SKALAR METHODS

ANALYSIS: TOTAL PHENOL

RANGE: 0.5 - 100 g C6H5OH/liter
SAMPLE: DRINKING, SURFACE, GROUND,
WASTE & SEA WATER
SAN++
Catnr. 497-001 issue 051811/MH/99266564

PRINCIPLE

The automated procedure for the determination of ortho- and meta substituted Phenols is based on the
following reaction; the sample is distilled. The distillate reacts with alkaline hexacyanoferrate(III) and
4-aminoantipyrine to form a red coloured complex, which is measured at 505 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA. Check voltage at the
back of the instrument before installation.
2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

PROCEDURE SAMPLE PREPARATION

water no. 1.2.8

Field of application

Sample preparation for the determination of phenols in water.

Principle preservation

Pretreatment to eliminate possible interferences in the determination of phenols, such as oxidising agents
and sulfur compounds.

Reagents

o-Phosphoric acid (H3PO4 85%)

Copper(II) sulfate (CuSO4.5H2O)

Ammonium iron(II) sulfate (FeSO4.(NH4)2SO4.6H2O)

1. Weigh 1.1 g of ammonium iron(II) sulfate in a liter beaker.

2. Add 500 ml of distilled water.
3. Add 1 ml sulfuric acid (H2SO4 95-97%) and dissolve.
4. Dilute to 1 liter with distilled water and mix.

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Procedure

1. Add, immediately after sampling, an excess of ammonium iron(II) sulfate solution if oxidising agents are
present (detected by the liberation of iodine upon acidification in the presence of potassium iodine).
2. Acidify the sample to pH <4 with phosphoric acid (85%).
3. If sulfur compounds are present, aerate briefly by stirring.
4. Add the copper(II)sulfate (1 g/liter sample).
5. The sample must be kept at 4C in glass bottles and analysed within 24 hours after collection.

Remarks

1. When no interferences are present, add phosphoric acid till pH < 4 and 1 g copper(II) sulfate per liter
sample.

Principle preservation

Reagents

Sulfuric acid solution (H2SO4 1:1)

Procedure

1. Add 1 ml sulfuric acid solution (1:1), or more, per 1 liter sample to pH < 2.
2. The sample must be kept refrigerated (4C) and analysed within 28 days.

Note : The sample preparation in this method is only meant to be an example procedure. It requires
validation on its correct procedure for all types of samples to be analyzed, which is the responsibility of the
user.

REAGENTS

A. Distillation reagent

Required chemicals: o-Phosphoric acid......... 50 ml. Preparation: Dilute the o-phosphoric acid in 100 ml
H3PO4 (85%) distilled water. Fill up to 200 ml with
Distilled water *......... 150 ml. distilled water and mix.
H2O
Note: Prepare fresh weekly.

B. Distilled water

Required chemicals: Distilled water * Note: Refresh weekly.

H2O

C. Stock buffer solution

Required chemicals: Sodium dihydrogen Preparation: Dissolve the sodium dihydrogen

phosphate...................... 14 g.
NaH2PO4.H2O
Distilled water *....... 1000 ml.
H2O
phosphate in 800 ml distilled water. Fill
up to 1 liter with distilled water and mix.
Note: Solution is stable for 1 month. Store at
4C when the solution is not used.

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D. Buffer solution pH 10

Required chemicals: di-Sodium hydrogen Preparation: Dissolve the disodium hydrogen

phosphate................... 25.0 g.
Na2HPO4.2H2O
Stock buffer C .............. 10 ml.
Sodium hydroxide
solution ....................... 5 ml.
NaOH (1M)
Distilled water *......... 985 ml.
H2O
Brij 35 (30%)................... 3 ml.
phosphate in 800 ml distilled water. Add
the stock buffer solution (C) and check the
pH. Add dropwise 1M sodium hydroxide
solution to reach pH 10.0. Fill up to 1 liter
with distilled water, add the Brij 35 and
mix.
Note: Solution is stable for 1 week. Store at 4C
when the solution is not used. Check the
pH every 2 days.

E. 4-Aminoantipyrine solution

Required chemicals: 4-Amino antipyrine ..... 1.25 g. Preparation: Dissolve the 4-amino antipyrine in 800
C11H13N3O ml distilled water. Add the stock solution
Stock solution 10 mg phenol/liter and fill up to 1 liter
10 mg phenol/liter...... 1.0 ml. with distilled water and mix.
Distilled water *......... 999 ml.
H2O Note: Stable for 3 days. Store in a dark coloured
bottle. Prepare 24 hours in advance.

F. Hexacyanoferrate(III) solution

Required chemicals: Potassium hexacyano- Preparation: Dissolve the potassium hexacyano-

ferrate(III) ..................... 0.5 g.
K3[Fe(CN)6]
Distilled water *......... 200 ml.
H2O
ferrate(III) 150 ml distilled water. Fill up
to 200 ml with distilled water and mix.
Note: Prepare fresh daily. Store in a dark
coloured bottle.

