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A useful microscopic screening estimate compares thermal energy with the buoyant gravitational potential energy over one particle radius. Write $\Delta \rho =50\mathrm{kg}{\mathrm{m}}^{-3}$ and $m_b=(4\pi /3)a^3\Delta \rho$. The sedimentation length is ${\ell }_g=k_{B}T/(m_{b}g)$, and the gravitational Peclet number isUsing $T=298\mathrm{K}$ and requiring ${\mathrm{Pe}}_g\lesssim 1$ givesEquivalently, combining Stokes law speed $v_s=2\Delta \rho g{a}^2/(9\eta )$ and diffusion coefficient $D=k_{B}T/(6\pi \eta a)$ gives $v_{s}a/D={\mathrm{Pe}}_g$. Buoyancy, not the full polystyrene density, is essential to the estimate. Taking a diameter rather than radius as the comparison length changes the numerical cutoff by an order-one factor.

There is no universal largest particle that stays uniformly suspended indefinitely. If uniformity over a vessel height $H$ is required, use ${\ell }_g\gtrsim H$, not merely ${\ell }_g\gtrsim a$. For example $H=1\mathrm{cm}$ gives a much smaller radius, about 58 nm. Observation time, density matching, convection, particle polydispersity and shape also matter. Independent of sedimentation, electrostatic/steric barriers must prevent aggregation; low ionic strength, suitable surface chemistry and clean solvent can be as important as size.

For this self-assembly into a colloidal crystal, four useful elements are building blocks, interactions, environment and reversible dynamics. The building blocks here are nearly monodisperse polymer spheres; their shape and size distribution constrain the accessible packing. Their interactions include excluded volume, Van der Waals forces and electrostatic or steric stabilization, which set the free energy landscape. The aqueous environment supplies temperature, viscosity, ionic strength, confinement and concentration controls. Finally, Brownian motion and rotation must let particles encounter one another and rearrange or detach from incorrect contacts. Attractions that bind every collision irreversibly favour an amorphous aggregate rather than an annealed colloidal crystal. These four categories include both thermodynamic selection and the transport/kinetic access needed to realize it.

Flocculation indicates that the attraction/kinetic balance has changed: electrolyte may screen a double-layer barrier; pH may reduce surface charge; multivalent ions can promote aggregation; added polymer can bridge particles or cause depletion force; poor solvent conditions can collapse stabilizing layers. Rapid evaporation concentrates both particles and salts and can force contacts faster than they anneal. Aggregates then sediment much faster because their effective buoyant size has grown.

Check ionic strength, pH, surface charge and the particle size distribution before and during assembly. Dialyse or reduce salt where appropriate, restore surface charge, add a nonbridging steric stabilization with a compatible solvent, and slow the concentration step. Compare untreated and deliberately salt-screened controls. Restoring repulsion after irreversible aggregation may not redisperse the particles, so prevention is preferable to assuming all settled material can be recovered.

The Debye–Hückel approximation linearizes the hyperbolic sine, giving $({\nabla }^2-{\kappa }^2)\psi =0$ withThe planar Debye-Hückel double layer has $\psi (s)={\psi }_0{e}^{-\kappa s}$. For an isolated sphere of radius $a$, the Debye-Hückel potential of a charged sphere is $\psi (r)={\psi }_0(a/r)e^{-\kappa (r-a)}$ outside the sphere; the factor $a/r$ accounts for spherical spreading as well as screening.

The Debye–Hückel screening length is the characteristic distance over which the ion cloud suppresses a weak surface potential. Adding salt increases ionic strength and therefore increases $\kappa$, shortening the interaction range. Linearization requires $|q\psi |\ll k_{B}T$, particularly near the surface where the potential is largest. It means the ionic electrostatic energy is small compared with thermal energy; for monovalent ions near room temperature, $k_{B}T/e\simeq 26\mathrm{mV}$. This condition alone does not remove other limitations of point-ion mean-field theory at high concentration or strong correlations.

