Recent Trends in Methods & Calculations of Adjusting Isotonicity

Ajay Kumar*, V.K. Maurya

R. K. Pharmacy College, Kashipur, Surai, Sathiaon, Azamgarh, India

Received: 01-Mar-2018 , Accepted: 15-Aug-2018

Keywords: Lachrymal fluid, Ophthalmic solution, Isotonicity, Hypertonic, Colliery, NaCl solution

Full-Text PDF      

Google Scholar  

How To Cite       


Lachrymal fluid is isotonic with blood having Associate in nursing isotonicity value like that of a 0.9% NaCl resolution. Ideally, Associate in nursing ophthalmic resolution ought to have this isotonicity value; however the attention will tolerate isotonicity values as low as that of a 0.6% NaCl resolution and as high as that of a two NaCl resolution while not marked discomfort. Some ophthalmic solutions square measure essentially hypertonic so as to reinforce absorption and to supply a level of the active ingredients robust enough to exert a prompt and effective action. The quantity of such resolution used is little as a result of, on administration; the dilution with lachryma fluid takes place apace with stripped discomfort from the hyper tonicity that is merely temporary. However, any adjustment toward isotonicity by dilution with tears is negligible wherever massive volumes of solutions square measure used as colliery to clean the eyes. It is, therefore, vital that solutions used for this purpose be around isotonic.

1 Introduction

Tonicity could be a live of effective osmolarity in cell biology. Osmolarity and osmolarity area unit properties of a specific answer, freelance of any membrane. Osmolarity could be a concentration scale to precise the entire concentration of matter particles and is directly associated with any of the four colligative properties. it`s derived from concentration by resolution within the dissociation ofelectrolytic solutes tension could be a property of an answer in respect to a specific membrane, Associate in Nursingd is capable the total of the concentrations of the solutes that have the capability to exert an diffusion force across the membrane tension depends on matter porosity. The permeable solutes don`t have an effect on tension. If a semi-permeable membrane is employed to separate solutions of various matter concentrations, a development referred to as diffusion happens to determine concentration equilibrium. The pressure driving this movement is named force per unit area and ruled by the quantity of particles of substance in a very resolution. If substance could be a non-electrolyte, then range of particles is decided alone by the substance concentration. If the substance is a solution, the amount of particles is ruled by the concentration and degree of dissociation of the substance.

The distinction between the isotonic and isotonic terms comes with the belief that red somatic cell membrane don`t seem to be good semi-permeable membranes however enable passage of some solutes, like alcohol, sal ammoniac,   glycerin, water-soluble vitamin, acid etc. As carboxylic mentioned  earlier  a pair of resolution of element acid, once physically  measured,  is  found  to be isotonic (containing a  similar range of  particles)  with blood  and not  isotonic(exerting  equal pressure  or  tone)  with blood however is  isotonic with tears. This distinction doesn`t have any nice significance and, therefore, isotonicity values are calculated on the premise of the amount of particles in answer. The clinical significance of all this can be to insure that isotonic or isotonic solutions don`t harm tissue or turn out pain once administered.

2 Tonicity is usually classified in 3 types


A solution having higher osmotic pressure than the body fluids (0.9% NaCl) is known as hypertonic solution. These solutions draw water from the body tissues to dilute and establish equilibrium. An animal cell in a hypertonic environment is surrounded by a higher concentration of impermeable solute than exists in the inside of the cell.

For example, if 2% NaCl solution is added to blood (defibrinated),osmotic pressure directs a net movement of water out of the cell, causing it to shrink (the shape of the cell becomes distorted) and wrinkled (crenated),as water leaves the cell. This movement is continued until the concentrations of salt on both sides of the membrane are identical. Hence, 2% NaCl solution is hypertonic with the blood.


Solutions that have identical force per unit (area) as that of bodily fluids are afore said to be isotonic with the body fluid. Body fluids like blood and tears have pressure comparable to that of zero.9% NaCl or glucose binary compound solution; therefore, a 0.9%NaCl or five the troubles, glucose resolution is named as isotonic or  isotonic.  The term isotonic means that equal tone, and is employed interchangeably with isotonic with regard to specific body fluids. For example, a 0.9% w/v resolution of NaCl in water is taken into account to be isotonic in relevancy RBC’s and their semi-permeable membrane.

Requirements of isotonic solutions square measure that they need to not cause any contraction or swelling of the tissues.

The product should not turn out discomfort once put in within the eye, nasal tract, blood, or alternative body tissue. On addition of zero.9gm NaCl/100ml (0.9%) in to blood (defibrinated), the cells retain their traditional size. solution ought to be restricted to solutions having equal force per unit area with relation to a specific membrane.

The addition of any compound to an answer affects its isotonicity, inflicting changes in force per unit area of an answer. It shouldn`t be affected solely by medication however additionally by any buffer parts value-added within the formulation. Therefore, it`s necessary to feature extra Nacl to bring the answer to isotonicity. Adjustment of isotonicity is needed for many indefinite quantity forms like epithelial duct solutions, e.g., IV infusions, irritating solutions, lotions for open wounds, body covering injections, preparations meant for diagnostic applications, solutions meant for intra thecal injections, nasal drops and ophthalmic drops.


