Amino Acid Racemization Dating

Sean D. Pitman M.D.

© August,2004

All living things use proteins as building

blocks in the construction of their physical

forms. In turn, proteins are composed of

folded strands of 20 different smaller

subunits called "amino acids". All amino

acids, except for one (glycine), come in two

different forms known as the levoratory (L -

left) and dextrorotary (D - right) forms.

These two forms are called

"enantiomers", "chirals", or

"stereoisomers", which basically means

that they have the same molecular and

structural formula but cannot be

superimposed on each other no matter

how they are oriented in space. In other words, they are like one's left and right hands,

which are mirror images of each other, but cannot be superimposed onto one another.
 What is especially interesting about these two L- and D-forms, at least for the

purposes of this topic, is that the vast majority of living things only use the L-form.

However, as soon as the creature dies, the L-amino acids start to spontaneously convert

to the D-form through a process called "racemization". If the rate of conversion can be

determined, this process of racemization might be useful as a sort of "clock" to determine

the time of death.

 Basic Assumptions

In order to use the rate of

racemization as a clock to

accurately estimate when a

living thing died, one must

know how various

environmental factors may have affected the rate of change from the L- to the D-form. As

it turns out, this rate, which is different for each type of amino acid, is also exquisitely

sensitive to certain environmental factors. These include:

1. Temperature

2. Amino acid composition of the protein

3. Water concentration in the environment

4. pH (acidity/alkalinity) in the environment

5. Bound state versus free state

6. Size of the macromolecule, if in a bound

state
 7. Specific location in the macromolecule, if

in a bound state

8. Contact with clay surfaces (catalytic

effect)

9. Presence of aldehydes, particularly when

associated with metal ions

10. Concentration of buffer compounds

11. Ionic strength of the environment

Of these, temperature is generally thought to play the most significant role in

determining the rate of racemization since a 1o increase in temperature results in a 20-

25% increase in the racemization rate.1,2 Clearly, this factor alone carries with it a huge

potential for error. Even slight ranges of error in determining the "temperature history" of a

specimen will result in huge "age" calculation errors.

Calibrating for even a known temperature history also seems to be rather problematic.

Consider that the rate of racemization for various amino acids is determined by placing a

protein into a very high temperature environment (between 95o and 150o C) and then

extrapolating these results to low temperature environments.1

Such extrapolations have been fairly recently (1999) called into question by

experiments showing that models based on high temperature kinetics fail to predict

racemization kinetics at physiologic temperatures (i.e., 37o C). The authors of this

particular paper went on to suggest that, "As conformation strongly influences the rate of

Asu [cyclic succinimide] formation and hence Asx [aspartic acid + asparagine]
racemization, the use of extrapolation from high temperatures to estimate racemization

kinetics of Asx in proteins below their denaturation temperature is called into question . . .

We argue that the D:L ratio of Asx reflects the proportion of non-helical to helical collagen

"3 Others, such as Collins and Riley have commented in the press that, "racemization in

free amino acids has, unfortunately, little bearing on observed racemization of

archaeological biominerals."1

Other experiments have shown that the L- and D-forms of the same amino acid do not

racemize at the same rate. This means that the equilibrium ratio may be off from "50:50"

by as much as 25%.4

As far as pH is concerned, it seems that the temperature of a solution also affects the

pH of that solution. So, the amino acid racemization (AAR) rates not only change with the

effects of temperature, but also with the concurrent effects of pH changes, which are

themselves affected by temperature.1 This can only increase the potential range of error

for age determinations. The local buffering effects of bone and shell matrixes are

supposed to limit this effect, but it is still something to consider as potentially significant

when acting over the course of tens of thousands to millions of years.

Also, the actual physical structure of an intact protein significantly affects the rate of

racemization of various amino acids. In fact, in many cases this may even be a more

significant factor than the temperature history. As it turns out, the N-terminal amino acids

racemize faster than the C-terminal amino acids of the same types. Also, the surface

amino acids racemize much faster than the interior amino acids. And, interestingly

enough, free amino acids have the slowest racemization rate of all. Studies with short

peptides have shown that, "replacement of the asparagine residue with aspartic acid
resulted in a 34-fold decrease in the rate of succinimide (Asu) formation. In the position

carboxyl to asparagine in the peptide the replacement of glycine with a bulky amino acid

such as proline or leucine resulted in a 33-50-fold decrease in the rate of deamidation"1

[Emphasis added] This clearly emphasizes the rather dramatic importance that amino

acid position and overall protein amino acid sequence has on the racemization rate.

