PHENOLS

Method - Direct Photometric

Principle

Steam- distillable phenolic compounds react with 4 aminoantipyrine at pH 7.9 0.1 in the
presence of potassium ferric cyanide to form a colored antipyrine dye. This dye is kept in
aqueous solution and the absorbance is measured at 500 nm.

Interference

Interference are eliminated or reduced to minimum by using the distillate from the
preliminary distillation procedure.

Reagents

(1) Stock Phenol Solution: - Dissolve 100-mg phenol in freshly boiled and cooled
distilled water and dilute to 100 ml.

(2) Intermediate Phenol Solution: - Dilute 1.0-ml stock phenol solution in freshly
boiled and cooled distilled water to 100 ml.

(3) Standard Phenol Solution: - Dilute 50.0-ml intermediate phenol solution to 500 ml
with freshly boiled and cooled distilled water.
1 ml = 1.0 g phenol Prepare within 2 hrs. of use

(4) Bromate-bromide Solution: - Dissolve 2.784 gm an hydrous KbrO3 in water, add 10
gm Kbr Crystals, dissolve, and dilute to 1000 ml.
(5) Hydrochloric Acid (HCL) :- Concentrated

(6) Standard Sodium Thiosulphate Titrant) 0.025 M): - Dissolve 6.205 gm Na2S2O3
5H2O in distilled water. Add 1.5 ml 6 N NaOH or 0.4 gm solid NaOH and dilute to
1000 ml. Standardize with bi-iodate solution.
(7) Starch Solution: - Use either an aqueous solution or soluble starch powder mixtures.

To prepare an aqueous solution, dissolve 2 gm laboratory grade soluble starch and
0.2 gm salicylic acid, as a preservative, in 100 ml hot distilled water.

(8) Ammonium Hydroxide (NH4OH 0.5): - Dilute 35 fresh concentrate NH4OH to 1 ltr.
with distilled water.

(9) Phosphate Buffer Solution: - Dissolve 104.5 gm K2HPO4 and 72.3 gm KH2 PO4 in
water and dilute to 1 ltr. The pH should be 6.8.

(10) 4-Aminoantipyrine Solution; - Dissolve 2.0 gm 4 aminoantipyrine in water and
dilute to 100 ml. Prepare daily.

(11) Potassium Ferricyanide Solution: - Dissolve 8.0 gm K3 Fe (CN) 6 in water and
dilute to 100 ml. Filter if necessary. Store in a brown glass bottle. Prepare fresh
weekly.

(12) Chloroform (CHCl3)

(13) Sodium Sulphate: - Anhydrous Na2SO4, granules

(14) Potassium Iodide: - K I, Crystals.
Procedure

Place 100-ml distillate or a portion containing not more than 0.5-m phenol diluted to 100 ml,
in a 250-ml beaker. Prepare 100-ml. Distilled water blank and a series of 100 ml. Phenol
standards containing 0.1, 0.2,0.3, 0.4 and 0.5 mg phenol. Treat sample, blank and standards
as follows: Add 2.5 ml. 0.5 N NH4OH solution and immediately adjust to pH 7.9 0.1 with
phosphate buffer. Add 1.0 ml 4-aminoantipyrine solution, mix well, add 1.0 ml K3Fe (CN)6
solution, and mix well. After 15 min. transfer to cells and read absorbance of sample and
standards against at 500 nm.
Calculation

Mg phenol/L = A x 1000 / B

Where,

A = mg phenol in sample, from calibration curve and
B = ml original sample