Diesel Exhaust Fluid Characterization

White Paper
Using METTLER TOLEDO Instruments

Content
1. Abstract 2
2. Introduction 3
2.1 ANNEX C: Refractive index and determination of urea content by refractive index 4
2.2 ANNEX D: Determination of alkalinity 5
2.3 ANNEX E: Determination of biuret content 5
2.4 ANNEX F: Determination of aldehyde content 7
2.5 ANNEX G: Determination of insoluble matter content by gravimetric method 8
3.  Setup and Materials 9
3.1 Materials 9
3.2 Chemicals 11
3.3 Preparation 11
4.  Results and Discussion 12
4.1 Refractive index and urea content 12
4.2 Alkalinity 13
4.3 Biuret content 14
4.4 Aldehyde content 16
4.5 Insoluble matter content 18
5. Summary 19
6. References 20
 1. Abstract
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Several quality parameters of AUS 32 solutions were tested using METTLER TOLEDO instruments. Using refrac-
tometry, titration, UV/Vis spectroscopy, gravimetric methods, balances and analytical balances, the contents of
urea, alkalinity, biuret, aldehydes and insoluble matter were determined. These determinations are specified by
the norm ISO 22241-2 (Annexes C, D, E, F and G). Other parameters, described in Annexes B, H, I and J were
not tested in this study. However, several of these test methods require equipment like UV/Vis spectrophotometers
(Annex H) and balances (Annexes B and I) that can be provided by METTLER TOLEDO. Furthermore, Annex B
relies on the biuret content, which can be determined using METTLER TOLEDO UV/Vis spectrophotometers.

ISO 22241-1 gives the acceptance limits for all the tests described in ISO 22241-2. The results obtained in this
study were compared to these acceptance limits. Both the instruments and the PC software LabX can automati-
cally highlight results that are not within specifications. Furthermore, the obtained repeatabilities were compared
to the repeatabilities estimated in ISO 22241-2.

METTLER TOLEDO instruments can be used to determine most of the requested quality parameters of AUS 32
solutions. A single software, LabX, can manage all the obtained data. This PC software stores all the data and
automatically performs overall tolerance checks delivering documented pass/fail results.

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 2. Introduction

Where once there was just aThe environmental compatibility of diesel engines as compared to gasoline engines
is a matter of great dispute in the discussion about the cause of and measures against anthropogenic climate
change. While the combustion of gasoline produces more of the greenhouse gas CO2, diesel engines generally
exhaust more nitrogen oxides. The nitrogen oxide molecules NO and NO2, which are included in the generic term
NOX, are relevant for air pollution as they cause smog and acid rain in high concentrations and contribute to the
depletion of the ozone layer.

Hence, there are ambitions to reduce the NOX exhaust in diesel engines on different levels. In particular, many
countries have agreed on emission norms that include upper limits for NOX discharge in diesel vehicles.

One of the most promising approaches towards emissions in diesel engines is the so-called selective catalytic
reduction (SCR) of NOX to nitrogen gas and water after the combustion process [1]. NOX exhaust reduction relies
on reactions with the molecule urea ((NH2)2CO) that is introduced into the exhaust by the means of a 32% urea
solution in water (AUS 32) that is sprayed from a separate tank into the exhaust. The reactions inside the SCR
catalyst compartment convert up to 90% of the NOX in the exhaust to environmental friendly compounds. This
globally established solution is registered under the brand name AdBlue®, which is held by the German Association
of the Automotive Industry (VDA). Figure 1 shows a schematic overview on the NOX reduction mechanism in
diesel vehicles while Figure 2 further explains the processes that occur in the NOX reduction process.

Figure 1. NOx reduction in diesel vehicles following the selective catalytic reduction (SCR)
mechanism with AdBlue.

Figure 2. Chemicals and reactions involved in the NOx reduction in diesel vehicles. NOx is ultimately
converted to nitrogen gas (N2) and water (H2O) by reactions involving urea ((NH2)2CO).

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The quality assessment of AUS 32 solutions is described in the ISO standard 22241-2 [2]
and the corresponding acceptance limits are given in ISO 22241-1 [3].