Hydrolysis, or the process of breaking down a protein into smaller and smaller

fragments, clearly affects the rate of racemization. The rate itself of hydrolysis "depends

on the strength of the individual peptide bonds, which in turn is determined by the

characteristics of the amino acids on either side of the bond, the presence of water and

the temperature."1 With increased hydrolysis, the overall rate of the whole specimen

would increase since there are more terminal end and surface amino acids to undergo

higher rates of racemization. All of these are confounding factors, which, if not known

exactly over extended periods of time, would play havoc with any sort of age

determinations. Even the process of preparing a specimen for racemic dating can affect

the D/L ratio.

In this light, consider that age determinations are usually performed on specimens for

which the amino acid order of the original proteins is unknown. For example, consider

that neither the structure nor the proportion of the amino acids used for dating coral,

ostrich eggshell, or snail shells is known. Again, those like Goodfriend and Hare (1995)

have pointed out the "difficulties in estimating the amount of asparagine in proteins and

reminded researchers of the dangers of extrapolating from the behaviour of pure Asn in

solution at high temperature, to the behaviour of Asn in proteins associated with

biominerals at ambient temperature . . . Using a simplified model of 'racemization kinetics'

of bone collagen over geological time Collins shows that almost any value of D-Asx can

be obtained by varying the rates of collagen denaturation and leaching of the denatured

product."1

The Interaction of bacteria and fungi with organic specimens may also be

problematic. Such creatures have various enzymes that can digest various types of

proteins, such as collagen, that are commonly used for dating. Various studies, such as

those dealing with 18th and 19th century burials, showed "unexpectedly high levels of

aspartic acid racemization." The authors "suggested that either biological or chemical

degradation of the tooth collagen might have caused these results." 1 Also, certain types

of bacteria and other creatures actually produce the D-form instead of the L-form of

amino acids. So, special care is obviously needed in order to particularly avoid this sort

of contamination.

Calibrating the Amino Acid "Clock"

"Amino acid dating cannot obtain the age of the material purely from the data

itself. The rate of racemization can not be standardized by itself because it is too

changeable. Thus, because of the rate problem, this dating technique must rely on other

dating techniques to standardize its findings. As a matter of fact, the ages obtained from

racemization dating must rely on other techniques such as Carbon 14, and if the dating

of Carbon 14 is not accurate, racemization dating can never be certain."6

 "The potential variation in the racemization rate has led some paleoanthropologists

to consider this dating technique relative rather than chronometric. It is, perhaps, best

considered to be a calibrated relative dating technique which puts it somewhere

between relative and chronometric methods."7

Clearly, all of the above described variables for amino acid racemization rates create

great difficulty for AAR as a dating technique. In fact, the difficulties are so great that

this technique cannot be and is not used as any sort of "absolute" dating technique. So,

how is it thought to be at all helpful? Well, it is thought to be helpful as a "relative"

dating technique.

To overcome the various uncertainties inherent to amino acid dating, the method must

be "calibrated" based on other more reliable techniques such as radiocarbon dating

(carbon 14 dating). What happens is that a specimen from a site is chosen as the

"calibration sample" and both a radiocarbon date as well as a D/L amino acid ratio is

determined. These values are used to solve for a constant or "k" in the formula used to

estimate ages based on the calibration sample. Of course, the "major assumption

required with this approach is that the average temperature experienced by the

'calibration' sample is representative of the average temperature experienced by other

samples from the deposit."1

Much effort has gone into transforming the data in various ways to achieve linearity

between the D/L ratio and the calibrated age of the specimens in a given location. At first

"cubic transformations"' and then later "power function transformations" were used that

seemed to show a "strong correlation with time, but did not explain the observed kinetics."
 What this basically means is that amino acid dating is not based on any sort of

understanding about how racemization takes place, but is strictly a function of correlation

with other dating techniques, such as the radiocarbon technique. So, if there is any

problem with the basis of the correlation (i.e., radiocarbon dating) then there will also be

the same problem with amino acid dating.

