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Turbidimetry and Nephelometry

Chapter · March 2012

DOI: 10.1002/9783527645206.ch6

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 3.1 Turbidimetry and nephelometry
Angelo Chianese, M. Bravi and E. Fazio
University of Rome La Sapienza
via Eudossiana 18, I-00184 Rome, Italy, angelo.chianese@uniroma1.it

3.1.1 Introduction

Turbidimetry is a very simple technique to evaluate both the appearance and disappearance of a
solid in a solution or the overall surface of solid particles in a suspension. In the first case, it is
possible to determine the nucleation point of a crystallization process (first appearance of a solid)
and to measure the solubility point (disappearance of the last crystal).
In lab experiments visual detection of appearance and disappearance of crystals is an usual practice,
which is also nowadays applied [Moj2011], [Sah2007]. More accurate measurements are provided
by optical fibres probes [Lia2004]. Turbidimeter is the most widely used instrument to detect the
solution cloud and clear points, corresponding to nucleation and solubility points [Mos2000].
Avantium[1] has developed a system based on turbidimetric measurements to simultaneously
measure the solubility in solutions contained in 16 vials, operating a cooling crystallization.
[Avan2010]. This latter instrument is very helpful to make a quick choice of the suitable solvents
for a pharmaceutical cooling crystallization process.
Recently, a sensor based on the forward beam reflectance method (FBRM) has been adopted for
investigation on nucleation [Sho2006], [Mitc2001], it is accurate as a turbidimeter in the detection
of a nucleation point [He2010], but it is one order of magnitude more expansive.

3.1.2 Measurement of nucleation and solubility points

As above mentioned, the appearance of nuclei in a bench scale vessel made by glass may be done
by the human eye, but it is not any more the case for industrial equipment. In this case, in situ
sensors are used. Nucleation is detected by turbidimetry in correspondence to a sharp attenuation of
the light intensity transmitted through the solution. However, at the onset of nucleation the light
intensity acquired by the receiver is the sum of the direct attenuated light and of the forward
scattered light and the sum of the intensity of these two contributions may reduce the measurement
resolution.
Turbidimetry is popular, since it also provides the measurement of the suspension density up to a
rather high value, but its sensitivity in the detection of nucleation point has not yet been fully
ascertained. Most commercial turbidimeters designed for measuring low turbidities give
comparatively good indications of the intensity of light scattered in one particular direction,
predominantly at right angles to the incident light. Turbidimeters with scattered-light detectors
located at 90° to the incident beam are called nephelometers, they are relatively unaffected by small
differences in design parameters and therefore are specified as the standard instrument for
measurement of low turbidities.
The intensity of light acquired by a nephelometer is only due to the scattered light caused by the
appearance of crystals in suspension and a high sensitivity with respect to the detection of the
nucleation phenomenon is expected.
This work reports a comparison between nucleation measurements made by eye, a turbidimetric
sensor and a nephelometric one with respect to different model systems, exhibiting a very different
nucleation rate: potassium sulphate and dextrose monohydrate in aqueous solutions.

3.1.3 The developed turbidimetric and nephelometric instruments

Two different instruments were developed at the University of Rome for nucleation detection: a
turbidimeter and a nephelometer (Chianese, 2005), (Chianese, patent 2005).
Both instruments consists of a primary sensor, to be put in contact with the solution, and an optical
system to provide the light source and the light intensity measurement. This optical measurement
system is the same for the two instruments, whereas the primary sensor consists of two glass fibres,
1 mm in diameter, on the opposite side or on the perpendicular side (at 90°) for the turbidimeter and
the nephelometer, respectively (see Fig. 3.1.1) . The optical system worked as follows. Light
source, contained in the electronic block, is a laser diode (wavelength 650 nm), whereas the light
intensity measurement device is a digital photo detector which produces a digital signal of 50%
duty cycle at a frequency proportional to the brightness of the acquired light intensity. A further
digital photo detector is placed beside the laser diode to monitor the emitted light stability.
The fibres that connect the immersion probe to the electronic block are step-index multimode ones
with a 600 µm core.

Fig. 3.1.1: Schemes of turbidimeter and nephelometer

The photo detector has a dynamic of 120 dB which provides a sensibility (<1 NTU) capable of
detecting both the nucleation and solubility point with high accuracy.
A micro controller in the electronic device makes the following actions:
• Reading the signals from the digital photo detectors;
• Handling such signals in order to detect the state of the solution, that is absence or presence of
crystals in suspension.
• Sending information to a computer via RS-232 serial port or to an industrial control panel via 4-
20mA output port.
The accuracy of the measurement is determined by the sensitivity of the micro controller with
respect to the light flux, equal to 10 nanowatt/mm2.