The different test methods handled in ISO 22241-2 are described in annexes B – J.
Amongst these criteria, the following can be determined using METTLER TOLEDO instruments:
• Annex C: Refractive index and determination of urea content by refractive index
• Annex D: Determination of alkalinity
• Annex E: Determination of biuret content
• Annex F: Determination of Aldehyde content
• Annex G: Determination of insoluble matter content by gravimetric method
• Annex H: Determination of phosphate content by photometric method

Annex H was not tested in this study and is not treated further. However, with some additional equipment,
annex H could be conducted using a METTLER TOLEDO UV/Vis spectrophotometer.

Annex B: determination of urea by total nitrogen, Annex I: determination of trace elements (Al, Ca, Cr, Cu, Fe,
K, Mg, Na, Ni, P and Zn) by ICP-OES method, and Annex J: determination of identity by FTIR spectrometry
method cannot be integrally conducted with METTLER TOLEDO instruments.

2.1 ANNEX C: Refractive index and determination of urea content by refractive index
Annex C of ISO 22241-2 describes the determination of the refractive index of AUS 32 solutions. The content
of urea as the main component of AUS 32 solutions is a function of the refractive index and can be calculated
according to the following equation:

Where wU is the mass fraction of urea given in %, nD 20 is the refractive index at 20 °C with the sodium D line
as the reference wavelength and B is the mass fraction of biuret (see 2.3 and 4.3: Biuret content). B is usually
small and biuret has the same refractive index per unit of mass as urea.

Figure 3. R5 refractometer used for AUS 32 refractive index measurement.

AUS 32 solutions typically contain 32% urea. The urea content can vary in the range of approximately
31.5 – 35%. Corresponding refractive indices vary in the range of approximately 1.379 – 1.385.

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 2.2 ANNEX D: Determination of alkalinity
Ammonia and other amines are a minor component of AUS 32 solutions with a typical combined content
of less than 0.5% (mass fraction). These alkaline constituents account for the alkalinity of the sample solution.

Figure 4. T9 autotitrator with InMotion autosampler used for AUS 32 alkalinity determination.

The alkalinity is determined via a potentiometric titration with 0.1 M HCl to the endpoint pH 5.7 and
represented in terms of percent ammonia (NH3). The alkalinity determination of AUS 32 samples is
further described in [4].

2.3 ANNEX E: Determination of biuret content

Concentrated urea solutions always contain small amounts of biuret. This compound shows the same
refractive index per unit of mass as urea. Therefore, the biuret content must be known in order to determine
the urea concentration by refractive index measurement.

0 Δ 0 0
2
H2N NH2 – NH3 H2 N N NH2
H
Figure 5. Formation of biuret from urea.

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A UV/Vis spectrophotometer with a path length of 5 cm and a wavelength of 550 nm is used for the biuret
determination.

Figure 6. UV7 spectrophotometer used for AUS 32 biuret determination.

Prior to the sample measurement, a calibration based on a series of biuret standard solutions with known
concentrations is performed. The calibration mainly served as a linearity check and should be performed once
a year. A calibration factor FC is determined using the following equation:

Where mB,i is the biuret content of the corresponding standard i in milligrams and Ai is the (blank compensated)
absorbance of standard solution i.

The biuret content is calculated using a so-called day-factor FD, which is based on the measurement of a single
biuret standard solution. The day factor is obtained according to the following equation:

Where A is the measured absorbance of the standard solution. The day-factor FD should be compared to
the calibration factor FC. The two factors should not deviate from each other by more than 5%.

The biuret content in the sample can then be determined based the day-factor using the following equation:

Where FD is the day-factor, A is the absorbance of the measured sample and mS is the sample size in grams
that was used for the sample preparation. The conversion factor of 2.5 serves to account for the sample
preparation and to get the right result unit, which is given in % (mass fraction). The procedure is further
described in [5].

The biuret content determined via UV/Vis is also needed for the determination of urea according to Annex B
or Annex C.

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 2.4 ANNEX F: Determination of aldehyde content
AUS 32 solutions typically contain 0.5 – 10 mg/kg of free or bound aldehydes, which are represented as
formaldehyde. The amount of free and bound aldehydes in can be determined using a UV/Vis spectrophotometer
at a wavelength of 565 nm.

The content determination relies on a 6-point calibration using a dilution series prepared from a formaldehyde
standard solution. The exact concentration of the formaldehyde standard solutions should be determined
potentiometrically using the sulfite procedure [6] [7].

Figure 7. CuvetteChanger used for the automation of UV/Vis measurements.

