Current Medicinal Chemistry, 2010, 17, ????-????

Infrared-Spectroscopy: A Non-Invasive Tool for Medical Diagnostics and

Drug Analysis
H. Hahn, J.D. Pallua, C. Pezzei, V. Huck-Pezzei, G.K. Bonn and C.W. Huck*
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 52a, 6020 Innsbruck,
Austria
Abstract: Constant development enabled Infrared (IR) spectroscopy to become a widely used, non-invasive tool for fast
sample analyses with less to no pre-preparation. Furthermore, computational data handling is no more a limiting factor
and hence, IR measurements are predestined for clinical diagnostics and drug analysis. Within this review the focus was
put on clinical topics of high interest. One example is Alzheimers disease, where the exact metabolism is still not clarified, or blood glucose monitoring for high throughput screening of patients without taking any drop of blood. The second
section of this manuscript was focused on the analysis of drugs. The detection of physico-chemical parameters in pharmaceutics and the improvement of industrial proceedings allowed a dramatic increase of quality of produced medicine. In
pharmaceutical industries problems with the equable allocation of agents occurs especially in scaling up processes. IRanalyzing-techniques serve as fast and precise indicators for the detection of active components and their distribution in
tablets. In combination with statistical factors and medical investigations pharmaceuticals can be improved from their development until their application, and every step can be easily controlled by IR spectroscopy.

Keywords: Infrared spectroscopy, medicine, allergens, alzheimers disease, pharmaceuticals, drugs, glucose monitoring.
1. INTRODUCTION
IR-spectroscopy is a widely used technique for fast, noninvasive and reproducible results in various application
fields. Many clinical fields utilize this analyzing tool not
only for research but also for routine diagnostics. The importance of diagnostic setups increased dramatically which can
be observed all over the world.
In industrial fields like pharmaceutics this vibrational
spectroscopic technique has become a standard tool for quality control since the 1970s [1-7]. In the last 40 years data
processing and prediction became faster and more precise
because of high performance computational systems. But
with this development other problems occurred. Especially
data processing models became very complicated and adaption of such models for the quantification of any substance
needs high verification of reproducibility and accuracy [8,
9]. Further challenges appeared with the measurement of
pharmaceuticals because physico-chemical parameters differ
between the product synthetized in laboratory and industrial
scale [10].
In clinical diagnostics IR spectroscopy serves as a robust
tool for various diagnostics. A review of Wang and Mizaikoff [11] reported about biomedical diagnostics of tissues,
cells and bio-fluids using mid-infrared (MIR) for analyzing
e.g. atherosclerotic plaques, glioblastoma, a.o., and the significance of multivariate data analysis. Data interpretation
with principal component analysis (PCA), cluster analysis
(CA), linear discriminant analysis (LDA) and other techniques were explained and compared. Additionally analytical
approaches with MIR on the diagnostics of diseases like
atherosclerosis, leukaemia and brain tumors were accomplished.

*Address correspondence to this author at the Head of Spectroscopy Group,

Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens
University, Innrain 52a, 6020 Innsbruck, Austria; Tel: +43/0512-507-5195;
Fax: +43/0512-507-2965; E-mail: christian.w.huck@uibk.ac.at
0929-8673/10 $55.00+.00

Cancer is a hot and permanently demanding topic

worldwide and researchers are focussing on more specific
and sensitive diagnostic tools [12]. Non-invasive IR spectroscopy serves as a perfect tool for in-vivo cancer diagnostics. Kondepati et al. [13] published a review in 2008 focussing on this specific issue. The authors gave a brought overview on researches in fields of breast cancer, tumors, and
distinct other carcinogenic symptoms. Furthermore correlations between cancer and characteristic bands in infrared
spectra are widely discussed.
Within this article IR spectroscopy is introduced as a tool
for manifold medicinal diagnostics and drug analysis applications. The focus was knowingly set on the hot spots of
nowadays science and development. This range begins with
routine analyses like blood glucose monitoring and spans to
special investigations like fetal observations and Alzheimers
disease (AD). A further point of interest is the investigation
of the detection of allergens in food samples and for pharmaceutical approaches too. All together this article underlines
the importance and diversity of nowadays IR spectroscopy in
diagnostical fields of high interest.
2. CLINICAL APPLICATIONS AND DISEASES
This part is divided into 4 sections which were selected
due to highly promising fields in the medical diagnostics in
the last 3 years.
2.1. Allergens
Allergens are becoming more and more a problem for the
worldwide population. Allergens in food, vaccines or plants
are influencing peoples life and even compromise their
health. Allergens can cause serious reactions of the human
body, e.g. asthma or eczema [14-18]. Therefore many tests
and diagnostical tools for allergens in food, nuts, and blood
were established in the last few years [19, 20].
For the detection of allergens in food samples, IRspectroscopy supplies fast and reliable results [21]. One of
2010 Bentham Science Publishers Ltd.

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

the latest approaches was introduced by Gaudin et al. in

2008 [22]. A protein-micro assay for the detection of immune response of patients with cow milk allergy, which is
known to be a common allergy in children, was developed.
This microarray was created by immobilizing milk-specific
proteins, namely
-S1-caseine (Cas),
-S2-Cas, -Cas and Cas, which are approximately 80% of the protein content in
milk. Further proteins were
-lactalbumin, -lactoglobulin,
lactoferrin, human serum albumin (HSA) and lysozyme. All
of them were immobilized on porous nitrocellulose placed in
96-spots well plates. After washing and blocking steps with
phosphate buffered saline (PBS) containing 0.1% Tween 20
and 2% polyvinyl alcohol, sera of milk-allergic patients and
controls (healthy people) were incubated on the surfaces.
After removing unbound serum, fluorescent antibodies (Ab),
human immunoglobulin E (hIgE) and immunoglobulin G
(IgG) were added to the slides. Then fluorescent intensities
were scanned at 700 nm with infrared imaging. Results
showed comparable performance of this test to an enzymelinked immune sorbent assay (ELISA) with sensitivities of
0.8 and 1.2 kU/L, respectively. Data showed a high responsibility of bovine lactoferrin for allergic reactions and bovine
caseins were found to be the main allergens in milk.
Allergic reactions to hazelnuts are also quite prevalent.
Main allergen proteins are lipid transfer and seed storage
globulins. For the detection of three responsible allergens
Rigby [23] introduced an extraction method for three globulins that are responsible for allergic reactions. In detail, 7S-,
11S-globulin and lipid transfer protein (LTP) were extracted
out of hazelnut flour using 50 mM tris(hydroxymethyl)aminomethanhydrochloride (tris-HCL) containing sodium
chloride for 7S and 11S and 20 mM 2-morpholinoethanesulfonic acid (MES) containing 3% polyvinylpolypyrrolidone (PVPP) for LTP. Further separation using gel- and
ion chromatography was followed by gel-filtration and gelelectrophoresis. After tryptic digestion of the separated proteins at 37 C for 3 hours, LC-ESI/MS and MALDI-TOF/MS
investigations were performed. Enclosed database search
(MASCOT) identified the isolated proteins with sequence
coverages (SC) of 70%, 45% and 52% for 7S, 11S and LTP,
respectively. For the confirmation of the retained secondary

Hahn et al.

