DFT analysis of nitrogen isotopic reduction partition function ratios in proton exchange equilibria of glutamic acid species in water and its application to the estimation of nitrogen isotope fractionation.

L-Glutamic acid (Glu) plays a pivotal role in amino acid metabolism in living organisms. Theoretical knowledge of how metabolism propagates the nitrogen isotope composition (δ15N) is essential for elucidating the mechanism of amino acid metabolism in vivo. In this paper, to estimate the nitrogen isotope fractionation involving Glu and its protonated/deprotonated species, their nitrogen isotopic reduction partition function ratios (RPFRs) were calculated at the apfd/6-311+G(2d,p) level of theory by the SCRF method. The results show that the RPFR values of the eight possible Glu species at 25 °C are: cation > zwitterion with α-COOH and γ-COO- > zwitterion with α-COO- and γ-COOH > anion with -NH3+ ≫ uncharged molecule > anion with -NH2 and γ-COO- > anion with -NH2 and α-COO- > di-anion. Several correlations between RPFR and bond distances were found. Most importantly, it was found that the shorter the distance of the (C-)N⋯H(-O) hydrogen bond (HB), the greater the RPFR value for that species. This correlation indicates that the tri-coordinate nitrogen atom of the amino group may take on the character of the tetra-coordinate nitrogen atom to some extent by forming a HB of this type. The nitrogen isotope exchange equilibrium between Glu and glycine/serine was evaluated at a typical physiological pH of 7.4. Calculations suggest that the equilibrium constant for the former is an increasing function of temperature from 0 to 100 °C, while the latter is a decreasing function of temperature over the same temperature range.

Nitrogen Isotopic Reduced Partition Function Ratios of Glycine and Serine in Water.

The nitrogen isotopic reduced partition function ratio (RPFR) of glycine (Gly) and serine (Ser) in water has been evaluated by a couple of methods. The experimental fact that those amino acids are present in the zwitterionic form in water has been best reproduced when trihydrated Gly and Ser molecules were treated by the discrete microsolvation method coupled with the self-consistent reaction field (SCRF) approach. The RPFR values of Gly and Ser in water at 25 °C are evaluated as 1.11152 and 1.11070, respectively, and the equilibrium constant of the nitrogen isotope exchange reaction between the two amino acids is calculated as 1.0007 ± 0.0008 at 25 °C. Equivalent results may be obtained by the SCRF method for non- and monohydrated molecules in the zwitterionic form.

Chlorine isotopic compositions of deep saline fluids in Ibusuki coastal geothermal region, Japan: using B-Cl isotopes to interpret fluid sources.

We report chlorine stable isotopic compositions (δ(37)Cl, expressed in ‰ relative to the standard mean ocean chloride) as well as δ(2)H and δ(18)O values of deep saline fluids taken at eight drill-holes reaching from 73 to 780 m below sea level in the Ibusuki coastal geothermal region, Japan. Analytical results show that the δ(37)Cl values narrowly range between -0.26 and +0.21 ‰ with an analytical precision of ±0.06 ‰. Except for one sample, the samples examined are negative in δ(37)Cl value with varying Cl/B molar ratios from 117 to 1265. A correlation study between the Cl/B molar ratio and the δ(37)Cl/δ(11)B ratio indicates a hyperbola-type mixing of at least two Cl sources in the Ibusuki region. One of them depletes in (37)Cl with a higher value of Cl/B molar ratio; and the other one enriches in (37)Cl with a lower Cl/B molar ratio. The former is chemically identical to that of the deep brine, which is altered seawater through the seawater-hot rock interaction. The latter is chemically similar to gas condensate derived from the high-temperature (890 °C) vent of an island-arc volcano near the Ibusuki region.

Statistical analysis of rice samples for compositions of multiple light elements (H, C, N, and O) and their stable isotopes.

Stable isotopic compositions and elemental contents of the H, C, N, and O in 163 rice samples were analyzed. The samples were taken from three different farming countries; Japan (n = 103), United States of America (n = 30), and Australia (n = 21), in addition of Asian rice samples from Thailand (n = 2), Vietnam (n = 1), and China (n = 6) as comparison. They were mostly short grain samples known as "Koshihikari," with several samples of middle and long grains included. All samples were grown in the presence of either natural manure or artificial fertilizer. The climate of the rice farming environment was diverse, from arid to humid. Excluding deltaD data showing large uncertainty, according to the statistical analysis of the principal components based on the stable isotopic compositions such as delta(13)C, delta(15)N, and delta(18)O of rice samples, the Japanese rice samples were clearly distinctive from the Australian and the American rice samples. This fact may be explained by the regional differences in isotopic signatures of the climate, utilized nutrition, and/or quality of irrigation water among the farming countries. This statistical distinction could be one of the useful tools to extract the rice samples grown in Japan from those grown in the other countries.

