusion 1) Alternative to Dextran or Hydroκyethyl-starch as a plasma expander. 2) As dialysis 2) As dialysis fluid fluid 3) Tissue 3) As tissue culture media culture 4) Contac ing 4) Contancting cells living ceils (1) Bovine serum albuminate is prepared as described and carries about 20 - / mole. For human use, human albumin is used. (2) A 70,000 HH starch, with β D-glucose primarily in β 1 - 4 linkage or alternatively with the a 1 - 6 linkage is prepared chloroacetate to give about 24 Na-carboxylate groups/mole. -27- When either of these two solutions is used as a plasma expander for the treatment of a severely wounded man who is hemorrhaging, it is found that the patient's blood osmotic pressure normalizes as does the distribution of water between intravascular and intracellular space. Administration of an equivalent molar amount of a presently used plasma expander such as dextran or hydroxye hy1st rch (but no sodium albuminate) is found not to elevate the patient's blood pressure and not to maintain the osmotic pressure of the patient's blood plasma. The laws and relationships controlling the technology and the effects described therein are presented in the following equations. There is no intent to be bound by theory. Various changes, improvements, alternatives, uses, and the like, will be apparent to those skilled in the art from the present description, and no undue limitations are to be inferred or implied. Although the teachings of my invention have been herein.disclosed with reference to certain embodiments and illustrations, it is to be understood that such are by way of illustration only and that others may wish to utilize my invention in different designs, applications, or the like. 0. Eqn 0 - The Second Law J. illard Gibbs. On the equilibrium of heterogeneous substances. J Conn Ac ad Sci 1876; III : 343. 0 - 1 Definition of Gibbs Free Energy and Other Properties of State": B « H - TS where: 6 ~ Gibbs free energy H ~ Enthalpy or heat content T ~ absolute temperature S "" Entropy, or state of randomness or disorder 0 - 1 a Entropy may be more rigorously defined' by statistical and quantum mechanics in the Boltsmann Equations where: S " Entropy 1, ./ Bol t∑ ann constai B Avagadro's number = i4 1.3B :< 10~ 2"' J/° n ■. Degeneracy 0 _ _> - G ΔH - T Δ S where * change i n 0 - 3 Standard Free Energy ~ Δ G l ΔG - ΔG° + RT In [products] . reactantsD where: R "** gas constant = 1.987 calories/°K /mole and ° - 273 + °C T * °K In • 2.303 log 10 where • • 0 Keq ~ Cproductε] Creactants] 0 - 4 At equilibrium, AG *= 0 , so in A + B •.- ■ * C •« ■ D ΔG - - RT In K + RT In CC3CD3 q CA. CB] where: C ] * activity or ~ concentration "A theory is the more impressive the greater the simplicity of its premises, the more different are the kinds of things it relates, and the - more extended is its range of applicability... It is the only physical theory of universal content.which I am convinced, that within the framework of applicability of its basic concepts, will never be overthrown. Eqn 1 - The Henderson-Hasselbalch Equation The major buffer and controller of e«tra- and intracellul r pH. Henderson LJ. Blood, A Study in General Physiology. Sil liman Lecture , Yale University Press, 1928 CHCO., 3 1. a pH s pK , + log - a CC0 2 where: o pK , = ό.10 at 38 C and serum concentrations of electrolytes 1. b The solubility of C0 in fluid, i.e. dissolved CO^ gas plus H 2„CQ -_r. from: * " *~ C0 + H„0 ^ -^ H^CCL. pCO- in mmHg °( l C0 /ml of H„0 lOOOmmol CC0 in mol/liter = . . * 7< 0 mmHg 22.26 L/ mole mole * °\ = 0.553 / ml serum H 0 at 3Θ°C from: C0 Van Sl e DD. J Biol Chem 73: 765-799, 192B. 1. c The pH of a bicarbonate containing solution to which has been added a carbocylic acid such as acetic, lactic, acetoacetic acid with a pK' in the 3 to 4 range and where the concentration of HC0-, is much larger that the concentration of carboxcylic acid: Thus adding 1.8 mM Hlactate and 0.2 ml. Hpyruvate to 25 ml NaHCO ^ , yeilds what pH? C253 pH = pK - log 2 ( CHCO ^ - CHAD 6.1 (1.36) 7.46 II Donnan Equilibrium Equation Donnan FG.Z Electrochem 17: 572, 1911 Donnan FG. Chem Rev 1: 73-90, 1924. ΔE Δp 1. From Gibbs (Eqn 0) [Na+] lNa÷] : CC1~ CNa 3 [CI-] ? RT In RT In 0 ICI-], * CC1 3, CNa + 3, υr . CC1~3 CNa + D., [ ] = activity = concentration l.a CC1~. , CNa 3 A = non-diffusable polyanion Z = valance of polyanion CC1 3 CC1 3, CNa + 3, Therefore: CC1~3 CC1 3 ZCA 2 ~ CNa + 3 and for polyvalents: 1/s anions 1/z cations CAnions "" _ ECations3, CAnions] ? [Cations], 2. From the Law of Electrically Neutrality: CNa + 3 = CC1~3^ a + 3 j = CCl " 3 1 + Z 2 ~3 1 3. Quadratic equation: 2a E mple: Consider albumin dialysed against 100% CO^, / 3.13 NaHCO, buffer with 1.17 M albumin (i.e. 7. solution). Hypothetically keep charge on albumin at -20/mole. 1.17 x 10 ^" CHCO, " . = 0.4 x 10~ -J> M II Eqn 2 Hulticotponcnt Donnan Equilibriui Syitβa for Solutions Such as the Heaodialysis oi Blood Plasaa Electrolytes: where Ap = 0 and all coaponents but albuain are peraeant. Subscript * in dialysis fluid, subscript . * in patient's plasaa, Ap * difference in pressure. ° 2.1. Stateaent of electrical neutrality on two sides of an uncharged aeabrane 2.b.l. INa*l ♦ tK*l ♦ 2ICa 2 *] ♦ 2IHg 2+ l « ICl " ] ♦ IHCQ " ) ♦ l.βt. Pi "1,8 ] ♦ Ilac " ] ♦ Ipyr ' l ♦ lacac " ] ♦ IBHB " ] ♦ [acet " ] 0 0 0 0 0 3 0 0 0 0 0 0 0 2.b.2. INa*], ♦ IK*]. ♦ 2ICa 2 *l. ♦ 2Hg 2 *]. ■ ICl'l, ♦ IHCO."]. ♦ l.βttPi! 1,8 ]. ♦ Ilac " ]. ♦ Ipyr " ]. ♦ lacac " ]. ♦ IBHB-]. ♦ lacβt ' l. ♦ i i ι * i i 3 i i ι r ' ι i l ι Ztprot* 1 Distribution of cations on two sides of the aeabrane: Distribution of Anions: 2.d ICl ' lj • } "«"l HB " ]. Now solving for INa l./INa ] for a dialysis fluid of known coaposition: 2.t and: . 2 INa 1 ♦ IK 1 2(lCa* 2 l o ♦ IHg 2 *]^ tprot 1" ] 0 0 ♦,] O — INa*]. 2 ♦ m j 3 . , Zi r| *j . ( 1#β cepij tNa* °'. 8 B)»l r N u a*♦],.(-0.B ) i 1 I 0 0 / i IN.*] 2 IN.*! 3 INa*] 0 0 ICl ] ♦ IHCO,"] ♦ lacet 1 • (lact ' l ♦ Ipyr " ] ♦ lacac 1 ♦ IBHB 3 0 3 0 0 r ' o 0 c Plasaa [concentration] * 0.935 x plasaa HO [concentration] Ill Eqn 3. Nernst Equation ΔE Nernst W. Theoretical Chemistr y 4th Edition, 1904, McMillan, London. See also Si l l iman Lectur , 1906, Yale U. Press, New Haven. RT Can on Doutβide 3. Δ E - - — In nF Canion Dinside ors RT Ccation 3inside ΔE In nF Ccation Doutβide where: at 38°C T • 311 °K R, the gas constant * * • 8.314 joules degree/mole n * * number of equivilents F, the Faraday, ~ 96,494 coulombs ΔE "" potential in volts To convert In to log.., multiply be 2.303 From Cornell N, Anal