FTIR-Spectroscopic Analysis of Kidney Stones Bruker Optics

Infrared Light 12. 02. 2018 2

Infrared (IR) Spectroscopy: Principle Interaction between light and matter Matter incident light beam Absorption Transmission Reflection February 12, 2018 3

Infrared (IR) Spectroscopy: Principle Interaction between light and matter Symmetric stretching vibration Anti-symmetric stretching vibration Deformation vibration Infrared light is transformed into vibrations 12. 02. 2018 4

Infrared (IR) Spectroscopy: Principle Infrared light is transformed into vibrations Example: water spectrum 12. 02. 2018 5

IR Spectroscopy: Analytical Value Examples of IR-spectra of different substances Lactose Magnesium carbonate Magnesium sulfate Urea Ø IR-Spectrum: Chemical fingerprint of the sample February 12, 2018 6

IR Spectroscopy: Analytical Value IR-spectrum: Qualitative analysis 1. Identity check of a known substance e. g. for control of incoming goods Analytical approach: Comparison of the „spectral fingerprint“ of a sample with the spectrum of a reference material February 12, 2018 7

IR Spectroscopy: Analytical Value C=O CH IR-spectrum: Qualitative analysis 2. Identification of unknown substances e. g. to characterize new materials or to determine contaminations Analytical approaches: a. Assignment of bands to functional groups b. Search for similar spectral fingerprint in spectral libraries February 12, 2018 8

IR Spectroscopy: Analytical Value Calibration IR-spectrum: Quantitative analysis 3 Absorbance Concentration proportional to absorption (Lambert-Beer‘s law) 4 X 2 1 1. Measurement of reference samples with known composition. Wavenumber 3. Analysis of samples using the calibration function Analysis 4 3 Absorbance 2. Determination of a calibration function between measured spectra and reference data (concentration or sample properties) X 2 1 Concentration 12. 02. 2018 9

FTIR-Spectroscopic Analysis of Kidney Stones: Benefits IR • Short analysis time (typically <1 minute) • No sample preparation (when using the ATR-technique) • Little sample amount (~1 mg) • Easy to use, no technical know-how required • No chemicals, solvents or disposables required • Clear identification of the kidney stone independent from visual appearance • Reliable and accurate results • Identification of other substances (solids and liquids) than kidney stones, e. g. pharmaceutical and other chemical substances, packaging materials, etc. February 12, 2018 10

Bruker Kidney Stone Analyzer: System Components • Robust and portable ALPHA-P FT-IR spectrometer system with ATR measurement interface February 12, 2018 + • Intuitive software user interface with dedicated work-flow for kidney stone analysis + • IR-database of >5, 000 authentic kidney stones for sample identification at the push of a button 11

Bruker Kidney Stone Analyzer: ALPHA-P FT-IR Spectrometer • Spectral range: Mid-Infrared (7. 500 – 375 cm-1) • Measurement principle: Attenuated Total Reflectance (ATR) • Measurement spot: 1. 5 x 1. 5 mm • Open access to sampling area • Quality components with long lifetime: ü Most robust sample interface: Pure diamond mounted in ceramics ü Rock. Solid Interferometer ü Diode laser February 12, 2018 12

Bruker Kidney Stone Analyzer: ALPHA-P FT-IR Spectrometer • Compact size: 22 x 33 x 26 cm (W x D x H) • Footprint of a laboratorytextbook! • Weight < 7 kg February 12, 2018 13

Bruker Kidney Stone Analyzer: Principle of ATR Method ATR: Attenuated Total Reflectance • IR-light penetrates sample that is in touch with the crystal surface sample Requirements on ATR-crystal material: • IR-transparent • High refractive index ATR crystal • Robust Ø ALPHA-P utilizes diamond as ATR-crystal ATR-benefits: • Defined penetration depth of IR-light into the sample surface no dilution of sample required • Measurement of virtually all types of solid and liquid samples allowed without sample preparation February 12, 2018 14

