Polish J. of Environ. Stud. Vol. 15 No.

4A (2006), 37-40

Invited article

Infrared Spectroscopy in Medical Diagnostics

– Advances in Instrumentation for Glycemic
Control and Applications
H. M. Heise

ISAS – Institute for Analytical Sciences, Bunsen-Kirchhoff-Str. 11, D-44139 Dortmund, Germany

Abstract
Applications of infrared spectroscopy for clinical chemistry and non-invasive diagnostics with the focus on
glucose are presented and reviewed. Principally, reagent-free multi-parameter assays for various biofluids can
be set up using the “fingerprint” mid-infrared spectra. Dry film measurements of nanoliter sample volumes can
be used for patient self-monitoring or point-of-care instrumentation, for which fibre-optic accessories can assist
in device miniaturisation. Continuous blood glucose monitoring is another recent application for critically ill
patients. Near-infrared radiation is essential for assays using diffuse reflectance spectroscopy of skin for non-
invasive glucose quantification and diabetes screening based on glycation effects. Recent results for both fields
are presented.

Keywords: mid- and near-infrared spectroscopy, glucose, clinical chemistry, non-invasive diagnostics

Introduction mentation design is important for reaching a specific skin

Molecular spectroscopy has brought much progress for
medical diagnostics, and particularly the marriage
of vibrational spectroscopy with clinical chemistry will
enable the implementation of several methods into point of
care analytics, patient monitoring or screening methods for
diabetes mellitus. Infrared spectroscopy is a powerful
method for the study of various biomedical samples for in-
vitro analysis in the laboratory and non-invasive
diagnostics. The reagent-free analysis of biofluids such
as whole blood, plasma or serum takes advantage of the fact
that a multitude of analytes can be quantified simul-
taneously. The main focus is on glucose as the most
interesting analyte, since the monosaccharide plays
an important role in the body metabolism for energy supply.
The provision of body fluids such as blood requires
invasive sampling techniques, whereas the penetration of
near-infrared radiation and its back-scattered fraction can be
used for non-invasive assays, probing the integral skin
tissue for glucose and protein glycation effects. Instru-
depth and the required signal-to-noise level for glucose
quantification and detection of skin alterations under
hyperglycaemic conditions.
Mid- and near-infrared instrumentation
and applications
An overview on the applications of IR-spectroscopy in
the clinical chemistry environment has been published four
years ago [1]. Since then, further developments can be
reported. Whereas previously the attenuated total reflection
(ATR) technique was preferred for liquid body fluids, the
measurement as dry-films proved to be promising for
routine analysers. Here, especially fibre-optic devices have
been developed and tested for clinical applications (see also
Fig. 1A) [2]. For example, micro-prisms coupled efficiently
to wave-guiding silver halide optical fibres or fibre-only
devices can be used for the analysis of microscopic samples
(also applicable for remote sensing). For the latter, sample
volumes of 100 nl have been successfully quantified.
38 Heise H. M.

(a) (b)

(c) (d)

Fig. 1. Mid-infrared instrumentation for micro-analysis of dry-films from body fluids (a) and for continuous measurements of fluids
using the combination of microdialysis and transmission spectroscopy (b); near-infrared accessories used for skin reflectance
spectroscopy aiming at non-invasive methods for medical diagnostics: reflection optics for ultimate spectral signal-to-noise
ratios with device for lip measurements (c) and fibre-optic probes with different fibre arrangements for realization of different
probing depths with demonstration of forearm skin measurements (d).