structure of purified proteins in-circular dichroism with far

ultraviolet light and Fourier-transform infrared (FT-IR) spectroscopy were conducted. Both spectroscopic methods confirmed to be successfully deployable for the structural analysis of allergens. The folded structures were clearly identified
and hence, the purified proteins were in native state, compared to previously observed spectra [24].
2.2. Blood
Blood is the most important but also one of the most
complex body fluid for clinical diagnostics and allows to
receive widespread information about the health condition of
a human. Therefore blood has to be taken usually with a syringe for any laboratory diagnostics. Hence many approaches
with IR spectroscopy came up for quantifying various parameters in blood partly avoiding any withdrawal in order to
improve patient compliance.
Since the determination of glucose in blood was enabled
in 1989 [25] the idea was further developed [26-28] and different data processing models were tested and discussed
[29]. An approach of Fischbacher et al. [30] investigated
partial least-square regression (PLS) and radial basis function (RBF) on their prediction performance concerning noninvasive blood glucose monitoring from the finger. For reference testing a standardized Beckman glucose analyzer was
used (http://www.beckmancoulter.com/). The study was carried out with three patients having type-1 diabetes and obtaining insulin therapy. Glucose levels in blood were manipulated by tiny insulin injections and special nutrition. It
was found out that very small deviation of glucose levels
created high errors on the prediction of real value. Therefore
a special quality control model was created using Pearsons
correlation coefficient and a second method based on the
leverage variables from the RBF. All measurements were
carried out in diffuse reflection mode at 850 1350 nm.
With these two methods prediction accuracy down to 1.2
mmol/l was achieved, which can be deduced from Fig. (1).
The detailed effect of correlation onto accuracy is shown in
Fig. (1a-c). Without any selection criteria obtained accuracy
was 1.9 mmol/l (Fig. 1a), 1.7 mmol/l at a correlation of

Fig. (1). Bias plots of calibration models using an object selection algorithm for evaluating the similarity of the spectra based on Pearsons
correlation coefficient. (a) without selection criterion; RMSP = 1.9 mmol/L; (b) rG = 0.995; RMSP = 1.7 mmol/L; (c) rG = 0.997; RMSP =
1.2 mmol/L. LS regression line; line y = x (Reproduced from reference [30], with permission).

Infrared-Spectroscopy

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

0.995 (Fig. 1b). Considering a correlation of 0.997 the corresponding accuracy was about 1.2 mmol/l. Finally, this
achieved accuracy was very high compared to routine glucose monitoring, introduced with 1.1 mmol/l [31].
Similar results were achieved by an experiment of Heise
et al. [32]. NIR spectra recorded directly from the oral mucosa, outer lip, index finger and forearm skin enabled glucose quantification. As a reference glucose concentrations of
individual probands were measured with a routine laboratory
method, enzymatic methodology (Test Combination Glucoquant Glucose, Boehringer, Mannheim, Germany) with the
aid of hexokinase/G6P-DH on an ACP 5040 analyzer from
Eppendorf (Hamburg, Germany). For calibration, a PLS algorithm programmed in MATLAB (The Mathworks,
South Natik, MA, USA) was used. In order to make the system more robust, glucose levels of diabetes patients were
manipulated with insulin and glucose doses. Best results
were achieved with oral mucosa because of the largest blood
volume and fewest fat amount compared to the other sites
(outer lip, finger, forearm skin). The root-mean-square error
of prediction (RMSEP) was taken for establishing the accuracy of the calibration models, as suggested by Marbach et
al. in the 1990s [33, 34]. RMSEP values for the calibration
model was 36.4 mg/dl with a regression of calibration curve
of R2 = 0.952. A comparison to Arnolds et al. work [35] was
carried out in detail. Within this work a RMSEP of 74.9
mg/dl for standards and almost 127.6 mg/dl for the glucose
studies were found. The same author summarized all available perspectives of non-invasive glucose monitoring in humans, introducing direct and indirect non-invasive glucose
sensing [36]. As direct measurements are based on the monitoring of the glucose molecule itself, indirect measurements
are focussed on investigating the physiological effects of
glucose in individuals.
Since blood-rich body sites like oral mucosa (see above)
are known as preferable points for glucose monitoring the
Table 1.

data processing methods were advanced. Du et al. [37] introduced region orthogonal signal correction (ROSC) for this
application. The idea for this correction model goes back to
Wold et al. [38] and was developed by Wise [39], Fearn
[40], and Sjoblom [41]. This chemometric tool was designed
for removing strong variations out of spectra which are not
correlated to a specific concentration of a compound. In the
work of Du et al. [37] this tool was performed for glucose
monitoring to remove the absorbance band of water, in the
range between 1404 and 1454 nm, for improving in vivo
glucose measurements in the wavelength window of 1212 to
1889 nm. Results obtained are depicted in Table 1 and spectra are shown in Fig. (2). Informative spectral regions are
compared to the whole wavelength region based on their
prediction performances of the PLS models. The advantage
of this approach, compared to conventional orthogonal signal
correction (OSC), was the better interpretability because the
new ROSC method used special regions in the spectra for the
prediction of glucose scores while OSC only used a fixed
region. Finally, ROSC with PLS allowed to improve the prediction performance significantly. Correlation coefficients of
whole spectra and selected spectral regions increased the
extracted spectral information throughout. For example in
the region of 1240 to 1320 nm R was 0.8751 applying the
OSC and 0.9173 the ROSC model. In the same wavelength
region the root mean square error of cross validation
(RMSECV) was about 20.1 mg/dL with ROSC compared to
almost 23.1 mg/dL with OSC. Concluded it was stated that
ROSC was established quite successfully for removing interferences during glucose monitoring.
A newly developed method for the measurement of oxygenation in blood was introduced by Lehr and Wickramasinghe [42]. The special approach of Lehr was to develop a four-channel NIR prototype out of a single-channel
instrument. The advantage was to achieve information of
blood oxygenation from four regions of the neonatal brain.
Further, only one instrument was used for measuring all re-

Prediction Performance of PLS Models Developed by Using Different Spectral Regions of OSC- (a) and ROSC- (b) Pretreated Spectra. [Reproduced from Reference, 37, p. 1344, with permission]
Spectral region/nm

Orthogonal component number

PLS component number

RMSECV/mg dL-1

Whole region

1212 - 1889

0.9819

19.9144

Whole region

1212 - 1889

0.9818

19.9068

Whole region

1212 - 1889

0.9816

19.9025

Info. region 1

1240 - 1320

0.8751

23.1437

Info. region 2

1600 - 1730

0.9539

17.1703

Info. region 2
Info. region 2

1600 - 1730
1600 - 1730

2
3

2
1

0.9536
0.9382

17.1688
17.3408

Whole region

1212 - 1889

0.9780

20.2300

Whole region

1212 - 1889

0.9808

18.7406

Whole region

1212 - 1889

0.9811

18.9237

Info. region 1

1240 - 1360

0.9173

20.3942

Info. region 1

1240 - 1360

0.9338

20.1154

Info. region 1

1240 - 1360

0.9427

20.0794

Info. region 2

1600 - 1730

0.9660

15.8911

Info. region 2

1600 - 1730

0.9696

16.9939

Info. region 2

1600 - 1730

0.9688

16.7170

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

Hahn et al.