FTIR-Spectroscopic Analysis of Kidney Stones: Workflow of ATR Measurement Workflow: 1. Background measurement of empty ATR-crystal Measurement time: ~30 seconds February 12, 2018 15

FTIR-Spectroscopic Analysis of Kidney Stones: Workflow of ATR Measurement Workflow: 2. Press “Measure Sample” & enter sample information 3. Start Sample Measurement: Display of spectrum preview February 12, 2018 16

$FTIR-Spectroscopic Analysis of Kidney Stones: Workflow of ATR Measurement Workflow: 4. Place fraction of$

FTIR-Spectroscopic Analysis of Kidney Stones: Workflow of ATR Measurement Workflow: 4. Place fraction of pure kidney stone sample on the diamond ATR-crystal of the Platinum ATR unit Live spectrum preview Measurement time: ~30 seconds 5. Press sample on ATR crystal using clamping mechanism 6. Spectrum preview shows sample spectrum Start Measurement February 12, 2018 17

FTIR-Spectroscopic Analysis of Kidney Stones: Workflow of ATR Measurement Workflow: 7. Identification of the kidney stone sample by automated library search 8. Generation of a report (hardcopy, pdf, etc. ) 9. Clean ATR crystal with paper wipe + isopropanol 10. Proceed with next sample February 12, 2018 18

FTIR-Spectroscopic Analysis of Kidney Stones: Spectra Analysis 1. Dedicated spectra library: • Bruker Library BLG 2 containing ~5000 spectra of authentic kidney stones • Created by the renowned expert for kidney stone analysis Michel Daudon (at Laboratoire Cristal, Service de Biochimie A, Hôpital Necker, Paris, France) • Allows determination of up to three main components in kidney stone samples at the push of a button February 12, 2018 19

FTIR-Spectroscopic Analysis of Kidney Stones: Spectra Analysis 2. Powerful OPUS spectrum search: • Integrated in analytical workflow • Spectrum search with Bruker’s reliable peak based standard algorithm • Mixture analysis*: Identifies & quantifies single components in mixture spectra • Residual search: Finds minor components by automatic calculation and search of the difference spectrum between sample and library spectrum • Information search*: Powerful and intuitive search of terms and numbers in the complete information matrix of one or several spectral libraries • Direct internet search* of any info term stored with the library spectrum • Set-up and editing of user spectra libraries • Convenient documentation: Easy reporting of search and mixture analysis results by OPUS Analysis Report Generator (to clipboard, hardcopy, pdf or csv). *Requires software option O/SR-N February 12, 2018 20

FTIR-Spectroscopic Analysis of Kidney Stones: Evaluation of ATR Spectra Identification of a kidney stone sample as Cystine (>90%) Sample spectrum Library spectrum February 12, 2018 21

FTIR-Spectroscopic Analysis of Kidney Stones: Evaluation of ATR Spectra Identification of a kidney stone sample as Uric acid (>85% anhydrous, ~5 -15% dihydrate) Sample spectrum Library spectrum February 12, 2018 22

FTIR-Spectroscopic Analysis of Kidney Stones: Evaluation of ATR Spectra Identification of a kidney stone sample as brushite (50 -70%) and whewellite (20 -40%) Sample spectrum Library spectrum February 12, 2018 23

FTIR-Spectroscopic Analysis of Kidney Stones: Evaluation of ATR Spectra Identification of a kidney stone sample as Xanthine Sample spectrum Library spectrum February 12, 2018 24

ALPHA Kidney Stone Analyzer: Easy FTIR-Spectroscopic Analysis of Kidney Stones Summary: Ø Integrated IR-Analyzer system: • Robust and compact ALPHA FT-IR spectrometer system with diamond-ATR measurement interface • Intuitive software user interface, dedicated work-flow for the analysis of kidney stones • Extensive spectral library containing reference spectra of >5, 000 authentic kidney stones Ø No sample preparation or dilution Ø Faster analysis compared to the classical transmission method resulting in a higher sample throughput February 12, 2018 25