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1.
High-temperature isothermal heating of biominerals has commonly been used to artificially accelerate protein degradation in order to extrapolate kinetic parameters to the lower temperatures experienced in vivo and in the burial environment. It is not easy to test the accuracy of these simulations due to the difficulty of finding samples of known age held at a known temperature. We compare protein degradation in the intra-crystalline organic matrix of heated (80 °C, 110 °C, and 140 °C) massive Porites sp. coral to that directly measured in the skeleton of colonies growing at ∼26 °C and deposited over the last five centuries. This provides the opportunity to critically evaluate the underlying assumption that high-temperature experiments accurately mimic degradation processes and kinetics occurring in a ‘naturally aged’ biomineral. In all samples the intra-crystalline protein fraction was isolated and the L- and D- concentration of multiple amino acids measured using reverse-phase high-performance liquid chromatography (RP-HPLC). There was no evidence of a failure of the closed system in the high-temperature experiments (assessed by X-ray diffraction, thermogravimetric analyses and determination of leached amino acid concentration). We compared conventional methods for estimation of kinetic parameters with a new ‘model-free’ approach that makes no assumptions regarding the underlying kinetics of the system and uses numerical optimisation to estimate relative rate differences. The ‘model-free’ approach generally produced more reliable estimates of the observed rates of racemization in ‘naturally aged’ coral, although rates of hydrolysis (as estimated from the release of free amino acids) were usually over-estimated. In the amino acids for which we were able to examine both racemization and hydrolysis (aspartic acid/asparagine, glutamic acid/glutamine and alanine), it was clear that hydrolysis was less temperature sensitive than racemization, which may account for the differences in degradation patterns observed between the ‘naturally aged’ coral and high-temperature data. It is clearly important to estimate the individual temperature dependence of each of the parallel reactions. 相似文献
2.
《Quaternary Geochronology》2008,3(1-2):2-25
When mollusc shells are analysed conventionally for amino acid geochronology, the entire population of amino acids is included, both inter- and intra-crystalline. This study investigates the utility of removing the amino acids that are most susceptible to environmental effects by isolating the fraction of amino acids encapsulated within mineral crystals of mollusc shells (intra-crystalline fraction). Bleaching, heating and leaching (diffusive loss) experiments were undertaken on modern and fossil Corbicula fluminalis, Margaritifera falcata, Bithynia tentaculata and Valvata piscinalis shells. Exposure of powdered mollusc shells to concentrated NaOCl for 48 h effectively reduced the amino acid content of the four taxa to a residual level, assumed to represent the intra-crystalline fraction. When heated in water at 140 °C for 24 h, only 1% of amino acids were leached from the intra-crystalline fraction of modern shells compared with 40% from whole shell. Free amino acids were more effectively retained in the intra-crystalline fraction, comprising 55% (compared with 18%) of the whole shell after 24 h at 140 °C. For fossil gastropods, the inter-shell variability in D/L values for the intra-crystalline fraction of a single-age population was reduced by 50% compared with conventionally analysed shells. In contrast, analysis of the intra-crystalline fraction of C. fluminalis does not appear to improve the results for this taxon, possibly due to variability in shell ultrastructure. Nonetheless, the intra-crystalline fraction in gastropods approximates a closed system of amino acids and appears to provide a superior subset of amino acids for geochronological applications. 相似文献
3.
The inter- and intra-crystalline fractions of Patella vulgata limpets recovered from archaeological sites in Northern Spain (covering Neolithic, Mesolithic, Magdalenian, Solutrean, and Aurignacian periods) were examined for amino acid composition and racemisation over time. The calcitic apex and rim areas of the shells were found to probably be composed of similar proteins, as the D/L values and amino acids were comparable and varied in the same way with increasing age; however, the mineral structures present in these areas differed. The aragonitic intermediate part of the shell showed a distinctly different amino acid composition and mineral structure. The main protein leaching from the inter-crystalline fraction occurred within the first 6000 yr after the death of the organism. In contrast, the intra-crystalline fraction — comprised of a different protein composition than the inter-crystalline fraction — appeared to behave as a closed system for at least 34 ka, as reflected by the lack of a significant decrease in the amino acid content; however, changes in the amino acid percentages occurred during this period. The concentration of aspartic acid remained almost constant with age both in inter- and intra-crystalline proteins, and its contribution to the total amino acid content increased with age at the expense of other amino acids such as glutamic acid, serine, glycine and alanine. Temperature is thought to play a key role in the amino acid racemisation of P. vulgata and could explain why in the localities belonging to the Gravettian and Solutrean period, which formed during relatively cold conditions, D/L values were similar to those detected in shells from sites formed during the Magdalenian. 相似文献
4.