embryos for single transfer were selected by morphological

criteria and the culture media were screened with NIR.
Based on these metabolomic profiles predictions concerning
the viability of transferred embryos were carried out. The
accuracy of assessing viable embryos on the third day by
NIR profiling was 53.6% and 38.5% based on the morphological selection. Furthermore, the positive predictive value
of profiling was 0.365 and the negative predictive value was
0.830. Hence, embryo viability after transfers could be predicted with higher accuracy using NIR metabolomic profiling of culture media than using morphological parameters.
Additionally results showed that the diagnostic NIRS tool
was not suitable to perform any prognosis concerning the
pregnancy outcome. NIRS was used as additional method for
the selection of any morphologically equal embryos for single embryo transfer but not as single instrument for the estimation of pregnancy outcome.
Fig. (2). NIR spectra in the 1212 -1889 nm region of blood glucose
in skin and a NIR spectrum of glucose powder (Reproduced from
reference [37], with permission).

gions at once and therefore this application was very cost

effective compared to four single-channel instruments operated in parallel. The four-channel sensor was tested on forearms of adults as depicted in Fig. (3). Transmitter and receiver sensors were sites as shown in the figure. The venous
blood stream was regulated by a cuff placed on the upper
arm. Spectra showed clearly differences in the optical density depending on the venous occlusion. The LOD for haemoglobin detection was 0.06
mol per 100 mL blood and
even 0.01
mol per 100 mL for baseline drifts in one hour.
These results confirmed the high performance of the four
channel NIR prototype for the adaption for neurological injuries and abnormalities in newborns, e.g. periventricular leucomalacia. This brain injury is characterized by death of the
white matter in infantile brains and causes delayed motor
development and epilepsy.
2.3. Reproduction Medicine and Fetal Investigation
Hence, many couples in the United States of America
(USA) population were affected by infertility in the 1990s
[43, 44] with increasing trend towards now, reproduction
medicine has been an upcoming field of interest. Approximately 1% of all pregnancies in the USA were assisted by
reproduction technologies [45, 46]. In Europe the percentages were between 0.2% and 3.9% in 2000 and 2001 with an
increase of 1% to 4%, depending on the country [47, 48].
IR spectroscopy seems to be a predestined tool for the
investigation of embryos in-vitro conditions. Metabolomic
profiles can be detected within minutes and, with the help of
statistical methods, predictions of the pregnancy outcome
indicated by in-vitro fertilization (IVF) became possible. In
2008 two articles were published [49, 50] reviewing the latest techniques for the metabolomic profiling of culture media
as useful additive to conventional, morphological tool for the
estimation of an embryos survival potential. In the same
year Vergouw et al. [51] stated that NIR spectroscopy is a
fast and precise method for testing the reproductive assessment of embryos. In this study 333 embryo culture media
were investigated on their metabolomic profile. In parallel

Additionally, pre-implantation genetic investigation of

blastocytes has been carried out since 1989 [52]. A review
by McArthur et al. [53] published in 2005 summarized laboratory and clinical experiences including infrared technological applications with laser technique.
During the nine months of pregnancy an observation of
the embryos status is feasible with IR spectroscopy. Based
on the guidelines on the standard observation of fetal health
in labour many various investigations were published on the
topic of fetal observation [54]. Mozurkewich and Wolf reviewed investigations of fetal assessment using NIRS [55]
already in 2000. In the perinatal phase complications can
occur which can cause damage of the newborn brain, e.g.
hypoxic-ischiaemic brain injury, as Wyatt et al. reviewed in
the late 80s [56]. Since that NIR has become a widely used
tool for perinatal diagnostics and of course data processing
became much easier since then, stated by Nicklin et al. in
2003 [57], who also reviewed technical developments and
the most important clinical applications from 1985 to 2003.
Focussing the time between fertilization and nativity, different occurrences happening to the mother can affect the
fetus or embryo. One example is the influence of anaesthetic
agents on the developing brain like investigated by McClaine
et al. in 2005 [58]. Four hours of anesthetization with common drugs like midazolam, sodium thiopental and isoflurane
were carried out with pregnant sheep. In-utero recording of
fetal cerebral oxygenation was carried out using NIRS. Results showed no effects on the oxygenation during anaesthesia. Additional histological findings of non-instrumented
sheep brain confirmed the innocence of inhalational anaesthesia on pregnant women.
Another example for the highly useful approach of NIRS
is fetal monitoring. Since the 1990s control of fetal cerebral
blood oxygenation was introduced as one of the first investigations in the field of fetal observation [59-61]. Most important approaches in this investigation field until 1996 were
summarized in the review of DAntona [62]. Since that, further more detailed NIRS investigations were carried out. The
screening of cerebral oxygenation of preterm infants with
NIRS was introduced by Watkin et al. [63]. A comparative
study to conventional pulse oximetry was carried out focussing on the variations of cerebral oxygenation and brain hypoxia related to apnoea. NIR spectra were recorded from the

Infrared-Spectroscopy

infants heads at wavelengths of 775, 845 and 904 nm with 1

second sampling time in reflection/backscatter mode. The
sensor of the pulse oximeter was placed on the right foot for
measuring arterial oxygen saturation.
Results showed that baseline variability in oxygenation
was from -0.12 to +0.13
mol per 100 g brain. Only in 20%
of all measuring cycles no decrease of cerebral oxygenation
was detected although the level of oxygenated hemoglobin
decreased; in 8% the inverse phenomenon was recorded.
Concluded it was stated that no close correlation between the
two parameters, cerebral oxygenation and oxygenated hemoglobin, was observed. Only large falls of more than 1.5
mol
per 100 g brain in cerebral oxygenation were related to large
changes in hemoglobin concentration.
2.4. Neurological Investigation
IR is predestined also for non-invasive optical neuroimaging. A review by Strangman [64] published in 2002

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

described instrumentation enabling optical measurements of

changes in brain functions and diseases. Furthermore,
physiological and clinical applications were introduced.
Taillefer [65] published a review discussing the efficiency of
IR spectroscopy as a tool for cerebral scanning during heart
surgeries.
A special field of interest is the Alzheimers disease
(AD). Within the last years much research was done, also in
the field of IR. This chronic neurodegenerative disease is the
most common cause of dementia in the aging population [66]
and develops along with the building of plaques in the brain,
which are characteristic for patients with Alzheimers disease [67]. A review by Hillman [68] published in 2007 introduced the most important approaches in the field of optical
brain imaging in vivo and focused on AD too. But since that
several novel technological approaches have been introduced. More or less each study concentrated on the two biochemical markers, namely amyloid- and tau () proteins,

Fig. (3). Setup for the measurement on a resting adult forearm. Transmitter fibres Txl and Tx2 and receiver fibres Rrl and tr2 are attached
dose to the wrist of the left forearm (a). Vertical axis shows changes in concentration of deoxyhaemoglobin (AHb), oxyhaemoglobin
(AHb02), total haemoglobin (AHb,o,) and cytochrome aa3 (A Cvt aa3) in the four regions during venous occlusion. Regional differences are
observed (b). (Reproduced from reference [42], with permission).