Biochem 1980; 102s 326 - 331, for isolated hepatocytes from starved rats incubated in Krebs- Henseleit. CO.128 M Cl Doutεide ΔE = -0.0617 log -- CO. 41 M Cl . inside = -0.0305 V or - 30.5 V and for cat brain, from Eccles JC. The Physiology of Ner ve Cel l , 1957, Johns Hopkins U Press, Baltimore. CO.125 M Cl 3outside E = -0.0617 log CO.009 M Cl 3inside ΔE = -0.0705 V or - 70.5 V 3. Redoκ Potential of Half Reactions RT Coκidized3 E h - E° + — In nF Crβduced3 where: R * 8.31431 J/°K/mole T - °K n * * • number of electrons F * Faraday ~ 96,494 coulombs In * " 2.303 log 10 IV Eqn 4 Radox State Equation!. [NAD*]/ [NADH] or [NADP*]/ tNADPH]. Near equilibriui reactions are given a nuaber depending upon location. The E of the [NAD 1/lNftDHl couple at pH 7 is -0.32V. That of the [NADP 1/CNADPH] couple, -0.33S V. Abbreviation Definition of Value of Value of E at E at eq K at pH = 0 K at pH 7 at pH 7.0 at pH 7.0 Enzyae No. eq eq oxidized 0.- = 1.5 all reduced or 0.5 aH NH.+ V or I ιH Pi V Cytoolasaic NAD - Linked Dehydrogenases [pyruvate ' HNADHHH*] 4 c 1 nH 1.11x10 H 1.11x10 -0.201 Lϋ U-lactate-HNAD EC 1.1.1.27 , [oxaloacetate ItNADHJtH ] 4 c 2 K N . H 2.86x10 H 2.86x10 -0.184 n [l-aalate2-][NAD EC 1.1.1.37 , W-glycerol-P 3CHADH1CH ] 4 c 3 K RPΠH * 1.3»lθ "I ! H 1.3x10 -0.203 [DHAP2-HNAD+] EC 1.1.1.94 . [1 ,3 DiP6 HNADHHH 1 . 4 c * cήpnu = " 5.3x10 H 5.3x!0 -0.302 -0.222 Here, Pi is a reactant Wm I6AP2-][ Pi2-][NAD * EC .1.2.1.12 [acetaldehydeHNADHHH ] K ΔΠH = 1-94x10 M 1.9x 10 -0.209 [ethanol HNAD+l EC 1.1.1.1 [d-fructose][NADH][H ] K HΠH = 1.14x10 H 1.14x10 -0.262 iαU [d-βorbitol][NAD+] Hitochondrial NAD - Linked Dehydrogenases [acetoacetate ' KNADHHH*] 4 ' ! K H B DH s 4.93x10 H 4.93xlθ "2 -0.281 [d-B-hydroxybutyrate-HNAD EC 1.1.1.30 W-KG ][NH4 ][NADH]tH ] 4 ' 2 K G1DH * 3.87xl0 _1 V 3.87x10 H -0.158 -0.257 [l-glutaaateHNAD+] EC 1.4.1.3 [acetate " ][NADH][H ] K Λ1DH s 1.45xl0 * V 1.45xl0 +9 -0.596 [acetaldehydeHNAD+J EC 1.2.1.3 ' Cytoplasaic NADP - Linked Dehydrogenases [*- KB 2" ][C0 2 1[NADPH] 4 T 1 * m * m 1<17H ~ 0 ' 337 "°- 422 Here, CO, is a reactant " [1 -isocitrate3-][NADP+] l EC 1.1.1.42 * [pyruvate~][C02][NADPHl 4 T 2 HillC E E n n , z " [•a te2-][NADP*l "" 3 ' 44 * l ° « EC 1.1.1.40 . [ribulose 5-P 1EC02HHADPH1 4 T 3 K 6PBDH r [6-phosphogluconate3-][NA 1 D- 7 P 2 + x ] , ° " EC 1.1.1.43 • See ref. Linking Isoaerases - [<.- K6 2" ][J-aspartate " ] 4 L 1 K ωτ - 6.61 [l-glutaate-][oxaloacetate-l EC 2.6.1.1 W- K6 ][l-alanine] 4 L 2 ^ ■ ■ 1.52 [l-glutaιate-][pyruvate-] EC 2.6.1.2 2 _ [dihydroxyacetone-P ] 4 L 3 IC • 22 [glyceraldehyde 3-P2-J EC 5.3.1.1 References for Values of Near-Equilibriua Reactions in Equation 4 Equation Abbreviation Reference 4 C 1 K ιnu Nilliaason DH, Lund P, Krebs HA. Bi.cht. J 103: 514-527, 1967 4 C 2 IL DH 6uynn R, βelberg H, Veech RL. J hoi Chi.248: 6957-6965, 1973 4 C 3 K . pDH Russaan H. Ifcesij, Munich, 1969. 