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USP  – simple automated analysis of ultrapure water

USP <645> – simple automated analysis of ultrapure water

H2O – two simple elements, oxygen (O) and hydrogen (H), fuse together to form one of the most important molecules in the world: water. Water is everywhere on Earth and it is vital for our health and survival. It often contains other ions like calcium, magnesium, and chloride which are essential for the human body to function. However, in specific situations, ultrapure water (UPW) is needed to prepare e.g., injections or other solutions used in hospitals. How is the quality of UPW ensured so that it is always suitable for such medical purposes? The answer to this comes from USP <645>. This standard explains how the water quality can be determined and how this analysis must be performed.


For this analysis, a measuring device capable of measuring the conductivity and the pH value is required. If a combined device is not available, then using two separate ones is also fine. Then, a pH electrode that is especially suitable for the determination of the pH value of water and a conductivity cell for measurement of low conductivities is necessary. In this case, the Aquatrode plus and the stainless steel conductivity cell are recommended.

The Aquatrode plus responds very quickly in ion-deficient matrices (such as ultrapure water) and thanks to its double junction system, the bridge electrolyte can be chosen freely.
The stainless steel conductivity cell has been specially designed for measurements in samples with low conductivity. With a cell constant of c = 0.1 cm-1, it is ideal for conductivities ranging from 0–300 µS/cm. The built-in temperature sensor makes handling even easier as no additional sensor is needed for the temperature measurement.

USP <645> procedure

Now that the necessary instrumentation has been introduced, it’s time to take a look at the standard procedure itself. At first glance this looks a bit difficult as it is a three-step analysis, or actually a four-step analysis if you count the calibration as well.

Step 1: First, calibrate the pH electrode and the stainless steel conductivity cell (sensor). The pH electrode is calibrated with pH 4 and 7 solutions, whereas the stainless steel sensor is calibrated with a 100 µS/cm standard.

Find the standard solutions you need here:

Step 2: After calibrating the sensors, both the temperature of the water and the conductivity are measured without temperature compensation. If the measured conductivity is lower than the value mentioned in the table of USP <645>, then the requirement for the conductivity is met and the water can be used for medical purposes. If this is not the case, then step 3 must be performed.

Step 3: 100 mL water is transferred to an external titration vessel where its temperature is adjusted to 25 ± 1 °C. The water is stirred vigorously to incorporate carbon dioxide present in the air. If the conductivity does not change by more than 0.1 µS/cm per 5 minutes, the value is noted for further evaluation. If this value is below 2.1 µS/cm, then the water is usable for medical purposes. If not, then proceed with step 4.

Step 4: The solution is tempered to 25 ± 1 °C. Once the temperature is stable, 0.3 mL of saturated KCl solution is added and the pH of the water is measured. The pH value must lie between pH 5 and 7. If this is not the case, the water does not meet the requirements and must be discarded. If the pH is measured between 5 and 7, then the conductivity must additionally be lower as mentioned in the USP <645> table. If this is the case, the analysis passes and the water can be used for medical purposes.

Automation of USP <645>

The analysis can be quite time-consuming and therefore Metrohm has provided a solution to make this process much easier. Our system combines all of these steps into one method, allowing you to perform walk-away automation and focus on more important tasks.

To expand the capabilities of the measuring system, the 856/867 modules can be exchanged with a modular OMNIS Titrator which can also be used for other standard potentiometric titrations.

Take a closer look at our automated analysis solution

Proper electrode immersion depth
  • Aquatrode plus for accurate pH measurement in ion-deficient matrices
  • Stainless steel conductivity cell for low conducting samples
  • Special holder for performing step 2 of the standard procedure
  • Thermostated vessel (for step 2)
  • DIS-Cover lid to prevent the sample from CO2 uptake (before step 3)
  • Rod stirrer to saturate the solution with carbon dioxide (for step 3)


With this complete system, standard analysis of UPW quality according to USP <645> is performed in a fully automatic and reliable manner. At the end of each analysis a clear message is received if the deionized water (UPW) has passed the test or not. Handling is very easy and allows users to check if an analysis passes or not with just a glance.

Download our Application Bulletin

Automatic conductometry in water samples with low electrical conductivity in accordance with USP<645>
Post written by Iris Kalkman (Product Specialist Titration) and Heike Risse (Product Manager Titration – Automation), Metrohm International Headquarters, Herisau, Switzerland.
Nonaqueous acid-base titrations – Common mistakes and how to avoid them

Nonaqueous acid-base titrations – Common mistakes and how to avoid them

Nonaqueous acid-base titrations are widely used in several industries, including the petrochemical  and pharmaceutical sectors. Whether you are determining the acid or base number (AN or BN) in oils or fats, titrating substances that are insoluble in water, or quantifying products with different strengths of acidity or alkalinity separately, nonaqueous acid-base titration is the method of choice.

