inStableIsotopeMethods,MassSpectrometeryIsotopology.pptVIP

Converting delta units to atom%, of the heavier isotope: Switching between delta units and atom % + 1 100 1 * RR atom% = On a spreadsheet: atom% = 100/((1/(((d/1000)+1)*RR))+1) For 13C: RR = 0.0112372 15N: RR = 0.0036765 SMOW 2H RR = 0 18O PDB RR = 0.002067 18O SMOW RR = 0.0020052 34S RR = 0.045005 (Colorado Plateau Stable Isotope Lab Web Site) (Europe Sci. Handout) (Finnigan MAT Isodat Manual) + 1 d 1000 Calculating RA from atom% You’ve now calculated Atom% 18O Atom% 16O = 1 – Atom% 18O RA = Atom% 18O / Atom% 16O In our kinetic example: CO2 + H2O ? carbohydrate What is the a-constant of the enzyme? If the CO2 being used has a value of –12.4‰ and the carbohydrates have a value of -28‰, we can calculate that they have atomic ratios of 0.01110 and 0.01092 respectively. Next: (13C/12C)carbon dioxide RA 0.01110 (13C/12C)carbohydrate RB 0.01092 Fractionation Factors = = 1.01605 = There are many other terms which tell you the ‘per mil’ difference between compound A and compound B. ? Some are used in the biological literature and others in the geological literature. ? Although the numbers they yield are not identical, they are close approximations of one another. ? When you work up numbers it is very important that you indicate which calculation you are using. More fractionation terminology More fractionation terminology By definition: DAB = dA - dB ? and you should therefore use the previous equations to obtain dA and dB from a to calculate DAB Isotopic enrichment a. In the geological literature: eAB = (aAB – 1)?1000 b. In the biological literature: eAB = 103lnaAB ? Isotopic separation (big Delta, D) a.?In the geological literature: DAB = 103lnaAB ? da – db Isotope discrimination a.?Used in the biological literature and refers specifically to enzyme-mediated fractionation where A is the source and B is the product b.?DAB = (aAB – 1)?1000 It is much better to use the (aAB – 1)?1000 calculation.