In this light, it is interesting to consider what happened in 1974 when some of the major

proponents of amino acid dating (Bada et al) decided to analyze the Paleo-Indian skeletal

material from Del Mar, California. Their estimated age of 48,000 years before present

(BP) "stunned" the archaeological community who generally believed these bones to be

less than 10,000 years old. Bada went on to date other skeletal specimens between the

35,000 and 48,000 year range with one specimen from Sunnyvale being dated at an

astonishing 70,000 years BP. Then, in the 1980s, something very interesting happened.

"The Sunnyvale skeleton and the Del Mar tibia were re-dated

using uranium series dating. This resulted in dates of 8,000 to 9,000

years BP for Sunnyvale and 11,000 to 11,500 for Del Mar.

Conventional plus accelerator mass spectrometry (AMS) radiocarbon

dating (Taylor et al. 1983) was carried out on the Sunnyvale skeleton

and results of between 3,600 and 4,850 years BP were obtained. The

original amino acid extractions from the racemization studies of the

Paleo-Indian remains were independently dated by the AMS

radiocarbon method at the Oxford Radiocarbon Accelerator Unit of

Oxford University and the NSF Accelerator Facility for Radioisotope

Analysis, University of Arizona. Bada et al., (1984) published the

Oxford results and Taylor et al., (1985) published a paper combining

the results from both laboratories. The Oxford dates were all between

4,500 and 8,500 years BP and the Arizona dates were between

3,000 and 6,600 years BP. Bada et al., (1984) stated that the Oxford

AMS results reveal no clear relationship between the radiocarbon

ages of the various skeletons and the extent of the aspartic acid

racemization. They did note that there appeared to be a direct

relationship between the extent of racemization and the level of

preservation of collagen in the bones. Those samples with the most

racemization had the lowest amino acid content and this poor

preservation of protein would contribute to anomalous AAR results.

Later, based on AMS radiocarbon dates, Bada (1985) calculated a

new value for kasp for the Californian samples. He used the Laguna

skull and the Los Angeles Man skeleton as 'calibration' samples for

this. Using the revised value for kasp he recalculated the AAR dates

of the other Paleo-Indian samples. They all fell within the Holocene

but had much larger error estimates than those of the AMS values.

Although Bada claimed consistency between AAR and AMS dates

others (Pollard and Heron 1996, p. 228) argue that the dates only

appear to be consistent with one another because of the

unacceptably large error range associated with the AAR dates.

Pollard and Heron also point out that there is poor concordance

between the conventional and the AMS radiocarbon dates and there
is no concordance between the uranium series dates and any of the

other dates either. At best three of the four methods put the bones in

the Holocene."1

Because of these problems AAR dating of bone and teeth (teeth in different locations

in the same mouth have been shown to have very different AAR ages) is considered to be

an extremely unreliable practice even by mainstream scientists. That is because the

porosity of bones makes them more "open" to surrounding environmental influences and

leaching. Specimens that are more "closed" to such problems are thought to include

mollusk shells and especially ratite (bird) eggshells from the emu and ostrich. Of course,

even if these rather thin specimens were actually "closed" systems (more so than even

teeth enamel) they would still be quite subject to local temperature variations as well as

the other above-mentioned potential problems. For example, even today "very little is

known about the protein structure in ratite eggshell and differences in primary sequence

can alter the rate of Asu formation by two orders of magnitude [100-fold] (Collins, Waite,

and van Duin 1999). Goodfriend and Hare (1995) show that Asx racemization in ostrich

eggshell heated at 80 oC has complex kinetics, similar to that seen in land snails

(Goodfriend 1992). The extrapolation of high temperature rates to low temperatures is

known to be problematic (Collins, Waite, and van Duin 1999). A pilot study would be

necessary and a reliable relationship between racemate ratio and time could remain

elusive."1
 Also, there is a potential problem with radiocarbon correlations that is quite interesting.