If you already have some experience performing nonaqueous acid-base titrations, you may remember that there are several challenges to overcome in comparison to aqueous acid-base titrations.

In this blog post, I would like to cover some of the most typical issues that could pop up during nonaqueous acid-base titrations and discuss how to best avoid them. An important point to note is that there is no single solution regarding how to perform any nonaqueous acid-base titration correctly. The right procedure depends highly on the solvent and titrant used.

What is a nonaqueous acid-base titration?

Before discussing nonaqueous titrations, first let’s talk a little bit about aqueous acid-base titrations.

Here, a sample is dissolved in water, and depending of the nature of the sample (whether it is acidic or basic) a titration is performed either using aqueous base or aqueous acid as titrant. For indication, a glass pH electrode is used.

However, sometimes due to the nature of the sample, aqueous titration is not possible. Nonaqueous acid-base titration is used when:

  • the substance of interest is not soluble in water
  • samples are fats or oils
  • components of mixtures of acids or bases have to be determined separately by titration

In these cases, a suitable organic solvent is used to dissolve the sample instead of water. The solvent:

  • should dissolve the sample and not react with it
  • permits the determination of components in a mixture
  • if possible, should not be toxic

The solvents that are most often used include ethanol, methanol, isopropanol, toluene, and glacial acetic acid (or a mixture of these). Titrants are not prepared with water but rather in solvent. Frequently used nonaqueous basic titrants are potassium hydroxide in isopropyl alcohol or sodium hydroxide in ethanol, and a common nonaqueous acidic titrant is perchloric acid in glacial acetic acid.

Due to the nature of nonaqueous solvents, they are normally poor conductors and do not buffer well. This makes indication a bit challenging because the electrode must be suitable for such sample types. Therefore, Metrohm offers the Solvotrode which is developed specifically for nonaqueous titrations.

This pH electrode offers the following advantages over a standard pH electrode:

  • Large membrane surface and a small membrane resistance for accurate reading, also in poorly buffered solutions
  • A flexible ground-joint diaphragm which can easily be cleaned even when contaminated with oily or sticky samples, additionally it offers a symmetrical outflow for outstanding reproducibility
  • The electrode is shielded and is therefore less sensitive to electrostatic interferences
  • It can be used with any nonaqueous electrolyte such as lithium chloride in ethanol

In the following sections I will discuss the most common mistakes when performing potentiometric nonaqueous acid-base titrations and how you can avoid them.

Electrostatic effects

The influence of electrostatic effects during analysis is normally negligible. However, maybe you have once seen a curve like the one below which looks relatively normal until suddenly a spike occurs.

Figure 1. Titration curve with a spike which might have occurred from an electrostatic interference.

This is then an indication of an electrostatic effect. However, where does it come from and how can we overcome this?

Electrostatic charge can be generated from many sources, such as friction. For example, while walking across a surface you will generate an electrostatic charge which will be stored in your body. You have probably touched the doorknob after walking across a carpeted space in your socks and obtained a small electric shock—this is the discharge of built up electrostatic charge. If we now assume that you are electrostatically charged and then you approach an electrode that is currently measuring (in use), this will result in a spike (Figure 1). Therefore, it is essential to make sure that you are either properly discharged or that you do not approach the electrode during measurement. You can avoid this issue by wearing the appropriate clothes. ESD (electrostatic discharge) clothes and shoes are mostly recommended when performing nonaqueous titrations.

Blocked diaphragm

A blocked diaphragm is another point which occurs more regularly during nonaqueous titrations. Due to the oily and sticky sample, you might have seen that the electrode diaphragm is clogged and cannot be opened anymore. What should you do then?

In most cases, you can place the electrode in a beaker of warm water overnight. This treatment often helps to loosen the diaphragm. To completely prevent the diaphragm from clogging, a Solvotrode with easyClean technology should be used. With this electrode, electrolyte is released by pressing the head ensuring that the diaphragm is not blocked.

Choice of electrolyte and storage solution

We recommend two types of electrolyte for nonaqueous titrations.

For titrations with alkaline titrants: tetraethylammonium bromide c(TEABr) = 0.4 mol/L in ethylene glycol

For titrations with acidic titrants: lithium chloride c(LiCl) = 2 mol/L in ethanol

Please make sure to store the electrode in the same electrolyte with which it is filled.

