Comparison of Chlorophyll a Extractions with Ethanol and Dimethyl Sulfoxide/Acetone, and a Concern about Spectrophotometric Phaeopigment Correction

1992 ◽  
Vol 49 (11) ◽  
pp. 2331-2336 ◽  
Author(s):  
D. J. Webb ◽  
B. K. Burnison ◽  
A. M. Trimbee ◽  
E. E. Prepas

Chlorophyll a (Chl a) in water samples from three mesotrophic to eutrophic lakes in north-central Alberta was extracted with one of three solvents (95% ethanol, 90% ethanol, or a 2:3 mixture of dimethyl sulfoxide and 90% acetone (DMSO/acetone)) and analyzed by two techniques (spectrophotometry and high pressure liquid chromatography (HPLC). The dominant phytoplankton were blue-green algae and diatoms. Total Chl a concentrations (i.e. no correction for phaeopigments (Pha)) were not significantly different among solvents (P > 0.5). Total Chl a concentrations from spectrophotometric analyses were significantly higher than those from HPLC analyses (4.2 ± 0.88 and 2.6 ± 0.50 μg∙L−1 respectively, P < 0.05). Pha concentrations derived by spectrophotometry were 64 times higher than those derived by HPLC (1.7 ± 0.52 and 0.025 ± 0.01 μg∙L−1 respectively, P < 0.005). Thus, spectrophotometry appears to dramatically overestimate Pha concentrations and may overestimate total Chl a (i.e. no correction for Pha). Therefore, ethanol and DMSO/acetone are equally suitable for Chl a extraction from natural populations dominated by blue-green algae and/or diatoms, but if information on Pha and/or accessory pigments is required, HPLC analyses are the appropriate route rather than spectrophotometry.

1984 ◽  
Vol 41 (10) ◽  
pp. 1519-1522 ◽  
Author(s):  
Barbara J. Speziale ◽  
Stephen P. Schreiner ◽  
Paul A. Giammatteo ◽  
James E. Schindler

We evaluated the suitability of N,N-dimethylformamide (DMF) for extraction and measurement of chlorophyll in phytoplankton and compared the extraction efficiencies of DMF, dimethyl sulfoxide (DMSO)–acetone, and 90% acetone. Absorbance spectra and specific absorption coefficients of chlorophylls a, b, and c in DMF and 90% acetone are similar. Acidification of 100% DMF extracts of pure chlorophyll a and healthy algal cultures produced acid ratios approximating 1.7. Thus, equations derived from the extinction coefficients of chlorophylls in 90% acetone can be applied to DMF extracts. DMF and DMSO–acetone each extract chlorophyll from green and blue-green algae more effectively than does 90% acetone. DMF is a better extractant than is DMSO–acetone for chlorococcalean species, especially when pigment concentrations are high, but the difference, although significant, is slight. The three solvents are equally efficient in extracting chlorophyll from samples composed of chrysophytes and flagellated chlorophytes. The lack of grinding when using either DMF or DMSO–acetone is a considerable convenience. The potential for destabilized spectrophotometer readings and contamination of the investigator due to the incomplete mixing of DMSO and acetone is eliminated when pure DMF is used.


2008 ◽  
Vol 57 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Kazuo Taki ◽  
Tatsuhiro Seki ◽  
Sakiyori Mononobe ◽  
Kohichi Kato

Any kind of blue-green alga produces metabolites of musty substances and toxins. Therefore, it is necessary to remove the blue-green algae, and processing also including nutrient removal is desired for the water quality improvement of eutrophic lakes. The purpose of this study has been to investigate the possibility of a flotation system using a hybrid technique (chemical compounds and electrostatic bridge) applied to raw water containing phytoplankton with high pH of water, and to examine the zeta potential value of phytoplankton surface and the removal efficiency for phytoplankton, ammonia, nitrogen, and phosphoric acid. The results were as follows: firstly, zeta potential of M. aeruginosa particles was observed to achieve charge neutralization on their surface by adhesion of magnesium hydroxide precipitation with increasing pH. Secondly, maximum removal efficiency concerning chlorophyll-a was observed as 84%, and this efficiency was obtained in the condition of pH &gt; 10, and magnesium hydroxide precipitation was observed. Thirdly, in the pH condition that the maximum removal efficiency of chlorophyll-a was obtained, the removal efficiency and the amount of decrease of NH4-N and PO4-P before and after the change of pH values were observed as 6.7% (0.04 mg-P/L) and 63.6% (0.07 mg-N/L), respectively.


