Food Measure (2017) 11:1909–1918
DOI 10.1007/s11694-017-9573-7
ORIGINAL PAPER
Determination of total phenolic content and antioxidant capacity
of blueberries using Fourier transformed infrared (FT-IR)
spectroscopy and Raman spectroscopy
Xiaozhen Zheng1 · Yaxi Hu1 · Elfi Anggreani1 · Xiaonan Lu1 
Received: 9 January 2017 / Accepted: 7 June 2017 / Published online: 9 June 2017
© Springer Science+Business Media, LLC 2017
Abstract  This study investigated the feasibility of using
Fourier-transform infrared (FT-IR) and Raman spectros-
copies to quantify total phenolic content and antioxidant
capacity of blueberries. Blueberry extracts were applied to
determine total phenolic content (Folin–Ciocalteu assay)
and antioxidant capacity [oxygen radical absorbance capac-
ity (ORAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH)],
while pressed juices were used for spectroscopic analysis.
Six partial least squares regression models with cross-val-
idation were developed using FT-IR and Raman spectra
of blueberries from 11 locations and their corresponding
chemical testing values, followed by prediction tests using
samples from another five locations. FT-IR prediction
models show relatively good prediction power for ORAC,
DPPH, and Folin–Ciocalteu values ­(R2-prediction = 0.6359,
0.6580, and 0.8092; RMSE-prediction  = 6.19 and
0.71  µmol trolox equivalents/g fresh weight, and 0.14  µg
GAE/g fresh weight), but Raman prediction models did not
yield a satisfactory result. FT-IR spectroscopy may be used
to rapidly determine blueberry antioxidants.
Keywords  Blueberry · Phenolics · Antioxidant · Infrared
spectroscopy · Raman spectroscopy
Xiaozhen Zheng and Yaxi Hu have equally contributed as co-first
authors.
Electronic supplementary material  The online version of this
article (doi:10.1007/s11694-017-9573-7) contains supplementary
material, which is available to authorized users.
* Xiaonan Lu
xiaonan.lu@ubc.ca
1
Food, Nutrition and Health Program, Faculty of Land
and Food Systems, The University of British Columbia,
Vancouver V6T 1Z4, BC, Canada
Introduction
The benefits of antioxidants to humans are mainly asso-
ciated with the suppressing effects on free radicals [5].
Excessive production of reactive oxygen species (ROS)
may result in oxidative stress to cellular lipids, proteins,
and nucleic acids, which play a significant role in the devel-
opment of cancer, cardiovascular disorder, diabetes, central
nervous system diseases, and chronic obstructive pulmo-
nary diseases [5, 20]. Blueberries are well known as “super
fruit” with a rich resource of minerals, phenolics, vitamins,
and dietary fibers that can potentially quench ROS [28] and
show high level of antioxidant capacity [5, 6]. The antioxi-
dant properties of blueberries are due to the high content
of flavonoids, including flavones, isoflavones, flavanones,
anthocyanins, and catechins [6, 23, 28]. Flavonoids per-
form their antioxidant function through scavenging free
radicals, binding to metal ions, and inhibiting free radical-
generating enzymes [8]. Genotype, ripening stage, cultiva-
tion technique, climate condition, and storage condition can
affect antioxidants in blueberry [27].
Different chemical assays have been applied for the
examination of phenolic content and antioxidant capacity,
such as Folin–Ciocalteu (FC), 2,2-diphenyl-1-picrylhydrazl
radicals (DPPH), and oxygen radical absorbance capacity
(ORAC) assays. These assays can be categorized into two
major types: hydrogen atom transfer (HAT) and electron
transfer (ET). HAT involves a competitive reaction scheme
where the tested antioxidants will compete with the sub-
strate for the peroxyl radicals [10] while ET detects the
capability of antioxidants to transfer one electron to reduce
free radicals, metals, and/or carbonyls [14]. However, all
these chemical assays are time consuming, expensive, labor
intensive, and use a large amount of organic solvents.
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Coupled with multivariate statistical analysis, both
infrared and Raman spectroscopies have been applied for
the determination of phenolic and antioxidant profiles of
numerous food commodities, such as white wine [7], green
tea [29], onion [14, 19], and fruits [11]. Both spectroscopic
