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Vibrational Spectroscopy 55 (2011) 146–152

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Vibrational Spectroscopy
journal homepage: www.elsevier.com/locate/vibspec

Fourier transform near- and mid-infrared spectroscopy can distinguish between


the commercially important Pelargonium sidoides and its close taxonomic ally
P. reniforme
J.E. Maree, A.M. Viljoen ∗
Department of Pharmaceutical Sciences, Tshwane University of Technology, Private Bag X680, Pretoria 0001, South Africa

a r t i c l e i n f o a b s t r a c t

Article history: Pelargonium sidoides is indigenous to South Africa and abundant in the Eastern Cape Province. Several
Received 27 July 2010 herbal products have been formulated using P. sidoides of which Umckaloabo® is probably the most
Received in revised form 13 October 2010 popular and successfully marketed in Germany. The objective of this study was to discriminate between
Accepted 18 October 2010
P. sidoides and Pelargonium reniforme by FT-IR spectroscopy. Absorbance spectra were collected for
Available online 25 October 2010
P. sidoides (n = 96) and its close taxonomic ally P. reniforme (n = 57) in the near infrared (NIR) and mid
infrared (MIR) regions. The spectroscopic data were analysed using chemometric computations including
Keywords:
principal component analysis and orthogonal projections to latent structures discriminant analysis.
Pelargonium sidoides
Pelargonium reniforme
Phytochemical variation of 5.79% in the NIR dataset (R2 X(cum) = 0.962; Q2 (cum) = 0.918) and 9.22%
Fourier transform infrared spectroscopy variation in the MIR dataset (R2 X(cum) = 0.497; Q2 (cum) = 0.658) was responsible for the separation
Fourier transform near infrared of the two species. Seven absorption areas were identified as putative biomarkers responsible for the
Fourier transform mid infrared differences between the two species. These results indicate that FT-NIR and FT-MIR spectroscopy can be
Quality control used to discriminate between these two closely related species which occupy a sympatric distribution in
South Africa.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction used by consumers internationally [3,4]. The safety and efficacy


of the herbal medicine has been clinically researched and recom-
Pelargonium sidoides DC and Pelargonium reniforme Curt., mem- mended as a curative herbal medicine for a range of diseases such as
bers of the Geraniaceae family, are indigenous to South Africa with sinusitis, acute bronchitis and common colds. From 2001 to 2006
a centre of diversity in the Eastern Cape Province [1]. P. sidoides is the product experienced a 900% growth in turnover in Germany
morphologically similar to P. reniforme. The two species can only be (D 8–80 million) [3].
visually distinguished by the leaf shape and colour of their flowers. Quality control is vital in the pharmaceutical industry to guar-
P. sidoides has dark maroon to almost black flowers with crowded antee authenticity and quality of consumer products [5,6]. A major
velvety heart shaped leaves while P. reniforme has pink to magenta challenge in quality assurance of herbal material is the vast varia-
flowers with velvety kidney shaped leaves with crenate or finely tion of constituents in plants from the same species. This variation
lobed margins [1,2]. is due to a wide range of factors including locality, climate, age,
The enthnobotanical value of P. sidoides became known to harvesting season, storage conditions and soil type [7]. As a result
the world and led to the development of herbal products, like of this variation, the selection of only a few compounds as criteria
Umckaloabo® . This phytomedicine is successfully marketed and for quality control purposes is inadequate [8]. Correct identifica-
tion of raw materials is one of the first steps in the quality and
safety assurance of herbal products. Identification tests should be
able to discriminate between related species and or potential adul-
Abbreviations: A, number of components; FT-IR, Fourier transform infrared;
terants [7]. Some methods that have been used for identification
FT-MIR, Fourier transform mid infrared; FT-NIR, Fourier transform near infrared;
PC, principal component; PCA, principal component analysis; Pp, predictive com- such as visual inspection, physical and chemical properties may
ponent; Po, orthogonal component; OPLS-DA, orthogonal projections to latent often be inaccurate and even subjective [9]. High performance
structures discriminant analysis; R2 (cum), cumulative modelled variation in X liquid chromatography (HPLC) is conventionally used to estab-
matrix; Q2 (cum), fraction of the total variation of X that can be predicted by the
lish quality control protocols. HPLC is time-consuming, costly and
extracted components.
∗ Corresponding author. Tel.: +27 12 382 6360; fax: +27 12 382 6243. requires skilled personnel. A fast, non-destructive alternative is FT-
E-mail address: viljoenam@tut.ac.za (A.M. Viljoen). IR spectroscopy. This paper focuses on the application of FT-NIR

