Chlorophyll a Experiment

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(A) INTRODUCTION- Chlorophyll a The most accurate method for determining the total quantity of phytoplankton in seawater is to compute the amount of chlorophyll which usually as chlorophyll a. This value can be connected either to the total biomass of plant material using a factor or ot can be employed in determining assimilation index in conjunction with photosynthetic rate measurements. An already known volume of seawater is filtered through a filter membrane. The pigments contained in the phytoplankton are extracted in acetone and their concentration is estimated spectrophotometrically. If the value of spectrophotometer show reading more than 20 mg/m3 is found, the eutrophication henomenon might occur. OBJECTIVE 1) To determine the amount of chlorophyll a content in water sample. APPARATUS AND REAGENT * UV-1601 SHIMADZU UV-Visible Spectrophotometer * Filtering set * Vacuum pump * 0.47 mm GFC filter paper * Centrifuge tube * Quartz 1 cm curvette * 90% acetone * Magnesium carbonate (MgCO3) METHODOLOGY 1) A polyethylene bottle containing the seawater sample is inverted into the Milipore filtering equipment containing a membrane of fiberglass filter. The sample is allowed to filter under half atmospheric pressure vacuum. 2) Several drops of MgCO3 solution is added to the seawater as it is being filtered. The MgCO3 will acts a s a buffer to the sample to prevent premature phaeophytinization. 3) If added prematurely, it may greatly increase the filtration time. 4) The filter is drained thoroughly with the suction and is stored or extracted as necessary. 5) The moist filter is fold into quarters, wrapped in aluminium foil and is stored frozen at -10 °C 6) The filtrate (GFC filter paper ) is placed in a 15 ml centrifuge tube. 10 ml of 90% acetone is added to the volume and is shaked
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