techniques measure the vibrational modes of molecules
and can fingerprint the chemical composition of a complex
matrix, such as foods. Specifically, infrared spectroscopy
records the absorption signals while Raman spectroscopy
collects the scattering signals [13].
To date, no study has been conducted to perform Raman
spectroscopy for the quantification of phenolic content
and antioxidant capacity in berries or to comprehensively
evaluate the performance of Raman and infrared spectro-
scopic methods. In this study, we determined the total phe-
nolic content and antioxidant capacity of blueberries from
different countries through the use of both infrared and
Raman spectroscopic models developed based on partial-
least squares (PLS) regression algorithm. In addition, the
functional groups that contribute to antioxidants in blue-
berry were studied using spectroscopic based PLSR load-
ing plots.
Materials and methods
Chemical and reagents
FC reagent, DPPH, 6-hydroxy-2,5, 7,8-tetramethyl-2-car-
boxylic acid (trolox), gallic acid, 2,2′-azobis (2-methyl-
propionamidine) dihydrochloride (AAPH), and fluorescein
sodium salt were obtained from Sigma-Aldrich (St. Louis,
MO, USA). Sodium carbonate ­ (Na2CO3), sodium phos-
phate monobasic anhydrous ­(NaH2PO4), potassium phos-
phate dibasic (­K2HPO4), and methanol were purchased
from Fisher Scientific (Ottawa, Ontario, Canada). All rea-
gents and solvents used were analytical or HPLC grade.
Deionized water (18.2  MΩ/cm) was received from a Mil-
lipore system at The University of British Columbia.
Blueberry sample preparation
A total of 16 boxes of blueberry samples were received
during the period from 2014 to 2015. These blueberries
were produced from different farms originated at various
locations in Argentina, Chile, Canada, and USA. All sam-
ples were stored in the dark at −80 °C until analysis.
Blueberry samples were thawed and ground with mor-
tar and pestle. A total of 2.0  g (±0.02  g) of each sample
was extracted with 70% aqueous methanol (15  mL × 3)
in an ultrasonic water bath (Fisher Scientific) for 15  min
each time for three times at 20 °C. The remaining ground
blueberry samples were stored in the dark at −80  °C
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X. Zheng et al.
until further analysis. The extracts were centrifuged at
13,000 × rpm (rotor: fiberlite f15-8 × 50  cy, Sorvall Leg-
end X1R, Thermo Scientific) for 20 min. Each supernatant
was obtained through filtration with syringe and Millex
­Express® PES filter membrane (0.22  µm) and stored in
the dark at −80 °C for further analysis. The end product of
extraction was referred to as blueberry extract.
The remaining ground blueberry samples were homog-
enized and pressed through sterile gauze to receive crude
juices. The crude juices were filtered with syringe and
Millex ­Express® PES filter membrane (0.22 µm) and stored
in the dark at −80 °C for further analysis. The end product
was referred to as blueberry juice.
Determination of total phenolic content using FC assay
Total phenolic content of each blueberry extract was deter-
mined using the procedures of the FC assay reported in a
previous work with minor modification [9]. Briefly, 50 μL
blueberry extract, 3  mL deionized water, and 500  μL FC
reagent were mixed for 8  min at 20 °C, followed by the
addition of 1.5  mL saturated sodium carbonate. The mix-
ture was incubated in the dark at 20 °C for 2 h before meas-
uring the absorbance at 765 nm using a Shimadzu UV–Vis-
ible spectrophotometer. Blank sample was deionized water.
Total phenolic content values were determined using a cali-
bration curve prepared with gallic acid standards (0, 0.05,
0.1, 0.15, 0.30, 0.45, 0.60, 0.75, and 1.00  mg/L). Results
were expressed as mg of gallic acid equivalents/g fresh
weight (mg GAE/g FW) blueberry. Each extract was deter-
mined at least in triplicate.
Determination of total antioxidant capacity using
DPPH Assay
The antioxidant capacity of blueberry extracts was deter-
mined using a DPPH assay as described by Hu and col-
leagues in a previous study [9] with minor revisions. DPPH
stock solution was prepared by dissolving 15  mg DPPH
in 30  mL 70% methanol and stored at −20 °C until use.