0924-2031/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.vibspec.2010.10.005
J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152 147

spectroscopy (13 333–4000 cm−1 or 750–2500 nm), and FT-MIR 2.3. FT-NIR and FT-MIR spectroscopy measurements
(4000–400 cm−1 or 2500–25 000 nm) spectroscopy in the quality
assessment of this commercially important ethnomedicinal plant. The vials containing the samples were introduced to the FT-
FT-IR spectroscopy is often the chosen spectroscopic method due NIR spectrophotometer (Büchi NIRFlex N500 Fourier transform NIR
its superior speed, wavelength accuracy and sensitivity. It is used as spectrophotometer, Buchi, Labortechnik AG, Flawil, Switzerland)
a routine method in the pharmaceutical industry and is a popular and the reflectance spectra of the samples were collected from
method for the identification of medicines in the Pharmacopoeia of 1000 to 2500 nm (10 000–4000 cm−1 ), with NIRWARE 1.2.3000
many countries [10–13]. It is rarely used in quality control of tradi- ADVANCED Edition software, yielding 1501 data points. Each spec-
tional medicine and other complicated systems due to the lack of trum was the average of 32 scans. Before chemometric analysis
integrated interpretation of the spectra. A great deal of effort and the reflectance spectral data were converted to absorbance using
accomplishments has been made since 1998 to use this method log 1/R (R = reflectance).
for quality control purposes and in the identification and stability Dried powdered samples were scanned, positioned on the
prediction of traditional Chinese medicines [12,13]. FT-IR spec- horizontal diamond crystal of the Alpha-P FT-IR spectrometer
troscopy provides a characteristic fingerprint (absorption spectra) (Bruker Optik GmbH, Germany) equipped with a diamond ATR.
of the sample [11]. Physical and chemical parameters of a sample The absorbance spectra of the samples were collected from 500
can be predicted from a single spectrum. to 4000 cm−1 (MIR region). The spectra were obtained with 2407
Qualitative and quantitative data on herbal medicines safety and data points from the accumulation of 32 scans. The sample platform
efficacy are not adequate to meet the criteria needed to support its was cleaned between every sample using a 30% ethanol solution.
international use. Not only do health care policies contribute to
the lack of research data but also the lack of adequate or accepted
2.4. Multivariate data analysis
methodology for the evaluation of herbal medicine. Vast variation
of active constituents in plants from the same species exists. This
SIMCA-P (11.0.0.0, Umetrics, Sweden) was used to perform
variation is due to a wide range of factors including time of har-
chemometric computations including cluster and classification
vest, locality, age, storage conditions and soil type [7,14]. To ensure
analysis. Spectral filters which were used to remove additive and
the reliability and repeatability of pharmacological and clinical
multiplicative differences in the spectral dataset were first deriva-
research and to improve final product quality almost all phyto-
tive (quadratic polynomial order, gap of 5), second derivative (cubic
chemical constituents of the herbal material must be determined
polynomial order, gap of 5), Multiplicative scatter correction (MSC)
[14]. Therefore it is of atmost importance to formulate quality con-
and standard normal variate (SNV). The first derivative removes
trol protocols based on the entire metabolome.
the constant additive background effects while second derivative
In the case of P. sidoides, most of the plant material used in the
removes the baseline linear slope variation and additive effects.
preparation of the phytomedicine is still being wildcrafted by locals
Standard normal variate is used to reduce multiplicative effects of
from rural communities [4]. Due to an increase in the demand of
scattering, particle size and multicollinearity changes while MSC
P. sidoides it has suffered from unsustainable harvesting practices.
corrects both multiplicative and additive scatter effects [15]. Spec-
The decrease in natural populations of P. sidoides as resulted in
tral data were centred scaled by subtracting the average of the
the closely related P. reniforme being harvested to substitute for
dataset from each spectrum.
P. sidoides which has several undesirable consequences in product
Data analysis was done in two phases. In the first phase, unsu-
formulation and quality assurance protocols [3]. As both species
pervised principal component analysis (PCA) was performed to
are annuals, the defining taxonomic characters (flowers and leaves)
provide an overview of all observations and samples. Scatter score
may not be present during the time of harvesting the roots (autumn
plots of the first two PCs were constructed to identify and eval-
and winter). It is imperative for manufacturers to comply to phar-
uate groupings, trends and strong outliers. Orthogonal projection
macopeia standards which stipulates the use of P. sidoides. The
to latent structures discriminant analysis (OPLS-DA) was the sec-
objective of this study was to investigate infrared spectroscopy as
ond phase in data analysis. It is a supervised pattern recognition
a rapid method for the discrimination between raw plant material
method where the Y-matrix contained qualitative values. The dis-
of P. sidoides and P. reniforme and to construct and compare their
tinct dummy Y-variables indicated the class of the sample namely
spectral profiles.
P. sidoides as 1 and P. reniforme as 0. The main purpose of an OPLS
model is to separate the systematic variation in the X-matrix into
two parts, one linearly related to the Y-matrix and one that is
orthogonal (unrelated) to the Y-matrix [16]. Loading line plots of
2. Materials and methods
the first predictive component (Pp) were constructed to explain
the chemical differences responsible for the separation of the two
2.1. Sample collection
species. S-plots were constructed to filter out putative biomarkers.
An S-plot is an interactive visual representation of both magnitude
A total of 153 fresh root samples of P. sidoides (n = 96) and
(intensity) and reliability (modelled covariance vs. modelled corre-
P. reniforme (n = 57) were collected in January 2007 from different
lation). The quality of the models was evaluated based on the cross
localities in South Africa. Retention samples and voucher material
validation results such as the cumulative modelled variation in X
is retained in the Department of Pharmaceutical Sciences (TUT).
matrix (R2 X(cum)) and the fraction of the total variation of X that
can be predicted by the extracted components (Q2 (cum)).