Checking the electrode according to ASTM D664

To check whether the Solvotrode is still in good working condition, perform a test according to ASTM D664 using aqueous buffer solutions of pH 4 and 7. The procedure is as follows:

  • Measure the potential of buffer pH 4.0 while stirring and note the value after 1 minute
  • Remove the electrode and rinse it well with deionized water
  • Measure the potential of buffer pH 7.0 while stirring and note the value after 1 minute
  • Calculate the mV difference between the reading of buffers 4.0 and 7.0
  • The difference must be larger than 162 mV (20–25 °C) to indicate an electrode in good shape

If the measured potential difference is less than 162 mV, the electrode requires maintenance. Lift the flexible sleeve of the ground-joint diaphragm to let some electrolyte flow out. Repeat the measurement according to the steps above. If the value is still less than 162 mV, clean the electrode or replace it.

Proper rinsing and cleaning

Proper rinsing is essential if you want to obtain reliable results. Otherwise, the curve might flatten and the equivalence points are no longer recognizable. Figure 2 illustrates this phenomenon well.

Figure 2. Different determinations according to ASTM D664. With time, the start potential of the curves shifts which indicates an unsuitable cleaning procedure.

The sample is the same, however, you see that the equivalence point and starting potential begin to shift and the curves become flatter. This indicates an improper cleaning procedure between measurements. The corresponding electrode is shown in Figure 3.

Figure 3. Appearance of the electrode used in Figure 2 after five measurements.

This electrode was certainly not cleaned properly! Anyone who performs a nonaqueous titration must consider which solvent might best dissolve the residue—this is not an issue that other analysts can easily solve due to the nature of each individual sample. However, do not ignore an electrode with such an appearance.

Conditioning the glass membrane correctly

As you may remember from our previous blog post about pH measurement, it is essential that the hydration layer of the glass membrane stays intact. Nonaqueous solvents dehydrate the glass membrane rather quickly. A change in the hydration layer can have an impact on the measured potential, therefore it is important that the hydration layer is always in the same state before starting a titration to achieve the most reproducible results.

Proper electrode immersion depth

This can be established with a conditioning step of the glass membrane to rebuild the hydration layer. However, if the solvent is able to remove the hydration layer faster than it takes to perform a titration, this can lead to ghost equivalence points. Therefore, the electrode should be completely dehydrated and kept like this for all further titrations.


Polar solvents (e.g., ethanol, acetone, isopropyl alcohol, or mixtures with toluene)

Water-free solvents (e.g., dimethylformamide, acetonitrile, acetic anhydride, or mixtures of these)

Preparation of electrode

Store only the pH membrane (not the diaphragm) in deionized water overnight to build up a proper hydration layer.

Lift the flexible sleeve to allow some electrolyte to flow out.

Dehydrate the pH membrane by placing only the pH membrane (not the diaphragm) in the solvent you will use afterwards for titration.

Lift the flexible sleeve to allow some electrolyte to flow out.

Conditioning of glass membrane Place the pH membrane (bulb only) into deionized water for 1 minute. Place the pH membrane (bulb only) into the corresponding solvent for 1 minute.
Rinsing procedure Rinse electrode with 50–70% ethanol. If this does not help, use a suitable solvent to rinse the electrode and then clean afterwards with 50–70% ethanol. Rinse electrode with glacial acetic acid. If this does not help, use a suitable solvent to rinse the electrode and then clean afterwards with glacial acetic acid.
Remarks Make sure to always keep the bulb of the electrode in deionized water for the same time duration, otherwise the thickness of the hydrated layer (and therefore the response) may vary. Avoid any contact of the electrode with water as this can induce a reaction with the solvent causing ghost equivalence points and irreproducible results.

Maintenance of burets

It is not only the electrode that needs some special attention when performing nonaqueous titrations, but also the electrical buret. Some special maintenance is required since alkaline nonaqueous titrants are especially aggressive and they tend to crystallize, therefore leakage of the buret is likely.

The buret must be maintained on a regular basis according to the manufacturer’s instructions. Metrohm recommends the following procedure:

  • For shorter titration breaks, it is recommended to refill the cylinder with titrant (especially with OMNIS)
  • Clean the buret with deionized water at the end of the day
  • Lubricate the cylinder unit on the centering tube and on the cylinder disc

Also check the corresponding manual of the buret. The most important points are mentioned there which will lead to a longer working life of the buret.

Thermometric titration as an alternative

One alternative to using potentiometric nonaqueous acid-base titration is thermometric titration (TET), depending on the sample and analyte to be measured. Thermometric titration monitors the endothermic or exothermic reaction of a sample with the titrant using a very sensitive thermistor.

