TY - JOUR
T1 - Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy Sensitivity to the Thermal Decay of Bone Collagen
AU - Thomas, Brian
AU - Anderson, Kevin
AU - De Silva, Imesha
AU - Verbeck, Guido
AU - Taylor, Stephen
N1 - Funding Information:
We thank Joshua Jacobi and Ryan Anderson for their assistance in the laboratory. Abhi Bugde and Kate Phelps of the Live Cell Imaging Core Facility at UT Southwestern Medical Center assisted with SHG imaging, supported by NIH S10 RR029731-01. We specially thank Tom Kiselak from Guido Verbeck’s Laboratory of Imaging Mass Spectrometry at the University of North Texas for their assistance with ATR-IR.
Publisher Copyright:
© The Author(s) 2022.
PY - 2023/1
Y1 - 2023/1
N2 - The analysis of collagen stability is of interest in forensics, archaeology, and molecular paleontology. Collagen decay rates are often measured by thermal kinetic studies that employ liquid chromatography mass spectrometry (LC-MS) to assay collagen quantities. However, these kinetic studies generally focus on measuring the decreasing levels of collagen instead of an exact molecular concentration of each sample. Thus, attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy can offer a simpler and less expensive alternative to LC-MS. The application of a new protocol to determine decreasing amounts of bone collagen in artificially decayed porcine and bovine bone was assessed. The protocol uses a forensic application of ATR FT-IR spectroscopy on size-restricted bone powder from three uniformly high temperature conditions. Also, for the first time, collagen-specific second-harmonic generation (SHG) imaging was also applied to artificially aged bone to add an independent, qualitative perspective to parallel FT-IR assessments. SHG images and ATR FT-IR spectra together reveal the same orderly bone collagen decay as found in previous thermal kinetic studies. Resulting Arrhenius plots with r2 values > 0.95 suggest that the ATR FT-IR-based protocol has potential as a precise and simple tool for measuring bone collagen decay rates. The results are significant for applications of thermal kinetic studies, and our protocol can serve as an inexpensive, precise, and pragmatic means of evaluating bone collagen stability within an array of conditions.
AB - The analysis of collagen stability is of interest in forensics, archaeology, and molecular paleontology. Collagen decay rates are often measured by thermal kinetic studies that employ liquid chromatography mass spectrometry (LC-MS) to assay collagen quantities. However, these kinetic studies generally focus on measuring the decreasing levels of collagen instead of an exact molecular concentration of each sample. Thus, attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy can offer a simpler and less expensive alternative to LC-MS. The application of a new protocol to determine decreasing amounts of bone collagen in artificially decayed porcine and bovine bone was assessed. The protocol uses a forensic application of ATR FT-IR spectroscopy on size-restricted bone powder from three uniformly high temperature conditions. Also, for the first time, collagen-specific second-harmonic generation (SHG) imaging was also applied to artificially aged bone to add an independent, qualitative perspective to parallel FT-IR assessments. SHG images and ATR FT-IR spectra together reveal the same orderly bone collagen decay as found in previous thermal kinetic studies. Resulting Arrhenius plots with r2 values > 0.95 suggest that the ATR FT-IR-based protocol has potential as a precise and simple tool for measuring bone collagen decay rates. The results are significant for applications of thermal kinetic studies, and our protocol can serve as an inexpensive, precise, and pragmatic means of evaluating bone collagen stability within an array of conditions.
KW - ATR FT-IR
KW - Attenuated total reflection Fourier transform infrared spectroscopy
KW - collagen degradation
KW - second harmonic generation
KW - SHG
KW - thermal kinetics
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U2 - 10.1177/00037028221135634
DO - 10.1177/00037028221135634
M3 - Article
C2 - 36253880
AN - SCOPUS:85140117058
SN - 0003-7028
VL - 77
SP - 53
EP - 61
JO - Applied Spectroscopy
JF - Applied Spectroscopy
IS - 1
ER -