1979 ◽  
Vol 36 (2) ◽  
pp. 152-157 ◽  
Author(s):  
R. E. Stauffer ◽  
G. F. Lee ◽  
David E. Armstrong

A simplified chlorophyll analytical procedure is described that requires 4 min/sample and features an overall precision (measured by the coefficient of variation, C.V.) of ~ 3% at the 15 mg/m3 chlorophyll a level. The C.V. varies inversely with concentration for spectrophotometric determinations. The molar extinction coefficients (665 nm) for chlorophyll a were estimated to be 2.5–6.0% greater in acetone (90%) compared to methanol (100%) and 2.5% greater in a 1:1 mixture (v/v) of methanol-dimethyl formamide compared to methanol. Acetone (90%) extractions of naturally occurring mixtures of the common bloom-forming blue-green algae Microcystis, Oscillatoria, Aphanizomenon, and Anabaena yielded erratic recoveries of chlorophyll a. The lowest recovery (34%) occurred with an Oscillatoria population. On the one occasion when a four-way solvent extraction comparison was performed, acetone (90%) failed to extract 29 ± 2% of the chlorophyll a in a mixture of Anabaena, Microcystis, and Aphanizomenon, whereas the other three solvent preparations yielded identical (complete) extractions. Methanol or acetone-dimethyl sulfoxide are recommended for chlorophyll extraction from blue-green algae, while acetone-dimethyl sulfoxide is recommended for diatoms or Chlorophyta populations. Key words: chlorophyll, Cyanophyta, Chlorophyta, chlorophyll analysis, methanol, acetone, methanol-dimethyl formamide, acetone-dimethyl sulfoxide


2013 ◽  
Vol 64 (4) ◽  
pp. 303 ◽  
Author(s):  
M. Bresciani ◽  
M. Rossini ◽  
G. Morabito ◽  
E. Matta ◽  
M. Pinardi ◽  
...  

Eutrophic lakes display unpredictable patterns of phytoplankton growth, distribution, vertical and horizontal migration, likely depending on environmental conditions. Monitoring chlorophyll-a (Chl-a) concentration provides reliable information on the dynamics of primary producers if monitoring is conducted frequently. We present a practical approach that allows continuous monitoring of Chl-a concentration by using a radiometric system that measures optical spectral properties of water. We tested this method in a shallow, nutrient-rich lake in northern Italy, the Mantua Superior Lake, where the radiometric system collected data all throughout the day (i.e. every 5 min) for ~30 days. Here, specifically developed algorithms were used to convert water reflectance to Chl-a concentration. The best performing algorithm (R2 = 0.863) was applied to a larger dataset collected in September 2011. We characterised intra- and inter-daily Chl-a concentration dynamics and observed a high variability; during a single day, Chl-a concentration varied from 20 to 130 mg m–3. Values of Chl-a concentration were correlated with meteo-climatic parameters, showing that solar radiance and wind speed are key factors regulating the daily phytoplankton growth and dynamics. Such patterns are usually determined by vertical migration of different phytoplankton species within the water column, as well as by metabolic adaptations to changes in light conditions.