The benefit of TET over potentiometric titration is clearly the maintenance-free sensor which does not require any conditioning nor electrolyte refilling. More information about thermometric titration can be found in our previous blog posts below.


Hopefully this article has provided you with information about the main problems encountered during nonaqueous titrations. First, make sure that all electrostatic influences are eliminated. This will save a significant amount of troubleshooting. Then prepare and treat your electrode correctly before, during, and after titration. Make sure to condition the electrode right before your first measurement!

Of special importance here is the solvent you plan to use. If it is a polar solvent, the electrode should be conditioned in deionized water. If nonpolar solvents like acetic anhydride are used, the electrode should be dehydrated first. Between measurements, the electrode should be cleaned with a suitable solvent and the diaphragm should be opened on occasion.

Last but not least, take care of your buret. Maintain it regularly and replace it whenever necessary. With this advice, performing nonaqueous titrations should be a breeze!

For more information

about nonaqueous titrations, download our monograph:

Nonaqueous titration of acids and bases with potentiometric endpoint indication

Post written by Iris Kalkman (Product Specialist Titration at Metrohm International Headquarters, Herisau, Switzerland) and Dr. Sabrina Gschwind (Head of R&D at Metroglas, Affoltern, Switzerland).

Photometric complexometric titration

Photometric complexometric titration

…and how to choose the right wavelength for indication

Complexometric titration was discovered in 1945 when Gerold Schwarzenbach observed that aminocarboxylic acids form stable complexes with metal ions, which can change their color by addition of an indicator. From the 1950’s on, this technique gained popularity for the determination of water hardness. Soon it was clear that aside from magnesium and calcium, other metal ions could also be titrated in this way. The use of masking agents and new indicators gave further possibilities to determine not only the whole amount of metal ions present in solution, but also to separate and analyze them. A new titration type was born: complexometric titration.

Dear readers, have you ever performed a complexometric titration? According to my assumption, quite a lot of you will respond with “yes” as it is one of the most frequently used types of titration. However, I assume you probably struggled over the detection of the endpoint and over the titration itself. In contrast to other types of titrations, the boundary conditions such as pH and reaction time play an even bigger role in complexometry since the complex binding constant is very pH dependent and the reaction might be slow. This article presents the most common challenges and how to overcome them when carrying out complexometric titrations.

For a complexometric titration analysis, it is very important to know the qualitative composition of your sample. This determines the indicator, the complexing agent, and the masking agent you need to use.

Due to the length of the article, I have provided an easy legend of the topics so you can click and jump directly to the area that interests you the most.

Complexometric titration and complex-forming constant

Complexometric reactions always consist of a metal ion which reacts with a ligand to form a metal complex. Figure 1 shows an example of such a chemical reaction of a metal ion Mn+ with Ethylene diamine tetra acetic acid (EDTA). EDTA is the most commonly used titrant for complexometric titrations and reacts in a stoichiometric ratio of 1:1. As shown on the right side of Figure 1, EDTA can form six coordinational bonds, in different words: EDTA has a denticity of six. The more coordinational bonds a ligand can form, the more stable is the formed complex.

Figure 1. Example complexation reaction of a metal M with charge n+ with EDTA.

As with most chemical reactions, this type of reaction stays in an equilibrium. Depending on the metal ion used, this equilibrium can shift more to the left (reactants) or on the right (products) of the equation. For a titration, it is mandatory that the equilibrium is on the right side (complex-forming). The equilibrium constant is defined as shown in Equation 1.

Equation 1. Equilibrium constant, where c = concentration of the individual substances.

Equation 1 also illustrates why it is so important to keep the pH value constant. The concentration of hydronium ions influences the complex-forming constant by a factor of the square of its concentration (e.g., if one titrated with H2Na2EDTA). This means if the pH value of the reaction is changed, its complex-forming constant is also changed, which influences your titration.

Generally, the higher the concentration of the complex in comparison to the free metal / Ligand concentration, the higher the Kc and also the log(Kc) value. Some log(Kc) values are shown later on in Table 2 and can give you a hint regarding which titrant is most suitable for your titration.

Complexometric reactions are often conducted as a photometric titration. This means an indicator is added to the solution so that a color change at the endpoint can be observed. 

Color Indicator

As in acid–base titration, the color indicator is a molecule which indicates when the end of titration (the endpoint, or EP) is reached by a change in the solution color. For acid–base titration, the color change is induced by a change of pH, whereas in complexometric titration the color change is induced by the absence/presence of metal ions. Table 1 gives you an overview of different color indicators and the metals which can be determined with them.

Table 1. List of color indicators for different kinds of metal ions.