2.2. Sample preparation 3. Results and discussion

The samples were air-dried at room temperature and milled to 3.1. FT-IR fingerprints
a powder (Pore size: 0.75 mm, Dietz Motoren GmbH) and sieved
(500 ␮m, Endecotts Ltd., England) to obtain homogenous particle Fig. 1 illustrates the characteristic NIR spectrum of P. sidoides and
size. Approximately 0.5 g of the powder was placed in chromacol P. reniforme. Spectral differences between the species FT-NIR spec-
vials (Chromacol Ltd., United Kingdom). tra were not apparent. Typical average FT-MIR spectra of P. sidoides
148 J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152

Fig. 1. Characteristic FT-NIR spectroscopy fingerprint of P. sidoides (red) and P. reni-


Fig. 2. Characteristic FT-MIR spectroscopy fingerprint of P. sidoides (red) and P. reni-
forme (green). (For interpretation of the references to colour in this figure legend,
forme (green). (For interpretation of the references to colour in this figure legend,
the reader is referred to the web version of the article.)
the reader is referred to the web version of the article.)

and P. reniforme are illustrated in Fig. 2. No apparent spectral dif- Table 1


ferences between the two spectra could be observed, therefore Statistics of spectral filters evaluated.
spectral filtering methods were required to enhance the discrete
Spectral filter A R2 X(cum) Q2 (cum)
spectral features.
FT-NIR 1st derivative 4 0.909 0.898
2nd derivative 3 0.604 0.576
3.2. Principal component analysis MSC 2 0.956 0.952
SNV 2 0.37 0.932

The full dataset of 96 P. sidoides and 57 P. reniforme samples FT-MIR 1st derivative 3 0.535 0.505
were analysed by PCA. Multiplicative scatter correction proved to 2nd derivative 3 0.274 0.236
MSC 3 0.731 0.731
be the best spectral filter modelling the most variation in the X
SNV 3 0.753 0.737
matrix and had the highest predictive power (FT-NIR: R2 X(cum):
0.956, Q2 (cum) = 0.952. FT-MIR: R2 X(cum): 0.753, Q2 (cum) = 0.731)
(Table 1). A scatter score plot of the first two PCs, which accounts
for 96.6% of variation (R2 X(cum) = 0.956) in the NIR region, was shown) confirmed that all four samples are strong outliers occur-
constructed and evaluated (Fig. 3). Two main clusters can be ring above the D-crit(0.05) value. The outliers were removed and
observed along the second PC (Y-axes). Intra-species variation can a new model was created. The new model (N = 149, K = 1501) was
be observed along PC1 (X-axes). Four strong outliers were identi- fitted with three components and yielded an R2 X(cum) of 0.928 and
fied (PS 032, PS 081, PS 030 and PS 024). The DMod-X plot (not Q2 (cum) of 0.933.