Note what happens to the correlation constant (k) with assumed age of the specimen in

the following figures.

 Interestingly enough, the racemization constant or "k" values for the amino acid dating

of various specimens decreases dramatically with the assumed age of the specimens

(see figures).5 This means that the rate of racemization was thousands of times (up to

2,000 times) different in the past than it is today. Note that these rate differences include

shell specimens, which are supposed to be more reliable than other more "open system"

specimens, such as wood and bone.

Is this a reasonable assumption? Well, this simply must be true if radiocarbon dating

is accurate beyond a few thousand years. But, what if radiocarbon gets significantly

worse as one moves very far back in time beyond just a few thousand years? In other

words, what would it mean for one to assume that the k-values remained fairly constant
over time as would seem intuitive? Well, with the k-values plotted out horizontally on the

graph, the calculated ages of the specimens would be roughly affected as follows:5

Current Fossil Age

40,000 100,000 350,000 1,000,000
Assignment
Adjusted Fossil Figure 1 6,000 11,000 18,000 8,000

Age Assignment

with horizontal k- Figure 2 5,000 14,000 18,000 14,000

values

Clearly this is a dramatic adjustment that seems to suggest that amino acid

racemization may be more a reflection of the activities of local environmental differences

than any sort of differences in relative ages. This seems especially likely when one

considers that each type of specimen and each different location have different k-values

meaning that the radiocarbon-derived constant in one region or with one type of

specimen cannot be used to calculate the age of any other specimen or even the same

type of specimen in a different location.1

Add to this the fact that radiocarbon dating is also dependent upon the state of

preservation of the specimen.

 "Stafford et al., (1991) discussed AMS radiocarbon

dating in bone at the molecular level. They dated a

number of fractions (ranging from insoluble collagen to

individual amino acids) from each of a selection of

differentially preserved mammoth and human bone.

Age estimates from the fractions within a bone were

consistent if it was well preserved. They concluded that

a poorly preserved Pleistocene-age fossil >11,000

years in age would go unrecognised because it would

yield a Holocene 14C date. Thus the final irony is that

the poorly preserved Californian Paleo-Indian bones

would return Holocene 14C dates even if they were

actually Pleistocene. The state of preservation of the

bone appears to be as important an issue for

radiocarbon dating as it is for AAR dating."1

So, what do we have? In short, it seems like the claims of some scientists that amino

acid racemization dating has been well established as reliable appears to be wishful

thinking at best. The huge number of confounding factors and a complete inability to

explain the calibrating k-values in terms of amino acid kinetics leaves those with even a

tiny pessimistic bone in their bodies just a bit underwhelmed.

 1. Judith Robins, Martin Jones and Elizabeth Matisoo-Smith, Amino Acid
Racemization Dating in New Zealand: An overview and Bibliography, Auckland
University, Auckland, New Zealand, March 20, 2001 ( Link )

2. Mike Brown, Amino Acid Dating, Molecular History Research Center and Mikes
Origins Resource, Accessed July, 2004 ( Link )

3. Collins MJ, Waite ER, van Duin AC, Predicting protein decomposition: the case of
aspartic-acid racemization kinetics. Philos Trans R Soc Lond B Biol Sci 1999 Jan
29;354(1379):51-64 (Fossil Fuels and Environmental Geochemistry (Postgraduate
Institute), NRG, University of Newcastle-upon-Tyne, UK. ( m.collins@ncl.ac.uk )

4. Meyer MW, Meyer VR, Ramseyer S, The kinetics of diastereomeric amino acids
with o-phthaldialdehyde, Chirality 1991;3(6):471-5 PMID: 1812958, UI: 92256092
(Institute of Organic Chemistry, University of Bern, Switzerland)

5. R. H. Brown, Amino Acid Dating, Origins 12(1):8-25 (1985). ( Link )

6. Minnesota State University, Racemization essay, accessed December 2, 2007 (

Link )

7. Dennis O'Neil, "Chronometric Techniques - Part 1," Palomar.edu, June 7, 2007 (

Link )

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Amino Acid Racemization Dating

Sean D. Pitman M.D.