It is very important to choose the right indicator, especially when analyzing metal mixtures. By choosing an appropriate indicator, a separation of the metal ions can already take place.

As an example, consider a mixture of Zn2+ and Mg2+, which is titrated with EDTA. The log(Kc) value for the zinc ion is 16.5, and 8.8 for the magnesium ion. If we choose to titrate this sample with PAN-indicator then the indicator will selectively bind to the zinc, but not magnesium. As zinc has the higher complex-formation constant, the zinc ion will react first with EDTA, which will lead to a color change, and the endpoint can be detected. In such a case, the separation of the ions is possible. If this is not the case, the choice of a more suitable complexing agent might help you to obtain a separation of metal ions.

Complexing agent

At the beginning of your titration, the metal ions are freely accessible. By addition of the complexing agent (your titrant), the metal ions become bound. The prerequisite for that is a higher complex-formation constant of the metal with the complexing agent than with the indicator. In 95% of cases, this does not pose a problem. Some complexing agents are mentioned in Table 2. In general, ions with higher charges will have a higher complex-formation constant. 

However, what can you do if you are still not able to separate your metal ions sufficiently and determine them individually? The answer to that is: use a masking agent to make the second metal ion “invisible” to the titrant.

Table 2. Complex-formation constants log(Kc) of different complexing agents with various metal ions. The higher the number in the table the higher the binding strength between metal ion and ligand. As an example: aluminum binds stronger to DCTA than to EDTA.

Masking agents

In general, masking agents are substances which have a higher complex-formation constant with the metal ion than the complexing agent. Metal ions which react with the masking agent can no longer be titrated, and therefore the metal ion of interest (which does not react with the masking agent) can be determined separately in the mixture using the complexing agent. Table 3 shows a small selection of common masking agents. There are many more masking agents available which can be used for the separation of metal ions.

Table 3. A selection of different masking agents.

Complexometric titration is still often carried out manually, as the color change is easily visible. However, this leads to several problems. My previous post “Why your titration results aren’t reproducible: The main error sources in manual titration” explains the many challenges of manual titration.

Subjective color perception and different readings lead to systematic errors, which can be prevented by choosing a proper electrode or using an optical sensor, which accurately indicates the color change. This optical sensor changes its signal depending on the amount of light reaching the photodetector. It is usually the easiest choice when switching from manual titration to automated titration, because usually it does not require any changes to your SOP.

Which wavelength is optimal for indication?

Figure 2. The Optrode from Metrohm can detect changes in absorbance at 470, 502, 520, 574, 590, 610, 640, and 660 nm.

If you choose to automate your complexometric titration and indicate the color change with a proper sensor, you should use the Optrode. This sensor offers eight different wavelengths enabling its use with many different indicators.

Perhaps you’re asking yourself “why do I need eight different wavelengths”? The answer is simple. This sensor monitors the absorbance of a certain wavelength in the solution. Each wavelength change is best detected when the light is strongly absorbed by the color of the sample solution, either before or after the endpoint is reached. For example, during a color change from blue to yellow, it is recommended to select the wavelength 574 nm (yellow) for the detection of the color change, as it is the complementary color of blue. For even more accuracy, the optimal wavelength can be chosen by knowing the UV/VIS spectra of the indicator before and after complexation.

Figure 3. Left: spectra of complexed (purple) and uncomplexed (blue) Eriochrome Black T are shown. Right: the difference in absorption of the two spectra is shown.

On the left side of Figure 3 is a graph with the spectra of complexed and uncomplexed Eriochrome Black T. The uncomplexed solution has a blue tint, whereas the complexed one is more violet. On the right, another graph shows the difference of both spectra. According to this graph, the maximum difference in absorption is obtained at a wavelength of 660 nm. Therefore, it is recommended to use this wavelength for the detection of the color change.

For more examples of indicators and their spectra, check out our free monograph “Complexometric (Chelometric) Titrations”.

Challenges when performing complexometric titrations

As mentioned in the introduction, complexometric titrations are a bit more demanding compared to other types of titration.

First, the indicators themselves are normally pH indicators, and most complexation reactions are pH-dependent as well. For example, the titration of iron(III) is performed in acidic conditions, while the complexation of calcium can only take place under alkaline conditions. This leads to the fact that the pH has to be maintained constantly while performing complexometric titrations. Otherwise, the color change might not be visible, indicated incorrectly, or the complexation might not take place.

Second, complexation reactions do not occur immediately, as with e.g. precipitation reactions. The reaction might take some time. As an example, the complexation reaction of aluminum with EDTA can take up to ten minutes to be completed. Therefore it is also important to keep this factor in mind.

