Isolation of chlorophyll a in Microsystis spp. for determination of stable isotopes of carbon and nitrogen, and variation in Suwa lake.

KATASE Takao, WADA Eitaro

doi:10.2116/bunsekikagaku.39.8_451

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炭素及び窒素安定同位体比測定のためのアオコ中クロロフィルａの単離と諏訪湖におけるそれらの変動
タンソオヨビチッソアンテイドウイタイヒソクテイノタメノアオコ
炭素及び窒素安定同位体比測定のためのアオコ中クロロフィルαの単離と諏訪湖におけるそれらの変動

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We studied on an isolation of chlorophyll a for determination of stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) and applied it to determine in Microcystis spp. from Suwa lake, Japan. The algae were collected four times between June and September, 1987 by using a 20l plankton net (40μm, mesh size). Collected freezed-samples (7.320.6 g, wet base) ground with anhydrous Na₂SO₄ using a mortor and a pestle were extracted with acetone (200 ml). The extracts were precipitated with 1, 4-dioxane (crude chlorophyll) and consequently purified chlorophyll a (2.38.1 mg) was obtained quantitatively by successive chromatography with DEAE-Toyopearl 650 M (2288μm) and Sepharose CL6B (45165μm) packed in glass tubings. The ratio, (δ¹³C, ‰ ) of chlorophyll a in the algae had changed from -33.8±0.05 in June 19 to -27.5±0.3 September with different growth. Difference (Δ¹³C) between δ¹³C of chlorophyll a and that of a whole body of the algae had also changed from -9.4±0.05 to -3.4±0.1.Comparing to the carbon ratios, the both ratios of δ¹⁵N and Δ¹⁵N slightly changed from 22.0 to 23.5 and 12.9 to 16.0, respectively. The change of heavy isotopes in Δ¹³C and Δ¹⁵N was reverse: The heavy carbon was depleted and heavy nitrogen was enriched.

Journal

BUNSEKI KAGAKU

BUNSEKI KAGAKU 39 (8), 451-456, 1990

The Japan Society for Analytical Chemistry

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