In the planar Debye–Hückel approximation, a surface potential decays as $\psi (x)={\psi }_s{e}^{-\kappa x}$, with $\kappa \propto \sqrt{I}$ at fixed temperature and solvent permittivity; $I$ is the ionic strength. For a monovalent salt, reducing its concentration by ten reduces $\kappa$ by $\sqrt{10}$. Calibrating the first decay from the supplied potential ratio, and assuming unchanged ${\psi }_s$, givesHere the printed meV unit is interpreted as the electrostatic potential energy of a singly charged probe; the same ratio calculation applies to potentials in mV.

This numerical answer uses an effective exponential decay calibrated from the data. The inferred initial Debye length is $5/\log (35/6.6)=3.00\mathrm{nm}$, whereas the standard dilute aqueous-electrolyte expression at room temperature gives about $0.96\mathrm{nm}$ for $0.1\mathrm{M}$ monovalent salt. The numbers therefore are not consistent with that absolute ideal-electrolyte model. Moreover a spherical colloid has an additional radius-dependent geometric factor, and its surface potential need not remain constant when salt is changed. Without a surface boundary condition and geometry, the new potential is not uniquely specified beyond the stated planar, fixed-potential approximation.

For monovalent mobile ions, the linearized Debye–Hückel screening length isHere $n_i$ is a number density in ${\mathrm{m}}^{-3}$, not a molarity. Use the question's idealized neutral-water convention, $c_{+}=c_{-}=10^{-7}\mathrm{mol}{\mathrm{L}}^{-1}$, and $T=293.15\mathrm{K}$. Both ion species contribute:For added salt concentration $c_s$ in $\mathrm{mol}{\mathrm{L}}^{-1}$, the ionic strength in this model is $I=c_s+10^{-7}\mathrm{mol}{\mathrm{L}}^{-1}$. With the numerical molarity $I_M=I/(1\mathrm{mol}{\mathrm{L}}^{-1})$,The water contribution is negligible at that salt level. The factor $10^3$ converts molarity into $\mathrm{mol}{\mathrm{m}}^{-3}$; omitting it or counting only one ion species gives a wrong screening length. Exact water autoionization and activities vary with temperature, so the first estimate uses the simplified neutral $10^{-7}\mathrm{M}$ concentrations implied by the question. At the higher salt level, ion-size effects, nonideal activities and changes in permittivity limit the numerical accuracy of this continuum estimate.

At fixed temperature and solvent permittivity, the Debye–Hückel screening length satisfies ${\lambda }_D^{-2}\propto {\sum }_i{c}_i{z}_i^2=2I$, where $I=\frac{1}{2}{\sum }_i{c}_i{z}_i^2$ is ionic strength. Fully dissociated sodium chloride has two monovalent ion species, giving $I_1=0.01$ M. Fully dissociated magnesium sulfate has two divalent species, givingThe multivalent-electrolyte screening-length ratio is thereforeThe squared valences are essential: counting only the total ion concentration misses a factor of four in the screening strength per formula unit. This is the comparable linear-screening estimate, assuming unchanged dielectric response and full dissociation. ion pairs and concentrated-electrolyte nonideality can make a real divalent-salt solution depart from it; no such correction is specified.

For an ideally dissociated electrolyte, the Debye–Hückel screening length isHere $n_i$ is ionic number density, $z_i$ the charge in units of the elementary charge, and $c_i$ is molar concentration. Conversion between $I$ and number density cancels in a ratio. At fixed relative permittivity, ${\lambda }_D\propto \sqrt{T/I}$.

In $0.01\mathrm{M}$ sodium chloride, $c_{\mathrm{N}{\mathrm{a}}^{+}}=c_{\mathrm{C}{\mathrm{l}}^{-}}=0.01\mathrm{M}$, giving $I_1=0.01\mathrm{M}$. Complete dissociation of $0.1\mathrm{M}$ aluminium chloride gives $c_{\mathrm{A}{\mathrm{l}}^{3+}}=0.1\mathrm{M}$ and $c_{\mathrm{C}{\mathrm{l}}^{-}}=0.3\mathrm{M}$, soThe factor is sixty, not ten: the squared charge and the three counterions per formula unit both matter. Using the given $5\mathrm{nm}$ reference length directly, and $T_1\simeq 298\mathrm{K}$, $T_2=323\mathrm{K}$,The modest temperature increase raises the screening length by only about $4\%$ at fixed concentration; the large increase in ionic strength shortens it by almost an order of magnitude overall.

