Vibrational Spectroscopy 114 (2021) 103244

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

Discriminant analysis and quantitative study of antibiotics in infant milk

powder based on hyperspectral detection
Jun Hu a, Zhen Xu a, Maopeng Li a, Yong He b, Yande Liu a, *, 1
a
School of Mechatronics & Vehicle Engineering, East China Jiaotong University, Nanchang, Jiangxi, 330013, PR China
b
School of Mechanical Engineering, Zhejiang University, Hangzhou, 310027, PR China

A R T I C L E I N F O A B S T R A C T

Keywords: Objective: Food quality and safety has become the focus of attention for people from all walks of life. As antibiotic
Hyperspectral residues in food will cause serious harm to human health, it is necessary to realize the rapid and non-destructive
Infant milk powder detection of antibiotic residues in food. The problem of antibiotic residues is among the most urgent problems to
Antibiotic
be tackled in the quality problems of milk powder, so it is very important to conduct accurate qualitative
RF
LS-SVM
identification and quantitative detection of antibiotics in milk powder.
Method: Based on hyperspectral technology and combined with chemometrics, this research took the three
common residual antibiotics (doxycycline, chlortetracycline and oxytetracycline) in milk powder as the research
objects to monitor the quality of milk powder. Firstly, Samples were prepared by grinding, drying, weighing,
mixing and performing successively according to the designed concentration gradient. Then, the spectral of pure
sample (infant milk powder and pure antibiotic) and samples containing three types of antibiotic residues were
acquired characteristics and compared. Thirdly, to establish a qualitative discriminant model for different
antibiotic residues in infant milk powder, the Partial Least Squares Discriminant Analysis (PLS-DA) and Random
Forest (RF) models were established to identify antibiotic residues in milk powder. Fourthly, to establish a
quantitative discriminant model for antibiotic residues in infant milk powder, to simplify the models and reduce
the computational complexity, three methods, namely, Successive projection Algorithm (SPA), Uninformative
Variable Elimination (UVE), and Competitive Adaptive Reweighted Sampling (CARS) were used to select the
wavelengths for the optimal method. Then the Least Squares Support Vector Machine (LS-SVM) model was
established to conduct quantitative detection of residual antibiotics.
Result: In the qualitative analysis, PLS-DA model can roughly identify three antibiotics, with an accuracy rate of
96.2 %. RF model has better effect, with an identification accuracy reaching 100 %. In the establishment of
quantitative detection model, after the spectrum wavelengths of three types of milk powder samples was selected
by CARS algorithm, the CARS-LS-SVM model which was established by using only 7% of the data showed good
effect. Among them, the prediction set correlation coefficient Rp and Root Mean Square Error of Prediction Set
(RMSEP) of milk powder samples containing aureomycin, doxycycline and oxytetracycin residues were 0.9990
and 0.08 %, 0.9996 and 0.05 %, 0.9997 and 0.04 %, respectively. The LOD(Limit of Detection) of aureomycin,
doxycycline, and oxytetracycline were 2.44 × 10− 3, 1.51 × 10− 3, 1.2 × 10-3, respectively.
Conclusion: The identification of infant milk powder can be well realized by using hyperspectral technology
combined with RF algorithm. The LS-SVM models were established by hyperspectral technology combined with
CARS algorithm can then be used to set up better quantitative determination models of antibiotic residues in
infant milk powder. This research can provide a theoretical basis for the detection of antibiotics in other types of
food and can guarantee food safety to a certain extent.

* Corresponding author at: School of Mechatronics & Vehicle Engineering, East China Jiaotong University, 330013, PR China.
E-mail addresses: hujun_ecjtu@163.com (J. Hu), jxliuyd@163.com (Y. Liu).
1
Received: Oct 5, 2020.