DPPH working solution was obtained by diluting stock
solution with 70% methanol to obtain an absorbance of
1.1 ± 0.1 at 515  nm using a spectrophotometer [27]. An
aliquot (0.1  mL) of each blueberry extract was added to
2  mL DPPH working solution. Then, the absorbance was
measured at 515 nm after incubation for 30 min in the dark
at 20 °C. Trolox dilutions in 70% methanol in a range of
0–800 μmol/L were used as the calibration standards. Total
antioxidant capacity of blueberry was expressed as μmol
trolox equivalents/g fresh weight (μmol trolox/g FW) blue-
berry. Each extract was determined at least in triplicate.
Determination of total phenolic content and antioxidant capacity of blueberries using Fourier…
Determination of total antioxidant capacity using
ORAC Assay
Phosphate buffer (75  mM, pH 7.4) prepared by ­NaH2PO4
and ­K2HPO4 was used to dilute samples and reagents.
Samples and trolox with a series of dilutions (100 μL) was
mixed with 60  μL fluorescein (200  nmol/L) and 40  μL
AAPH (60 mmol/L) in a 96-well plate. Fluorescence with
the excitation of 485 nm and the emission of 527 nm was
read every minute for 60 min by using a fluorescent micro-
plate reader (Tecan Austria GmbH, Tecan infinite M200
Pro). A linear relationship between the area under the curve
for blueberry sample or trolox and its corresponding con-
centration was determined. The value of ORAC is equiva-
lent to the slope of sample divided by the slope of trolox
and the result was expressed as μmol of trolox equivalents/g
fresh weight (μmol trolox/g FW) blueberry. Each blueberry
extract was determined at least in triplicate.
FT‑IR spectral collection
FT-IR spectra of each blueberry juice sample were col-
lected at 20 °C using a Perkin Elmer FT-IR spectrometer
equipped with an attenuated total reflectance sensing sys-
tem (PerkinElmer, Waltham, MA, USA). Blueberry juice
samples were scanned over the wavenumber region of
4000–550  cm−1 at a resolution of 4  cm−1 against a back-
ground of air. Each spectrum was an average of 16 scans,
which was collected using SPECTRA software (Perki-
nElmer, Waltham, MA, USA). An aliquot of 10 µL of blue-
berry juice was applied onto the attenuated total reflectance
sensing system of the FT-IR spectrometer and a total of 10
spectra of were collected for each sample.
Raman spectral collection
A Renishaw confocal Raman spectroscopic system
equipped with a 785  nm near-infrared laser (maximum
30 mW incident laser power) and a charge coupled device
(CCD) array detector was used for collecting Raman spec-
tra. The settings of the Raman spectrometer include the
entrance aperture at 50  µm, focal length at 300  mm, and
a 1200 line m­ m−1 grating. The CCD array detector (578-
pixel by 385-pixel) has a pixel size of 22 µm. Raman spec-
tra were collected from 400 to 1800 cm−1 at 20 °C. A total
of ten spectra were collected from different locations from
each blueberry juice sample.
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Multivariate statistical analysis
FT-IR spectral fingerprinting region (2000–900  cm−1)
was selected for further analysis because the target func-
tional groups in blueberry juice can generate spectral
bands at this wavenumber region while noises domi-
nate other wavenumber regions. The whole wavenumber
region of Raman spectra (400–1800  cm−1) was selected
for further analysis. Automatic baseline correction was
applied to both FT-IR and Raman spectra using OMNIC
v7.0 (Thermo-Nicolet, Madison, WI, USA).
A total of 16 blueberry samples from different geo-
graphical regions were divided into both calibration set
and prediction set. Eleven samples were assigned as
the calibration set and the remaining five samples were
assigned as the prediction set for evaluating the perfor-
mance of the calibration models. Specifically, samples
containing the highest and lowest values of the chemical
profiles as determined by FC, DPPH, and ORAC assays
were allocated into the calibration set and the remaining
samples were randomly allocated into both calibration set
and prediction set.
The calibration models were constructed to correlate
FT-IR or Raman spectra and the values of total phenolic
content or antioxidant capacity determined by traditional
chemical-based assays using PLS regression algorithm in