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

which are associated with the building of the histopathological plaques and tangles [69-71]. Additionally, other
approaches dealt with the association of apolipoprotein E in
case of Alzheimers disease [72, 73], especially using NIRS
[74].
Hintersteiner et al. [75] introduced a technique for the invivo detection of the same peptide in mice. In this work the
synthesis and characterization of near-IR fluorescence oxazine dye is described. NIR fluorescence imaging confirmed
the interaction between oxazine and amyloid plaques in
mice, in-vivo as well as in brain samples post-mortem. The
intensity of the fluorescence signal was taken for quantification. Therefore the advancement of Alzheimer disease could
be observed as well as effects of drugs on the plaque amount;
both applyomg non-invasive measurements.
Griebe et al. [76] compared a FT-IR method to conventional enzyme-linked immune sorbent assays (ELISA) for
amyloid- and  proteins in cerebrospinal fluid. Samples
were taken from 71 patients with clinically diagnosed AD
and 66 control samples. Results showed a decreased amyloid- and an increased tau level in AD samples. A combination of these two parameters enabled the diagnostic discrimination between healthy and AD patients with a specificity
and sensitivity of 86% and 99%, respectively. Infrared spectra were recorded in a wave number range of 3900 to 600
cm-1. The differentiation between healthy and diseased patients using FT-IR was successfully carried out with 80%
and 88% achieved for specificity and sensitivity. A disadvantage of this approach was the collection of cerebrospinal
fluid although it is difficult to collect from patients routinely.
Therefore it was easier to obtain plasma from AD patients
for any analysis. Meanwhile several biomarkers in serum
samples were reported to be associative with AD [77, 78] but
tests were mainly carried out with ELISA which is time- and
cost intensive. Therefore an FT-IR method for the differentiation between AD and healthy patients was introduced by
Peuchant et al. in 2009 [79]. The authors combined fast and
cost-efficient IR measurements with the advantage of fast
and easy sample collection. FT-IR was used for the identification of biochemical moderations of plasma components.
Therefore 40 plasma samples from AD patients and 112 controls were investigated. In detail, the contents of glucose,
total cholesterol, malondialdehyde (MDA), 8-epiprostaglandine F2
, 8-hydroxydeoxyguanosine and the two
vitamins A and E were quantitatively determined with
ELISA and high-performance liquid chromatography
(HPLC) in order to point out differences between AD and
healthy samples. Based on these results, FT-IR spectra were
recorded in the range between 1480 and 910 cm-1. Wards
algorithm and cluster analysis were carried out for data processing. With this setup samples from AD patients were identified with an accuracy of almost 95%.

Hahn et al.

drugs [83] without violation of the packing [84, 85]. Especially the NIR region is preferable because no extensive
sample preparation is necessary to obtain a spectrum and,
combined with statistical methods, NIR spectroscopy is an
excellent tool for quality control in production processes [8688].
3.1. Quantitative Investigations and Statistical Analysis
Amigo and Ravn [8] focused on two models for the
quantitative and spacious determination of pharmaceutical
tablets based on a previous work of Ravn et al. [89]. Classical least squares and multivariate curve resolution models
were taken for the investigation of five ingredients of a
pharmaceutical tablet. The expected advantage of this technique was to omit calibration before measurements but
knowing the weight-percentage of ingredients and total
weight of the tablet. Main components, cellulose and lactose,
were quantitatively determined with a standard deviation of
approximately 2.5 to 10%, depending on the used model.
Other ingredients, e.g. magnesium stearate and talc, were
detected with far higher errors of more than 40%. Best results were obtained by using multivariate curve resolution
showing reliable information concerning major components.
Most interesting application was the controlling of homogenous distribution of ingredients in tablets during production
and not the quantitative analysis, especially for the major
components with more than 20 weight-percent.
Powder blends are commonly used in therapy and the
homogeneity of agents is an important factor to be considered. NIRS combined with different data processing methods
served a useful tool for the qualitative and quantitative determination of components, also in blends [90].
Quantitative analyses in the pharmaceutical field contain

3. DRUG INVESTIGATIONS
In the last few years IR spectroscopy has become a robust
method for the determination of pharmaceutical products on
their chemical and physical parameters [80-82]. As IRspectroscopy is a non-invasive tool, pharmacological ingredients are not chemically modified while measuring. Hence
IR spectroscopy can widely be used for the identification of

Fig. (4). Strategy used to study the impact of concentration correlation on the PLS model (Reproduced from reference [9], with permission).

Infrared-Spectroscopy

chemometric pitfalls as pointed out by the group of Xiang

and coworkers [9]. Within this article main disadvantages
and mistakes of statistical methods for quantitative component analysis was worked out on the basis of a pharmaceutical example. The strategy of this study is shown in Fig. (4).
One active pharmaceutical ingredient (API) and three excipients (out of seven) were measured in this experiment. Two
concentration designs with seven tablet batches were created
for calibration. For the random design the three excipients
were mixed randomly concerning the concentration ratio to
the API. Calibration was carried out with tablets from seven
batches of each design. PLS models for multivariate data
analysis were calculated. These models, basing on the
chance correlation, were tested on their prediction accuracy,
selectivity and robustness. Results showed clearly that randomization is the method of choice for calibration of NIR
spectra in case of chance correlated data processing models.
The cross-prediction accuracy showed doubtless predominance of the PLS model produced by the randomized calibration design. Selectivity was tested by using 30 placebo tablets and again the prediction of the first PLS model was only
14.1% for API content, the second model predicted nearly
90%. For testing the robustness of the models, particle sizes
and hardness impacts, 110-190 N, of the tablets were varied.
The standard errors were again smaller for the predicted API
content using the randomized calibration model (all lower
3%, overall 2.2%) compared to the correlated designed
model (all significantly higher than 2.6%, overall error
4.4%). The variation of the hardness affected higher differences than the different particle sizes. Randomized design
showed overall errors of 3.5% and correlated design 3.9%.
3.2. Physical Parameters
Another important approach is thinking about the transfer
of a developed strategy from laboratory-scale to industry
application because scaling up processes causes changes in
physical parameters. An example was shown by Sarraguca et
al. [10]. Paracetamol was taken as API and mixed with cellulose, talc and magnesium stearate. For calibration and validation of the developed method small charges of powders were
mixed in various ratios of the ingredients. In parallel, semiscale charges of these mixtures were produced and tablets
were pressed out of these charges. First analytical step was
the investigation of the pure components and creating a fingerprint of the calibration powder followed by applying a
developed PCA model to ensure conformation. Then the
semi-scale samples, powder and tablets, could be assigned to
that model. Statistical data processing was carried out using
PLS with cross validation. Applying these techniques calibrations were arranged and semi-scale produced samples
could be recorded. Results showed few differences concerning the samples produced in the laboratory and the semiscale products. The errors concerning the quantitative analysis of the four ingredients were approximately 1% lower for
powders than for the tablets. The differences in the quantification of the four components between control powders produced in laboratory scale and those produced semi-industrial
were up to 5%, between tablets even 9%. With this experiment it was clearly shown that a developed model for NIR
analyses based only on calibration samples produced in the
laboratory need further improvement for industrial applica-

