METHODS

Alkenones

Long-chain alkenones were analyzed to estimate paleo-SSTs. The analytical method is based on Yamamoto et al. (2000) with slight modifications. A 1-g sample of freeze-dried sediment was ultrasonic extracted with dichloromethane:methanol (6:4) for 6 min then concentrated and passed through a short bed of Na2SO4 to remove water, yielding a solvent extraction.

An aliquot of the lipid extract was separated into four fractions by column chromatography (eluants are as follows: F1 = 3 mL n-hexane, F2 = 3 mL n-hexane:toluene [3:1], F3 = 4 mL toluene, and F4 = 3 mL toluene:methanol [3:1]). The column was 5.5 mm in diameter x 45 mm long and was filled with 5% deactivated SiO2. C36 n-alkane was added as an internal standard into fraction F3 (alkenone and alkenoates). Fraction F3 was dissolved in 100 µL of n-hexane, and a 1-µL aliquot was then analyzed by gas chromatography at Hokkaido University, on a Hewlett-Packard Model 6890 gas chromatograph equipped with a 6-m x 0.25-µm (0.25 µm film thickness) CP-Sil 5CB (Chrompack) capillary column. For the analysis of the F3 fraction, the oven temperature was programmed from 70° to 310°C at 20°C/min ramp rate and then held at 310° for 40 min. Identification of alkenone compounds was based on comparison of mass spectra and retention time with those in the literature (e.g., de Leeuw et al., 1980; Volkman et al., 1980). The unsaturation index of alkenone was calculated as

Uk´37 = (C37:2)/[(C37:2) + (C37:3)],

where [C37:2] and [C37:3] are the concentration of C37 in each compound (Prahl and Wakeham, 1987). The calculation of paleotemperature was conducted according to the equation

Uk´37 = 0.034 SST + 0.039,

based on an experimental result for cultured Emiliania huxleyi (Prahl et al., 1988).

Organic Carbon, Nitrogen, and Sulfur in Acid-Treated Samples

Organic carbon (Org-C), nitrogen (N), and sulfur (S) contents in acid-treated samples were determined for all 75 samples. Approximately 0.2-0.3 g of powdered sample was treated with 1-M HCl for ~12 hr, then was washed two times with 50 mL deionized filtered water (DIFW) to remove HCl and sea salt. These samples were then dried at 70°C in a glass centrifuge tube for ~24 hr. The samples were analyzed using a LECO-CNS2000 elemental analyzer at the Department of Earth and Planetary Science, Tokyo University. Approximately 0.1 g of sample was weighed in a ceramic crucible and oxidized at 1350°C. The evolved C and S were measured in an infrared absorption analyzer, and N was measured in a thermal conductivity analyzer. The standard deviations for six duplicate analyses are C = 0.251, N = 0.064, and S = 0.081.

Major Element Oxides

The concentrations of 10 major element oxides (SiO2, TiO2, Al2O3, Fe2O3, MnO, CaO, Na2O, K2O, and P2O5) were determined for all 75 samples by X-ray fluorescence (XRF) analysis of fused glass beads using a Philips-PW1480 at the Department of Earth and Planetary Science, University of Tokyo. Powdered samples were dried at 110°C for >4 hr and were then ignited at 1000°C for 6 hr. Loss on ignition (LOI) was calculated from the weight loss. Approximately 0.4 g of ignited sample was mixed with ~4 g of Li2B4O7 flux with an exact 1:10 mixing ratio and was fused in a platinum crucible to make a glass bead. The accuracy of these analyses are ±0.7% for SiO2, ±1.2% for TiO2, ±1.3% for Al2O3, ±2.0% for Fe2O3, ±3.8% for MnO, ±1.5% for MgO, ±2.9% for CaO, ±3.9% for Na2O, ±3.9% for K2O, and ±4.6% for P2O5 (Yoshida and Takahashi, 1997).

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