Rapid and Efficient Extraction Method of Chlorophyll by Bead Beating
Oral Presentation
Prepared by M. Citriglia, D. Bhattacharyya
Northeast Ohio Regional Sewer District, 4747 East 49th Street, Cuyahoga Height, OH, 44125, United States
Contact Information: CitrigliaM@neorsd.org; 216-641-6000
ABSTRACT
Estimation of chlorophyll a has been routinely applied for determination of algal and phytoplankton levels in marine and freshwater to assess the eutrophic status and thereby monitor water quality. The existing method of extracts chlorophyll a from glass fiber filters by manual mastication in presence of cold 90% acetone. The manual mastication of the filters is a prolonged and arduous process that allows processing of only two samples (in triplicates) in 20 minutes i.e. about 3 minutes on average for processing each glass fiber filter. An alternative method of bead beating the glass fiber filters using an Omni bead ruptor 24. The new method significantly reduced the time of processing of two samples analyzed in triplicates. Comparative analyses of the new extraction method and the existing method were done keeping all other processing parameters constant and the chlorophyll a content was determined. Comparison of the results from both the methods were statistically analyzed by Passing and Bablok regression which suggests that there were no significant systematic and no proportional differences between the two methods of extraction. It was also noted that the standard deviations amongst the data obtained by the bead beating method were less compared to the existing method, which probably is due to less handling required. We conclude that the application of bead beating of the glass fiber filters can be used with better efficiency to routinely process chlorophyll samples.
Oral Presentation
Prepared by M. Citriglia, D. Bhattacharyya
Northeast Ohio Regional Sewer District, 4747 East 49th Street, Cuyahoga Height, OH, 44125, United States
Contact Information: CitrigliaM@neorsd.org; 216-641-6000
ABSTRACT
Estimation of chlorophyll a has been routinely applied for determination of algal and phytoplankton levels in marine and freshwater to assess the eutrophic status and thereby monitor water quality. The existing method of extracts chlorophyll a from glass fiber filters by manual mastication in presence of cold 90% acetone. The manual mastication of the filters is a prolonged and arduous process that allows processing of only two samples (in triplicates) in 20 minutes i.e. about 3 minutes on average for processing each glass fiber filter. An alternative method of bead beating the glass fiber filters using an Omni bead ruptor 24. The new method significantly reduced the time of processing of two samples analyzed in triplicates. Comparative analyses of the new extraction method and the existing method were done keeping all other processing parameters constant and the chlorophyll a content was determined. Comparison of the results from both the methods were statistically analyzed by Passing and Bablok regression which suggests that there were no significant systematic and no proportional differences between the two methods of extraction. It was also noted that the standard deviations amongst the data obtained by the bead beating method were less compared to the existing method, which probably is due to less handling required. We conclude that the application of bead beating of the glass fiber filters can be used with better efficiency to routinely process chlorophyll samples.
