Method-Dependent Variations in Oxygen Isotope Compositions Obtained for Structural Carbonate in Bone Bioapatite

The carbon and oxygen isotope compositions of structural carbonate contained in bioapatite can be obtained by reaction with ortho-phosphoric acid at various temperatures and reaction times, using off-line or automated sample preparation, and continuous-flow or dual-inlet isotope-ratio mass spectrometry (IRMS). Here, we compare the isotopic compositions obtained for structural carbonate in bone bioapatite using (1) conventional off- line gas extraction (25°C) and dual-inlet IRMS, (2) GasBench automated sampling (50°C) and continuous-flow IRMS, and (3) MultiPrep automated sampling (50°C, 90°C) and dual-inlet IRMS. On average, the stable carbon isotope compositions obtained for the same sample using different methods are within ±0.33 per mil (n=29). The reproducibility of oxygen isotope compositions using the different methods is much poorer (±3.18 per mil; n=29). The differences among these methods were most pronounced for samples analyzed using the MultiPrep at 90°C and dual-inlet IRMS. In the latter case, some samples consistently had extremely low oxygen isotope compositions, and also yielded a contaminant gas containing masses 47, 48, and 49, which was not separated from the normal reaction product (carbon dioxide) during cryogenic processing. Normal and anomalous samples do not systematically differ in their crystallinity indices, C/P ratios, gas yields, or total organic content. Anomalous samples have an additional peak in their deconvoluted FTIR spectra at 866 cm-1 and their ignition products lack β-TCP. They also contain slightly more Al and Si cations, and slightly fewer Na cations. It is not clear how these subtle structural and chemical differences relate to the production of the contaminant gas.