P. reniforme
P. sidoides

0.30 PS081

0.25

0.20
PR 032
0.15 PR 049PR 050
PR PR
PR 044 045 005
PRPR024006 PR 002
PR PR
PR 046
055PR 009 PR PR 008
PR043PR
PR
PRPR
036 048
007
056
041 PR
PR PR 025
053
034 022PR 003
0.10 PR 047 PRPR
PRPR
PR PR
054026
051
012
PR
PR 052
030 PR 004
042
PR 021 PR029
035
PR 057
38
028
PRPR PR
031 PR014
PR 011
018 PS 030
PR 020
010
PR40 PR 019 PRPS 047
001
PC 2

0.05 PRPR PR
3701339
PRPR 015
023 PR
PR027033
PR 016PS 025 PS 041
PS 033PRPS017 PS
040 PS 052PS 027 PS 024
-0.00 PS 094 PS 037 PS035
PS
PS 050
082
PS 083
088
PS 046 PS 031 PS PSPS
PS
087 PS 064
045
023
043 PS 059 PS 060
PS 092 PSPS PS
PS
051
PS
PS
048
PS 067
039
PS034
PS
036
078 079 PS 084
086
-0.05 PS 044 PS PS
026011
PS028
PS 005 PSPSPS 049
015
PS
038
PS
006 PS 091
014053 PS 054
PS 080
PS
PS PS
PSPS
PS 089
013
029001
090
010
076 PS
PS 071PS 075
PS
PS012
PS PS
PS
PS
004
074
PS 093
070
061
PS
042
096 PSPS
PS 073
057021
-0.10 PS 002 PS 085
PS
PS
PS PS
065PS
063
008
009 062077 PS 032
PS 003 PS
PS 007
066 PS095
068 PS 019
PS
PS 072 PS 056
PSPS055
020
PSPS 017069
PS 058
-0.15 PS 018 PS 022
PS 016
-0.20

-0.25
-1.2 -1.0 -0.8 -0.6 -0.4 -0.2 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0
PC 1
R2X[1] = 0.915294 R2X[2] = 0.0407863
Ellipse: Hotelling T2 (0.95) SIMCA-P+ 12 - 2010-04-05 10:00:42 (UTC+2)

Fig. 3. PCA overview of FT-NIR spectroscopy data of all samples (P. sidoides: red, P. reniforme: green). The ellipse represents a 95% confidence interval for the two PCs based
on the Hotelling T2 . (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of the article.)
J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152 149

Fig. 4. PCA overview of FT-MIR spectroscopy data of all samples (P. sidoides: red, P. reniforme: green). (For interpretation of the references to colour in this figure legend, the
reader is referred to the web version of the article.)

The full FT-MIR dataset of the two species was also analysed 3.3. Identification of species using orthogonal projections to
by PCA. A two dimensional scatter score plot of the first two latent structures discriminant analysis
PCs (PC1 vs. PC2), with an R2 X(cum) of 0.753, was constructed
and evaluated (Fig. 4). Two groups, although overlapping, can be Orthogonal projections to latent structures discriminant anal-
observed along PC1 (X-axes) that accounts for 36.6% of the vari- ysis (OPLS-DA) was performed to extract and visualise the main
ation in X (R2 X = 0.366). Phytochemical variation within species information in the FT-IR spectra and examine the qualitative dif-
can be observed along PC2 (Y-axes). Clusters can be observed ferences between samples. A five component OPLS-DA model, one
in both FT-NIR and FT-MIR spectroscopy models. This cluster- predictive component (Pp) and four orthogonal components (Po),
ing is difficult to explain due to the overlapping as well as the was constructed for the FT-NIR spectroscopy data. The accumu-
distribution over more than two quadrants, hence, OPLS-DA was lative reliability of the five PCs was 0.962 with a cross-validated
performed in order to simplify interpretation of cluster differ- predictive ability of 91.8% (Q2 (cum) = 0.918). The predictive infor-
ences. mation is concentrated in the first predictive component that