4 C 4 K GApDH Cornell N, Lcadbetter H, Veech RL.3 liøJ Chit 254: 6522-6527, 1979 4 H 1 K Nilliaason DH, Lund P, Krebs HA. Biochn ] 103: 514-527, 1967 " 4 H 2 Engel P, Dalziel K. liochaa J 105: 691-695, 1967 4 T 1 K ]cDH Londesbourgh J, Dalziel K. fiioch» J 110: 217-222, 1968 T 2 Veech R, Eggleston LV, Krebs HA. Biochi. J 115: 609-619, 1967 .I.E. T 3 Villet R, Dalziel K. fiiocλ.a J 115: 633-638, 1969 6P6DH L 1 Krebs HA. Mr hi fttg 13: 449-472, 1975 Λ 60T L 2 6PT Krebs HA. Mv lit fteα 13: 449-472, 1975 L 3 Veech RL, Raijaan L, Dalziel K, Krebs HA. liøcAti 3115: 837-842, 1969 Λ TPI The enzyae aldose reductase EC 1.1.1.21 aay be redox active during fructose infusion in certain tissues. The reaction is: td-sor bitol HNADPHHH ] 2x10 -11 H.» Hy estiaate Aldose R [d-glucoseHNADP*] For description,-see Hayaan S, Kinoshita JH.3 tiol Che» 240: 877, 1965 V Eqn 5 Phoiphorylation State Equations - I. ATP1/IΣADPH∑Pi] Veech RL, Laitson JR, Cornell NH, Krebs HA. J Biol Cftta 254: 6538-6547, 1979 5a. The equilibriua constant of the glycer ldehyde ' 3-phosphte dehydrogenase (EC 1.1.1.29) and 3 phosphoglγcerate kinase reactions (EC 2.7.2.3) at 3B°C, I = 0.25, and free [Hg ] = 1 βH is: - tϊ3PGl [CATPl [NADHHH*] . K B*B ' ' * * l«β3 » 10 (C6AP) IfADPl Pi] tNAO 5b. Coabining the above reaction Nith K.. H and substituting [ DHAPl = I 6AP1/22 K 6*6 6t 3P6] tiATP] [1-lactate] .. . . . , « i.tfuto H" 1 K, . H I. 6AP] I. ADPlfcPi l [pyruvate] 5c. Or: CATPI fcDHAPl [pyruvate] .. , Free Cytoplisaic ■ — . ———— x 7.5x10 ♦5H„-l ADPIK-Pi] I. 3PB] [1-lactate] 5d. Alternatively, frαa the creatine phosphokinase reaction (EC 2.7.3.2 ) For the Pyrαphosphoryl tion State or tr Pil/fir Pil: Laitson JNR, Guynn RH, Cornell NH, Veech RL. In 6lucow.ogtt.isis (Hanson RH, Hehlaan HA eds) pp 481-511 , John Niley, Nen York, 1976 5e. Froa the UDP6 Pyropfwisphoryl. se reaction (EC 2.7.7.9) : [iglucoie 1-PlttUTP] Frte Cytoplasaic [. PPil ■ RUDPglucostl'K, UDPBPPi it " here K UDPGPPiase ' >55 5f. For liver and blood glucose: Pβlucosf 6-P][tPi ] K 6-PPi Trans Pan ' J^^ 5g. [free F 1 ,4 diPlfcPil - ' -P-PPi Trani Paie „ . . . « .„,- » ., ['fructOK 6-rø*PFιl ■ 36- Determination o-F Osmotic Pressure - "H * Van't Ho-ff JH. Arch Neer 1 Sci 20: 239-303, 138ώ if = 2CC3 RT where: 17 ^ osmotic pressure in atmospheres (relative to pure H^O) Z LC1 "" "Σ. [concentrations] o-f solutes in mole/liter R "* gas constant = 0.082 liter atmospheres/ mole/ degree K T " 273 + °C VII Eqn 7 The Equation of State of the Cell Relating theΔE across the cell aeabrane, the distribution of [Na*], IK*], [Cl * ], and [C fluid and cytoplasaic H*D and hence cell voluae to the cytoplasaic UATPl/t∑ADP][«Pi] Δ to/X A Ϊ Pase " Since _G * 0, then: 0 * -7.73 kcal/aole - ' . + 1-6.3 kcal/aole) ■ B.5 kcal/aole ♦ 5.5 kcal/aole 0.082 liter ataos/aole/ a K 1 As 1 kcal /aole = x * 1.85 ataospheres 1.98x10-' kcal/aole/ a K 22.4 1/aole then the US tera * 5.5 x 1.85 = 10.2 ataospheres. ( And further froa V.n't Hoff (Eqn 6) Eqn 7 states that since u τ H.O outside = u τ M *o inside, the cell is prevented froa swelling by the Na*/K* ATPase which "electroneutrally puaps out 2 aOsaoles/ ATP hydrolysed. The ΔE across the cell (aeabrane) is reflected by the distribution of ICl * ], / [Cl-], in accordance with the Nernst equation (Eqn 3). The TΔS or decreased entropy nithin the living cell represents the increase "order* characteristic of the living cell. See Eqn 0. 7b. From the high capacity Na /Ca " ' 1' exchanger written in an electroneutral manner reflecting the free permeability of Cl in accordance with the dictates of the Nernst equation, (Eqn 3): 3 Na o + Ca *1" l. + Cl~o <%, • 3 Na I. + Cs-^ o + Cl~l. The net osmσlar movement of eqn 7a is 2 osmoles > outside. In contrast, the net movement of eqn 7b is 3 osmqles > inside, requiring the Na /K ATPase to cycle 3 times for each 2 times the Na +/Ca2+ exchange mechanism operates in order to maintain osmotic equilibrium. * ■ >+ *?■+- The gradient CCa" *" 1l/ CCa' 1' Do is thus a direct function of the CNa + 3 Λ O° CNa " * " ]1. 0 , (the CC1~3O/ CC1~]1. >, and a function of the phosphorylation and entropy state of the cell.

