METHOD (A) Step 1: Sample Filtration

Filter paper

Filter paper was placed on filter holder (using forcep)

100ml of sample 1 were filtered. Suction were continued for 3 minutes.

Filter paper was taken and was placed in the centrifuge tube.

The filtration was repeated with sample 2 (200ml)

(B) Step 2: Chlorophyll-a Extraction

10ml of 90% Acetone was added Sample 1 Sample 2

Sonicator and cold water bath. - The samples were sonicated for 15 minutes.

The samples were put in room temperature for 30 minutes. Then shake well.

Then, the samples were centrifuged for 5 minutes (maximum speed)

Ambient temperature

(C) Step 3: Chlorophyll-a Quantification

Sample extract Sample 1

cuvette

90% Aceton ee 1 Sample 2 Blank

Absorbance of the samples were measured (spectrophotometer)

750 nm (E750o) VS Blank 665 nm (E665o)

0.2ml 1% v/v hydrochloric acid Sample extract 1

0.2ml 1% v/v hydrochloric acid Sample extract 2 Blank

90% Acetone

Absorbance of the samples were measured (spectrophotometer)

750 nm (E750o) VS Blank 665 nm (E665o)

Chlorophyll-a were calculated using Lorenzo (1967) equation and trophic state index (TSI) were calculated using Carlsons TSI