A calibration factor FC is calculated from the calibration results according to the following equation:

Where mHCHO,i is the content of formaldehyde of the corresponding dilution step and Ai is the measured
absorbance of the corresponding dilution step.

The formaldehyde content in the sample can then be determined based on the calibration factor FC using
the following equation:

Where wA is the aldehyde content represented as % formaldehyde (mass fraction), A is the measured
absorbance and mS is the sample size in grams that was used for the sample preparation. The procedure
is further described in [8].

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2.5 ANNEX G: Determination of insoluble matter content by gravimetric method

AUS 32 can contain small insoluble particles, usually originating from insoluble impurities in urea. These particles
can compromise the exhaust reduction by blocking the injection nozzle or accumulate and cover the solid catalyst.

Figure 8. XPR205 analytical balance with filter weighing tray.

 
The insoluble matter content is determined by weighing the residuals of a portion of AUS 32 that has been
vacuum filtered through a membrane with 0.8 µm pore size. A detailed procedure is given in [9]. The insoluble
matter content in mg/kg is calculated according to the following equation:

Where wIns is the insoluble matter content in mg/kg, mFL is the mass of the dry, empty filter in milligrams,
mFR is the mass of the dry filter with sample deposit in milligrams and mS is the sample size in grams.

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 3. Setup and Materials

3.1 Materials
Table 1 and Table 2 contain all the instruments and accessories that are used for AUS 32 testing.
The corresponding Annexes show for which type of analysis the material is used.

Used in Annex
Instruments and accessories Part number
C D E F G
Refractometry Excellence R5 30474904 •
Titration Excellence T5 30252672
T7 30252675 • •
T9 30252676
UV/VIS Excellence Spectrophotometer 30254725
• •
UV5/UV7 30254726
XPR205DR Analytical balance 30355415 • • • •

XPR203SN Precision balance 30316540 •

HX204 Halogen moisture analyzer 30019875 •

Aluminum sample pans 00012865 •

InMotion Flex Base 30094121 • •

InMotion Flex Rack Kit 100 mL 30094126 • •

Manual titration kit 51109220 •

2 × Dosing units 51109220 •

1 × Burette DV1010 10 mL 51107501 •

1 × Burette DV1020 20 mL 51107502 •

DGi111-SC Combined pH glass electrode 51109500 •

DM143-SC Platinum double pin electrode 51109220 •

Pt1000 Temperature probe 51109220 •

Thermostatable beaker 51107497 •

Titration beakers PP 100 mL 00101974 • •

Pipette LTS L-5000XLS 17011790 • •

Cuvettes, quartz glass, 10 mm 30258738 •

Cuvette holder long path lengths 30254738 •

Cuvettes, quartz glass, 50 mm 30258739 •

CuvetteChanger 30236313 •

Rainin pipette and tips L-10ML 27060051 • •

Table 1. Specific instruments and accessories used for AUS 32 testing according to ISO 22241-2.

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Used in Annex
Instruments and accessories
A B C D E
Plastic Pasteur pipettes •
Glass microfiber filter pads, diameter = 70 mm,
•
particle retention = 0.7 µm
Tissue wipes •
Equipment for vacuum filtration, suitable for 70 mm
•
filter pads
Flat-tipped tweezers •

Spatula • • •

Erlenmeyer flask, 500 mL •

Volumetric flasks and glass beakers • •

Water bath •

Any pH meter or autotitrator •

External thermostat for 0 °C applications •

Drying oven •

Table 2. Non-specific instruments and accessories used for AUS 32 testing according to ISO 22241-2.
 

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 3.2 Chemicals
Table 3 contains all the chemicals that are used for AUS 32 testing. The corresponding Annexes show
for which type of analysis the substance is used.

Substance Annex

Hydrochlochloric acid (HCl), c = 0.01 mol/L

Tris(hydroxymethyl)aminomethane, THAM, c = 0.01 mol/L D

DIN(19266)/NIST buffer solutions pH 4.006, 6.865 and 9.180

Biuret reference standard

Copper sulfate pentahydrate (CuSO4 ∙ 4 H2O)

Potassium sodium tartrate tetrahydrate (KNaC4H4O6 ∙ 4 H2O)

E
Sodium hydroxide (NaOH)

Hydrochloric acid (HCl), c = 0.1 mol/L

Potassium hydroxide (KOH)

Sulfuric acid (H2SO4), 96%

Formaldehyde standard solution

Chromotropic acid

Iodine (I2) solution, c = 0.05 mol/L

F
Sodium sulfite (Na2SO3) solution, c = 1 mol/L

Acetic acid (CH3COOH), c = 1 mol/L

Sodium carbonate (Na2CO3) solution, c = 1 mol/L

Sodium thiosulfate (Na2S2O3) solution, c = 0.1 mol/L

Deionized water C, D, E, F, G

Acetone C

Table 3. Substances used for AUS 32 testing according to ISO 22241-2.