Intra-crystalline protein diagenesis (IcPD), a recent development of amino acid racemization dating (AAR), is now established as a reliable geochronological tool for the Quaternary. However, extending the method to new biominerals requires extensive testing in order to provide evidence for the closed-system behaviour of the intra-crystalline proteins and to assess the temporal span that can be covered.Here we present results from high-temperature experiments on the IcPD of the bivalve Pecten, demonstrating that a fraction of proteins can be isolated from a bleach-resistant mineral matrix, which effectively operates as a closed system under conditions of accelerated diagenesis in the laboratory. Analyses of Pecten from the well-dated terrace system of the Gulf of Corinth (Greece) provided a pilot test for the integrity of the intra-crystalline fraction in subfossil shells. The small sample sizes in this preliminary study preclude a full assessment of the aminostratigraphic power of Pecten IcPD, but a concordance is observed between the extent of IcPD and sites dating from between MIS 5 and MIS 11.We conclude that Pecten is a potentially good substrate for IcPD dating in the Mediterranean, and that the temporal limit of the technique in this area lies beyond MIS 11. 相似文献
5.
Artificial diagenesis of the intra-crystalline proteins isolated from Patella vulgata was induced by isothermal heating at 140 °C, 110 °C and 80 °C. Protein breakdown was quantified for multiple amino acids, measuring the extent of peptide bond hydrolysis, amino acid racemisation and decomposition. The patterns of diagenesis are complex; therefore the kinetic parameters of the main reactions were estimated by two different methods: 1) a well-established approach based on fitting mathematical expressions to the experimental data, e.g. first-order rate equations for hydrolysis and power-transformed first-order rate equations for racemisation; and 2) an alternative model-free approach, which was developed by estimating a “scaling” factor for the independent variable (time) which produces the best alignment of the experimental data. This method allows the calculation of the relative reaction rates for the different temperatures of isothermal heating.High-temperature data were compared with the extent of degradation detected in sub-fossil Patella specimens of known age, and we evaluated the ability of kinetic experiments to mimic diagenesis at burial temperature. The results highlighted a difference between patterns of degradation at low and high temperature and therefore we recommend caution for the extrapolation of protein breakdown rates to low burial temperatures for geochronological purposes when relying solely on kinetic data. 相似文献
6.
This study successfully isolates a fraction of intra-crystalline proteins from shells of the marine gastropod Patella vulgata and assesses the suitability of these proteins for IcPD (Intra-crystalline Protein Diagenesis) geochronology. We discuss the mineralogical composition of this gastropod, investigated for the first time by X-ray diffraction mapping, and use the results to inform our sampling strategy. The potential of the calcitic rim and of a bulk sample (containing both apex and rim) of the shell to act as stable repositories for the intra-crystalline proteins during diagenesis is examined. The composition and the diagenetic behaviour of the intra-crystalline proteins isolated from different locations within the shell are compared, highlighting the necessity of targeting consistent sampling positions.We induced artificial diagenesis of both intra-crystalline and whole-shell proteins by conducting high-temperature experiments in hydrous environment; this allowed us to quantify the loss of amino acids by leaching and therefore evaluate the open- or closed-system behaviour of the different fractions of proteins. The results obtained provide further confirmation that patterns of diagenesis vary according to the protein sequence, structure, and location within or outside the intra-crystalline fraction. As Patella is frequently found in the fossil record, both in archaeological and geological contexts, the application of IcPD geochronology to this biomineral opens up the possibility to obtain reliable age information from a range of sites in different areas of the world. 相似文献
7.