© August,2004

All living things use proteins as building

blocks in the construction of their physical

forms. In turn, proteins are composed of

folded strands of 20 different smaller subunits called "amino acids". All amino acids,

except for one (glycine), come in two different forms known as the levoratory (L - left) and

dextrorotary (D - right) forms. These two forms are called "enantiomers", "chirals", or

"stereoisomers", which basically means that they have the same molecular and structural

formula but cannot be superimposed on each other no matter how they are oriented in

space. In other words, they are like

one's left and right hands, which are

mirror images of each other, but cannot

be superimposed onto one another.

What is especially interesting about

these two L- and D-forms, at least for

the purposes of this topic, is that the

vast majority of living things only use the L-form. However, as soon as the creature dies,

the L-amino acids start to spontaneously convert to the D-form through a process called

"racemization". If the rate of conversion can be determined, this process of racemization

might be useful as a sort of "clock" to determine the time of death.

 Basic Assumptions

In order to use the rate of

racemization as a clock to

accurately estimate when a

living thing died, one must

know how various

environmental factors may have affected the rate of change from the L- to the D-form. As

it turns out, this rate, which is different for each type of amino acid, is also exquisitely

sensitive to certain environmental factors. These include:

1. Temperature

2. Amino acid composition of the protein

3. Water concentration in the environment

4. pH (acidity/alkalinity) in the environment

5. Bound state versus free state

6. Size of the macromolecule, if in a bound

state
 7. Specific location in the macromolecule, if

in a bound state

8. Contact with clay surfaces (catalytic

effect)

9. Presence of aldehydes, particularly when

associated with metal ions

10. Concentration of buffer compounds

11. Ionic strength of the environment

Of these, temperature is generally thought to play the most significant role in

determining the rate of racemization since a 1o increase in temperature results in a 20-

25% increase in the racemization rate.1,2 Clearly, this factor alone carries with it a huge

potential for error. Even slight ranges of error in determining the "temperature history" of a

specimen will result in huge "age" calculation errors.

Calibrating for even a known temperature history also seems to be rather problematic.

Consider that the rate of racemization for various amino acids is determined by placing a

protein into a very high temperature environment (between 95o and 150o C) and then

extrapolating these results to low temperature environments.1

Such extrapolations have been fairly recently (1999) called into question by

experiments showing that models based on high temperature kinetics fail to predict

racemization kinetics at physiologic temperatures (i.e., 37o C). The authors of this

particular paper went on to suggest that, "As conformation strongly influences the rate of

Asu [cyclic succinimide] formation and hence Asx [aspartic acid + asparagine]
racemization, the use of extrapolation from high temperatures to estimate racemization

kinetics of Asx in proteins below their denaturation temperature is called into question . . .

We argue that the D:L ratio of Asx reflects the proportion of non-helical to helical collagen

"3 Others, such as Collins and Riley have commented in the press that, "racemization in

free amino acids has, unfortunately, little bearing on observed racemization of

archaeological biominerals."1

Other experiments have shown that the L- and D-forms of the same amino acid do not

racemize at the same rate. This means that the equilibrium ratio may be off from "50:50"

by as much as 25%.4

As far as pH is concerned, it seems that the temperature of a solution also affects the

pH of that solution. So, the amino acid racemization (AAR) rates not only change with the

effects of temperature, but also with the concurrent effects of pH changes, which are

themselves affected by temperature.1 This can only increase the potential range of error

for age determinations. The local buffering effects of bone and shell matrixes are

supposed to limit this effect, but it is still something to consider as potentially significant

when acting over the course of tens of thousands to millions of years.

Also, the actual physical structure of an intact protein significantly affects the rate of

racemization of various amino acids. In fact, in many cases this may even be a more

significant factor than the temperature history. As it turns out, the N-terminal amino acids

racemize faster than the C-terminal amino acids of the same types. Also, the surface

amino acids racemize much faster than the interior amino acids. And, interestingly

enough, free amino acids have the slowest racemization rate of all. Studies with short

peptides have shown that, "replacement of the asparagine residue with aspartic acid
resulted in a 34-fold decrease in the rate of succinimide (Asu) formation. In the position

carboxyl to asparagine in the peptide the replacement of glycine with a bulky amino acid

such as proline or leucine resulted in a 33-50-fold decrease in the rate of deamidation"1

[Emphasis added] This clearly emphasizes the rather dramatic importance that amino

acid position and overall protein amino acid sequence has on the racemization rate.