Quantum cascade lasers, miniaturized fibre-optic acces-
sories, and pyroelectric detectors appears well suited for the
continuous monitoring of glucose concentrations at
physiological levels [3]. Further assessment of the limits to
be reached for minimal-invasive biofluid sample harvesting
yielded the result that samples down to 10 nl could be
quantified with a coefficient of variation below 5 % [4].
This certainly needs appropriate methods for skin
perforation and sample transfer.
Another hot topic is the development of blood glucose
monitoring technology for critically ill patients as part of an
artificial pancreas system using a subcutaneously implanted
microdialysis probe and spectroscopy, but also for diabetic
patients – with their requirement of frequent glucose
measurements – continuous monitoring is desirable to
improve therapy. Investigations were carried out on
microdialysates of blood and interstitial fluid using the most
simple transmission technique with flow-through micro-
cells (see Fig. 1B), in contrast to previously favoured ATR-
measurements [6]. A comparison of spectra of different
body fluids is shown in Fig. 2A. Whereas the plasma
spectra are dominated by protein absorption bands, the
dialysates only show contributions from low molecular
mass compounds such as bicarbonate, glucose, lactate, urea
and others. A fluidic system was implemented and the
simultaneous determination of additional compounds has
been reported including micro-dialysis recovery rates [5].
Near-infrared spectroscopy is mostly reported for non-
invasive application and has a promising potential for
patients [6]. Owing to the optical properties of skin, the
diffuse reflection technique can be favourably used. Several
accessories for diffuse reflectance spectroscopy have been
employed, e.g., bifurcated fibre-optic probes or rotational
ellipsoidal mirrors for the collection of backscattered
radiation (see Figs. 1C and D). However, low optical
throughput of the skin and the minute glucose signals with
Infrared Spectroscopy in Medical…
overwhelming other tissue components and variables is
further a major hurdle with near infrared spectroscopy. Skin
spectra are shown in Fig. 2B, recorded using different
accessories, illustrating also the different penetration depths
according to the dominating water overtone and combi-
nation bands [7]. An obstacle is certainly the existence
of the glucose in the intravascular, interstitial and
intracellular compartments of the skin, so that the
correlation to the capillary glucose concentration values, as
seen as the “golden standard” for glycemic control, is
weakened. Calibration problems with accidental
correlations and data overfitting have been reported by us
previously [6]. However, recent progress with regard to
robust statistical PLS calibration models can be noted that
were based on Monte Carlo simulations of photon transport
within skin tissue [8].
0,15 (a)
A composition and provide the absorbance fingerprints for the
biofluid measurements
0,10 ATR
absorbance
0,05
0,00
-0,05 transmission
x2
1800 1600 1400
4 (b)
B probe 2
diffuse reflectance
skin measurements
- log (reflectance)
3
2 322, 2001.
Fibre-optic probes
forearm
1 inner lip
earlap
forearm
0 Reflection optics (x 2)
-1
10000 8000 6000
-1
wavenumber [cm ]
Fig. 2. ATR-spectra of blood plasma spectra recorded using
a µ-Circle cell (upper traces) and absorbance spectra
of different dialysates using a transmission cell of 32
µm pathlength (all spectra were recorded with water
absorbance compensation); glucose spectral features are
evident in the spectral interval of 1200 to 1000 cm-1 (a);
diffuse reflectance spectra of different skin tissues
using different accessories; the quartz fibre-optic probes
allowed only the measurement of spectra down to 4000
cm-1, whereas for the mirror-based device measure-
ments up to 1000 cm-1 were possible, but dependent on
the used photodetector cut-off (b).
39
Another application is diabetes screening, which is
based on the detection of epidermal and dermal skin
changes due to alterations in collagen structure and protein
glycation that are observable in diabetic subjects with
significant deviations from euglycemia. For successful
classification, linear discriminant analysis based on near-
infrared skin spectra was used and a prediction accuracy
of 85 % could be achieved [9]. However, glycation effects
proceed in parallel to normal skin ageing effects, so that age
dependent classification is required for improving the
diabetes screening assay.
Conclusions
The mid-infrared spectroscopic analysis of body fluids
can be successfully carried out, either using minimal
invasive or continuous monitoring approaches. The spectra
of skin tissues are shedding light on their chemical
development of gentle non-invasive medical diagnostic
methods. However, a blood glucose assay based on
plethysmographic measurements [6], i.e., using the blood
EDTA blood plasma volume changes in the dermal arterial and capillary vascular
system emerging from pulsatile blood pressure changes,
EDTA plasma dialysate will provide in future a true non-invasive blood analysis.
serum ultrafiltrate
interstitial fluid dialysate
1200 1000
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