1. Diagenetic Affects of Ground Water on Bone By: Kristyn Voegele NDSU Geol 428 Geochemistry 2010

2. Diagenesis = Fossilization Process Organism dies in a depositional environment Organism dies in a depositional environment Buried by sediment Buried by sediment Spends millions of years under ground Spends millions of years under ground Ground water brings ions Ground water brings ions Ions form new crystal in open space of bone Ions form new crystal in open space of bone Organism become a fossil with little to no soft tissue Organism become a fossil with little to no soft tissue

3. Importance Not exactly what happens… Not exactly what happens… Discovery of tissues including original proteins Discovery of tissues including original proteins Discovery that REE composition is not stable (Moses 2010) Discovery that REE composition is not stable (Moses 2010) Used PHREEQC to model diagenesis of bone in the presence of ground water Used PHREEQC to model diagenesis of bone in the presence of ground water

4. Data Altered states: Effects of diagenesis on fossil tooth chemistry, Kohn et al 1999. Elemental brake down of modern and fossil teeth from Africa. A Black Hills-Madison Aquifer Origin for Dakota Aquifer Groundwater in Northeastern Nebraska, Stotler et al 2010 Ground water recharged from similar time period as fossils

5. Possible Errors Assumed teeth are mostly Hydroxyapitite and can use basic formula of Hydroxyapitite Assumed teeth are mostly Hydroxyapitite and can use basic formula of Hydroxyapitite Ca 5 (PO4) 3 OH Ca 5 (PO4) 3 OH “Created” minerals in Phreeqc “Created” minerals in Phreeqc

9. Process 1. Convert documented element concentrations to oxide concentrations 1. Convert documented element concentrations to oxide concentrations 2. Convert oxide concentrations to the subscripts in a chemical formula 2. Convert oxide concentrations to the subscripts in a chemical formula 3. Create input for Phreeqc 3. Create input for Phreeqc

10. 1. Convert element concentrations to oxide concentrations Used Andy Tindle’s website spread sheets Used Andy Tindle’s website spread sheets Element concentration in ppm * Compound Ratio Element concentration in ppm * Compound Ratio Compound Ratio (CPR) – molecular weight of oxide divided by molecular weight of element Compound Ratio (CPR) – molecular weight of oxide divided by molecular weight of element Multiply by 10,000 to get % Oxide Multiply by 10,000 to get % Oxide

11. Sum elements that substitute into the same structural position Sum elements that substitute into the same structural position Used Andy Tindle’s website spread sheets Used Andy Tindle’s website spread sheets Take % Oxide and convert to Atomic Ratios Take % Oxide and convert to Atomic Ratios Based on Ca 5 (PO4) 3 OH formula Based on Ca 5 (PO4) 3 OH formula 2. Convert oxide concentrations to the subscripts in a chemical formula

12. Step 1: % Oxide / Molecular weight * # of Oxygens = X Step 2: Repeat for each element Step 3: Sum X’s for each element Step 4: Divide # of Oxygens by the total Step 5: Based on the chemical formula sum all X’s that substitute into the same structural postion Step 6: Divide X by the total from step 5 for its substitute group and multiply by the amount of the group in the basic formula = Amount of element in mineral

13. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Needed to set bone as supersaturated so it will exist as a solid. Needed to set bone as supersaturated so it will exist as a solid. Needed to add ground water solution Needed to add ground water solution 3. Create input for Phreeqc