Amino acid racemization (AAR) is a geochronological method that uses the ratio of D- to L-configurations in optically active amino acids from carbonate fossils to determine the time elapsed since the death of an organism. Although AAR techniques have been widely applied to foraminiferal tests, there have been limited dedicated assessments of the potential of isolating a bleach-resistant, intra-crystalline fraction of proteins to improve the reliability of AAR in this biomineral system. In this study, we evaluate the effect of two oxidative pre-treatments (hydrogen peroxide and bleach) on amino acid concentrations and D/L values in sub-modern benthic foraminifers (Ammonia spp. and Haynesina germanica) and well-preserved mid Holocene and mid Pleistocene planktic foraminifers (Pulleniatina obliquiloculata, Globorotalia truncatulinoides, and Globorotalia tumida). The oxidative pre-treatments successfully reduced the amino acid content of the foraminiferal tests to a residual fraction, and with the exception of Ammonia spp., neither pre-treatment substantially affected the relative proportion of individual amino acids. The bleaching pre-treatment does not consistently alter D/L values when compared to peroxide pre-treatment, but it does tend to reduce the subsample variability in D/L values, albeit only to a small degree in an inconsistent fashion. Therefore, we recommend that a relatively weak oxidative pre-treatment with 3% hydrogen peroxide is sufficient for foraminifera-based AAR applications. 相似文献
8.
Here we quantified the aspartic acid and glutamic acid racemization rates of the four main ostracode species (Herpetocypris reptans, Candona neglecta, Ilyocypris gibba and Cyprideis torosa) present in several Iberian Peninsula localities covering a wide chronological range (ca. 1 Ma to present). At low D/L values (at Asp D/L < 0.40; and Glu D/L = 0.09–0.18), H. reptans racemized at higher rates than C. neglecta, C. torosa and I. gibba. In contrast, for Asp D/L > 0.4 and Glu D/L > 0.18, H. reptans, C. neglecta and C. torosa showed similar racemization rates. I. gibba exhibited the lowest D/L values in old samples (Middle and Lower Pleistocene). We attribute these differences in amino acid racemization rates mainly to variations in valve protein composition. We found that the microstructure of the valves of each species (size, morphology, and arrangement of crystals) differed, but did not appear to change over time (at least for the last ca. 1 Ma). Such differences may also be linked to the type of proteins involved in the respective calcification processes of these organisms. On the basis of our results, and given that other studies have demonstrated that the majority of inter-crystalline proteins are leached early after death (a few centuries or millennia), we propose that the degradation rates of the most resistant inter- and intra-crystalline proteins in each species differ depending on the protein composition of the valves. Although further research is required, we suggest that amino acid racemization in each ostracode species might be related to valve microstructure. 相似文献
9.
Rob Westaway 《Quaternary Geochronology》2009,4(3):241-259
The concentration ratio of the amino acids alanine and serine in calcite opercula of freshwater gastropods of genus Bithynia has been assessed as an age proxy for British Middle and Late Pleistocene sites. Due to decomposition of peptide-bound serine within intra-crystalline protein molecules, alanine/serine ratios increase with sample age, enabling resolution of the marine oxygen-isotope stage of almost all sites and substage resolution for some sites. However, slight overlaps are evident in alanine/serine ratios between some sites from late in one temperate stage and early in the next; this effect results from the low decomposition rates during cold stages, combined with an evident slight dependence of decomposition rates on site conditions. The technique, which can be extended to the Early Pleistocene without technical refinements, is amenable to quantitative calibration using a first-order rate equation, making it feasible in future to present results as numerical ages, subject to an assumed temperature history. Most Pleistocene sites investigated are shown using this technique to have the same ages as previously deduced using biostratigraphy. However, the serine–alanine ages obtained preclude glaciation of southeast England during MIS 16 or of central England during MIS 10. 相似文献
10.