P. reniforme
P. sidoides

0.7

0.6

0.5

0.4

0.3

0.2
Po1

0.1

-0.0

-0.1

-0.2

-0.3

-0.4

-0.25 -0.20 -0.15 -0.10 -0.05 -0.00 0.05 0.10 0.15 0.20 0.25
Pp1

R2X[1] = 0.161168 R2X[0.36 Comp. 1] = 0.704092


Ellipse: Hotelling T2 (0.95) SIMCA-P+ 12 - 2010-04-06 12:33:18 (UTC+2)

Fig. 5. OPLS-DA score plot (Pp1 vs. Po1), of FT-NIR spectroscopy data, for the classification of P. sidoides (red) and P. reniforme (green) (R2 X for Pp1 = 0.161; Po1 = 0.704). (For
interpretation of the references to colour in this figure legend, the reader is referred to the web version of the article.)
150 J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152

0.08 3

0.07

0.06

0.05

0.04

0.03

0.02

0.01
Pp 1

-0.00

-0.01

-0.02

-0.03

-0.04

-0.05

-0.06
1
-0.07

-0.08 2
0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500
Num
R2X[1] = 0.161168 SIMCA-P+ 12 - 2010-04-06 08:57:36 (UTC+2)

Fig. 6. FT-NIR spectroscopy loading line plot of weights against wavelengths for the predictive component (Pp1) responsible for the clustering of P. sidoides and P. reniforme.

explains 16.1% (R2 X = 0.161) of the variation in X correlated with Table 2


Spectra-structure correlation summary of the first predictive component.
Y. The remaining four orthogonal components described the sys-
tematic variation in the data that are uncorrelated to Y. The four Peak no. Wavelength Bond vibration Structure Correlated
orthogonal components had an R2 X (cum) of 0.801 (Fig. 5). Only 1 1407 nm O–H str. first overtone ROH P. reniforme
0.36 of 16.1% variation in X is responsible for the separation of the 2 1899 nm O–H str. + 2xC–O str. Starch P. reniforme
two species. 3 2151 nm 2x amide I + amide III CONH2 P. sidoides
A loadings line plot of the first predictive component was con-
structed to determine which wavelengths correlate with which
species (Fig. 6). The positive loadings correlate with P. sidoides while while one peak was highly correlated with P. sidoides (peak no. 3,
negative loadings correlate with P. reniforme. The higher the peak Fig. 7). These absorbance peaks can be ascribed to certain identifi-
intensity the more it contributes to the separation between the able chemical structures as listed by Osborne et al. [17]. A summary
species. An interactive S-plot was constructed to ensure that the of the main chemical moieties possibly responsible for the separa-
correct variables responsible for interspecies variation (biomark- tion of the species are summarised in Table 2.
ers) were identified (Fig. 7). These probable biomarkers were A two component OPLS-DA model was constructed for the FT-
selected as the spectral regions with high correlation and covari- MIR spectroscopy data. The accumulative reliability of the two
ance to the y-variable. Variables selected as possible biomarkers components was 0.479 with a cross-validated predictive ability
can be observed in Fig. 7 where variables highly correlated with P. of 65.8% (Q2 (cum) = 0.658) where the predictive component (Pp1)
reniforme were selected in blue and P. sidoides in red. Two peaks explained 28.8% (R2 X = 0.288) of the variation in X correlated with Y.
(peak nos. 1 and 2, Fig. 6) were highly correlated with P. reniforme The remaining orthogonal components had an R2 X (cum) of 0.191

0.8

0.6

0.4

0.2
p(corr)[1]

-0.0

-0.2

-0.4

-0.6

-0.8

-0.08 -0.07 -0.06 -0.05 -0.04 -0.03 -0.02 -0.01 -0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08
p[1]
R2X[1] = 0.161168 SIMCA-P+ 12 - 2010-04-06 08:58:29 (UTC+2)

Fig. 7. S-plot from OPLS-DA model of FT-NIR spectroscopy data showing modelled covariance (p[1]) vs. modelled correlation (p(corr)[1]).
J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152 151