Under charge stabilization of colloids, the electrostatic double-layer repulsion now has a much shorter range. Attractive Van der Waals forces can then dominate at closer separations, reducing the barrier to particle aggregation in DLVO theory. Thus an electrostatically stabilized colloid is likely to be destabilized, with aggregation more likely than in the original dispersion.

This is a conditional physical prediction, not a proof of instability from a length alone. No particle radius, surface potential, attractive interaction strength or steric coating is specified. Steric stabilization could preserve stability, and trivalent-ion adsorption can alter the surface charge. Moreover, ideal screening theory is only an estimate at $I\simeq 0.6\mathrm{M}$, where ion association, hydrolysis and activities can matter. The numerical result uses the supplied reference length and the stipulated unchanged permittivity, rather than replacing either with an unprovided calibration.

Stabilization and screening. Charge stabilization of colloids means suppressing aggregation through repulsion between similarly charged particles and their electric double layers. In DLVO theory, this repulsion competes with attractive Van der Waals forces. A substantial repulsive barrier compared with thermal energy $k_{B}T$ can make aggregation slow, even when the dispersed state is only metastable.

The Debye–Hückel screening length ${\lambda }_D={\kappa }^{-1}$ is the characteristic range over which a weak electric potential is attenuated by rearrangement of mobile ions. It measures the thickness of a diffuse screening cloud, not a molecular diameter. For a plane, a weak potential decays as $e^{-\kappa x}$ into the bulk.

Derivation and the source convention. Put a planar interface at $x=0$, with electrolyte in $x>0$, bulk potential zero, and absolute permittivity $\epsilon ={\epsilon }_0{\epsilon }_r$. For a symmetric $z:z$ electrolyte, let $n_{\infty }$ be the bulk number density of each ionic species. The Boltzmann distribution givesso the mobile charge density isSubstitution into the SI Poisson equation, ${\nabla }^2\psi =-{\rho }_e/\epsilon$, gives the Poisson-Boltzmann equationFor $|ze\psi |\ll k_{B}T$, linearize the hyperbolic sine. The Debye–Hückel approximation is thenHere $z$ is the ionic valence magnitude, $e$ the elementary electric charge, $k_B$ the Boltzmann constant, $T$ absolute temperature, ${\epsilon }_0$ the vacuum permittivity, and ${\epsilon }_r$ the solvent's relative permittivity. For several ionic species, the numerator $2z^2{e}^2{n}_{\infty }$ is replaced by $e^2{\sum }_i{z}_i^2{n}_{i,\infty }$.

The original PDF includes an additional $4\pi$ in the denominator. That factor does not follow from SI Poisson equation with number densities and the stated permittivities. The Bjerrum length provides a useful check: ${\ell }_B=e^2/(4\pi \epsilon k_{B}T)$ and the consistent result is ${\kappa }^2=8\pi {\ell }_B{z}^2{n}_{\infty }$. A Gaussian-unit convention moves factors of $4\pi$ together with the charge and permittivity conventions; it does not justify inserting this extra factor while retaining all the given SI quantities. Both numerical interpretations are reported below rather than silently changing the source.

Conditions favoring stability. Increasing the surface-potential or surface-charge magnitude usually strengthens the electrostatic barrier. Lower ionic strength increases ${\lambda }_D$ and generally makes repulsion longer ranged; added salt and especially multivalent counterions shorten it and can promote aggregation. Suitable pH can keep an ionizable surface charged, while conditions near an isoelectric state weaken charge repulsion. The solvent-dependent Van der Waals forces, charge regulation and particle geometry also affect the barrier, so a long screening length is not by itself sufficient. At fixed density and valence, the linear formula gives ${\lambda }_D\propto \sqrt{\epsilon T}$, but actual temperature changes can also alter the permittivity and surface chemistry.