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

tions. Physical parameters were considerably changed by

process up-scaling.
3.3. Combination of Medical Diagnosis and Pharmaceutical Analysis
As quality control in the production process of vaccines
can be easily handled with IR spectroscopy, the detection of
the drugs and metabolites in individuals is a further point of
interest. An example is the distribution of polymeric
nanoparticles which are often used as carriers for fluorescence dyes as introduced by Yang et al. [91]. The aim of the
study was the investigation of the particles size and surface
chemistry on their pharmacokinetics and bio-distribution
applying a NIR-fluorescence (NIRF) spectroscopy. This approach combines clinical analysis with pharmaceutical investigations underlining the increasing predominant role of
NIRS as described in the following paragraph. In detail, latex
nanoparticles, methoxyethylmethacrylate, cross-linked with
divinylbenzene, with diameters of 20, 30 and 60 nm were
compared to gel nanoparticles with a diameter of 20 nm,
composed of hydroxymethyl methacrylate, cross-linked with
methylene-bis-acrylamide. These particles were radiolabeled with 111Indium and added to human breast cancer
cells, mouse macrophages and a monocyte cell line, all incubated at 37C in Dulbeccos Eagles medium. Three times
washing with Hanks balance salt solution and cell lysis using CelLytic reagent (Sigma) was carried out after 1 and 2
hours, respectively. After transferring the remaining solution
to 96 well plates the fluorescent intensity was measured operating a TECAN micro-plate reader at 755 nm (excitation)
and 781 nm (emission). The kinetic- and distribution studies
were carried out by injecting 5 x 1014 particles to mice to
achieve a radiation activity of 65-80
Ci per mouse. Two
days after injection blood, lung, muscle, spleen, kidney,
liver, and tumor tissues were removed, weighed, and counted
for radioactivity with a Packard Cobra gamma counter
(Downers Grove, IL). Results showed 4 times higher emission intensity of the latex particles than of the gel nanobeads, both with a diameter of 20 nm but there was little
difference in the cell uptake between the two kinds of particles. Concerning the kinetics the 20 nm beads showed a decrease from approximately 45 to 10 % of the injected dose
within 8 hours, the latex beads with 30 and 60 nm decreased
from 75 to 38 % within the same period. The found amounts
of particles are summarized in Fig. (5). Comparing again the
20 nm particles (A) and the latex particles (B) the highest
amount could be observed in the liver tissues with more than
20% (A) an 10% (B) of injected dose, respectively. Summarized, the study stated clear predominance of the 20 nm particles concerning the pharmacokinetics and bio-distribution
controlled by NIRF. Again, IR-spectroscopy was successfully established as a feasible tool for the investigation of
drug distribution in individuals.
CONCLUSION AND PROSPECTS
IR spectroscopy has been a fast upcoming analysing tool
for various applications since years. Fast results and improved sensitivity as well as specificity can be performed
using this non-invasive technique. Nearby low or even no
sample preparation is necessary for measurements and prices

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

Fig. (5). Biodistribution of 111In-labeled nanogel and 111In-labeled

nanolatex at 48 h after intravenous injection of particles in nude
mice bearing subcutaneousMDA-MB-468 tumors. Data were plotted as percentage of injected dose per gram of tissue (%ID g
1).
All data are expressed as mean SD (n = 3). Significant differences, p < 0.05
and p < 0.005

, were observed between different
groups (Reproduced from reference [91], with permission).

for hard- and software became affordable. Blood glucose

monitoring, allergens, fetal observation, quality control for
pharmaceuticals and neurological diagnostics are far developed. Nevertheless, further improvements of data processing
models are necessary for highly reproducible and specific
results. Changes and variations of physico-chemical parameters during up-scaling of processes and between patients
need highly precise data processing to ensure precise results
and highest reproducibility. However, IR spectroscopy as
analytical tool or alternative method is on a high level but
need further improvement for special diagnostics.
ACKNOWLEDGEMENTS
The authors thank the Leopold-Franzens University,
Innsbruck, Austria for financial support (Nachwuchsfrderung).
ABBREVIATIONS
MES

= 2-Morpholinoethanesulfonic acid

AD

= Alzheimers disease

Ab

= antibody

a.o.

= and others

API

= active pharmaceutical ingredient

CA

= cluster analysis

Hahn et al.

Cas

= casein

ELISA

= enzyme-linked immune sorbent assay

FT-IR

= Fourier-transform infrared

hIgE

= human immunoglobulin E

HPLC

= high-performance liquid chromatography

HSA

= human serum albumin

IgG

= immunoglobulin G

IR

= infrared

IVF

= in-vitro fertilization

LDA

= linear discriminant analysis

LC-ESI/
MS

= liquid chromatography coupled to mass

spectrometry via electrospray-ionization

MALDITOF/MS

= matrix-assisted laser-desorption/ionisation
time-of-flight mass spectrometry

MDA

= malondialdehyde

MIR

= mid-infrared

MSC

= multiplicative scattering correction

NIR

= near infrared

NIRF

= NIR-fluorescence

OSC

= orthogonal signal correction

oxy-Hb

= oxy-haemoglobin

PBS

= phosphate buffered saline

PVPP

= polyvinylpolypyrrolidone

PCA

= principal-component analysis

PLSR

= partial least-square regression

RBF

= radial basis function

ROSC

= region orthogonal signal correction

RMSEP

= root-mean-square error of prediction

RMSECV = root mean square error of cross validation

SC

= sequence coverages

S/N

= signal-to-noise

= tau

tris-HCL) = tris(hydroxymethyl)aminomethanhydrochloride
REFERENCES
[1]

[2]

[3]
[4]

Vendeland, D.; Pakholkov, G.V.; Arzamastsev, A.P. Prospects for

using IR spectroscopy in the pharmaceutical analysis of penicillin
group preparations. II. IR spectroscopy as a method of identifying
oxacillins and phenoxymethl penicillins. Farmatsevtychnyi zhurnal, 1975, 30(3), 33-38.
Vendeland, D.; Arzamastsev, A.P. Prospects for using IR spectroscopy in the pharmaceutical analysis of penicillin group preparations. I. IR spectroscopy as a method of identifying benzylpenicillins and ampicillins. Farmatsevtychnyi zhurnal, 1975, 1, 50-56.
Shoemaker, M.J.; Sideman, L. Letter: Use of infrared spectroscopy
in drug analysis. Clin. Chem., 1976, 22(8), 1411.
Mynka, A.F.; Liuta, M.L. Use of IR spectroscopy for the quantitative determination of drug preparations containing the carbonyl

Infrared-Spectroscopy

[5]

[6]
[7]

[8]

[9]

[10]

[11]

[12]

[13]

[14]

[15]

[16]

[17]

[18]

[19]

[20]

[21]
[22]

[23]

[24]

[25]

group. Farmatsevtychnyi zhurnal, 1976, 4, 34-38.