P. reniforme
P. sidoides

0.14

0.12

0.10

0.08

0.06

0.04

0.02

-0.00
Po 1

-0.02

-0.04

-0.06

-0.08

-0.10

-0.12

-0.14

-0.16
-0.24 -0.22 -0.20 -0.18 -0.16 -0.14 -0.12 -0.10 -0.08 -0.06 -0.04 -0.02 -0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16 0.18 0.20 0.22
Pp 1
R2X[1] = 0.288041 R2X[XSide Comp. 1] = 0.191394 Ellipse: Hotelling T2 (0.95)

0.32 SIMCA-P+ 12 - 2010-04-20 13:07:41 (UTC+2)

Fig. 8. OPLS-DA score plot (Pp1 vs. Po1) for the classification of P. sidoides (red) and P. reniforme (green) (R2 X for Pp1 = 0.288; Po1 = 0.191). (For interpretation of the references
to colour in this figure legend, the reader is referred to the web version of the article.)

(Fig. 8). A 9.22% (0.32 of 28.8%) variation in X is responsible for the Table 3
Spectra-structure correlation summary of the first predictive component.
separation of the two species.
Biomarkers responsible for the separation of the two species Wave number Type of vibration Structure Correlated
were identified by evaluating Fig. 9 (S-plot). The selected vari- 1606 cm−1 Bending N–H P. sidoides
ables with high correlation and covariance to the y-variable are 1314 cm−1 Stretch C–O P. sidoides
illustrated in Fig. 10. Variables selected as possible biomark- 1055 cm−1 Stretch C–O P. reniforme
ers can be observed in Fig. 9. Variables highly correlated with 963 cm−1 Bending C–H P. reniforme

P. reniforme were selected in blue and P. sidoides in red.


Four spectral regions namely 1621–1617 cm−1 , 1321–1298 cm−1 ,
1060–1048 cm−1 and 970–944 cm−1 were identified as possible Kolodziej et al. [1–3] have made considerable progress in unravel-
biomarkers (Figs. 9 and 10). Table 3 is a summary of the chemi- ling the phytochemistry of P. sidoides a large section of the complex
cal moieties identified as possible biomarkers. It is important to chemical composition remains unresolved. Furthermore, the active
mention that no attempt has been made to identify the phyto- constituents in the herbal extract remains unknown and it gen-
chemicals corresponding to the specific spectral region. Although erally accepted that there are several phytoconstituents which

0.8
0.7

0.6

0.5
0.4

0.3

0.2
0.1
p(corr)[1]

0.0
-0.1

-0.2

-0.3
-0.4

-0.5

-0.6
-0.7

-0.8

-0.9

-0.08 -0.07 -0.06 -0.05 -0.04 -0.03 -0.02 -0.01 -0.00 0.01 0.02 0.03 0.04 0.05 0.06
w[1]
R2X[1] = 0.288041 SIMCA-P+ 12 - 2010-04-20 13:05:11 (UTC+2)

Fig. 9. S-plot from OPLS-DA model of FT-MIR spectral data showing modelled covariance (p[1]) vs. modelled correlation (p(corr)[1]).
152 J.E. Maree, A.M. Viljoen / Vibrational Spectroscopy 55 (2011) 146–152

0.06

0.05

0.04

0.03

0.02

0.01

-0.00
Pp 1

-0.01

-0.02

-0.03

-0.04

-0.05

-0.06

-0.07

0 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400
Num
R2X[1] = 0.288041 SIMCA-P+ 12 - 2010-04-20 13:06:39 (UTC+2)

Fig. 10. FT-MIR spectroscopy loading line plot of weights against wave numbers for the predictive component (Pp1) responsible for the clustering of P. sidoides and
P. reniforme.

collectively contribute to the pharmacological activity of P. sidoides. Acknowledgements


Searching in the complex metabolic pools for the specific molecules
responsible for the discrimination between the species at this stage We wish to thank Mr. Ulrich Feiter (Parceval, South Africa) and
remains unwarranted and a daunting challenge for the future. Dr. Egon Koch (Schwabe, Germany) for providing logistical support.
Dr. Claire Gavaghan, application specialist at Umetrics (Sweden) is
4. Conclusions thanked for providing valuable advice on the chemometric analy-
sis.
Fourier transform near- and mid-infrared spectroscopies com-
bined with the appropriate spectral filters, show potential as rapid References
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