Surface charge at small potential. Take the usual one-sided planar-interface model, with negligible electric displacement on the nonaqueous side. The solution decaying into the solution isThe electric field is $E_x=-{\psi }^{\prime }=\kappa {\psi }_0{e}^{-\kappa x}$. The electrostatic interface boundary conditions then give the surface charge densityOne can also derive it by integrating the diffuse charge density:Thus the diffuse layer neutralizes the surface charge. This is the planar Debye-Hückel double layer. If instead the object is an isolated sheet wetted by identical electrolyte on both sides, each side screens half its total charge and ${\sigma }_{\mathrm{sheet}}=2\epsilon \kappa {\psi }_0$. The one-sided assumption is the conventional local model for the exterior of a large charged colloid; it must be stated because the question does not specify the material on the other side.

Numerical ion density. For $T=298.15\mathrm{K}$, $z=1$, $\kappa =10^9{\mathrm{m}}^{-1}$ and $\epsilon =80{\epsilon }_0=7.0834\times 10^{-10}\mathrm{F}{\mathrm{m}}^{-1}$, the consistent SI result isThis is $\boxed{0.0943\mathrm{mol}{\mathrm{L}}^{-1}}$ for each ion species, and hence also the salt concentration for a monovalent $1:1$ salt. The combined number density of both signs is $2n_{\infty }$.

Literal substitution into the formula printed in the PDF instead givesThese values are distinct consequences of the source's extra factor; they cannot both describe the same physical screening length in SI with the same definition of $n_{\infty }$.

Surface potential and validity. The requested linear estimate in the one-sided model isHowever, the thermal voltage is $k_{B}T/e=25.7\mathrm{mV}$, so $e{\psi }_{0,\mathrm{linear}}/(k_{B}T)\simeq 13.7$. Thus the small-potential approximation is not self-consistent for the supplied surface charge. The boxed value is its formal extrapolation, not a reliable prediction of the actual surface potential.

For a valid nonlinear comparison within the ideal one-sided planar Poisson-Boltzmann equation, multiply the differential equation by ${\psi }^{\prime }$ and use zero potential and field in the bulk. This givesUsing the surface boundary condition yields the Grahame equation and its inverse:With the SI screening length, the latter gives $\boxed{{\psi }_{0,\mathrm{PB}}\simeq 0.135\mathrm{V}}$. This is a nonlinear mean-field estimate under explicit assumptions, not a unique experimentally determined potential: compact interfacial layers, finite ion size and surface chemistry were not supplied. The source's normalization problem and its use of a linear formula far outside the small-potential regime are separate issues.

One method is to give every particle a sufficiently large surface charge density of the same sign. For example, attach ionizable sulfonate groups covalently to the surface. Dissociation leaves negatively charged surface groups and releases mobile positive counterions into the water. A surfactant with a strongly bound head group can provide another charging route, provided it does not desorb during the experiment.

Each charged particle is surrounded by an electric double layer. As two such particles approach, overlap of these ionic clouds raises the electrostatic and ionic free energy, producing repulsion. In a dilute electrolyte, the range is set by the Debye–Hückel screening length,For identical weakly charged spheres a representative screened Coulomb pair potential isIts sum with the Van der Waals force attraction is the DLVO theory pair potential. Choose the charging level so that the repulsive barrier is many $k_{B}T$ high; particles then rarely reach the primary attractive well during the observation time. This is kinetic stabilization rather than removal of all attraction.

Deionized water is useful because low added ionic strength gives a long-ranged repulsion. It does not mean that the charged dispersion contains no ions: the surface's own counterions are unavoidable and contribute to screening. Surface dissociation, contamination and particle concentration therefore matter even without added salt. An alternative valid route is a dense, firmly attached layer of water-soluble polyethylene glycol, producing steric stabilization without relying on a large electrostatic barrier.

Let $\epsilon ={\epsilon }_0{\epsilon }_r$ be the permittivity, $\beta =(k_{B}T{)}^{-1}$, $e>0$ the elementary charge, and $c$ the bulk salt formula-unit number density. The Boltzmann distribution givesConsequently the exterior Poisson-Boltzmann equation isFor small $Q$, meaning $2\beta e|\varphi (a)|\ll 1$, linearize the exponentials. The Debye–Hückel approximation givesThis is also ${\kappa }^2=\beta e^2{\sum }_i{z}_i^2{n}_{i,\infty }/\epsilon$: the divalent cation contributes $4c$ and the two monovalent anions contribute $2c$. In terms of molar salt concentration $C$ in $\mathrm{mol}{\mathrm{m}}^{-3}$, replace $c$ by $N_{A}C$; if $C$ is in $\mathrm{mol}{\mathrm{L}}^{-1}$, use $c=10^3{N}_{A}C$. The molar ionic strength is $3C$ in the same concentration units.