Perhaps a back-titration needs to be performed in such a case to increase accuracy and precision. Please have a look at our blog post “What to consider during back-titration” for more information about this topic.


Complexometric titrations are easy to perform as long as some important points are kept in mind:

  • If more than one type of metal is present in your sample, you might need to consider a masking agent or a more suitable pH range.
  • Reaction duration of your complexation reaction might be long. In this case, a back-titration or titration at elevated temperatures might be a better option.
  • Make sure that you maintain a stable pH during your titration. This can be achieved by addition of an adequate buffer solution.
  • Switching from manual to automated titration will increase accuracy and prevent common systematic errors. When using an optical sensor, make sure that you choose the right wavelength for the detection of the endpoint.

For a general overview of complexometric titration, have a look at Metrohm Application Bulletin AB-101 – Complexometric titrations with the Cu ISE.

For more detailed information

Download our free monograph:

Complexometric (Chelatometric) Titrations

Post written by Iris Kalkman, Product Specialist Titration at Metrohm International Headquarters, Herisau, Switzerland.

Moisture Analysis – Karl Fischer Titration, NIRS, or both?

Moisture Analysis – Karl Fischer Titration, NIRS, or both?

In addition to the analysis of the pH value, weighing, and acid-base titration, measurement of water content is one of the most common determinations in laboratories worldwide. Moisture determination is important for nearly every industry, e.g., for lubricants, food and feed, and pharmaceuticals.

Figure 1. Water drops in a spider web

For lubricants, the water concentration is very important to know because excess moisture expedites wear and tear of the machinery. For food and feed, moisture content must be within a narrow range so that the food does not taste dry or stale, nor that it is able to provide a breeding ground for bacteria and fungi, resulting in spoilage. For pharmaceuticals, the water content in solid dosage forms (tablets) and lyophilized products is monitored closely. For the latter, the regulations state that the moisture content needs to be below 2%.

Karl Fischer Titration

Karl Fischer (KF) Titration for water determination was introduced back in the 1930’s, and to this day remains one of the most tried and trusted methods. It is a fast and highly selective method, which means that water, and only water, is determined. KF titration is based on the following two redox reactions.

In the first reaction, methanol and sulfur dioxide react to form the respective ester. Upon addition of iodine, the ester is oxidized to the sulfate species in a water-consuming reaction. The reaction finishes when no water is left.

Figure 2. Manual sample injection for volumetric KF Titration

KF titration can be used for the determination of the water content in all sample types: liquids, solids, slurries, or even gases. For concentrations between 0.1% and 100%, volumetric KF titration is the method of choice, whereas for lower moisture content between 0.001% and 1%, coulometric KF titration is recommended.

Depending on the sample type, its water content, and its solubility in the KF reagents, the sample can either be added directly to the titration vessel, or would first need to be dissolved in a suitable solvent. Suitable solvents are those which do not react with the KF reagents — therefore aldehydes and ketones are ruled out. In case the sample is dissolved in a solvent, a blank correction with the pure solvent also needs to be performed. For the measurement, the sample is injected directly into the titration vessel using a syringe and needle (Fig. 2). The endpoint is detected by a polarized double Pt pin electrode, and from this the water concentration is directly calculated.

Insoluble or hygroscopic samples can be analyzed using the gas extraction technique with a KF Oven. Here, the sample is sealed in small vial, and the water is evaporated by heat then is subsequently carried to the titration cell.

Figure 3. Fully automated KF Titration with the Metrohm 874 KF Oven Sample Processor

For more information, download our free Application Bulletins: AB-077 for volumetric Karl Fischer titration and AB-137 for coulometric Karl Fischer analysis.

If you would like some deeper insight, download our free monograph: “Water determination by Karl Fischer Titration”. 

Near-infrared spectroscopy

Near-infrared spectroscopy (NIRS) is a technique that has been used for myriad applications in the areas of food and feed, polymers, and textiles since the 1980’s. A decade later, other segments began using this technique, such as for pharmaceutical, personal care, and petroleum products.

NIRS detects overtones and combination bands of molecular vibrations. Among the typical vibrations in organic molecules for functional groups such as -CH, -NH, -SH, and -OH, it is the -OH moiety which is an especially strong near infrared absorber. That is also the reason why moisture quantification is one of the key applications of NIR spectroscopy.

For a further explanation, read our previous blog entry on this subject: Benefits of NIR spectroscopy: Part 2.

NIR spectroscopy is used for the quantification of water in solids, liquids, and slurries. The detection limit for moisture in solids is about 0.1%, whereas for liquids it is in the range of 0.02% (200 mg/L), However, in special cases (e.g., water in THF), moisture detection limits of 40–50 mg/L have been achieved.