A method is described for the specific hydrolysis of amino acids from humic substances by using the proteolytic enzyme “Pronase E”. The amino acids are analyzed by a fluorescence detector after precolumn derivatization and HPLC-separation. The pattern and the concentration of amino acids after the enzymatic hydrolysis are set up for humic substances of different origin. Due to the results the enzymatic hydrolysis is suitable for the characterization of humic substances with a nitrogen content higher than 3%. As shown in a kinetic study of the hydrolysis of a fulvic acid the influence of humic substances on the enzyme activity increases in case of lower nitrogen content. Due to the specificity of the proteolytic enzyme the operationally defined method is limited as shown by a protein of a known amino acid content. The amino acids leucine, valine, tyrosine and histidine are released to the highest extent. The determined total extent for the hydrolysis of Insulin B is 26.34% and for Casein 38.37%. By comparison of the enzymatic hydrolysis with the classic acid hydrolysis the advantages of the biochemical method are the simple experimental handling and the mild reaction conditions. Amino acids which are very unstable like glutamine and asparagine can only be analyzed by the enzymatic hydrolysis. 相似文献
11.
It has been nearly three decades since the last systematic interlaboratory comparison of amino acid racemization (AAR) measurements among active laboratories. The advent of new methods and improved instrumentation for existing techniques requires that these comparisons be conducted more frequently than has occurred. The present study represents a first step in this process. Five homogeneous liquid samples were distributed to six participating laboratories that use one or more of the following analytical methods: Ion-exchange liquid chromatography (IEx), Reverse-phase liquid chromatography (RP), or Gas chromatography (GC). The five samples have been used in previous formal or informal interlaboratory comparisons: three are Pleistocene mollusk samples, two are Pleistocene eggshell samples. Use of homogeneous liquids eliminated variables involved in the majority of the sample preparative steps (sample cleaning, hydrolysis, desalting), so any observed variability between laboratories can be attributed to instrumental factors or possible small effects associated with the hydration procedures employed prior to instrumental analysis. Although most results indicate good agreement (within 10%) for all amino acid d/l values, there are some notable exceptions for certain amino acids or certain samples. For the five amino acids that are most commonly used in geochronological applications (Asx, Glx, Leu, Val, and A/I), inter-method comparisons reported here provide quantitative regressions that can be used when results from one method are compared with those from another. 相似文献
12.
In this study we compare the organic geochemistry of fossil and modern Glycymeris shells. Amino acids were preserved within the shells. The amino acid content of the shells was similar at all the sites studied. Amino acid racemization and epimerization of Glycymeris shells are suitable techniques for dating Pleistocene raised marine deposits. As reported in other studies, we found that isoleucine epimerization analysis has a greater capacity to discriminate between sites of different age than glutamic acid and aspartic acid. However, particular constraints regarding the use of amino acid dating concern intrashell variability, so to avoid divergent results, it is necessary to sample the same part of the shell, namely the complex cross lamellar region near the umbo. The dating of high-energy coastal marine deposits calls for extensive field work in order to ensure the collection of a large number of samples in order to obtain robust results and reject spurious values. Shell accumulations on the shore-line are conditioned by several factors. The high coefficients of variation for epimerization values and their distribution pattern can be attributed mainly to time-linked taphonomical processes (time-averaging) that gave rise to the shell-bearing bed. However, reworking from former highstand sea level deposits, which usually occupy higher topographic levels, is not a common occurrence. Therefore, as a result of time-averaging and post-depositional processes, it is difficult to identify substages in stacked shell beds in raised beach deposits belonging to the same marine oxygen isotope stage by means of amino acid racemization/epimerization. 相似文献
13.