The boundary conditions are $\varphi (\infty )=0$ and, by Gauss's law,Here the surface charge is uniform and the spherically symmetric interior field is zero. Set $u=r\varphi$; the radial equation reduces to $u^{\prime \prime }={\kappa }^{2}u$. Decay excludes the growing exponential, so write $\varphi =B{e}^{-\kappa (r-a)}/r$. Then ${\varphi }^{\prime }(a)=-B(1+\kappa a)/a^2$, givingIt solves the linearized Poisson-Boltzmann equation, not the full nonlinear equation at finite $Q$. The radial ion profiles, consistently to first order, areThe exponential Boltzmann distribution formulas above express the full mean-field relation, but substituting a linearized potential into them is accurate only to the retained order. A useful electroneutrality check isThe result is the Debye-Hückel potential of a charged sphere.

First hold the surface electric charge fixed and increase the bulk salt concentration. Since ${\kappa }^2\propto c$, the Debye–Hückel screening length decreases as $c^{-1/2}$. For surface gap $h=R-2a\ge 0$,Thus both the range and the strength of repulsion decrease. In the repulsive phase diagram, the sample moves downwards at fixed volume fraction and can melt from a crystal into a fluid. Because screening length is already a diagram axis, changing salt moves the state through that diagram rather than independently changing its fixed-charge coexistence curves. If Van der Waals forces become dominant, however, adding salt may instead destabilize the dispersion and cause aggregation. These are distinct physical regimes.

Second increase $\sigma$ at fixed salt concentration and particle size. The electric charge is $Q=4\pi a^2\sigma$, so the pair repulsion increases approximately as ${\sigma }^2$ within the Debye–Hückel approximation, while the reservoir screening length remains unchanged. Freezing then becomes possible at lower volume fraction or shorter screening length. At fixed ${\lambda }_D$, the coexistence curves shift towards lower $\eta$; a previously fluid sample may crystallize.

These statements assume fixed, uniform charge and a salt reservoir dominating the screening. Strong charging requires nonlinear screening and possibly charge regulation. At very low added salt or appreciable particle concentration, particle-released counterions also contribute to ionic strength, so the screening length is no longer independently fixed by the added salt alone.

In gel sieving of synthetic polymers, gel electrophoresis combines drift under an electric field with size- and conformation-dependent obstruction by a porous matrix. In weak-field free solution, force balance gives $v=\mu E$ and, in a simple effective-charge/effective-friction model, $\mu =q_{\mathrm{eff}}/\zeta$. The mobility is motion relative to the medium, so fluid motion from electro-osmosis must be accounted for separately. This is the operational mobility convention, not a claim that the bare structural charge is always the effective driving charge.^[ref]

For a chain with free-draining polymer friction with approximately constant effective electric charge per segment, both charge and friction scale with $N$. The free-solution mobility can therefore become nearly independent of length, making size separation poor. A gel adds steric constraints. Coils smaller than the mesh can pass relatively easily; larger coils require deformation and pore-to-pore rearrangement. A schematic weak-sieving law is $\mu (c_g)\simeq \mu (0)e^{-K_r{c}_g}$, where matrix concentration $c_g$ and retardation coefficient $K_r$ are defined by a calibration; it is not a universal size law.

When a long flexible charged polymer is confined to a tube of available paths, reptation supplies another regime. An additive-friction model gives contour diffusion $D_c\propto N^{-1}$ and renewal time $\tau \sim N^2/D_c\propto N^3$. For ideal large-scale chain statistics, $R^2\propto N$, so centre-of-mass diffusion scales as $N^{-2}$. With total effective driving charge proportional to $N$, weak-field mobility can then scale as $N^{-1}$. This assumes screened/ideal large-scale conformations and linear response; strong-field alignment, trapping and nonideal matrices can alter the scaling.