Lunardelli, M.; De Marchi, G.; De Martiis, F. Application of near
infrared reflectance spectroscopy (NIRS) to process control in the
pharmaceutical industry. Bull. Chim. Farm., 1988, 127(1), 13-19.
Toffel-Nadolny, P.; Paulig, G. Proof of drugs of minute quantities
using IR spectroscopy. Arch Krimino, 1980, 165(3-4), 111-119.
Gonzalez, F.; Pous, R. Quality control in manufacturing process by
near infrared spectroscopy. J. Pharm. Biomed. 1995, 13(4-5), 419423.
Amigo, J.M.; Ravn, C. Direct quantification and distribution assessment of major and minor components in pharmaceutical tablets
by NIR-chemical imaging. Eur. J. Pharm. Sci. 2009, 37(2), 76-82.
Xiang, D.; Berry, J.; Buntz, S.; Gargiulo, P.; Cheney, J.; Joshi, Y.;
Wabuyele, B.; Wu, H.; Hamed, M.; Hussain, A.S.; Khan, M.A.
Robust calibration design in the pharmaceutical quantitative measurments with near-infrared (NIR) spectroscopy: avoiding the
chemometric pitfalls. J. Pharm. Sci. 2009, 98(3), 1155-1166.
Sarraguca, M.C.; Lopes, J.A. Quality control of pharmaceuticals
with NIR: from lab to process line. Vib. Spectrosc. 2009, 49(2),
204-210.
Wang, L.; Mizaikoff, B. Application of multivariate data-analysis
techniques to biomedical diagnostics based on mid-infrared spectroscopy. Anal. Bioanal. Chem. 2008, 391, 1641-1654.
Petter, C. H.; Heigl, N.; Rainer, M.; Bakry, R.; Pallua, J.; Bonn, G.
K.; Huck, C. W. Development and application of fourier-transform
infrared chemical imaging of tumour in human tissue. Curr. Med.
Chem. 2009,16(3), 318-326.
Kondepati, V.R.; Heise, H.M.; Backhaus, J. Recent applications of
near-infrared spectroscopy in cancer diagnosis and therapy. Anal.
Bioanal. Chem. 2008, 390, 125-139.
Luijk, B.; van den Berge, M.; Kerstjens, H.A.M.; Postma, D.S.;
Cass, L.; Sabin, A.; Lammers, J.W. J. Effect of an inhaled adenosine A2A agonist on the allergen-induced late asthmatic response.
Allergy 2008, 63(1), 75-80.
Constantin, C.; Quirce, S.; Grote, M.; Touraev, A.; Swoboda, I.;
Stoecklinger, A.; Mari, A.; Thalhamer, J.; Heberle-Bors, E.; Valenta, R. Molecular and immunological characterization of a wheat
serine proteinase inhibitor as a novel allergen in baker's asthma. J.
Immunol. 2008, 180(11), 7451-7460.
Tovey, E.R.; Almqvist, C.; Li, Q.; Crisafulli, D.; Marks, G.B. Nonlinear relationship of mite allergen exposure to mite sensitization
and asthma in a birth cohort. J. Allergy Clin. Immunol. 2008,
122(1), 114-118.
Volozhin, A.I.; Babakhin, A.A.; Dubova, L.V.; Sorokin, D.A.
Allergy to metals used in orthodontics and methods for its diagnosis. Stomatologiya 2004, 5, 57.
Forte, G.; Petrucci, F.; Bocca, B. Metal allergens of growing significance: epidemiology, immunotoxicology, strategies for testing
and prevention. Inflamm. Allergy Drug Targets 2008, 7(3), 145162.
Mayer, C.; Verheijen, R.; Stich, N.; Schalkhammer, T.G.M.
Biomedical instrumentation based on micro- and nanotechnology.
Proc. SPIE, 2001, 5265, 134.
Immer, U.; Schmitt, K.; Johnson, K.; Fellinger, A. Easy analysis of
allergen residues in food: a new lateral flow test and an ELISA.
ACS Symposium Series 2008, 1001, 476-481.
Lehrer, S.B.; Ayuso, R.; Reese, G. Seafood allergy and allergens: a
review. Marine Biotechnol. 2002, 5(4), 339-348.
Gaudin, J.C.; Rabesona, H.; Choiset, Y.; Yeretssian, G.; Chobert,
J.M.; Sakanyan, V.; Drouet, M.; Haertle, T. Assessment of the
immunoglobulin E-mediated immune response to milk-specific
proteins in allergic patients using microarrays. Clin. Exp. Allergy
2008, 38(4), 686-693.
Rigby, N.M.; Marsh, J.; Sancho, A.I.; Wellner, K.; Akkerdaas, J.;
van Ree, R.; Knulst, A.; Fernndez-Rivas, M.; Brettlova, V.;
Schilte, P.P.; Summer, C.; Pumphrey, R.; Shewry, P.R.; Mills,
E.N.C. The purification and characterization of allergenic hazelnut
seed proteins. Mol. Nutr. Food Res. 2008, 52(2), 251-261.
Mills, E.N.C.; Marigheto, N.A.; Wellner, N.; Fairhurst, S.A.; Jenkins, J.A.; Mann, R.; Belton, P.S. Thermally induced structural
changes in glycinin, the 11S globulin of soya bean (Glycine max)
an in situ spectroscopic study. Biochim. Biophys. Acta 2003,
1648(1-2), 105-114.
Zeller, H.; Novak, P.; Landgraf, R. Blood glucose measurement by

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

[26]

[27]
[28]

[29]

[30]

[31]

[32]

[33]

[34]

[35]

[36]
[37]

[38]

[39]
[40]
[41]

[42]

[43]
[44]
[45]

[46]

[47]

[48]

infrared spectroscopy. Int. J. Artif. Organs 1989, 12(2), 129-135.

Marbach, R.; Koschinsky, T.; Gries, F.A.; Heise, H.M. Noninvasive blood glucose assay by near-infrared diffuse reflectance spectroscopy of the human inner lip. App. Spectrosc. 1993, 47(7), 875881.
Heise, H.M.; Marbach, R.; Koschinsky, T.; Gries, F.A. Artif. Organs 1994, 18(6), 439.
De Blasi, R. A.; Ferrari, M.; Natali, A.; Conti, G.; Mega, A.; Gasparetto, A. Noninvasive measurment of forearm blood flow and
oxygen consumption by near-infrared spectroscopy. J. Appl.
Physiol. 1994, 76, 1388-1393.
Mller, U.A.; Mertes, B.; Fischbacher, C.; Jageman, K.U.; Danzer,
K. Non-invasive blood glucose monitoring by means of near infrared spectroscopy: methods for improving the reliability of the calibration models. Int. J. Artif. Organs 1997, 20(5), 285-290.
Fischbacher, C.; Jagemann, K.U.; Danzer, K.; Mller, U.A.;
Papenkordt, L.; Schler, J. Enhancing calibration modles for noninvasive near-infrared spectroscopical blood glucose determination.
J. Anal. Chem. 1997, 359, 78-82.
Hovorka, R.; Kremen, J.; Blaha, J.; Matias, M.; Anderlova, K.;
Bosanska, L.; Roubicek, T.; Wilinska, M.E.; Chassin, L.J.; Svacina, S.; Haluzik, M. Blood glucose control by a model predictive
control algorithm with variable sampling rate versus a routine glucose management protocol in cardiac surgery patients: a randomized controlled trial. J. Clin. Endocrinol. Metabol. 2007, 92(8),
2960-2964.
Heise, H.M.; Bittner, A.; Marbach R. Near-infrared reflectance
spectroscopy for noninvasive monitoring of metabolites. Clin.
Chem. Lab. Med. 2000, 38(2), 137-145.
Marbach, R.; Heise, H.M. On the efficiency of algorithms for multivariate linear calibration used in analytical spectroscopy. Trends
Anal. Chem. 1992, 11(8), 270-275.
Marbach, R.; Heise, H.M. Calibration modeling by partial leastsquares and principal component regression and its optimization
using an improved leverage correction for prediction testing. Chemom. Intell. Lab. Systems 1990, 9(1), 45-63.
Arnold, M.A.; Burmeister, J.J.; Small, G.W. Phantom glucose
calibration models from simulated noninvasive human nearinfrared spectra. Anal. Chem. 1998, 70(9), 1773-1781.
Arnold, M.A.; Small, G.W. Noninvasive glucose screening. Anal.
Chem. 2005, 77, 5429-5439.
Du, Y.P.; Liang, Y.Z.; Kasemsumran, S.; Maruo, K.; Ozaki, Y.
Removal of interference signals due to water from in vivo nearinfrared (NIR) spectra of blood glucose by region orthogonal signal
correction (ROSC). Anal. Sci. 2004, 20, 1339-1345.
Wold, S.; Antti, H.; Lindgren, F.; hman, J. Orhtogonal signal
correction of near-infrared spectra. Chemom. Intell. Lab. Syst.
1998, 44, 175-185.
Wise, B.M.; Gallagher, N.B. http.//www.eigenvector.com/
MATLAB/OSC.html.
Fearn, T. On orthogonal signal correction. Chemom. Intell. Lab.
Syst. 2000, 50(1), 47-52.
Sjoblom, J.; Svensson, O.; Josefson, M.; Kullberg, H.; Wold, S. An
evaluation of orthogonal signal correction applied to calibration
transfer of near infrared spectra. Chemom. Intell. Lab. Syst. 1998,
44(1-2), 229-244.
Lehr, H.P.; Wickramasinghe, Y. New prototype NIRS to investigate multi-regional cerebral blood and tissue oxygenation and
haemodynamics. Med. Biol. Eng. Comput. 2000, 38, 281-286.
Mosher, W.D. Fecundity and infertility in the United States. Am. J.
Public Health 1988, 78(2), 181-183.
Mosher, W.D.; Pratt, W.F. Fecundity and infertility in the United
States: incidence and trends. Fertil. Steril. 1991, 56(2), 192-193.
Wright, V.C.; Chang, J.; Jeng, G.; Macaluso, M. Assisted Reproductive Technology Surveillance --- United States, 2003. Surveil.
Summ. 2006, 55(4), 1-22.
Wright, V.C.; Schieve, L.A.; Reynolds, M.A.; Jeng, G. Assisted
reproductive technology surveillance --- united states, 2002. Surveil. Summ. 2005, 54(2), 1-24.
Andersen, A.N.; Gianaroli, L.; Nygren, K.G. Assisted reproductive
technology in Europe, 2000. Results generated from European registers by ESHRE. Hum. Reprod. 2004, 19(3), 490-503.
Andersen, A.N.; Gianaroli, L.; Felberbaum, R.; de Mouzon, J.;
Nygren, K.G. Hum. Reprod. 2005, 20(5), 1158.