Matlab 2014b (MathWorks Inc., Natick, MA, USA). A
total of six PLS regression models were built in the end
with the combination of two instrumental datasets (i.e.
FT-IR and Raman spectra) and three chemical datasets
(i.e. FC, DPPH, and ORAC assays). Internal validation
within the calibration model was conducted using tenfold
cross-validation: one-tenth of the data in the calibration
set was sequentially removed and treated as external to
validate the model built by the remaining data [3]. Cross
validation aids in selecting the optimal number of the
latent variables (m) with two criteria, namely minimizing
the root mean square error of cross-validation (RMSECV)
and avoiding over-fitting. RMSECV is calculated using
� �2
the equation: RMSECV = N v=1 i=1 yiv − ŷ iv
1 ∑v ∑n �
[25], where N is the number of samples in the training
set; v is the number of segments; n is the number of sam-
ples in a given segment; ­yiv is the reference value for
sample i in segment v, and ŷiv is the predicted value for
sample i in segment v. After establishing the linear
regression models between the spectral features and the
chemical results with the optimized number of latent var-
iables, the prediction power of these calibration models
were evaluated using the five samples in the prediction
set. Performance of each PLS regression model was
assessed based upon the calculated correlation of deter-
mination ­(R2), root mean squares error of calibration
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(RMSEC), root mean squares error of cross-validation
(RMSECV), and root mean squares error of prediction
(RMSEP).
Results and discussion
Phenolic and antioxidant profiles of blueberries
The total phenolic content and antioxidant capacity of
blueberries from different geographical locations are
summarized in Table  S1. These results were compara-
ble to a few previous studies. Specifically, the ORAC
values ranging between 13.9 and 118.7  µmol trolox
equivalents/g FW for blueberry samples were reported
[11, 20, 23, 28]. In general, blueberries harvested from
different farms originated from different countries (i.e.,
Argentina, Chile, Canada and USA) have different pro-
files of phenolic content and antioxidant capacity. This
variation was due to the differences in genotypes, ripen-
ing age, cultivation methods, and climate conditions [23].
In addition, the application of different chemical assays
can result in variations in the values of phenolic content
and antioxidant capacity, and no correlation was discov-
ered between the results obtained from DPPA, ORAC
and FC assays (data not shown). Such variation might
be due to the different principles involved in the three
aforementioned chemical assays. First, the ORAC assay
is a HAT-based reaction providing information on radi-
cal chain-breaking capacity, while DPPH and FC assays
are based upon the ET reactions reflecting the reducing
capacity of the samples [21]. It has been reported that
many ORAC-active antioxidants are not sensitive to the
stable DPPH free radicals (e.g. chlorogenic acid), thus
DPPH assay could result in a lower antioxidant capac-
ity [10, 12]. Besides, being conducted under basic con-
ditions, simple phenolics could react with FC reagent
but lack the capacity to scavenge the free radicals (e.g.
DPPH). Therefore, even though involving the same reac-
tion mechanism, total phenolic results (i.e. FC assay)
might not have high correlation with antioxidant capacity
determined by DPPH assay. Second, ORAC assay meas-
ures fluorescence to determine both inhibition degree
and time, while DPPH and FC assays simply measures
the inhibition effect at a certain time, which might not be
representative [4, 12, 23]. Lastly, DPPH assay is a colori-
metric assay that is dependent upon the color change at
515  nm, which is the maximum UV–Vis absorption for
DPPH radicals [10]. However, anthocyanin compounds in
blueberries also exhibit absorption at 500–550 nm, which
could disturb the accuracy of DPPH assay [26].
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X. Zheng et al.
FT‑IR spectroscopy and PLS regression models
for the determination of total phenolic content
and antioxidant capacity of blueberries
A total of ten FT-IR spectra were collected per blueberry