Author(s):  
T. K. Kalenik ◽  
E. V. Dobrynina ◽  
V. M. Ostapenko ◽  
Y. Torii ◽  
J. Hiromi

The article presents a study of the process of isolation of natural blue pigment – phycocyanin from the biomass of blue-green algae Spirulina platensis by water extraction, followed using its water solution as a natural food colorant in the production of milk chocolate. Recently, modern food enterprises are pursuing their policy towards expanding the range of products, which is closely related to the increasing needs of the population in food of a new kind. One of the solutions to this problem is the use of food additives of both natural and synthetic origin. Among the similar components widespread found dyes synthetic origin, which have high coverage rates and relatively low cost. However, many of the permitted in our country synthetic food dyes are banned in several developed countries as potentially dangerous to health. Synthetic dyes of red, yellow and green color have many natural analogues – carotenoids, lutein, chlorophyll, etc., except for the blue dye, the analogue of which is only anthocyanins, which are unstable depending on the pH conditions. In this article were identified phycobiliproteins and chlorophyll a in a water extract of spirulina. The mass concentration of phycobiliproteins and chlorophyll a was determined by spectrophotometric method before and after the addition of ammonium sulfate. A comparative analysis of the effect of fractionation (salting out) on the degree of purification of the phycocyanin solution. Presented and described the technological scheme of extraction of phycocyanin which allows to use it in food technologies as an extract or a dry powder. Established the concentration of phycocyanin extract from blue-green algae spirulina to produce milk blue chocolate. Determined organoleptic and hygienic characteristics of the finished product


2011 ◽  
Vol 11 (2) ◽  
pp. 69-79 ◽  
Author(s):  
Ewa Dembowska

Cyanobacterial blooms in shallow lakes of the Iławskie Lake DistrictThe dominance of blue-green algae observed in many lakes is related to a high trophic level. Shallow eutrophic lakes are particularly often abundant in blue-green algae. The research on phytoplankton, the results of which are presented in this paper, was carried out between 2002 and 2005 in six lakes. These lakes differed considerably in their size and management methods applied in the catchment (drainage) area. A few types of water blooms were distinguished, which is related to the catchment area management, the intensity of mixing and the trophic level. Algal blooms of the Planktothrix type appeared in lakes situated in an open area of agricultural catchment basins. Algal blooms of the Limnothrix type were characteristic of lakes with a forest-agricultural catchment area but surrounded by high shores, which reduced the wind influence on the mixing. Sporadic mixed algal blooms were typical of lakes situated in forest catchment areas.


2019 ◽  
Vol 1 (2) ◽  
pp. 37
Author(s):  
Katarina Purnomo Salim ◽  
Rosita Dwi Chandra ◽  
Heriyanto Heriyanto ◽  
Dwi Susilaningsih ◽  
Leenawaty Limantara ◽  
...  

Current efforts to explore the potency of marine resources have been made to promote economic development in Indonesia, especially in the industrial sector. Marine cyanobacteria such as Arthospira sp. and Scytonema sp. are two potential candidates of the blue-green algae that can be used in the application for food and health industries. This study focused on the identification of the composition of the dominant chlorophylls and carotenoids. The results showed the presence of Zeaxanthin, Chlorophyll a, and β-carotene. The economic potency of those dominant pigments will be reviewed in this article.


2007 ◽  
Vol 58 (7) ◽  
pp. 634 ◽  
Author(s):  
X. L. Shi ◽  
F. X. Kong ◽  
Y. Yu ◽  
Z. Yang

The cyanobacterium Microcystis aeruginosa and the green alga Scenedesmus obliquus were incubated individually and together in the dark and under anaerobic conditions created by adding the reducing agent cysteine. Flow cytometry was used to monitor cell concentrations, fluorescence of chlorophyll-a (chl-a), and cell metabolic activity measured with an esterase-sensitive probe to detect fluorescein diacetate (FDA) hydrolysis of the two species. M. aeruginosa showed a slight increase in cell metabolic activity, no conspicuous death of cells, and absence of decay of chlorophyll-a fluorescence in individual and competition cases under dark anaerobic conditions. Cell metabolic activity and fluorescence of S. obliquus, on the contrary, decreased sharply, and cell concentrations fluctuated markedly with time in the unialgal cultures, but showed only a slight decline in the mixed cultures. M. aeruginosa appeared to be more tolerant to dark anaerobic conditions than S. obliquus, which may arise in eutrophic lakes beneath thick surface scums in the water column, or in the bottom sediments. Tolerance of these conditions may be important to the dominance of M. aeruginosa in eutrophic lakes.


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