10 Current Medicinal Chemistry, 2010 Vol. 17, No. 21

[49]

[50]

[51]

[52]

[53]

[54]

[55]

[56]

[57]
[58]

[59]
[60]

[61]

[62]

[63]

[64]
[65]

[66]

[67]

[68]

[69]

Bromer, J.G.; Sakkas, D.; Seli, E. Metabolomic profiling of embryo

culture media to predict IVF outcome. Expert Rev. Obstetrics Gynecol. 2008, 3(4), 441-447.
Nagy, Z.P.; Sakkas, D.; Behr B. Non-invasive assessment of embryo viability by metabolomic profiling to culture media (metabolomics). Reprod. BioMed. Online 2008, 17(4), 502-507.
Vergouw, C.G.; Botros, L.L.; Roos, P.; Lens, J.W.; Schats, R.;
Hompes, P.G.A.; Burns, D.H.; Lambalk, C.B. Metabolomic profiling by near-infrared spectroscopy as a tool to assess embryo viability: a novel, non-invasive method for embryo selection. Hum. Reprod. 2008, 23(7), 1499-1504.
Handyside, A.H.; Pattinson, J.K.; Penketh, R.J.A.; Delhanty,
J.D.A.; Winston, R.M.L.; Tuddenham, E.G.D. Biopsy of human
preimplantation embryos and sexing by DNA amplification. Lancet
1989, 333(8634), 347-349.
McArthur, S.J,. Leigh, D.; Marshall, J.T.; de Boer, K.A.; Jansen,
R.P.S. Pregnancies and live births after trophectoderm biopsy and
preimplantation genetic testing of human blastocysts. Fertil. Steril.
2005, 84(6), 1628.
Liston, R.; Crane, J.; Hamilton, E.; Hughes, O.; Kuling, S.;
MacKinnon, C.; McNamara, H.; Milne, K.; Richardson, B.; Trpanie, M.J. Fetal health surveillance in labor. J. Obstet. Gynaecol.
Can. 2002, 24(3), 250-276.
Mozurkewich, E.L.; Wolf, F. Near-infrared spectroscopy for fetal
assessment during labor. Cochrane Database Syst. Rev. 2000, 3,
CD002254.
Wyatt, J.S.; Edwards, A.D.; Azzopardi, D.; Reynolds, E.O.R. Magnetic Resonance and near infrared spectroscopy for investigation of
perinatal hypoxic-ischiamic brain injury. Arch. Dis. Child. 1989,
64, 953-963.
Nicklin, S.; Hassan, I.; Wickramasinghe, Y.; Spencer, S. Arch. Dis.
Child. Fetal Neonatal Ed. 2003, 88(4), F263.
McClaine, R.J.; Uemura, K.; de la Fuente, S.G.; Manson,
R.J.; Booth, J.V.; White, W.D.; Campbell, K.A.; McClaine, D.J.;
Benni, P.B.; Eubanks, S.; Reynolds, J.D. General anesthesia improves fetal cerebral oxygenation without evidence of subsequent
neuronal injury. J. Cerebr. Blood Flow Metab. 2005, 25, 10601069.
O'Brien, P.M.; Doyle, P.M.; Rolfe, P. Near infrared spectroscopy in
fetal monitoring. Br. J. Hosp. Med. 1993, 49(7), 483-487.
Rolfe, P.; Wickramasinghe, Y.A.; Thorniley, M.S.; Faris, F.; Houston, R.; Kai, Z.; Yamakoshi, K.; O'Brien, S.; Doyle, M.; Palmer, K.
Fetal and neonatal cerebral oxygen monitoring with NIRS: theory
and practice. Early Hum. Dev. 1992, 29(1-3), 269-273.
Faris, F.; Rolfe, P.; Thorniley, M.; Wickramasinghe, Y.; Houston,
R.; Doyle, M.; O'Brien, S. Non-invasive optical monitoring of
cerebral blood oxygenation in the foetus and newborn: preliminary
investigation. J. Biomed. Eng. 1992, 14(4), 303-306.
DAntona, D.; Aldrich, C.J.; OBrien, P.; Lawrence, S.; Delpy,
D.T.; Wyatt, J.S. Recent advances in fetal near infrared spectroscopy. J. Biomed. Opt. 1997, 2(1), 15-21.
Watkin, S.L.; Spencer, S.A.; Dimmock, P.W.; Wickramasinghe,
Y.A.; Rolfe, P. A comparison of pulse oximetry and near infrared
spectroscopy (NIRS) in the detection of hypoxaemia occurring with
pauses in nasal airflow in neonates. J. Clin. Monit. 1999, 15, 441447.
Strangman, G.; Boas, D.A.; Sutton, J.P. Non-invasive neuroimaging using near-infrared light. Biol. Psychiat. 2002, 52, 679-693.
Taillefer, M.C.; Denault, A.Y. Cerebral near-infrared spectroscopy
in adult heart surgery: systematic review of its clinical efficacy.
Can. J. Anesth. 2005, 52(1), 79-87.
Gough, K.; Rak, M.; Bookatz, A.; Del Bigio, M.; Mai, S.; Westaway, D. Choices for tissue visualization with IR microspectroscopy. Vib. Spectrosc. 2005, 38(1-2), 133-141.
Skoch, J.; Dunn, A.; Hyman, BT.; Bacskai, BJ. Development of an
opticla approach for noninvasive imaging of Alzheimers disease
pathology. J. Biomed. Opt. 2005, 10(1), 11007/1-011007/7.
Hillman, E.M.C. Optical brain imaging in vivo: techniques and
applications from animal to man. J. Biomed. Opt. 2007, 12(5),
051402.
Arnold, S.E.; Hyman, B.T.; Flory, J.; Damasio, A.R.; van Hoesen,
G.W. The topographical and neuroanatomical distribution of neurofibrillary tangles and neuritic plaques in the cerebral cortex of patients with Alzheimer's disease. Cereb. Cortex 1991, 1, 103-116.