3.1.4 The examined systems

The developed instruments were used to measure both the nucleation and the solubility point of two
different classes of aqueous solutions, containing as solute dextrose monohydrate and potassium
sulphate, respectively. Both these solutes exhibit a direct solubility in water. The used dextrose was
supplied by Roquette Italia and had a purity of 99.4% b.w.. Impurities consist mainly of maltose
and malto-triose deriving from the incomplete hydrolysis of starch. Potassium sulphate was an RPE
grade product supplied by Carlo Erba, with a content of impurities less than 0,2 %.
The adopted experimental apparatus consisted of a 500 mL jacketed glass vessel connected with a
thermostatic bath. The vessel was magnetically stirred at a constant speed of 200 rpm and lighted by
a lamp placed behind it, in order to visually detect the nucleation or the dissolution of the crystals.
The temperature, measured by a digital thermometer with a precision of ± 0.1 °C, was continuously
recorded.
The measurement of the nucleation point was made by the following procedure. A saturated
solution was prepared at a fixed temperature, putting water in contact with an excess of solid and
the slurry was stirred in a thermostated vessel for 24 hours. This slurry was then filtered and the
clear mother liquor was poured into a jacketed stirred vessel, 0.5 l in capacity, operating at the
saturation temperature. The temperature was then lowered at a constant rate, equal to 10 °C/h. The
nucleation temperature was identified by eye when a cloud of very small crystals inside the solution
appeared and by the instruments when a meaningful change of the acquired light intensity was
detected. For the runs concerning the dextrose solutions at the beginning of each experiment the
solution was seeded with 4-5 big crystals (>425 µm obtained by sieving) to induce nucleation. In
this case the secondary nucleation point by catalytic mechanism was measured.
The measurement of the solubility point was made on the basis of the detection of the
temperature at which the dissolution of the last crystal occurs. The measurement procedure consists
of firstly causing the nucleation process by cooling, then inverting the temperature change, that is
the suspension was heated up, in order to progressively dissolve the fine crystals. The equilibrium
temperature was detected when the last crystal dissolved.
The adopted procedure to measure solubility point is a little different from the usual one, consisting
of seeding a few big crystals in a supersaturated solution and by inducing their dissolution by a
continuous solution heating. The proposed procedure is more accurate than the traditional one since
the dissolution of fines, slightly grown from the nuclei, is very rapid in correspondence of the
solubility point.

3.1.5 Obtained results

The first series of experiments concern the measurement of the nucleation point determined by eye,
turbidimetry and nephelometry for the two examined aqueous solutions systems. For each system,
two series of experiments were performed by carrying out three runs at constant operating
conditions. The obtained results for potassium sulphate are reported in Table 3.1.1. The temperature
measurement range concerns data reproducibility.

Table 3.1.1: Primary nucleation points for aqueous solutions of potassium sulphate
Temperature of nucleation, °C
cK2SO4
Visual method ephelometry Turbidimetry
g / 100 g water
14,0 24,6 ± 0,8 26,0 ± 0,3 25,0 ± 0,6
15,7 34,6 ± 0,6 35,2 ± 0,2 35,3 ± 0,4

From the data in Table 3.1.1 we may observe that: i) the nucleation temperature detected by
nephelometry is higher than those ones detected by turbidimetry and by eye, thus indicating that
nephelometry is the most sensitive method; ii) the reproducibility of the measurement is again the
best one in case of nephelometry, since the experimental data measured at the same conditions
exhibit the smallest deviations.

Some experiments were then performed with dextrose monohydrate solutions. The nucleation rate
for this system is very low. In particular, when the experiments are performed in absence of seeding
the nucleation phenomenon is stochastic, thus the nucleation point may occur in a wide temperature
range. Moreover the first appearance of nuclei gives rise to a very weak cloud of crystals and the
detection of nucleation is difficult. For these reasons only runs with seed addition were performed
and the secondary nucleation temperature was measured.
The results obtained for this system are reported in Table 3.1.2.

Table 3.1.2: Secondary nucleation points for aqueous solutions of dextrose monohydrate
Temperature of nucleation, °C
cDX
Visual method ephelometry Turbidimetry
g / 100 g water
63.87 28,5 ± 0,5 28,7 ± 0,2 27,5 ± 0,5
57.41 18,2 ± 0,6 18,5± 0,3 18.0 ± 0,5

Again, the detection of the nucleation point by nephelometry takes place earlier with respect to
turbidimetry and the measurements are more reproducible. However, in this case the visual
measurement and the nephelometric one compare quite well each other, whereas by the
turbidimetric method a later detection of the cloud point is obtained. A further series of experiments
on secondary nucleation were then carried out by comparing the visual method with the
nephelometric one. The results reported in Fig. 4.1 show a good agreement between the temperature
values given by the two methods.

30

25
visual detection, °C

20

15

10
10 15 20 25 30
autom atic detection, °C

Fig. 3.1.1: Comparison between DX nucleation points by visual and nephelometric (automatic)
methods

The use of a sensor gives also the chance of continuously measuring the magma of the suspension
after the first crystals appearance. This information may concern either the increase of the magma
density due to the growth of the nuclei or the dissolution of the fine crystals present in suspension,
when the solution becomes more or less supersaturated, respectively. Since the mass of crystals
generated just after nucleation or disappearing just before the clear point is quite low, the change of
the mass suspension may be accurately detected by nephelometry.
An example of the mass density recording along an experiment consisting of a controlled cooling of
a saturated solution, followed by a two steps controlled heating of the same solution is reported in
Fig. 3.1.2. The examined system was a potassium sulphate aqueous solution with a concentration of
14 g of salt per 100 g of water. The solution, initially saturated, became progressively
supersaturated by cooling at a rate of 10 °C/h. After the detection of the nucleation point at around
26 °C the solution was cooled by 1 more °C in order to make evidence of the increase of the magma
density due to both further heterogeneous nucleation and nuclei growth. The solution temperature
was then increased in order to induce crystal dissolution at a heating rate of 10° C/h. The light
intensity detected by the instrument along this period of time firstly reached a maximum, then
progressively decreased. Just before the expected solubility point the heating rate was decreased to
6 °C/h in order to slightly dissolve the residual crystals. The same transmitted light intensity of the
saturated solution was measured by the instrument when the last crystal was dissolved, thus
showing the attainment of the solubility point, equal to 30 °C.

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9.30 9.58 10.27 10. 56 11.25 11.54 12.22 12. 51 13.20
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Fig. 3.1.2: Detection of the nucleation and solubility point by a nephelometer

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Proceedings. , vol. 2 ,pp.93-96

Chianese, A., Bravi, M. and Mascioletti, A. . Metodo e strumento optoelettronico per la misura
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