sample; therefore, 160 spectra were collected and they
are shown in Fig.  1a. Band assignment of FT-IR spectra
of blueberries are summarized in Table  S2 [17, 22, 23].
There was only one band (at 3360 cm−1) shown up between
the wavenumber region of 4000–2000  cm−1, which was
mainly due to the –OH stretching of water in sample.
Moreover, there was noise in the wavenumber region of
900–500  cm−1. As a consequence, only the fingerprinting
region (i.e. 2000–900 cm−1) was selected for baseline cor-
rection and further spectral analysis (Fig. 1b).
The baseline corrected fingerprinting spectra were
applied for the construction of PLS regression models with
the corresponding ORAC, DPPH, and FC values, respec-
tively. Eleven samples were selected to be the calibration
set (Fig.  2) while the remaining five samples as the test
set. The statistical parameters of PLS regression models
for the determination of total phenolic content and antioxi-
dant capacity of blueberries are summarized in Table 1. A
strong PLS regression model has a relatively high regres-
sion coefficient ­(R2 close to 1) with preferably a small
number of latent variables and root mean squares error
[14]. The calibrated models for all three chemical assays
(i.e. ORAC, DPPH, and FC) demonstrated high correla-
tion between FT-IR spectra and the chemical assays. How-
ever, the performance of the models for cross-validation
and prediction was less satisfactory, with R ­ 2 of cross-val-
idations to be 0.7397, 0.7459, and 0.8373, and predictions
to be 0.6359, 0.6580, and 0.8092 for ORAC, DPPH, and
FC assays, respectively. PLS regression calibration models
resulted in the best parameters for the FT-IR spectra against
FC assay and this could be attributed to the nature of the
FC reaction. ORAC and DPPH assays have relatively short
reaction/monitoring time (i.e. 60 and 30 min, respectively),
while FC assay requires 2  h to complete. The shorter the
reaction/monitoring time, the more random error could be
induced during operation, resulting in the reduced accu-
racy. Moreover, conducted under basic conditions, activ-
ity of antioxidants toward FC reagent increased [10], thus
the relative random error and systematic error could be
minimized and the results could be more representative to
reflect the antioxidant composition of the overall sample.
The loading plots of PLS regression models (Figure
S1) illustrated the contribution of each wavenumber in
FT-IR spectra to the correlation relationship. The band at
~1650  cm−1 (assigned to aromatic C=C vibration in phe-
nolics) in the ORAC model contributed to the highest
positive correlation. The band at 1020  cm−1 (assigned to
O–H group in carbohydrate) in both DPPH and FC models
Determination of total phenolic content and antioxidant capacity of blueberries using Fourier…
Fig.  1  a 160 raw FT-IR spectra
of blueberry (500–4000 cm−1);
b 160 baseline corrected
FT-IR spectra of blueberry at
fingerprinting region (900–
2000 cm−1)
contributed to the highest negative correlation, while the
band at 1080 cm−1 (assigned to aromatic CH bending and
rocking and C–OH bending) contributed to the highest pos-
itive correlation [17].
Raman spectroscopy and PLS regression models
for the determination of total phenolic content
and antioxidant capacity of blueberries
Similar to FT-IR spectroscopic analysis, ten Raman spectra
were collected per blueberry sample. A total of 160 Raman
spectra in the wavenumber region of 1800–400  cm−1 are
1913
shown in Fig. 3a and the baseline corrected Raman spectra
are shown in Fig. 3b. Band assignment of Raman spectra of
blueberries are summarized in Table S3 [2, 15, 16, 22, 24].
For the construction of PLS regression models, the
blueberry samples used for calibration set and prediction
set were the same as the ones used in FT-IR spectroscopic
models (Fig. 4). The statistical parameters of Raman spec-
troscopic-based PLS regression models for the determina-
tion of total phenolic content and antioxidant capacity of
blueberries are summarized in Table  2. The calibration
models for both DPPH and FC assays yield a relatively
high regression coefficient of 0.93 and 0.96, respectively,
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1914 X. Zheng et al.