Hydrolysis, or the process of breaking down a protein into smaller and smaller

fragments, clearly affects the rate of racemization. The rate itself of hydrolysis "depends

on the strength of the individual peptide bonds, which in turn is determined by the

characteristics of the amino acids on either side of the bond, the presence of water and

the temperature."1 With increased hydrolysis, the overall rate of the whole specimen

would increase since there are more terminal end and surface amino acids to undergo

higher rates of racemization. All of these are confounding factors, which, if not known

exactly over extended periods of time, would play havoc with any sort of age

determinations. Even the process of preparing a specimen for racemic dating can affect

the D/L ratio.

In this light, consider that age determinations are usually performed on specimens for

which the amino acid order of the original proteins is unknown. For example, consider

that neither the structure nor the proportion of the amino acids used for dating coral,

ostrich eggshell, or snail shells is known. Again, those like Goodfriend and Hare (1995)

have pointed out the "difficulties in estimating the amount of asparagine in proteins and

reminded researchers of the dangers of extrapolating from the behaviour of pure Asn in

solution at high temperature, to the behaviour of Asn in proteins associated with

biominerals at ambient temperature . . . Using a simplified model of 'racemization kinetics'

of bone collagen over geological time Collins shows that almost any value of D-Asx can

be obtained by varying the rates of collagen denaturation and leaching of the denatured

product."1

The Interaction of bacteria and fungi with organic specimens may also be

problematic. Such creatures have various enzymes that can digest various types of

proteins, such as collagen, that are commonly used for dating. Various studies, such as

those dealing with 18th and 19th century burials, showed "unexpectedly high levels of

aspartic acid racemization." The authors "suggested that either biological or chemical

degradation of the tooth collagen might have caused these results." 1 Also, certain types

of bacteria and other creatures actually produce the D-form instead of the L-form of

amino acids. So, special care is obviously needed in order to particularly avoid this sort

of contamination.

Calibrating the Amino Acid "Clock"

"Amino acid dating cannot obtain the age of the material purely from the data

itself. The rate of racemization can not be standardized by itself because it is too

changeable. Thus, because of the rate problem, this dating technique must rely on other

dating techniques to standardize its findings. As a matter of fact, the ages obtained from

racemization dating must rely on other techniques such as Carbon 14, and if the dating

of Carbon 14 is not accurate, racemization dating can never be certain."6

 "The potential variation in the racemization rate has led some paleoanthropologists

to consider this dating technique relative rather than chronometric. It is, perhaps, best

considered to be a calibrated relative dating technique which puts it somewhere

between relative and chronometric methods."7

Clearly, all of the above described variables for amino acid racemization rates create

great difficulty for AAR as a dating technique. In fact, the difficulties are so great that

this technique cannot be and is not used as any sort of "absolute" dating technique. So,

how is it thought to be at all helpful? Well, it is thought to be helpful as a "relative"

dating technique.

To overcome the various uncertainties inherent to amino acid dating, the method must

be "calibrated" based on other more reliable techniques such as radiocarbon dating

(carbon 14 dating). What happens is that a specimen from a site is chosen as the

"calibration sample" and both a radiocarbon date as well as a D/L amino acid ratio is

determined. These values are used to solve for a constant or "k" in the formula used to

estimate ages based on the calibration sample. Of course, the "major assumption

required with this approach is that the average temperature experienced by the

'calibration' sample is representative of the average temperature experienced by other

samples from the deposit."1

Much effort has gone into transforming the data in various ways to achieve linearity

between the D/L ratio and the calibrated age of the specimens in a given location. At first

"cubic transformations"' and then later "power function transformations" were used that

seemed to show a "strong correlation with time, but did not explain the observed kinetics."
 What this basically means is that amino acid dating is not based on any sort of

understanding about how racemization takes place, but is strictly a function of correlation

with other dating techniques, such as the radiocarbon technique. So, if there is any

problem with the basis of the correlation (i.e., radiocarbon dating) then there will also be

the same problem with amino acid dating.