3.3 Preparation
The following preparation steps must be followed prior to the analyses:
• Refractometers must be verified using deionized water and air.
• The DGi111-SC electrode should be calibrated using the DIN(19266)/NIST buffer solutions prior
to the alkalinity determination.
• The titrant HCl, c = 0.01 mol/L should be standardized against a 0.01 mol/L THAM standard solution.
The procedure can be found in METTLER TOLEDO application note M470 [10].
• The titrant I2, c = 0.05 mol/L should be standardized against a 0.1 mol/L Na2S2O3 standard solution.

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 4. Results and Discussion
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This chapter summarizes the obtained test results. The results are evaluated according to the acceptance
limits that are given in ISO 22241-1. Furthermore, repeatabilities of the individual test methods are estimated
and compared to the estimated repeatabilities given in ISO 22241-2.

The repeatability r was calculated according to ISO 4259 [11] using the following equation:

Where k is the coverage factor for the conversion to the confidence limit, in this case to a double-sided 95%
probability level, having a value of 1.96, and s is the standard deviation of the measured values.

4.1 Refractive index and urea content

The refractive indices of a series of 6 AUS 32 samples were measured using a R5 refractometer (METTLER TOLEDO).
The corresponding urea mass fractions were calculated according to the equation given in chapter 2.1 on page
4, including the mass fraction of biuret, which is determined in chapter 4.3: Biuret content. Table 4 gives an
overview on the obtained results.

Sample nD20 wU [%]

1 1.38235 32.081

2 1.38235 32.081

3 1.38237 32.093

4 1.38238 32.099

5 1.38239 32.105

6 1.38240 32.112

Mean 1.38237 32.095

s 0.00002 0.013

r 0.00006 0.036

Table 4. Measured refractive indices and urea weight fractions of an AUS 32 sample. A biuret weight
fraction of 0.205% (see 4.3 Biuret content) was included in the urea weight fraction calculation.

The conducted measurements and corresponding calculations yield a urea content, which is very close to
32.5%. Both the measured refractive index and urea content lie well inside the tolerated range given in ISO
22241-1. The estimated repeatability r of 0.036% (mass fraction) is clearly better than the expected repeatability
of 0.154% given in ISO 22241-2 for the urea content.

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 Figure 9. Measured refractive index and urea content of an AUS 32 sample. The average of the
measured values is indicated with a solid blue line. Dashed blue lines indicate the boundaries
of the 95% confidence interval and red lines are the limits given in ISO 22241-1.

In the presented series of refractive index measurements, a slight trend towards higher nD 20 values can
be observed. This could be attributed to the evaporation of water during the time of analysis. To avoid such
a trend, the refractive index should be measured quickly after the sampling or the sample should be protected
against the evaporation of water.

4.2 Alkalinity
The alkalinity of AUS 32 sample series consisting of 6 samples was determined by potentiometric titration to
an end point of pH 5.7 using a T9 autotitrator (METTLER TOLEDO). The result is given as the ammonia content
in % (mass fraction). Table 5 summarizes the obtained results for a series of 6 AUS 32 samples.
 

Sample Sample size [g] Content NH3 [%]

1 1.4708 0.064

2 1.5009 0.062

3 1.5029 0.062

4 1.4725 0.064

5 1.5198 0.063

6 1.4775 0.063

Mean 0.063

s 0.001

r 0.003

Table 5. Sample size and alkalinity given as content NH3 in % (mass fraction).

The ammonia content determined in this setup was as low as 0.065% (mass fraction). This conforms to
the maximum level of 0.2% given in ISO 22241-1. The estimated repeatability of 0.003% (mass fraction)
is clearly better than the expected repeatability of 0.077% given in ISO 22241-2.