Fig.  2  a FT-IR calibration

model for ORAC assay; b
FT-IR calibration model for
DPPH assay; c FT-IR calibra-
tion model for FC assay

13
Determination of total phenolic content and antioxidant capacity of blueberries using Fourier… 1915

Table 1  PLS regression models for the determination and prediction of total phenolic content and antioxidant capacity of blueberries using
FT-IR spectroscopy
N n LV RMSEC R2 RMSECV R2-CV RMSEP R2-P

ORAC 110 50 7 2.8125 0.9195 4.6676 0.7397 6.1854 0.6359

DPPH 110 50 9 0.4355 0.9576 0.9689 0.7459 0.7069 0.6580
FC 110 50 7 0.1933 0.9307 0.2857 0.8373 0.1434 0.8092

N number of spectra for calibration, n number of spectra for prediction, LV latent variables, RMSEC root mean squares error of calibration,
RMSECV root mean squares error of cross-validation (tenfold), RMSEP root mean squares error of prediction

Fig.  3  a 160 raw Raman

spectra of blueberry (400–
1800 cm−1); b 160 baseline
corrected Raman spectra of
blueberry (400–1800 cm−1)

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1916 X. Zheng et al.

Fig.  4  a Raman calibration

model for ORAC assay; b
Raman calibration model for
DPPH assay; c Raman calibra-
tion model for FC assay

13
Determination of total phenolic content and antioxidant capacity of blueberries using Fourier…
Table 2  PLS regression models for the determination and prediction of total phenolic content and antioxidant capacity of blueberries using
Raman spectroscopy
N n LV RMSEC
ORAC 110 50 5 4.1370
DPPH 110 50 5 0.5656
FC 110 50 6 0.1455
N number of spectra for calibration, n number of spectra for prediction, LV latent variables, RMSEC root mean squares error of calibration,
RMSECV root mean squares error of cross-validation (tenfold), RMSEP root mean squares error of prediction
with satisfactory number of latent variables. However,
none of the prediction models resulted in a good prediction
power on the basis of ­R2 for prediction and the RMSEP.
Furthermore, the loading plot (Figure S2) did not indicate
any specific wavenumber region that contributes to a posi-
tive or negative correlation to the results of the traditional
chemical assays. Taken together, Raman spectroscopic
based method was not feasible to determine antioxidant
capacity and total phenolic content in blueberry.
Only one study has been published using Raman spec-
troscopy to determine the concentration of anthocyanins in
blueberry at different stages of ripeness and concluded that
Raman spectroscopic method was feasible to quantify the
anthocyanins in blueberry [1]. However, the authors only
assessed the calibration and cross-validation models for
PLS regression and no external unknown sample was used
to validate the PLS regression models. Besides, the cross-
validation method used in the aforementioned study was
“leave-one-out”, which has higher possibility to overfit the
model, resulting in less robustness compared to “k-fold”
cross-validation method [18]. Therefore, no evidence has
been found to demonstrate the success of applying Raman
spectroscopy to quantify the antioxidants in blueberry.
The failure of using Raman spectroscopy to study the
antioxidant capacity and total phenolic compound could
be attributed to the interference of high florescent signals
developed from the biological samples (i.e. blueberry),
which is evidenced in Fig. 3. The 785-nm laser has higher
tendency to induce the florescent signals compared to the
1024-nm laser used by Arrobas et al. [1]. Although baseline
correction was applied to remove the florescent signals in
each spectrum, the weak Raman signals could be affected,
resulting in reduced sensitivity of Raman spectroscopy
towards compounds at low concentrations. Besides, the
laser power of the Raman spectroscopic system used in our
current study was 30 mW, which might be enough to have
high sensitivity towards compounds at low concentrations.
Moreover, as concluded by Arrobas et al. [1], large amount
of emerging compounds other than anthocyanins could be
presented during blueberry maturation, masking the Raman
signals of anthocyanins at relatively low concentrations,
thus Raman spectroscopy demonstrated worse performance
1917
R2 RMSECV R2-CV RMSEP R2-P
0.7998 5.9723 0.3507 4.3026 0.2874
0.9260 0.8059 0.6474 0.9192 0.1481
0.9620 0.2749 0.6627 0.3501 0.1066
to predict the anthocyanins in mature blueberry compared
to the prediction capability towards unripen fruits. How-
ever, in this current study, all the blueberry samples were
in their mature states and the failure of using Raman spec-
troscopy to quantify antioxidant capacity and total phenolic
content is in accordance with the results found by Arrobas
and the colleagues.
Conclusion
Both FT-IR and Raman spectroscopies were applied to
replace the traditional chemical assays (i.e. DPPH, ORAC
and FC assay) for the determination of antioxidant capac-
ity and total phenolic content of blueberry. Only FT-IR
spectroscopy was validated to be feasible to provide accept-
able accuracy for the prediction of the antioxidant values
determined by the three traditional assays and the overall
analysis can be finished in a few minutes instead of hours
or days required by traditional tests. Raman spectroscopy
failed to predict the antioxidant capacity and total phenolic
content of mature blueberries, but further research could be
conducted to apply Raman spectroscopy for assessing the
ripeness of blueberry.
Acknowledgements  This study was supported by Discovery,
Engage, and Collaborative Research and Development Grants
awarded to X. L. by Natural Sciences and Engineering Research
Council of Canada. X. L. also thanks the financial support by Mitacs
Accelerate Program in Canada.
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