In this light, it is interesting to consider what happened in 1974 when some of the major

proponents of amino acid dating (Bada et al) decided to analyze the Paleo-Indian skeletal

material from Del Mar, California. Their estimated age of 48,000 years before present

(BP) "stunned" the archaeological community who generally believed these bones to be

less than 10,000 years old. Bada went on to date other skeletal specimens between the

35,000 and 48,000 year range with one specimen from Sunnyvale being dated at an

astonishing 70,000 years BP. Then, in the 1980s, something very interesting happened.

"The Sunnyvale skeleton and the Del Mar tibia were re-dated

using uranium series dating. This resulted in dates of 8,000 to 9,000

years BP for Sunnyvale and 11,000 to 11,500 for Del Mar.

Conventional plus accelerator mass spectrometry (AMS) radiocarbon

dating (Taylor et al. 1983) was carried out on the Sunnyvale skeleton

and results of between 3,600 and 4,850 years BP were obtained. The

original amino acid extractions from the racemization studies of the

Paleo-Indian remains were independently dated by the AMS

radiocarbon method at the Oxford Radiocarbon Accelerator Unit of

Oxford University and the NSF Accelerator Facility for Radioisotope

Analysis, University of Arizona. Bada et al., (1984) published the

Oxford results and Taylor et al., (1985) published a paper combining

the results from both laboratories. The Oxford dates were all between

4,500 and 8,500 years BP and the Arizona dates were between

3,000 and 6,600 years BP. Bada et al., (1984) stated that the Oxford

AMS results reveal no clear relationship between the radiocarbon

ages of the various skeletons and the extent of the aspartic acid

racemization. They did note that there appeared to be a direct

relationship between the extent of racemization and the level of

preservation of collagen in the bones. Those samples with the most

racemization had the lowest amino acid content and this poor

preservation of protein would contribute to anomalous AAR results.

Later, based on AMS radiocarbon dates, Bada (1985) calculated a

new value for kasp for the Californian samples. He used the Laguna

skull and the Los Angeles Man skeleton as 'calibration' samples for

this. Using the revised value for kasp he recalculated the AAR dates

of the other Paleo-Indian samples. They all fell within the Holocene

but had much larger error estimates than those of the AMS values.

Although Bada claimed consistency between AAR and AMS dates

others (Pollard and Heron 1996, p. 228) argue that the dates only

appear to be consistent with one another because of the

unacceptably large error range associated with the AAR dates.

Pollard and Heron also point out that there is poor concordance

between the conventional and the AMS radiocarbon dates and there
is no concordance between the uranium series dates and any of the

other dates either. At best three of the four methods put the bones in

the Holocene."1

Because of these problems AAR dating of bone and teeth (teeth in different locations

in the same mouth have been shown to have very different AAR ages) is considered to be

an extremely unreliable practice even by mainstream scientists. That is because the

porosity of bones makes them more "open" to surrounding environmental influences and

leaching. Specimens that are more "closed" to such problems are thought to include

mollusk shells and especially ratite (bird) eggshells from the emu and ostrich. Of course,

even if these rather thin specimens were actually "closed" systems (more so than even

teeth enamel) they would still be quite subject to local temperature variations as well as

the other above-mentioned potential problems. For example, even today "very little is

known about the protein structure in ratite eggshell and differences in primary sequence

can alter the rate of Asu formation by two orders of magnitude [100-fold] (Collins, Waite,

and van Duin 1999). Goodfriend and Hare (1995) show that Asx racemization in ostrich

eggshell heated at 80 oC has complex kinetics, similar to that seen in land snails

(Goodfriend 1992). The extrapolation of high temperature rates to low temperatures is

known to be problematic (Collins, Waite, and van Duin 1999). A pilot study would be

necessary and a reliable relationship between racemate ratio and time could remain

elusive."1
 Also, there is a potential problem with radiocarbon correlations that is quite interesting.