Chlorine isotope fractionation associated with volcanic activity at the Kusatsu-Bandaiko hot spring in Japan.

Stable chlorine isotope compositions (delta(37)Cl, per-mil: per thousand, vs. a standard sample of sea water) of Kusatsu-bandaiko hot water samples, taken regularly in the years between 1974 and 1995 in the Kusatsu-Shirane volcanic region, Japan, were measured mass-spectrometrically. The results show that the delta(37)Cl values of the waters taken before 1984 were at around-0.12 per thousand, whereas those after 1984 were at around+0.18 per thousand. The delta(37)Cl values are thus distinct across 1984, which is consistent with the classification by the Cl to S molar ratio (Cl/S): the higher the Cl/S ratio, the larger the delta(37)Cl value. The delta(37)Cl value increased as much as 0.30 per thousand during 5 years between 1980 and 1984. This isotopic enrichment is likely correlated with increasing Cl/S ratios, suggesting that the heavier isotope ((37)Cl) may have preferentially increased in the original Cl source of the hot spring across 1984 when volcanic activity likely increased at Mt Kusatsu-Shirane.

Column chromatographic boron isotope separation at 5 and 17 MPa with diluted boric acid solution.

Boron isotopic fractionation factor (S) between boron taken up in strongly basic anion exchange resin and boron in aqueous solution was determined by breakthrough column chromatography at 5 and 17 MPa at 25 degrees C, using 0.1 mM boric acid solution as feed solution. The S values obtained were 1.018 and 1.012, respectively, which were smaller than the value reported by using the same chromatographic method at the atmospheric pressure at 25 degrees C with the boron concentration of 10mM, but were larger than the values under the same condition with much higher concentration of 100 and 501 mM. Calculations based on the theory of isotope distribution between two phases estimated that 21% (5 MPa) and 47% (17 MPa) of boron taken up in the resin phase was in the three-coordinated B(OH)(3)-form, instead of in the four-coordinated B(OH)(4)-form, at high pressures even with a very diluted boric acid solution. We discussed the present results by introducing (1) hydration and (2) a partial molar volume difference between isotopic molecules. Borate may have been partially dehydrated upon transfer from the solution phase to the resin phase at high pressures, which resulted in smaller S values compared with those at the atmospheric pressure. Instead, it may be possible that the difference in the isotopic partial molar volume difference between B(OH)(3) and B(OH)(4)(-) caused the S value to decrease with increasing pressure.

Anion-exchange chromatographic study of the chlorine isotope effect accompanying hydration.

The single-stage separation factor for chlorine isotopes ((35)Cl and (37)Cl) was determined to be 1.00034 by anion exchange chromatography on a 4.5-m column operated in reverse breakthrough manner at 25 degrees C. This value is in good agreement with those obtained in our previous works. It was confirmed that the lighter isotope ((35)Cl) was preferentially fractionated into the resin phase, whereas the heavier isotope ((37)Cl) partitioned into the aqueous phase. This observation, however, contradicts the experimental results for Cl isotope fractionation during NaCl precipitation and the recent theoretical results on Cl isotope fractionation: the (37)Cl isotope selectively enriched into the solid phase and not into the aqueous phase. This discrepancy is discussed based on the theory of isotope distribution between two phases. It is suggested that the chromatographic results reflect an isotope effect accompanying hydration rather than an isotope effect due to a phase change, whereas the reverse is the case for the results in the NaCl precipitation study.

An anion-exchange chromatographic study on boron isotopic fractionation at 2 MPa at 293 K.

To study boron isotopic fractionation at high pressure, column chromatography operated in the breakthrough manner was performed at 2.0 MPa at 25.0 degrees C. The fractionation factor (S) between boron adsorbed onto strongly basic anion-exchange resin and boron in solution was obtained as 1.013, which was smaller than the values at 0.1 MPa (atmospheric pressure) found in literature. The pressure dependence of S was discussed based on the polymerization of boron in the solution and resin phases and on the occurrence of the pressure dependent isotope effect relating to the molar volume changes of boron species upon isotope substitution.