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Figure 10. Measured alkalinity of an AUS 32 sample. The average of the measured values is indicated
with a solid blue line. Dashed blue lines indicate the boundaries of the 95% confidence interval and red
line represents the upper limit given in ISO 22241-1.

4.3 Biuret content

A calibration curve based on six biuret standard solutions with different concentrations was recorded on the
spectrophotometer. The six standard solutions had biuret contents of 1.6, 4, 8, 12, 16 and 20 mg, each in
a 50 mL volumetric flask containing copper sulfate and an alkaline solution of potassium sodium tartrate.
The results of the calibration factor determination are given in Table 6.

Biuret Amount of biuret Measured

standard number contained [mg] absorbance [A]
1 0.18795 0.06078

2 0.18523 0.10837

3 0.18575 0.20644

4 0.18395 0.30414

5 0.18594 0.40685

6 0.18799 0.49183

FC 39.0 mg

R 2
0.9992
Table 6. Contents in mg and corresponding absorbances of biuret standard solutions obtained
during the calibration.

Figure 11. Calibration curve for the biuret determination.

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Prior to the sample measurement, a day-factor FD was determined by measuring the absorbance of a
biuret standard solution. This standard solution contained 8 mg of biuret in 50 mL of an aqueous solution
containing copper sulfate and potassium sodium tartrate. The day-factor was taken directly before the
sample determination and the result is given in Table 7.

Biuret standard content [mg] Absorbance [A]

8 0.21163

FD 37.1 mg
Table 7. Day-factor determination using a 8 mg biuret standard solution.

Sample measurements were performed with the same setup as used for the calibration and day-factor
determination. The results are shown in Table 8. In order to minimize interferences from ammonia, the
sample solution was first neutralized to a pH value of 7.0 with hydrochloric acid.

Sample Absorbance [A] Biuret content [%]

1 0.18795 0.176

2 0.18523 0.174

3 0.18575 0.174

4 0.18395 0.173

5 0.18594 0.175

6 0.18799 0.176

Mean 0.18614 0.175

s 0.00158 0.001

r 0.00439 0.004

Table 8. Measured absorbances and resulting biuret contents given in % (mass fraction).

The calibration curve recorded in this study shows a good linearity with an R2 of 0.9992. The day-factor
deviates from the calibration factor by 3.1%, which is in accordance with ISO 22241-2. The estimated repeatability
of 0.004% (mass fraction) is better than the expected repeatability of 0.008% given in ISO 22241-2.

Figure 12. Measured biuret content of an AUS 32 sample. The average of the measured values is
indicated with a solid blue line. Dashed blue lines indicate the boundaries of the 95% confidence
interval and red line represents the upper limit given in ISO 22241-1.

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4.4 Aldehyde content

The concentration of the formaldehyde standard stock solution was determined using the sulfite procedure
in a series of 6 samples and amounted to 4.59 mg/L. A portion of chromotropic acid as well as 20 mL of
concentrated sulfuric acid were added to all the standards in the dilution series. The resulting formaldehyde
contents in micrograms as well as the measured absorbances of the diluted standards are given in Table 9.

Formaldehyde content [µg] Absorbance [A]

0.92 0.02974

2.30 0.04430

4.59 0.08202

9.18 0.15384

22.95 0.32732

45.90 0.64374

FC 67.01 mg/A
2
R 0.9995

Table 9. Contents in µg and absorbances of the formaldehyde standards in the dilution series.

Figure 13. Calibration curve for the aldehyde determination.

The calibration curve recorded in this study shows a good linearity with an R2 of 0.9995.

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The sample measurements were performed with the same setup as used for the calibration.
The results are shown in Table 10 and Figure 14.

Sample Absorbance [A] Aldehyde content [mg/kg]

1 0.06664 0.592

2 0.06794 0.603

3 0.06769 0.601

4 0.06580 0.584

5 0.06756 0.600

6 0.06544 0.581

Mean 0.06685 0.594

s 0.0011 0.009

r 0.0030 0.025

Table 10. Aldehyde determination in an AUS32 sample given in mg/kg Formaldehyde.

Figure 14. Measured aldehyde content of an AUS 32 sample. The average of the measured values
is indicated with a solid blue line. Dashed blue lines indicate the boundaries of the 95% confidence
interval and red line represents the upper limit given in ISO 22241-1.