Note what happens to the correlation constant (k) with assumed age of the specimen in

the following figures.

 Interestingly enough, the racemization constant or "k" values for the amino acid dating

of various specimens decreases dramatically with the assumed age of the specimens

(see figures).5 This means that the rate of racemization was thousands of times (up to

2,000 times) different in the past than it is today. Note that these rate differences include

shell specimens, which are supposed to be more reliable than other more "open system"

specimens, such as wood and bone.

Is this a reasonable assumption? Well, this simply must be true if radiocarbon dating

is accurate beyond a few thousand years. But, what if radiocarbon gets significantly

worse as one moves very far back in time beyond just a few thousand years? In other

words, what would it mean for one to assume that the k-values remained fairly constant
over time as would seem intuitive? Well, with the k-values plotted out horizontally on the

graph, the calculated ages of the specimens would be roughly affected as follows:5

Current Fossil Age

40,000 100,000 350,000 1,000,000
Assignment
Adjusted Fossil Figure 1 6,000 11,000 18,000 8,000

Age Assignment

with horizontal k- Figure 2 5,000 14,000 18,000 14,000

values

Clearly this is a dramatic adjustment that seems to suggest that amino acid

racemization may be more a reflection of the activities of local environmental differences

than any sort of differences in relative ages. This seems especially likely when one

considers that each type of specimen and each different location have different k-values

meaning that the radiocarbon-derived constant in one region or with one type of

specimen cannot be used to calculate the age of any other specimen or even the same

type of specimen in a different location.1

Add to this the fact that radiocarbon dating is also dependent upon the state of

preservation of the specimen.

 "Stafford et al., (1991) discussed AMS radiocarbon

dating in bone at the molecular level. They dated a

number of fractions (ranging from insoluble collagen to

individual amino acids) from each of a selection of

differentially preserved mammoth and human bone.

Age estimates from the fractions within a bone were

consistent if it was well preserved. They concluded that

a poorly preserved Pleistocene-age fossil >11,000

years in age would go unrecognised because it would

yield a Holocene 14C date. Thus the final irony is that

the poorly preserved Californian Paleo-Indian bones

would return Holocene 14C dates even if they were

actually Pleistocene. The state of preservation of the

bone appears to be as important an issue for

radiocarbon dating as it is for AAR dating."1

So, what do we have? In short, it seems like the claims of some scientists that amino

acid racemization dating has been well established as reliable appears to be wishful

thinking at best. The huge number of confounding factors and a complete inability to

explain the calibrating k-values in terms of amino acid kinetics leaves those with even a

tiny pessimistic bone in their bodies just a bit underwhelmed.

 1. Judith Robins, Martin Jones and Elizabeth Matisoo-Smith, Amino Acid
Racemization Dating in New Zealand: An overview and Bibliography, Auckland
University, Auckland, New Zealand, March 20, 2001 ( Link )

2. Mike Brown, Amino Acid Dating, Molecular History Research Center and Mikes
Origins Resource, Accessed July, 2004 ( Link )

3. Collins MJ, Waite ER, van Duin AC, Predicting protein decomposition: the case of
aspartic-acid racemization kinetics. Philos Trans R Soc Lond B Biol Sci 1999 Jan
29;354(1379):51-64 (Fossil Fuels and Environmental Geochemistry (Postgraduate
Institute), NRG, University of Newcastle-upon-Tyne, UK. ( m.collins@ncl.ac.uk )

4. Meyer MW, Meyer VR, Ramseyer S, The kinetics of diastereomeric amino acids
with o-phthaldialdehyde, Chirality 1991;3(6):471-5 PMID: 1812958, UI: 92256092
(Institute of Organic Chemistry, University of Bern, Switzerland)

5. R. H. Brown, Amino Acid Dating, Origins 12(1):8-25 (1985). ( Link )

6. Minnesota State University, Racemization essay, accessed December 2, 2007 (

Link )

7. Dennis O'Neil, "Chronometric Techniques - Part 1," Palomar.edu, June 7, 2007 (

Link )

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