Operationally, load narrow zones of independently characterized synthetic standards and sample into a controlled gel, apply a known field and detect migration positions after a measured time. Compare with a free-solution measurement, vary matrix concentration and field, and control temperature, ionic strength, pH and matrix charge. Larger gel pores or lower matrix concentration usually reduce sieving within a given calibrated material system. Excessive field causes Joule heating and can change conformation and mobility. adsorption or electro-osmotic drift can mimic a size effect; standards should match charge chemistry and topology as well as nominal mass.

Band centres move as $\mu Et$ while ordinary diffusion gives ${\sigma }_x^2(t)={\sigma }_x^2(0)+2Dt$. Resolution depends on separation of centres relative to band widths, not travel distance alone. A neutral tracer helps identify bulk flow, and multiple concentrations test adsorption or overloading. A mobility or band position does not uniquely determine size without a regime-specific calibration. TODO: biomolecular examples, sample preparation and biological applications are user deferred; the account here concerns synthetic charged polymers only.

Force peaks in membrane-protein extraction can reflect steric barriers, membrane anchoring and electrostatic interactions. Changing ionic strength changes screening and possibly specific ion binding; it need not weaken every barrier in the same direction. The rising branch of an exposed chain and rupture of a remaining structural barrier are different physical processes.

An authored manufacturer experiment used an Agilent 1260 Infinity Bio-inert Quaternary LC with Bio SEC-3 $300\mathring{\mathrm{A}}$ columns, $7.8\mathrm{mm}$ diameter and $150$ or $300\mathrm{mm}$ length. Four pump channels mixed water, sodium chloride, and two phosphate solutions into $20\mathrm{mmol}{\mathrm{L}}^{-1}$ phosphate at pH $7.0$, varying salt from $0.2$ to $0.6\mathrm{mol}{\mathrm{L}}^{-1}$. At $1.0\mathrm{mL}\mathrm{mi}{\mathrm{n}}^{-1}$ and $30{}^{\circ }\mathrm{C}$, the readouts were diode-array absorbance, differential refractive index, and scattering at $15^{\circ }$ and $90^{\circ }$. A supplied protein standard mixture calibrated retention; bovine serum albumin calibrated scattering constants and inter-detector delay. Increasing salt improved separation of two α-chymotrypsinogen A peaks; two $300\mathrm{mm}$ columns also resolved peaks that one column did not. At $0.5\mathrm{mol}{\mathrm{L}}^{-1}$ salt, the two scattering-derived estimates were about $26,100$ and $25,800\mathrm{g}\mathrm{mo}{\mathrm{l}}^{-1}$. The author inferred a possible size-changing conformation difference, rather than doubling of molar mass. The same poster shows delayed, tailing lysozyme peaks from unwanted interactions.^[ref]

The salt comparison and the independent mass readout constrain interpretation, but do not uniquely prove a particular protein structure. Salt can change protein folding, protein aggregation, and packing interactions together. In ideal SEC there is no adsorption; attraction from the hydrophobic effect or forces described by electrostatics can cause additional retention, tailing, and loss of recovered sample. Repulsion can instead reduce pore entry. A peak beyond $V_t$ contradicts the simple nonbinding model, rather than extending its size calibration.

Useful proposed controls compare a solvent blank, fresh calibration standards bracketing the sample, several injection amounts and volumes, and recovered sample quantity with injected quantity using an appropriately calibrated detector. Keep temperature and volumetric flow rate fixed while testing pH or ionic strength. Compare an independent mass readout before declaring an early peak an aggregate. Loss of an insoluble species during filtration, on-column association or dissociation, overloading, and broad unresolved peaks can all hide the original sample population. These are interpretation checks and limitations, not measurements invented for the figures.