The resulting aldehyde content conforms to the upper limit of 5 mg/kg that is given by ISO 22241-1.
The estimated repeatability of 0.025 mg/kg is clearly better than the expected repeatability of 0.109 mg/kg
given in ISO 22241-2.

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4.5 Insoluble matter content

The insoluble matter content was determined by filtering a portion of the AUS 32 sample and weighing the
filter pad before and after the filtration. The results of the insoluble matter determination are given in Table 11.

Sample mFL [mg] mFR [mg] mS [g] mINS [mg/kg]

1 297.20 298.90 124.530 13.7

2 294.25 298.80 127.040 35.8

3 294.69 296.60 129.900 14.7

4 293.02 296.80 126.960 29.8

5 295.70 297.50 135.340 13.3

6 298.93 301.60 135.650 19.7

Mean 21.2

s 9.5

r 26.3
Table 11. masses of the membrane filter before and after filtration in mg, sample size in g
and resulting insoluble matter content in mg/kg.

The insoluble matter content determination naturally shows a relatively large standard deviation, which can
be mainly attributed to the propagation of the error of the large values mFL and mFR, from which the result is
calculated. Knowing the acceptance limit, such as given in ISO 22241-1 enables the automatic out-of-limits
determination of measured values by the instrument. Figure 15 shows the measured results. Two of the results
lie outside the defined acceptance limits and are indicated as out-of-limits values on the instrument. The user
can then decide how to proceed with the measured sample, i.e. repeat the measurement or label the sample
as not conform to the acceptance criteria.

Figure 15. Measured insoluble matter content of an AUS 32 sample. The average of the measured
values is indicated with a solid blue line. Dashed blue lines depict the boundaries of the 95%
confidence interval and red lines are the limits given in ISO 22241-1.

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 5. Summary

The investigated sample of AUS 32 was determined according to ISO 22241-2, Annex C, D, E, F and G.
The test procedures included different analytical techniques such as refractometry, titration, UV/Vis spectroscopy
and gravimetry. A single PC software, LabX, managed all the measured data.

The test results were compared with the limits given in ISO 22241-1. All instruments can be set up to indicate
whether the measured results are within defined specifications. In the LabX software, all results are stored in
the same database and results that are outside the limits are highlighted for a simplified recognition. A summary
of all the results obtained in this study is given in Table 12.

Parameter Result average Within limits

Refractive index 1.38237 Yes

Urea content 32.095% Yes

Alkalinity 0.063% Yes

Biuret content 0.175% Yes

Aldehyde content 0.594 mg/kg Yes

Insoluble matter content 21.2 mg/kg No

Table 12. Averages of the measured results including a comparison with the limits given in
ISO 22241-1.

All but one of the obtained results were within the limits given in ISO 22241-1. The repeatabilities were
usually better than estimated in ISO 22241-2 except for the insoluble matter content.

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 6. References
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[1] M. S. K. S. F. Kuternowski, Catalysts, vol. 10(7), p. 749, 2020.

[2] “ISO 22241-2, Diesel engines – NOx reduction agent AUS 32”, 2019.
[3] “ISO 22241-1, Diesel engines – NOx reduction agent AUS 32”, 2019.
[4] “METTLER TOLEDO Titration application note. M848”, 2020.
[5] “METTLER TOLEDO Application note: Biuret Content in Diesel Exhaust Fluid According to ISO 22241-2”.
[6] “ISO 11402, Phenolic, amino and condensation resins – Determination of free-formaldehyde content”, 2004.
[7] “METTLER TOLEDO Titration application note: M789”, 2019.
[8] “METTLER TOLEDO Application Note: Aldehyde Content in Diesel Exhaust Fluid According to ISO 22241-2”, 2021.
[9] “METTLER TOLEDO Application note: Insoluble Matter Content of AdBlue according to ISO 22241-2”, 2020.
[10] “METTLER TOLEDO Titration application note: M470”, 2011.
[11] “ISO 4259, Petroleum and related products – Precision of measurement methods and results”, 2017.

Disclaimer
This report presents one or more examples of analysis. The experiments were conducted with the utmost
care using the instruments specified in the description of each application.

The results have been evaluated according to the current state of our knowledge. Different conditions in other
laboratories may entail slight modifications of the methods described in this study.

When chemicals, solvents and gases are used, the general safety rules and the instructions given by the
manufacturer or supplier must be observed.

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