A answer with low pressure than body fluids is thought as hypotonic solution. The consequences of administering a hypotonic answer square measure usually additional severe than with hypertonic solutions, since busted cells will repaired. Hypertonic solutions show the other impact compared to hypertonic solutions wherever cyber web movement of water into the cell causes them to swell. If the cell contains additional tight substance than its surroundings, water enters it just in case of animal cells, they swell till they burst; however this doesn’t happen to plant cells, i.e., they are do not burst as a result of the reinforcement their cyto membrane provides. If 0.2% NaCl answer is accessorial to blood (defibrinated), the cells swell and burst. Therefore, 0.2% NaCl answer is hypotonic with relevance the blood (Fig 1).

3 Method for determine tonicity value

Many chemicals and medicines area unit utilized in pharmaceutical formulations. These substances contribute to the tautness of the answer. Hence, strategies area unit required to verify the tautness and alter isotonicity. 2 strategies accustomed verify tautness valve area unit delineate below

Hemolytic Method

Isotonicity valve is calculated by exploitation the lysis methodology during which the result of varied solutions of drug is ascertained on the looks of red blood cells suspended in answer. During this methodology, RBC suspended in numerous solutions and also the look of RBC’s is ascertained for swelling, bursting, shrinking and wrinkling of the blood cells. In hypertonic solutions, the oxyhemoglobin discharged is proportional to the amount of cells haemolysed; just in case of hypertonic solutions, the cells shrink and become wrinkled or scalloped whereas just in case of isotonic solutions the cells don`t modification their morphology.

Cryoscopy Method

Isotonicity values are determined from the colligative properties of the solutions. For this purpose, melting point depression property is most extensively used. The melting point of water is 0 áµ’C, and once any substance like NaCl is added to that, the melting point of water decreases. The physical change point of depression of blood is -0.52 áµ’C. Hence, the drug resolution should be -0.52 áµ’C.

This solution shows vapor pressure level capable the blood.

4 Method adjustment of isotonicity

Several strategies are accustomed change the isotonicity of pharmaceutical solutions. Isotonicity may be calculated from the colligative properties of drug solutions. If solutions ar injected or introduced in to eyes and nose, these ar to be created isotonic so as to avoid hematolysis of RBC’s and to avoid pain and discomfort. This can be potential for either factory-made or impromptu ready solutions. By victimization the acceptable calculations supported colligative properties of solutions; it`s simple to work out the number of adjusting agents to be accessorial. It helps to beat the aspect effects caused from administering solutions that contain adjusting agents less or quite isotonic solutions. The 3 of times used strategies to calculate isotonicity of the solutions is delineated below.

Class-1 Methods

Nacl or another substance is accessorial to the answer of the drug to lower thetemperature of the answer to -0.52áµ’C and so build the answer isotonic.

Examples of this class-

1)   Cryoscopic technique

2)   Sodium chloride equivalent technique.

Class-2 Methods

Water is accessorial to the drug during a sufficient quantity to form it isotonic.Then the preparation is delivered to its final volume with associate degree isotonic or buffered isosmotic solution.

Cryoscopic technique

In this methodology, the number of every substance needed for associate degree solution is often calculated from the melting point depression values. An answer that is isotonic with blood encompasses a ∆Tf of 0.52ᵒC. Therefore, the melting point of drug answer should be adjusted to the present valve. Several pharmaceutical text books typically list the melting point of depression required to attain isotonicity from these values just in case of drug solutions, if it`s impracticable to regulate tension by sterilization the drug concentration, then associate degree adjusting substance is additional to attain desired tension (Table 1).

The weight (in grams) of adjusting substance is often calculated in manner represented below for instance, the drug concentration in 100ml answer could be grams, then

∆Tf  ( For drug solution) =ax

∆Tf  of 1% drug solution = x

If w be in grams of the adjusting substance to be accessorial to a hundred cubic centimeter of drug resolution to create it isotonic then:

∆Tf  (For adjusting solution) = w × âˆ†Tf of 1% adjusting substance

 = w × b

For isotonic solution


x + wb = 0.52

        w = 0.52 – x/b

If NaCl is used as adjusting substance whose ∆Tf of solution is 0.580C than w = 0.52 – x/0.58

Sodium chloride equivalent method

Tonicity equivalent or common salt equivalent methodology is employed to regulate the tautness of pharmaceutical solutions. Sodium chloride equivalent (E) of a drug is that the quantity of common salt that`s corresponding to one gram of the drug. The percent of binary compound needed for adjusting the isotonicity is calculated victimization the subsequent equation.

                PSA = 0.9 – (PSM × E of medicament)


PSM = Percent strength of medicament

PSA = Percent of sodium chloride for a adjustment of isotonosity

L ISO Method

The E NaCl worth of tonus adjusting substances may be calculated from the substances. The L iso values of the tonus adjusting substances square measure given in table and square measure mentioned as constants in several references.