G. Rinsing liquid sampler (Skalar)

Required chemicals: Phosphoric acid .............. 4 ml. Preparation: Dilute the phosphoric acid in 800 ml
H3PO4 (1M) distilled water. Add the copper(II) sulfate.
Copper(II) sulfate ............. 1 g. Fill up to 1 liter with distilled water and
CuSO4.5H2O mix.
Distilled water *......... 996 ml.
H2O Note: Solution is stable for 1 week.

H. Rinsing liquid sampler (Corplab)

Required chemicals: Distilled water * Note: For sea water samples; adjust the salinity
H2O of the rinsing water sampler to the same
level as the salinity of the sea water
samples when matrix peaks reach the
same level as the detection limit. Use
ultra pure sodium chloride or zero sea

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 water for adjustment.
Refresh weekly.

STANDARDS

Stock solution 1000 mg C6H5OH/liter

Required chemicals: Phenol ....................... 1.000 g. Preparation: Dissolve the phenol in 800 ml rinsing
C6H5OH (white crystals) liquid sampler. Fill up to 1 liter with
Rinsing liquid rinsing liquid and mix.
sampler .................... 1000 ml.
Note: Solution is stable for 1 week. Store at 4C
when the solution is not used.

Stock solution 10 mg C6H5OH/liter

Dilute 1 ml stock solution 1000 mg C6H5OH/liter to 100 ml with rinsing liquid.

Note: Prepare the stock solution 10 mg C6H5OH/liter fresh daily.

Working standards

100 g C6H5OH/liter: Dilute 1.00 ml stock solution 10 g C6H5OH/liter to 100 ml with rinsing liquid sampler.
80 g C6H5OH/liter: Dilute 0.80 ml stock solution 10 g C6H5OH/liter to 100 ml with rinsing liquid sampler.
60 g C6H5OH/liter: Dilute 0.60 ml stock solution 10 g C6H5OH/liter to 100 ml with rinsing liquid sampler.
40 g C6H5OH/liter: Dilute 0.40 ml stock solution 10 g C6H5OH/liter to 100 ml with rinsing liquid sampler.
20 g C6H5OH/liter: Dilute 0.20 ml stock solution 10 g C6H5OH/liter to 100 ml with rinsing liquid sampler.

Note:
1. Prepare the working standards fresh daily.
2. The water used for the reagents and standards must be degassed properly before making up the
reagents and standards. Especially water produced by reverse osmosis or ion exchange equipment
contains a lot of air, which must be removed by degassing (degassing procedure see operational
remarks and trouble shooting).

CATALOGUE NUMBERS REQUIRED CHEMICALS

Product Supplier and catnr. Danger classification

o-Phosphoric acid (85%) Merck 100573 corrosive

Copper(II) sulfate pentahydrate Merck 102790 harmful, irritant, dangerous
for the environment
Ammonium iron(II) sulfate hexahydrate Merck 103792
Sulfuric acid (95-97%) Merck 100731 corrosive
Sodium dihydrogen phosphate monohydrate Merck 106346
di-Sodium hydrogen phosphate dihydrate Merck 106580
Sodium hydroxide Merck 106498 corrosive
Brij 35 (30%) Skalar SC13900
4-Aminoantipyrine Merck 107293 harmful
Potassium hexacyanoferrate(III) Merck 104973
Phenol Merck 822296 toxic, corrosive
Hydrochloric acid (32%) Merck 100313 corrosive

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RECOMMENDED OPERATIONAL SETTINGS

1. System sample time: 120 sec., wash time: 120 sec., air time: 0 sec.
2. Module sample time: 90 sec., wash time: 90 sec., air time: 0 sec. (water)
3. Module sample time: 90 sec., wash time: 90 sec., air time: 0 sec. (sea water)
4. Calibration type: 1st order ISO 8466-1.

OPERATIONAL REMARKS AND TROUBLE SHOOTING

1. The stabilising time of the system is approximately 30 minutes.

2. The sensitivity of the highest standard 100 g C6H5OH/liter is 0.035 AU. (water)
3. The sensitivity of the highest standard 100 g C6H5OH/liter is 0.040 AU. (sea water)
4. Persons using this Skalar method should be familiar with normal laboratory practice. This Skalar method
does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices and to ensure compliance
with any national regulatory conditions.
5. The connection between the sampler and the sample pump tube is made of SA 5141 tube.
6. If a digital matrix photometer 28505901, is in use, a correction interference filter of 720 nm 10 nm is
advised.
7. Avoid any turbidity in the reagents, filter if necessary.
8. During the analysis it is important to keep the cover over the distillation unit, for safety reasons and
temperature stability.
9. Let the distillation unit heat up to preset the temperature before switching on the pump.
10. After 6 months the amount of Brij 35 can be reduced to 1 ml/liter.
11. The reagent bottles must be rinsed thoroughly with distilled water before refilling with fresh reagents.
This is done to remove precipitation of micro organism and other interferences.
12. If solvents or high concentrated acids (> 2M) must be controlled by a rinsing valve, use the solvent/acid
resistant type.
13. Do not place the phenol analysis in direct sunlight and protect from light with the supplied dustcover.
14. Degassing procedures;
a) Good: degas the water with a helium gas flow of approximately 20 liter/hour with aid of a sparger.
Degas 5 liter water for 15 minutes. This procedure removes all dissolved air or, degas the water by
boiling the water for 10 minutes. Cool down to room temperature before usage.
b) Medium: degas the water with a nitrogen gas flow of approximately 20 liter/hour with aid of sparger.
Degas 5 liter water for 15 minutes. This procedure will remove most of the dissolved air.
c) Bad: degas the water by ultrasonic waves. This procedure will remove a small part of the dissolved air.