14. Needed to create a formula for the bone Needed to create a formula for the bone Based on Ca 5 (PO4) 3 OH Based on Ca 5 (PO4) 3 OH Added as a Phase Added as a Phase 3. Create input for Phreeqc Phases Fossil Ca4.7896Al0.000067Fe0.0064Mn0.01137Mg0.02876Na0.1592 K0.0033Ba0.000669U0.000001Ce0.00000033Nd0.00000033(PO4)2.998 (SiO4)0.0018OH0.939F0.0347Cl0.0263 + 3.0582H+ 0.939H2O + 0.0347HF + 0.0263H+ 0.0263Cl- + 2.998HPO4-2 + 0.0018H4SiO4 + 0.000067Al+3 + 0.0064Fe+2 + 0.01137Mn+2 + 0.02876Mg+2 + 0.1592Na+ + 0.0033K+ + 0.000669Ba+2 + 0.000001U+4 + 0.00000033Ce+3 + 0.00000033Nd+3 + 4.7896Ca+2 log_k -3.421 delta_h -36.155 kcal

15. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Define SOLUTION_MASTER_SPECIES Define SOLUTION_MASTER_SPECIES Define SOLUTION_SPECIES Define SOLUTION_SPECIES 3. Create input for Phreeqc

16. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Define SOLUTION_MASTER_SPECIES Define SOLUTION_MASTER_SPECIES Name of element Name of element Charge Charge Molecular Weight Molecular Weight 3. Create input for Phreeqc SOLUTION_MASTER_SPECIES U U+4 0.0 238.0290 238.0290 U(4) U+4 0.0 238.0290 U(5) UO2+ 0.0 238.0290 U(6) UO2+2 0.0 238.0290

17. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Define SOLUTION_MASTER_SPECIES Define SOLUTION_MASTER_SPECIES Define SOLUTION_SPECIES Define SOLUTION_SPECIES Set of reactions that define the species Set of reactions that define the species Log K of each reaction Log K of each reaction Use log k = Δ G o / -1.364 Use log k = Δ G o / -1.364 Δ H o of each reaction in kcal Δ H o of each reaction in kcal Δ H o = products – reactants Δ H o = products – reactants Δ H o and Δ G o from Faure 1991 Δ H o and Δ G o from Faure 1991 3. Create input for Phreeqc

18. SOLUTION_SPECIES #primary master species for Ce #is also secondary master species for Ce(3) Ce+3 = Ce+3 log_k 0.0 Ce+3 + 3 H2O = Ce(OH)3 + 3 H+ log_k -45.496 delta_h -164.931 kcal Ce+3 + 4 H2O = Ce(OH)4- + 4 H+ log_k -60.662 delta_h -219.908 kcal #secondary master species for U(5) Ce+3 + 2 H2O = CeO2+ + 4 H+ + 2e- log_k 3.19 delta_h -5.8 kcal #secondary master species for U(6) 2Ce+3 + 3H2O = Ce2O3+ + 6 H+ + e- log_k 4.785 delta_h -8.7 kcal CeO2+ + CO3-2 = CeO2CO3- log_k 0 delta_h 0 kcal CeO2+ + 2CO3-2 = CeO2(CO3)2-3 log_k 0 delta_h 0 kcal Ce+3 + 3 H2O = Ce(OH)3 + 3 H+ log_k -45.496 delta_h -164.931 kcal

19. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Needed to set bone as supersaturated so it will exist as a solid. Needed to set bone as supersaturated so it will exist as a solid. Click Click Select solid Select solid Set SI and moles Set SI and moles 3. Create input for Phreeqc EQUILIBRIUM_PHASES 1 Fossil 1 1

20. Needed to create a formula for the bone Needed to create a formula for the bone Needed to add elements not in database Needed to add elements not in database Needed to set bone as supersaturated so it will exist as a solid. Needed to set bone as supersaturated so it will exist as a solid. Needed to add ground water solution Needed to add ground water solution Click Click Add pH, pe, and temperature in first tab Add pH, pe, and temperature in first tab Add element concentrations in second tab Add element concentrations in second tab 3. Create input for Phreeqc