Claims: -39- CLAIMS 1. An aqueous electrolyte solution comprising water having dissolved therein: (a) at least one inorganic cation or at least one inorganic anion, (b) at least one non-diffusible polyionic material selected from the group consisting of polyanionic materials, polycationic materials, and mixed polyanionic/ polycationic materials provided that: (1) the concentration of any one of (a) , and (b) always ranges from about 0.1 to 5,000 millimoles per liter, (2) the total positive charges equal the total negative charges, (3) the charge associated with a molecule of said polyionic material ranges from greater than 0 to infinity. 2. An electrolyte solution, for contacting living animal ceils comprising water which has dissolved therein the following components in the respective amounts indicated: -40- Table Quantity Component range mM/L Total Cations 1 - 2400 (1) sodium about 1 - 2400 (2) potassium about 0 - 90 2+ (3) calcium about 0 - 60 ( .4..) magnesium2+ about 0 - 15 Total Anions about 1 - 2400 (5) chloride about 0 - 2000 (6) bicarbonate about 0 - 465 z— (7) inorganic phosphate about o- - 22 (8) 1-lactate .+ pyruvate about 0 — 465 (9) d-B Hydroxybutyrate~+ acetoacetate 0 - 465 (10) non diffusible z- polyanionate in mEq/L 0. - 4200 Total Nonionics about 0 575 (11) carbon dioxide about 0 25 (12) osmotically active substance about 0 - 550 the interrelationships between components range from about: -mEq ratio of [bicarbonate ]/[C0 2 ] 0.1/1 ■ - 55/0.1 -mEq ratio of [1-lactate ]/[pyruvate-] 20/1 - 1/1 -mEq ratio of [d-hydroxybutyrate~]/ acetoacetate 3 6/1 - 0.5/1 -total of (6) , (8) , (9) , (10) 0.2 - 465 -milliosmolarity 260 - ' 5000 -pH 5 - 9 -41- 3. The solution of claim 2 additionally about 20 to 44 percent by volume of red blood cells. 4. The solution of claim 2 in admixture with from 5 to about 70 gms per liter of material selected from the group consisting of dried plasma, dried albumin, gamma globulin, fibrinogen, and anti hemophillic globulin. 5. The solution of claim 2 wherein said non-diffusible polyanionic material is selected from the group consisting of albumin, sodium albuminate, carboxy- methyl starch, carboxy methyl, cellulose, carboxy methyl dextran, gamma polyglutamate, red cells, polyacrylamides, and polysulfonates. •6. The solution of claim 2 wherein said non-diffusible polyanionic material is a naturally occuring proteinaceous substance. 7. The solution of claim 6 wherein said proteinaceous substance is one which occurs in blood and blood products. 8. The solution of claim 2 wherein said non- diffusible polyanionic material contains pendant acidic groups.

Inventors: VEECH RICHARD L, VEECH, RICHARD, L.
Applicants: VEECH RICHARD L, VEECH, RICHARD, L.
Application year: 1985
Main classification: A61K33
IPC codes: A61K33, A61M1