It is nearly thirty years since the last inter-laboratory study was carried out for amino acid racemization (AAR) analysis using powdered fossil material (Wehmiller 1984). Since then there have been major changes in sample preparation and instrumentation, and it was considered timely to coordinate a new inter-laboratory study in support of current methodologies. In 2010, two such studies were undertaken. The first of these, coordinated by Wehmiller (this edition), used homogeneous hydrolysates of Pleistocene mollusc and eggshell materials and focused on the agreement of analytical measurements between laboratories, without interference from differing sample preparation procedures. The second (this study) was designed specifically as a proficiency test. Unlike previous inter-laboratory comparisons that have focussed on precision estimates, the purpose of this study was to carry out an evaluation of measurement bias by comparing the measurement results of laboratories carrying out their routine methods, including extraction, against the consensus values. Participants were sent one dried sample of a mixed amino acid standards solution and five homogeneous powders: two Pleistocene mollusc test materials prepared from material (ILC-A) supplied and used by Wehmiller in previous inter-laboratory studies (1984; and this edition), one Pleistocene opercula test material from the terrestrial gastropod, Bithynia tentaculata, and two heat-treated modern ostrich eggshell test materials. Results from this study demonstrate that whilst individual laboratory precision may be excellent, suggesting good control of random error influences (less than 1% for replicate measurements by some individual laboratories), agreement between methods, or even between laboratories carrying out the same method, may be very different. Trueness evaluation (determined as the relative percentage bias) reveals the extent of the disagreement reflected by the inter-laboratory variability. Individual laboratory D/L value biases of 10–30% or more when compared to the consensus values are not uncommon. We demonstrate why bias contributions should also be included in AAR uncertainty estimation and recommend that the preparation of defined reference materials are seen as a priority in order to control and correct for systematic error influences in the analytical system. 相似文献
14.
Geochemical reaction rate laws are often measured using crushed minerals in well-mixed laboratory systems that are designed to eliminate mass transport limitations. Such rate laws are often used directly in reactive transport models to predict the reaction and transport of chemical species in consolidated porous media found in subsurface environments. Due to the inherent heterogeneities of porous media, such use of lab-measured rate laws may introduce errors, leading to a need to develop methods for upscaling reaction rates. In this work, we present a methodology for using pore-scale network modeling to investigate scaling effects in geochemical reaction rates. The reactive transport processes are simulated at the pore scale, accounting for heterogeneities of both physical and mineral properties. Mass balance principles are then used to calculate reaction rates at the continuum scale. To examine the scaling behavior of reaction kinetics, these continuum-scale rates from the network model are compared to the rates calculated by directly using laboratory-measured reaction rate laws and ignoring pore-scale heterogeneities. In this work, this methodology is demonstrated by upscaling anorthite and kaolinite reaction rates under simulation conditions relevant to geological CO2 sequestration. Simulation results show that under conditions with CO2 present at high concentrations, pore-scale concentrations of reactive species and reaction rates vary spatially by orders of magnitude, and the scaling effect is significant. With a much smaller CO2 concentration, the scaling effect is relatively small. These results indicate that the increased acidity associated with geological sequestration can generate conditions for which proper scaling tools are yet to be developed. This work demonstrates the use of pore-scale network modeling as a valuable research tool for examining upscaling of geochemical kinetics. The pore-scale model allows the effects of pore-scale heterogeneities to be integrated into system behavior at multiple scales, thereby identifying important factors that contribute to the scaling effect. 相似文献
15.
The deep-sea environment is among the most stable on Earth, making it well suited for amino acid geochronology. Foraminifera with calcareous tests are distributed across the World Ocean and are often recovered in sufficient abundance from sediment cores to derive robust mean amino acid D/L values of multiple replicates from each stratigraphic level. The extent of racemization (D/L) can be compared with independent age control, which in most cases is based on correlation with global marine oxygen-isotope stages and radiocarbon ages from the same stratigraphic levels. In this study, we report the results of amino acid racemization analysis of multiple foraminifera species from well-dated sediment cores taken from the Pacific, Atlantic, and Arctic oceans. The composite of results analyzed to date (179 samples, each composed of an average of 8.6 subsamples = 1531 analyses) show that D/L values generally increase systematically down core, and are similar for samples of comparable ages from different deep-sea sites. Previously published equations that relate D/L values of aspartic and glutamic acids to post-depositional temperature and sample age for Pulleniatina obliquiloculata generally conform to the D/L trends for species analyzed in this study. Laboratory heating experiments were used to quantify the difference in the rate of racemization between P. obliquiloculata and other taxa. For example, aspartic acid in P. obliquiloculata racemizes an average of 12–16% faster than in the common high-latitude species, Neogloboquadrina pachyderma (s). Apparently, the unexpectedly high D/L values previously reported for N. pachyderma (s) older than 35 ka from the Arctic Ocean cannot be attributed to taxonomic effects. 相似文献
16.