In this technique, the melting point depression equation is employed to calculate the quantity of the isotonicity.

Above equation is employed to calculate the number of adjusting substance (sodium chloride) needed for creating the answer isotonic. it`s valid for one hundred metric capacity unit resolution (Table 2).

5 Conclusion

Tonicity is a measure of the effective osmotic pressure gradient, as defined by the water potential of two solutions separated by a semipermeable membrane. In other words, tonicity is the relative concentration of solutes dissolved in solution which determine the direction and extent of diffusion. It is commonly used when describing the response of cells immersed in an external solution.Unlike osmotic pressure, tonicity is influenced only by solutes that cannot cross the membrane, as only these exert an effective osmotic pressure. Solutes able to freely cross the membrane do not affect tonicity because they will always be in equal concentrations on both sides of the membrane. It is also a factor affecting imbibition.

6 Conflict of interests


7 Author’s contributions

AK carried out literature review and draft the manuscript. VKM participated in collection of data. Both authors read and approved the final manuscript.

8 References

  1. United States Pharmacopeial Convention, Inc. United States Pharmacopeia 29- National Formulary 24. Rockville MD: U.S. Pharmacopeial Convention, Inc. 2005; 3261.
  2. Price JC. Glycerin. In: Rowe RC, Sheskey PJ and Owen SC, eds. Handbook of Pharmaceutical Excipients. 5th ed. Washington DC: American Pharmaceutical Association. 2005; 301-303.
  3. Day A. Dextrose. In: Rowe RC, Sheskey PJ and Owen SC, eds. Handbook of Pharmaceutical Excipients. 5th ed. Washington DC: American Pharmaceutical Association. 2005; 231-233.
  4. Armstrong NA. Mannitol. In: Rowe RC, Sheskey PJ and Owen SC, eds. Handbook of Pharmaceutical Excipients. 5th ed. Washington DC: American Pharmaceutical Association. 2005; 449-453.
  5. Owen SC. Potassium Chloride. In: Rowe RC, Sheskey PJ and Owen SC, eds. Handbook of Pharmaceutical Excipients. 5th ed. Washington DC: American Pharmaceutical Association. 2005; 600-602.
  6. Emmens CW. The motility and viability of rabbit spermatozoa at different hydrogen-ion concentrations. J Physiol. 1947; 106(4): 471–481.
  7. Pholpramool C, Chaturapanich G. Effect of sodium and potassium concentrations and pH on the maintenance of motility of rabbit and rat epididymal spermatozoa. J Reprod Fertil. 1979; 57(1): 245–251.
  8. Bagger PV, Byskov AG, Christiansen MD. Maturation of mouse oocytes in vitro is influenced by alkalization during their isolation. J Reprod Fertil. 1987; 80(1): 251–255.
  9. Downs SM, Mastropolo AM. Culture conditions affect meiotic regulation in cumulus cell-enclosed mouse oocytes. Mol Reprod Dev. 1997; 46(4): 551– 566.
  10. Leclerc C, Becker D, Buehr M, Warner A. Low intracellular pH is involved in the early embryonic death of DDK mouse eggs fertilized by alien sperm. Dev Dyn. 1994; 200(3): 257–267.
  11. Owen SC. Sodium Chloride. In: Rowe RC, Sheskey PJ and Owen SC, eds. Handbook of Pharmaceutical Excipients. 5th ed. Washington DC: American Pharmaceutical Association. 2005; 671-674.
  12. Lane M, Bavister BD. Regulation of intracellular pH in bovine oocytes and cleavage stage embryos. Mol Reprod Dev. 1999; 54(4): 396–401.
  13. Lane M, Lyons EA, Bavister BD. Cryopreservation reduces the ability of hamster 2-cell embryos to regulate intracellular pH. Hum Reprod. 2000; 15(2): 389–94.
  14. Edwards LJ, Williams DA, Gardner DK. Intracellular pH of the preimplantation mouse embryo: effects of extracellular pH and weak acids. Mol Reprod Dev. 1998; 50(4): 434–42.
  15. Squirrell JM, Lane M, Bavister BD. Altering intracellular pH disrupts development and cellular organization in preimplantation hamster embryos. Biol Reprod. 2001; 64(6): 1845–54.
  16. Zander-Fox D, Mitchell M, Thompson JG, Lane M. Alterations in mouse embryo intracellular pH by DMO during culture impair implantation and fetal growth. Reprod Biomed Online. 2010; 21(2): 219–29.
  17. Fitzharris G, Baltz JM. Granulosa cells regulate intracellular pH of the murine growing oocyte via gap junctions: development of independent homeostasis during oocyte growth. Development. 2006;133(4): 591–9.
  18. FitzHarris G, Siyanov V, Baltz JM. Granulosa cells regulate oocyte intracellular pH against acidosis in preantral follicles by multiple mechanisms. Development. 2007;134(23):4283– 95.
  19. Phillips KP, Baltz JM. Intracellular pH regulation by HCO3-/Cl- exchange is activated during early mouse zygote development. Dev Biol. 1999; 208(2): 392–405.