This technique does not require any sample preparation, which means that samples can be used as-is. Solid samples are measured in high quality disposable sample vials, whereas liquids are measured in high quality disposable cuvettes. Figure 4 displays how the different samples are positioned on the analyzer for a measurement.

Detailed information about the NIRS technique has been described in our previous blog article: Benefits of NIR spectroscopy: Part 1.

Figure 4. Solid (left) and liquid (right) sample positioning for NIR measurements

NIRS is a secondary technique, meaning it can only be used for routine analysis for moisture quantification after a prediction model has been developed. This can be understood by an analogy to HPLC, for which measuring standards to create a calibration curve is among the initial steps. The same applies to NIRS: first, spectra with known moisture content must be measured and then a prediction model is created.

The development of prediction models has been described in detail in our previous blog article: Benefits of NIR spectroscopy: Part 3.

The schematic outline is shown in Figure 5.

Figure 5. Workflow for NIR Method implementation for moisture analysis

For creation of the calibration set, around 30–50 samples need to be measured with both NIRS and KF titration, and the values obtained from KF titration must be linked to the NIR spectra. The next steps are model development and validation (steps 2 and 3 in Figure 5), which are quite straightforward for moisture analysis. Water is a strong NIR absorber, and its peaks are always around 1900–2000 nm (combination band) and 1400–1550 nm (first overtone). This is shown in Figure 6 below.

Figure 6. NIR Spectra of moisturizing creams, showing the absorptions related to H2O at 1400–1550 nm and 1900–2000 nm

After creation and validation of the prediction model, near-infrared spectroscopy can be used for routine moisture determination of that substance. The results for moisture content will be obtained within 1 minute, without any sample preparation or use of chemicals. Also, the analyst does not need to be a chemist, as all they need to do is place a sample on the instrument and press start.

You can find even more information about moisture determination by near-infrared spectroscopy in polyamides, caprolactam, lyophilized products, fertilizers, lubricants, and ethanol/hydrocarbon blends below by downloading our free Application Notes.

Your choice for moisture measurements: KF Titration, NIRS, or both!

As summarized in Table 1, KF Titration and NIR Spectroscopy each have their advantages. KF Titration is a versatile method with a low level of detection. Its major advantage is that it will always work, no matter if you have a sample type that you measure regularly or whether it is a sample type that you encounter for the first time.

Table 1. Overview of characteristics of moisture determination via titration and NIR spectroscopy

NIR spectroscopy requires a method development process, meaning it is not suitable for sample types that always vary (e.g., different types of tablets, different types of oil). NIRS however is a very good method for sample types that are always identical, for example for moisture content in lyophilized products or for moisture content in chemicals, such as fertilizers.

For the implementation of a NIR moisture method, it is required that samples are measured with KF titration as the primary method for the model development. In addition, during the routine use of a NIR method, it is important to confirm once in a while (e.g., every 50th or every 100th sample) with KF Titration that the NIR model is still robust, and to ensure that the error has not increased. If a change is noticed, extra samples need to be added to the prediction model to cover the observed sample variation.

In conclusion, both KF Titration and NIR spectroscopy are powerful techniques for measuring moisture in an array of samples. Which technique to use depends on the application and the individual preference of the user.

For more information

Download our free white paper:

Karl Fischer titration and near-infrared spectroscopy in perfect synergy

Post written by Dr. Dave van Staveren (Head of Competence Center Spectroscopy), Dr. Christian Haider (Head of Competence Center Titration), and Iris Kalkman (Product Specialist Titration) at Metrohm International Headquarters, Herisau, Switzerland.

Improving your conductivity measurements

Improving your conductivity measurements

Have you ever performed a conductivity measurement and obtained incorrect results? There are several possible reasons for this. In this post, I want to show you how you may overcome some of these issues.

By itself, conductivity measurement is performed quite easily. One takes a conductivity cell and a suitable measuring device, inserts the conductivity cell into the sample solution and reads the value given. However, there are some challenges such as choosing the right sensor, the temperature dependency of conductivity, or the CO2 uptake, which falsify your results.

The following topics will be covered in the rest of this post (click to jump to the topic):


So many measuring cells – which one to use?

The first and most important question about conductivity measurement is: which sensor is the most suitable for your application? The measuring range is dependent on the cell constant of your conductivity cell, and therefore this choice requires a few considerations:

  • What is the expected conductivity of my sample?
  • Do I have a broad range of conductivities within my samples?
  • What is the amount of sample I have available for measurement?