Jeffrey L. Bada E. Hoopes D. Darling Graham Dungworth Henk J. Kessels Keith A. Kvenvolden David J. Blunt 《Earth and Planetary Science Letters》1979,43(2):265-268
Enantiomeric measurements for aspartic acid, glutamic acid, and alanine in twenty-one different fossil bone samples have been carried out by three different laboratories using different analytical methods. These inter-laboratory comparisons demonstrate that D/L aspartic acid measurements are highly reproducible, whereas the enantiomeric measurements for the other amino acids show a wide variation between the three laboratories. At present, aspartic acid measurements are the most suitable for racemization dating of bone because of their superior analytical precision. 相似文献
17.
Filtration and extraction indicate that the bulk of the combined amino acids in Pacific Ocean waters are a constituent of a dissolved component, and that they cannot be efficiently extracted from seawater by the macroreticular XAD-2 resin. In deep waters of the Pacific Ocean, alanine accounts for～60–70% of the combined amino acids, while in surface waters alanine makes up only～30% of the combined amino acid fraction. Serine is present in the combined fraction in surface waters, but is essentially absent in deep waters. These differences in the serine and alanine abundances between surface and deep waters are possibly the result of the dehydration of serine, which produces racemic alanine. Threonine can also undergo dehydration producing racemicα-amino-n-butyric acid, a non-biological amino acid which has been detected in deep waters of the Pacific Ocean. Estimates of the rate of serine and threonine dehydration in the oceans suggest that these reactions may have half-lives at 0°C of less than 100 years. As a consequence of dehydration, the dissolved organic material in which this diagenetic reaction has occurred may have reduced biological degradability. 相似文献
18.
The kinetics of oxidation of lower aliphatic amines and pyridine derivatives by dichromate was studied. It has been found that lower aliphatic amines and pyridine derivatives are oxidized by dichromate in acid medium. However, the oxidation rates are very low so that within the reaction period of 2 h, prescribed by the standard dichromate method, only a part of a given compound is oxidized. The kinetics of oxidation can be approximately described by an equation of the first order reaction. The lowest values of rate constants, of the order of 10?3 h?1, were found for pyridine and methylamines. The rate constants of ethylamines and pyridine derivatives are much higher, of the order of 10?1 h?1. Propylamine and butylamine are oxidized easily with high rates. The rate of oxidation of pyridine derivatives increases with an increasing number of substituents on the ring. Dimethylpyridines are oxidized faster than methylpyridines but slower than 2,4,6-threemethylpyridine. Oxidation of methyl derivatives of pyridine proceeds in two steps. In the first, relatively fast one, methyl groups are oxidized to carboxylic groups. In the second one, originated pyridinecarboxylic acids are slowly oxidized to carbon dioxide and water. 相似文献
19.
20.
This paper deals with the influence of different humic substances (HS) on the enzyme activity (EA) of the proteolytic enzyme pronase E. The EA was determined by analyzing the concentration of the amino acid valine hydrolyzed from caseine. For amino acid analysis, a flourescence detector was used after precolumn derivatization and HPLC-separation of the hydrolysis products. The data show that the presence of HS has no adverse effects on amino acid analysis. Based on this result, a decrease of EA was observed in the presence of HS acting as enzyme inhibitors. Clear inhibition effects could be seen for different fulvic acids (HO3 FA, FBR FA and FBR 1.0 FA) at concentrations of 150 mg/L. In case of FBI-2 FA, higher concentrations were necessary to decrease the EA significantly. At an inhibitor concentration of 1500 mg/L the EA was mostly found to be less than 50%. The standard deviations of the results don't allow a differentiation between the HS of different origin and treatment. Specifying HS according to their influence on EA has to remain uncertain. As shown in a kinetic study of the hydrolysis of L-Leucine-2-naphthylamide-hydrochloride, the inhibitor HO3 FA decreases the maximum velocity of the reaction but has no effect on the Michaelis constant. These results indicate that this HS acts as a noncompetitive inhibitor. 相似文献