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The data show that all the Neanderthal remains are from a much earlier period ( Previous dating of the Vi-207 and Vi-208 Neanderthal remains from Vindija Cave (Croatia) led to the suggestion that Neanderthals survived there as recently as 28,000–29,000 B. Subsequent dating yielded older dates, interpreted as ages of at least ∼32,500 B. We have redated these same specimens using an approach based on the extraction of the amino acid hydroxyproline, using preparative high-performance liquid chromatography (Prep-HPLC).

This method is more efficient in eliminating modern contamination in the bone collagen. P., suggesting the Vindija Neanderthals did not live more recently than others across Europe, and probably predate the arrival of anatomically modern humans in Eastern Europe.

The latest data, both radiometric and genetic, suggest Neanderthals and modern humans coexisted or overlapped for up to several thousand years in Europe until Neanderthal disappearance at around 40,000 cal B. Our understanding of the biocultural processes involved in the transition have been greatly influenced by improved accelerator mass spectrometry (AMS) dating methods and their application to directly dating the remains of late Neanderthals and early modern humans, as well as artifacts recovered from the sites they occupied. (10) showed that, when redated using ultrafiltration methods, the bones that produced ages of ∼33,000 B. were in fact beyond the radiocarbon limit, suggesting the Neanderthal remains were unlikely to be as young as previously thought. For sample Vi-208, after ultrafiltration, the C/N atomic ratio was 3.4, which indicates collagen of acceptable quality.

It has become clear that there have been major problems with dating reliability and accuracy across the Paleolithic in general, with studies highlighting issues with underestimation of the ages of different dated samples from previously analyzed sites (6). At Mezmaiskaya, the AMS dates obtained for the Neanderthal excavated above the previously dated individual were substantially older (9). In both cases, revised radiocarbon dates produced with more robust chemical pretreatment methods have illustrated significant underestimates in the previous dates that cannot be reconciled with a hypothesis of late-surviving refugial Neanderthals. However, for Vi-207, the 30-k Da fraction obtained produced a C/N ratio of 4.3, which indicates the presence of a high molecular weight contaminant.

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Unfortunately, there was insufficient collagen remaining from this sample after pretreatment. The majority of the 383 samples we analyzed yielded poor collagen preservation, which prevented any identification to genus or taxon.

Only 101 samples produced identifiable spectra; a summary of all taxa identified by Zoo MS is shown in ), which again highlights the use of applying such techniques to groups of unidentified Paleolithic bone samples.

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This article presents revised radiocarbon dates of the human bones from this site obtained using a more robust purification method targeting the amino acid hydroxyproline.We also included the sample from a split-based bone point (Vi-3437) that had been analyzed in the laboratory previously, producing only a small collagen yield.We decided to attempt to redate it, using a larger starting mass of bone powder.The bone was analyzed using ancient DNA techniques to enable a formal species identification.) High-resolution photographs of the Vi-*28 Neanderthal bone found using Zoo MS.

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