21. SOLUTION_MASTER_SPECIES U U+4 0.0 238.0290 238.0290 U U+4 0.0 238.0290 238.0290 U(4) U+4 0.0 238.0290 U(4) U+4 0.0 238.0290 U(5) UO2+ 0.0 238.0290 U(5) UO2+ 0.0 238.0290 U(6) UO2+2 0.0 238.0290 U(6) UO2+2 0.0 238.0290SOLUTION_SPECIES #primary master species for U #primary master species for U #is also secondary master species for U(4) #is also secondary master species for U(4) U+4 = U+4 U+4 = U+4 log_k 0.0 log_k 0.0 U+4 + 4 H2O = U(OH)4 + 4 H+ U+4 + 4 H2O = U(OH)4 + 4 H+ log_k -8.538 log_k -8.538 delta_h 24.760 kcal delta_h 24.760 kcal U+4 + 5 H2O = U(OH)5- + 5 H+ U+4 + 5 H2O = U(OH)5- + 5 H+ log_k -13.147 log_k -13.147 delta_h 27.580 kcal delta_h 27.580 kcal #secondary master species for U(5) #secondary master species for U(5) U+4 + 2 H2O = UO2+ + 4 H+ + e- U+4 + 2 H2O = UO2+ + 4 H+ + e- log_k -6.432 log_k -6.432 delta_h 31.130 kcal delta_h 31.130 kcal #secondary master species for U(6) #secondary master species for U(6) U+4 + 2 H2O = UO2+2 + 4 H+ + 2 e- U+4 + 2 H2O = UO2+2 + 4 H+ + 2 e- log_k -9.217 log_k -9.217 delta_h 34.430 kcal delta_h 34.430 kcal UO2+2 + H2O = UO2OH+ + H+ UO2+2 + H2O = UO2OH+ + H+ log_k -5.782 log_k -5.782 delta_h 11.015 kcal delta_h 11.015 kcal 2UO2+2 + 2H2O = (UO2)2(OH)2+2 + 2H+ 2UO2+2 + 2H2O = (UO2)2(OH)2+2 + 2H+ log_k -5.626 log_k -5.626 delta_h -36.04 kcal delta_h -36.04 kcal 3UO2+2 + 5H2O = (UO2)3(OH)5+ + 5H+ 3UO2+2 + 5H2O = (UO2)3(OH)5+ + 5H+ log_k -15.641 log_k -15.641 delta_h -44.27 kcal delta_h -44.27 kcal UO2+2 + CO3-2 = UO2CO3 UO2+2 + CO3-2 = UO2CO3 log_k 10.064 log_k 10.064 delta_h 0.84 kcal delta_h 0.84 kcal UO2+2 + 2CO3-2 = UO2(CO3)2-2 UO2+2 + 2CO3-2 = UO2(CO3)2-2 log_k 16.977 log_k 16.977 delta_h 3.48 kcal delta_h 3.48 kcal UO2+2 + 3CO3-2 = UO2(CO3)3-4 UO2+2 + 3CO3-2 = UO2(CO3)3-4 log_k 21.397 log_k 21.397 delta_h -18.630 kcal delta_h -18.630 kcal SOLUTION_MASTER_SPECIES Ce Ce+3 0.0 140.116 140.116 Ce(3) Ce+3 0.0 140.116 Ce(4) Ce2O3+ 0.0 140.116 Ce(5) CeO2+ 0.0 140.116 SOLUTION_SPECIES #primary master species for Ce #is also secondary master species for Ce(3) Ce+3 = Ce+3 log_k 0.0 Ce+3 + 3 H2O = Ce(OH)3 + 3 H+ log_k -45.496 delta_h -164.931 kcal Ce+3 + 4 H2O = Ce(OH)4- + 4 H+ log_k -60.662 delta_h -219.908 kcal #secondary master species for U(5) Ce+3 + 2 H2O = CeO2+ + 4 H+ + 2e- log_k 3.19 delta_h -5.8 kcal #secondary master species for U(6) 2Ce+3 + 3H2O = Ce2O3+ + 6 H+ + e- log_k 4.785 delta_h -8.7 kcal CeO2+ + CO3-2 = CeO2CO3- log_k 0 delta_h 0 kcal CeO2+ + 2CO3-2 = CeO2(CO3)2-3 log_k 0 delta_h 0 kcal SOLUTION_MASTER_SPECIES Nd Nd+3 0.0 144.24 144.24 Nd(3) Nd+3 0.0 144.24 Nd(5) NdO2+ 0.0 144.24 Nd(6) NdO2+2 0.0 144.24 SOLUTION_SPECIES #primary master species for U #is also secondary master species for U(4) Nd+3 = Nd+3 log_k 0.0 Nd+3 + 3 H2O = Nd(OH)3 + 3 H+ log_k -45.496 delta_h -164.931 kcal 3. Create input for Phreeqc Nd+3 + 5 H2O = U(OH)5-2 + 5 H+ log_k -75.827 log_k -75.827 delta_h -274.885 kcal delta_h -274.885 kcal #secondary master species for U(5) #secondary master species for U(5) Nd+3 + 2 H2O = NdO2+ + 4 H+ + 2e- Nd+3 + 2 H2O = NdO2+ + 4 H+ + 2e- log_k 3.19 log_k 3.19 delta_h -5.8 kcal delta_h -5.8 kcal #secondary master species for U(6) #secondary master species for U(6) Nd+3 + 2 H2O = NdO2+2 + 4 H+ + 3 e- Nd+3 + 2 H2O = NdO2+2 + 4 H+ + 3 e- log_k 3.19 log_k 3.19 delta_h -5.8 kcal delta_h -5.8 kcal NdO2+2 + H2O = NdO2OH+ + H+ NdO2+2 + H2O = NdO2OH+ + H+ log_k -15.165 log_k -15.165 delta_h -54.977 kcal delta_h -54.977 kcal 2NdO2+2 + 2H2O = (NdO2)2(OH)2+2 + 2H+ 2NdO2+2 + 2H2O = (NdO2)2(OH)2+2 + 2H+ log_k -30.331 log_k -30.331 delta_h -109.954 kcal delta_h -109.954 kcal 3NdO2+2 + 5H2O = (NdO2)3(OH)5+ + 5H+ 3NdO2+2 + 5H2O = (NdO2)3(OH)5+ + 5H+ log_k -75.827 log_k -75.827 delta_h -274.885 kcal delta_h -274.885 kcal NdO2+2 + CO3-2 = NdO2CO3 NdO2+2 + CO3-2 = NdO2CO3 log_k 0 log_k 0 delta_h 0 kcal delta_h 0 kcal NdO2+2 + 2CO3-2 = NdO2(CO3)2-2 NdO2+2 + 2CO3-2 = NdO2(CO3)2-2 log_k 0 log_k 0 delta_h 0 kcal delta_h 0 kcal NdO2+2 + 3CO3-2 = NdO2(CO3)3-4 NdO2+2 + 3CO3-2 = NdO2(CO3)3-4 log_k 0 log_k 0 delta_h 0 kcal delta_h 0 kcalPhasesFossil Ca4.7896Al0.000067Fe0.0064Mn0.01137Mg0.02876Na0.1592K0.0033Ba0.000669U0.00 0001Ce0.00000033Nd0.00000033(PO4)2.998(SiO4)0.0018OH0.939F0.0347Cl0.0263 + 4 H+ = 0.939H2O + 0.0347HF + 0.0263HCl + 2.998HPO4-2 + 0.0018HSiO4-2 + 0.000067Al+3 + 0.0064Fe+2 + 0.01137Mn+2 + 0.02876Mg+2 + 0.1592Na+ + 0.0033K+ + 0.000669Ba+2 + 0.000001U+4 + 0.00000033Ce+3 + 0.00000033Nd+3 + 4.7896Ca+2 log_k -3.421 log_k -3.421 delta_h -36.155 kcal delta_h -36.155 kcalUraninite UO2 + 4 H+ = U+4 + 2 H2O UO2 + 4 H+ = U+4 + 2 H2O log_k -3.490 log_k -3.490 delta_h -18.630 kcal delta_h -18.630 kcal SOLUTION 1 pH 7.1 charge temp 12 pe units mg/L Ca 97.2 C 309 as HCO3 Cl 4 Mg 27.2 Mn(2).1 N 5.7 as N03- K 6.2 Si 11.3 Na 7.7 Sr.4 S 57 as SO4-2 EQUILIBRIUM_PHASES 1 Fossil 1 1 Fossil 1 1END