GENERAL REMARKS

1. In most analyses, the first peak coming from the baseline is lower than it should be. The first peak
coming from the baseline is rejected and will not be included in the calculation of the CV-value.
2. If the sample take up volume is less than 1.00 ml/min, a bypass is required to increase the sample
stream to approximately 1.00 ml/min. For SA 5210 debubbler, when the sample is 1.00 ml/min and
higher use 0.23 ml/min debubble tube.

MAINTENANCE

1. Daily; When daily analysis is finished rinse all reagent lines with distilled water for 30 minutes.

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2. Weekly; To decontaminate the system, rinse with 1M hydrochloric acid solution for 30 minutes and
with distilled water for another 30 minutes.
3. Monthly; Replace all pump tubing (or shift pump tube one colour bridge on pump).
MODULE CONSUMABLES

silicone tube catnr. SA 3150 sleeves catnr. SA 5400

polyethylene tube catnr. SA 3142 sleeves catnr. SA 5401
polyethylene tube catnr. SA 5141 sleeves catnr. SA 5406
polyethylene tube catnr. SA 5142 acid flex sleeves catnr. SA 5414
acid flex tube catnr. SA 5194 acid flex sleeves catnr. SA 5415
acid flex tube catnr. SA 5182 acid flex sleeves catnr. 2CA14321
pvc tube catnr. SA 5155 pump tube 0.16 ml/min. catnr. SA 3025
tygon tube catnr. SA 5133 pump tube 2.00 ml/min. catnr. SA 3035
pneumatic tube catnr. 90041092 pump tube 1.60 ml/min. catnr. SA 3034
shrink tube for glass catnr. 90015165 pump tube 0.10 ml/min. catnr. SA 3024
shrink tube catnr. 90015164 pump tube 0.23 ml/min. catnr. SA 3026
glass tube catnr. SA 5360 pump tube 1.00 ml/min. catnr. SA 3031

MODULE COMPONENTS

manifold holder catnr. 25200105 check valve nalge catnr. 2CA50022

manifold holder catnr. SA 5107 heat exchanger catnr. SA 5304
end block catnr. SA 5109 heat exchanger black catnr. SA 5302-B 2x
connector catnr. SA 5202 2x distillation unit catnr. SA 5268
inlet connector catnr. SA 5216 2x reactor catnr. SA 5521
inlet connector catnr. SA 5246 3x connector catnr. SA 5204
debubbler catnr. SA 5207 clamps large catnr. SA 5110 5x
glass coil catnr. SA 5325 2x clamps small catnr. SA 5111 3x
glass coil catnr. SA 5324 sinkers catnr. SA 5380
connector catnr. SA 5201 3x flow cell 50 mm catnr. SA 6425
flow meter catnr. SA 1330 filter 505 nm catnr. SA 6534

REFERENCES

Chemical method (no. 497)

1. Standard method for the examination of water and waste water, 14th edition, page 574-581, Method
510 through 510C, 1975.
2. ISO 3696:1987, Water for analytical laboratory use. Specification and test methods.
3. ASTM, D1193, Standard Specification for Reagent Water.

Sample preparation (water no. 1.2.8)

1. Environmental Protection Agency, methods for chemical analysis of water and wastes, 1983. Method
420.2.
2. International Organisation for Standardisation, ISO-6439.

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 waste
FLOW DIAGRAM
ml/min Flow cell 50 mm
Filter 505 nm
0.10
waste
4-Aminoantipyrine solution 0.16 5246
Hexacyanoferrate(III) solution## 0.16 5246 ## ##
Air

5302 5302
Resample 5216 5325
waste 1.00
5207 5201 5202 5202
Buffer solution 0.23 black black
5304 * built up in glass
sleeved with acidflex
cooling 5268
Distilled water 0.10 5246
1.60 5201
waste
1.60
5204 waste 5324

Nitrogen gas 70 - 80 units ** X

Air 1330
5325 #
Sample 2.00 5216
5201 5521
Distillation reagent 0.16
135C

1.00
waste

Sampler

*distillation unit is situated # if a distillation temperature of 135C is used

at the module support holder the interference of sulfide and calcium are less.
For 135C the nitrogen gas flow has to be
**check valve Nalge 2CA50022 70 units and the resample tube must be 1.00 ml/min
## black (shrink tube)

Revision: Rev 1 Rev 2 Rev 3 Method:

Date: Analysis: Total Phenol
Chemists Range: 0.5-100 g C6H5OH/liter
Authorization Sample: (Sea)water
First issued: 18-05-2011

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