There are different types of conductivity measuring cells available on the market. Two-electrode cells have the advantage that they can be constructed within a smaller geometry, and are more accurate at low conductivities. On the other hand, other types of measuring cells show no influences towards polarization, have a larger linear range, and are less sensitive towards contaminations.

Figure 1 below shows you the wide application range of sensors with different cell constants. As a general rule: Sensors with a low cell constant are used for samples with a low conductivity and sensors with high cell constants should be used for high conductivity samples.

Figure 1. Illustration of the range of applications for different conductometric measuring cells offered by Metrohm (click to enlarge).

To get more information, check out our Electrode finder and select «conductivity measurement».

Determination of the cell constant

Each conductivity cell has its own conductivity cell constant and therefore needs to be determined regularly. The nominal cell constant is dependent of the area of the platinum contacts and the distance between the two surfaces:

:  Cell constant in cm-1
Aeff :  Effective area of the electrodes in cm2
delectrodes :  Distance between the electrodes in cm

However, no sensor is perfect and the effective cell constant does not exactly agree with the ideal cell constant. Thus, the effective cell constant is determined experimentally by measuring a suitable standard. Its measured conductivity is compared to the theoretical value:

:  Cell constant in cm-1
ϒtheor. :  Theoretical conductivity of the standard at the reference temperature in S/cm
Gmeas :  Measured conductance in S

With increasing lifetime usage, the properties of the measuring cell might change. Changing its properties means also changing its cell constant. Therefore, it is necessary to check the cell constant with a standard from time to time and to perform a redetermination of the cell constant if necessary.

Temperature dependency of the conductivity

Have you ever asked yourself why the conductivity is normally referred to at 20 °C or 25 °C in the literature? The reasoning is that the conductivity itself is very temperature-dependent and will vary with different temperatures. It is difficult to compare conductivity values measured at different temperatures, as the deviation is approximately 2%/°C. Therefore, please make sure you measure in a thermostated vessel or you use a temperature compensation coefficient.

What is a temperature compensation coefficient anyway?

The temperature compensation coefficient is a correction factor, which will correct your measured value at a certain temperature to the defined reference temperature. The factor itself depends on the sample matrix and is different for each sample.

For example, if you measure a value of 10 mS/cm at 24 °C, then the device will correct your value with a linear correction of 2%/°C to 10.2 mS/cm to the reference temperature of 25 °C. This feature of linear temperature compensation is very common and is implemented in most devices.

However, the temperature compensation coefficient is not linear for every sample. If the linear temperature compensation is not accurate enough, you can also use the feature of recording a temperature compensation function. There, you will measure the conductivity of your sample at different temperatures and afterwards fit a polynomial function though the measured points. For future temperature corrections, this polynomial function will be used, and more accurate results will be obtained.

And… what about the conductivity standard?

Figure 2. The blue curve shows the actual conductivity (mS/cm) and the orange line is a linear temperature compensation. The temperature compensation here varies from 2.39–4.04 %/°C.

Which standard do I have to choose?

In contrast to pH calibration, the conductivity cell only requires a one-point calibration. For this purpose, you need to choose a suitable standard, which has a conductivity value in the same range as your sample and is inert towards external influences.

As an example, consider a sample of deionized water, which has an expected conductivity of approximately 1 µS/cm. If you calibrate the conductivity cell with a higher conductivity standard around 12.88 mS/cm, this will lead to an enormous error in your measured sample value.

Most conductivity cells will not be suitable for both ranges. For such low conductivities (1 µS/cm), it is better to use a 100 µS/cm conductivity standard. While lower conductivity standards are available, proper handling becomes more difficult. For such low conductivities, the influence of CO2 influence increases.

Last but not least: To stir or not to stir?

This is a controversial question, as stirring has both advantages and disadvantages. Stirring enables your sample solution to be homogeneous, but it might also enhance the carbon dioxide uptake from ambient air.

Either way, it does not matter if you choose to stir or not to stir, just make sure that the same procedure is applied each time for the determination of the cell constant, and for the determination of the conductivity of your sample. Personally, I recommend to stir slightly, because then a stable value is reached faster and the effect of carbon dioxide uptake is almost negligible.

To summarize, it is quite easy to perform conductometric measurements, but some important points should be considered thoroughly before starting the analysis, like the temperature dependency, choice of suitable conductometric measuring cell, and the choice of calibration standard. Otherwise false results may be obtained.

Curious about conductivity measurements?

Read through our free comprehensive monograph:

Conductometry – Conductivity measurement

Additionally, you can download our free two-part Application Bulletin AB-102 – Conductometry below:   

Post written by Iris Kalkman, Product Specialist Titration at Metrohm International Headquarters, Herisau, Switzerland.