The solubility product is the equilibrium constant for dissolution of a specified solid into its dissolved ions. For a pure bulk AB solid at the chosen temperature, pressure and standard states,The pure bulk solid's thermodynamic activity is one in this convention. Coexisting solid and chemical solution at chemical equilibrium give this equality. If all available solid dissolves, the final chemical solution may instead be undersaturated; do not impose equality merely because a salt was initially added.^[ref]

Using the thermodynamic activity definition $a_i={\gamma }_i{c}_i/c^{\circ }$, with activity coefficients ${\gamma }_i$ and reference molar concentration $c^{\circ }$, gives the independent conversionThus $K_{\mathrm{sp}}$ is dimensionless, while the conditional concentration form $K_{\mathrm{sp},c}$ has units ${\mathrm{M}}^2$ for this one-to-one model. Treating $K_{\mathrm{sp},c}$ as constant requires a justified fixed or approximately fixed activity coefficient product. Changing ionic strength can change that product even at fixed temperature; changing temperature can change the equilibrium constant itself. Specify the solid thermodynamic phase too: different crystal polymorphs need not have the same solubility product.

A spectator ion cancels from the chosen net ionic equation because its net inventory is unchanged. This label does not make it physically irrelevant: dissolved spectator ions contribute to ionic strength, which affects activity coefficients. Their concentrations can also change during dilution even though their amount of substance remains fixed.

A pH measurement becomes evidence for a weak acid chemical equilibrium when paired with a known analytical concentration and a calibrated glass electrode. For the pure-weak acid model, measured pH estimates $h=10^{-\mathrm{pH}}$ only in the ideal thermodynamic activity approximation. Then $o=K_w/h$, $x=h-o$, $y=c-x$, and $K_a=hx/y$. Require $0<x<c$; values outside that range signal inconsistent measurements or an inadequate model. Repeating this reconstruction across a dilution series tests whether one acid dissociation constant describes the data, rather than merely fitting one point.

The Analytical Sciences Digital Library's Potentiometric Titration of an Unknown Weak Monoprotic Acid provides a complementary acid-base titration experiment. Its apparatus includes a glass electrode and meter, a stirring magnet, a volumetric flask, pipettes, and a burette containing standardized sodium hydroxide. Prepare an aqueous acid stock of known analytical concentration, transfer a measured portion, and stir gently with the glass electrode clear of the magnet. Record added volume and stable pH after each addition; use a preliminary run to locate the steep region and smaller additions there on a fresh portion. The readout is the pH versus added volume curve, not a visual count of dissociated molecules.

Calibrate with fresh standard buffer solutions bracketing the pH range being interpreted. Use their assigned values at the actual temperature; a pair near pH $4$ and $7$ can cover the midpoint of many weak acid examples, but does not cover an entire alkaline endpoint region. Rinse between standards and samples to limit carryover, keep the sensing bulb and reference electrode junction immersed, and wait for a stable reading. These instrument practices are documented in HI5221 & HI5222 Instruction Manual: pH and mV and ISE and Temperature Bench Meters by Hanna Instruments (pp. 45–47, 49–50 and 73–74: calibration, measurement, rinsing and electrode conditioning).

Locate the acid-base titration equivalence region from the steepest change of pH with added volume. For one-to-one neutralization, standardized sodium hydroxide concentration times equivalence volume estimates the initial amount of substance of acid. Half that volume gives approximately equal $[\mathrm{HA}]$ and $[{\mathrm{A}}^{-}]$ only when subsequent chemical equilibrium redistribution is small. The Henderson-Hasselbalch equation then gives $\mathrm{pH}\approx \mathrm{p}{K}_a$. Unequal activity coefficients or appreciable hydronium or hydroxide inventories can shift this estimate. The full chemical equilibrium curve supplies a stronger consistency check than its midpoint alone.

Useful controls for this model test are a fresh independently prepared acid portion, a water blank, a check standard after measurement to detect instrument drift, and fixed recorded temperature. A blank diagnoses contamination but should not be subtracted directly from pH, which is a logarithm. Keep exposure to atmospheric carbon dioxide comparable, especially at high dilution, and record any background salt that changes ionic strength. These are experimental-design controls suggested by the model's assumptions. The observed curve and its uncertainty determine whether the model is adequate; the diagrams here contain no experimental readouts. An aqueous acid dissociation constant cannot be inferred unchanged from a sample requiring a different solvent mixture, and a nearly neutral dilute sample makes the difference $h-K_w/h$ particularly sensitive to pH error.