22. Results After several attempts Phreeqc would NOT recognize H+ and would not run my file After several attempts Phreeqc would NOT recognize H+ and would not run my file I was unable to find a solution to this problem I was unable to find a solution to this problem Here’s an example of what I was looking for… Here’s an example of what I was looking for…

23. Example Hydroxyapitite with an Fe substitution Hydroxyapitite with an Fe substitution Why Fe? Thought to be important in soft tissue preservation Why Fe? Thought to be important in soft tissue preservation

24. Example Input file Input file Phases Fossil Ca4.9936Fe0.0064(PO4)3OH + 4 H+ = H2O + 3 HPO4-2 + 4.9936 Ca+2 + 0.0064 Fe+2 log_k -3.421 delta_h -36.155 kcal SOLUTION 1 pH 7.1 charge temp 12 pe units mg/L Ca 97.2 C 309 as HCO3 Cl 4 Mg 27.2 Mn(2).1 N 5.7 as N03- K 6.2 Si 11.3 Na 7.7 Sr.4 S 57 as SO4-2 EQUILIBRIUM_PHASES 1 Fossil 1 1 END

25. Example Results Results Little to no iron entered solution Little to no iron entered solution Dissolved Ca and Mg relatively unchanged in solution Dissolved Ca and Mg relatively unchanged in solution Fossil, Hematite, Goethite, Hydroxyapatite, Dolomite, Calcite, Aragonite, Chrysotile, Sepolite, and Talc are supersaturated Fossil, Hematite, Goethite, Hydroxyapatite, Dolomite, Calcite, Aragonite, Chrysotile, Sepolite, and Talc are supersaturated

26. Example What does this mean? What does this mean? These results support a current theory (Schweitzer 2010) that when red blood cells break down their iron attaches to soft tissues in the bone to help preserve them so they would be a solid rather than in solution These results support a current theory (Schweitzer 2010) that when red blood cells break down their iron attaches to soft tissues in the bone to help preserve them so they would be a solid rather than in solution

27. References Andy Tindle – Free Software [Internet]. [cited 2010 Dec 6]. Available from: http://www.open.ac.uk/earth- research/tindle/AGTWebPages/AGTSoft.html Faure G. Principles and Applications of Geochemistry 2 nd Ed. Upper Saddle River, NJ: Prentice Hall; 1991. 520-574 p. Klein C, Dutrow B. The 23 rd Ed of the Manual of Mineral Science. Hoboken, NJ: John Wiley & Sons, INC; 2008. 99-104, 406, 428-433 P. Kohn MJ, Schoeninger MJ, Barker WW. Altered states: Effects of diagenesis on fossil tooth chemistry. Geochimica et Cosmochimica Acta 1999; 63: 2737– 2747. Moses R. Experimental Diagenisis of Bone: Implication for Rare Earth Elements uptake and stability. Presentation at 70 th SVP Annual meeting. 2010. Schweitzer M. Molecular Mechanisms for the Preservation of Soft Tissues and Oringinal Biomolecules in Fossils. Presentation at 70 th SVP Annual meeting. 2010. Stotler R, Harvey FE, Gosselin DC. A Black Hills-Madison Aquifer Origin for DakotaAquifer Groundwater in Northeastern Nebraska. Ground Water 2010; 48: 448–664.

28. Thank you!