Hahn et al.
[70]

[71]
[72]

[73]

[74]

[75]

[76]

[77]

[78]
[79]

[80]

[81]

[82]

[83]

[84]

[85]

[86]

[87]

[88]

Bierer, L.M.; Hof, P.R.; Purohit, D.P.; Carlin, L.; Schmeidler, J.;
Davis, K.L.; Perl, D.P. Neocortical neurofibrillary tangles correlate
with dementia severity in Alzheimer's disease. Arch. Neurol. 1995,
52, 81-88.
Galimberti, D.; Scarpini, E. New perspectives for the treatment of
Alzheimers disease. Open Geriatric Med. J. 2008, 1, 33-42.
Raygani, A.V.; Rahimi, Z.; Kharazi, H.; Tavilani, H.; Pourmotabbed, T. Association between apolipoprotein E polymorphism
and serum lipid and apolipoprotein levels with Alzheimer's disease.
Neurosci. Lett. 2006, 408(1), 68-72.
Aleong, R.; Blain, J.F.; Poirier, J. Pro-inflammatory cytokines
modulate glial apolipoprotein E secretion. Curr. Alzheimer Res.
2008, 5(1), 33-37.
Evans, K.C.; Berger, E.P.; Cho, C.G.; Weisgraber, K.H.; Lansbury,
P.T. Apolipoprotein E is a kinetic but not a thermodynamic inhibitor of amyloid formation: implications for the pathogenesis and
treatment of Alzheimer disease. Proc. Natl. Acad. Sci. 1995, 92,
763-767.
Hintersteiner, M.; Enz, A.; Frey, P.; Jaton, A.L.; Kinzy, W.;
Kneuer, R.; Neumann, U.; Rudin, M.; Staufenbiel, M.; Stoeckli,
M.; Wiederhold, K.H.; Gremlich, H.U. In vivo detection of amyloid-
deposits by near-infrared imaging using an oxazine-derivate
probe. Nat. Biotechnol. 2005, 23(5), 577-583.
Griebe, M.; Daffertshofer, M.; Stroick, M.; Syren, M.; AhmadNejad, P.; Neumaier, M.; Backhaus, J.; Hennerici, M.G.; Fatar M.
Infrared spectroscopy: a new diagnostic tool in Alzheimer disease.
Neurosci. Lett. 2007, 420(1), 29-33.
Ray, S.; Britschgi, M.; Herbert, C.; Takeda-Uchimura, Y.; Boxer,
A.; Blennow, K.; Friedman, L.F.; Galasko, D.R.; Jutel, M., Karydas, A.; Kaye, J.A.; Leszek, J.; Miller, B.L.; Minthon, L.; Quinn,
J.F.; Rabinovici, G.D.; Robinson, W.H.; Sabbagh, M.N.; So, Y.T.;
Sparks, D.L.; Tabaton, M.; Tinklenberg, J.; Yesavage, J.A.; Tibshirani, R.; Wyss-Coray, T. Classification and prediction of clinical
Alzheimer's diagnosis based on plasma signaling proteins. Nat.
Med. 2007, 13, 1359-1362.
Irizarry, M.C. Biomarkers of Alzheimer disease in plasma. NeuroRx. 2004, 1(2), 226-234.
Peuchant, E.; Richard-Harston, S.; Bourdel-Marchasson, I.; Dartigues, J.F.; Letenneur, L.; Barberger-Gateau, P.; Arnaud-Dabernat,
S.; Daniel, J.Y. Infrared spectroscopy: a reagent-free method to distinguish Alzheimers disease patients from normal-aging subjects.
Trans. Res., 2008, 152(3), 103-112.
Zhang, L.; Henson, M.J.; Sekulic, S.S. Multivariate data analysis
for Raman imaging of a model pharmaceutical tablet. Anal. Chim.
Acta 2005, 545(2), 262-278.
Shah, R.B.; Tawakkul, M.A.; Khan, M.A. Process analytical technology: Chemometric analysis of Raman and near infra-red spectroscopic data for predicting physical properties of extended release
matrix tablets. J. Pharm. Sci. 2007, 96(5), 1356-1365.
Furukawa, T.; Sato, H.; Shinizawa, H.; Noda, I.; Ochiai, S. Evaluation of homogeneity of binary blends of poly(3-hydroxybutyrate)
and poly(L-lactic acid) studied by near infrared chemical imaging
(NIRCI). Anal. Sci. 2007, 23(7), 871-876.
Dolbnev, D.V.; Dorofeev, V.L.; Arzamastsev, A.P.; Azimova, I.D.;
Vakhtel, A.V.; Stepanova, E.V. Use of near-infrared spectrophotometry (NIR) for identification of pharmaceutical drugs. Meditsinskoi i Farmatsevticheskoi Khimii 2008, 6, 27-30.
Ulmschneider, M.; Wunenburger, A.; Penigault, E. Using nearinfrared spectroscopy fort he noninvasive identification of five
pharmaceutical active substances in sealed vials. Analysis 1999,
27(10), 854-856.
Elizarova, T.E.; Shtyleva, S.V.; Pleteneva, T.V. Using nearinfrared spectrophotometry fort he identification of pharmaceuticals and drugs. Pharm. Chem. J. 2008, 42(7), 432-434.
Liu, F.; Zhao, W.; Liu, G.; Tang, Z.; Dou, Y.; Ren, Y. Nondestructive quality control of rutin pharmaceuticals by near infrared reflectance spectroscopy and artificial neural network. Huaxue Fenxi. Jiliang 2003, 12(3), 11-13.
Alvarenga, L.; Ferreira, D.; Altekruse, D.; Menezes, J.C.;
Lochmann, D. Tablet identification using near-infrared spectroscopy (NIRS) for pharmaceutical quality control. J. Pharm. Biomed.
2008, 48(1), 62-69.
Axon, T.G.; Brown, R.; Hammond, S.V.; Maris, S.J.; Ting, F.
Focusing near infrared spectroscopy on the business objectives of

Infrared-Spectroscopy

[89]

[90]

modern pharmaceutical production. J. Near Infrared Spec. 1998,

6(1-4), A13-A19.
Ravn, C.; Skibstedb, E.; Bro, R. Near-infrared chemical imaging
(NIR-CI) on pharmaceutical solid dosage forms comparing common calibration approaches. J. Pharm. Biomed. 2008, 48(3), 554561.
Storme-Parisa, I.; Clarot, I.; Esposito, S.; Chaumeil, J.C.; Nicolas,
A.; Brion, F.; Rieutord, A.; Chaminade, P. Near InfraRed Spectros-

Current Medicinal Chemistry, 2010 Vol. 17, No. 21

[91]

11

copy homogeneity evaluation of complex powder blends in a

small-scale pharmaceutical preformulation process, a real-life application. Eur. J. Pharm. Biopharm. 2008, 72(1), 189-198.
Yang, Z.; Leon, J.; Matrin, M.; Warder, J.W.; Zhang, R.; Liang, D.;
Lu, W.; Tian, M.; Gelovani, J.G.; Qiao, A.; Li, C. Pharmacokinetics and biodistribution of near-infrared fluorescence polymeric
nanoparticles. Nanotechnology 2009, 20(16), 165101.