https://doi.org/10.1016/j.vibspec.2021.103244
Received 26 November 2020; Received in revised form 15 February 2021; Accepted 10 March 2021
Available online 13 March 2021
0924-2031/© 2021 Published by Elsevier B.V.
J. Hu et al.
1. Introduction
Infant milk powder, as a necessity in the growing period of infants
and young children, is the main nutrition source for them. Therefore, it
is particularly important to ensure the safety of infant milk powder,
which is related to the future of the country [1]. Antibiotics are not only
used as veterinary drugs to treat animal diseases, but also as feed ad­
ditives to prevent animal diseases and promote animal growth. How­
ever, excessive use of antibiotics could lead to excessive antibiotic
residues in milk source, seriously threatening the health of people [2].In
order to seek economic benefits, some milk powder manufacturers
ignore the quality supervision in the process of milk powder production,
resulting in excessive antibiotic content in infant milk powder that pose
serious threats to the safety of infants and young children. Therefore, it
is necessary to strengthen the detection of antibiotic residues in milk
powder to ensure the quality and safety of milk powder.
The research of antibiotics in dairy products has always been the
focus of academic attention. Currently, most of the analytical methods
for antibiotics are based on the physical and chemical analysis, and the
biological determination method. Traditional physical and chemical
analysis methods mainly include High Performance Liquid Chromatog­
raphy (HPLC), Thin Layer Chromatography (TLC), Gas Chromatography
(GC), Mass Spectrometry (MS), and the combination of these techniques
[3]. The mentioned above methods require complex pretreatment, and
the detection process is tedious, time-consuming and expensive [4].
Traditional biological determination methods include immune analysis
method and biosensors method [5]. The biological determination
methods are all destructive detection methods with high cost and
complicated operation, which cannot meet the requirements of speed
and cost for quality detection of infant milk powder [6]. Although the
pure antibiotics have obvious characteristic absorption peaks in the
terahertz spectrum, the terahertz technology still has the disadvantages
of low laser source power and low detection sensitivity at this stage, and
it is currently difficult to use in actual milk powder safety testing [7].
Although some scholars have studied the use of metamaterials to
enhance detection sensitivity, the structure design of metamaterials is
relatively complex, the processing is difficult, and the metamaterials are
not universal [8,9]. Therefore, it is urgent to explore a rapid and
non-destructive method for detecting antibiotic residues in milk powder.
As a kind of non-destructive testing technology, hyperspectral
detection technology has been widely used in various fields in recent
years. Many researchers have applied it to the detection of food and
chemical substances. Jie Dengfei et al. [10] used hyperspectral tech­
nology to study the prediction models of soluble solid content in
different parts of citrus, with the correlation coefficient of the prediction
(Rp) of 0.950 and Root Mean Square Error of Prediction (RMSEP) of
0.636 %. Wang Guanghui et al. [11] used hyperspectral technology
combined with chemometrics to model and analyze the aflatoxin B1 and
gibberellin in moldy corn, with the prediction accuracy of aflatoxin B1
content was 98.74 %, and the correct rate of gibberellin content pre­
diction of 100 %.Yin Yong et al. [12] used hyperspectral technology to
classify the mildew degree of corn, and the identification accuracy of 6
grades of moldy corn was 98.6 %. Liu Zheng et al. [13] used hyper­
spectral technology to detect the nitrite content of sausages in different
storage periods, and the coefficient of determination and the root mean
square error of the accuracy of the established partial least square
regression model were 0.9829 and 0.0592, respectively. The results
showed that the spectral information at full wavelength was more
suitable for establishing hyperspectral detection model of nitrite content
in sausage storage. Hyperspectral technology is particularly suitable for
quality and safety research in food and agricultural products, but
hyperspectral imaging technology has rarely been reported in antibi­
otics detection.
In this paper, the infant milk powder and the widely used aureo­
mycin, doxycycline and oxytetracycline were used as the research ob­
jects to detect the antibiotic residues in milk powder. The PLS-DA and RF
2
Vibrational Spectroscopy 114 (2021) 103244
algorithm were established to find superior discrimination model of
antibiotic residues type. To optimize the model, SPA, UVE and CARS
algorithm were used to select the spectral wavelengths of the samples
[14–16]. Finally, different quantitative models were establish to detect
the antibiotic residues in milk powder, and to explore the optimal
model. This study can provide relevant basis and theoretical reference
for the subsequent research on antibiotics in food.
2. Materials and methods
2.1. Experimental materials and devices
The infant milk powder used in this experiment was under the brand
of FIRMUS; the antibiotic samples were purchased from the Aladdin
website (www.aladdin-e.com). Among them, the purity of aureomycin
hydrochloride was ≥80.0 % (USP grade), the purity of doxycycline
≥98.0 %, and the purity of oxytetracycline hydrochloride ≥95.0 %.
According to the designed doping ratio (quality ratio), the samples were
prepared according to the designed concentration gradient (concentra­
tion 0.07 %, 0.008 %, 0.01 %, 0.02 %, 0.04 %, 0.06 %, 0.08 %, 0.1 %, …,
5%), with the doping ratio of 0.007 % ~ 5.0 % for each type of antibi­
otics. A total of 21 groups were prepared, 5 samples for each concen­
tration group, with 1 group of pure antibiotic samples and 1 group of
pure milk powder samples. A total of 105 mixed samples were used to
simulate the milk powder samples containing antibiotic residues.
The samples in this experiment were prepared through steps such as
weighing, mixing, sample loading. The specific preparation process is as
follows: (1)In the process of sample preparation, all the samples
involved have been fully ground and passed through 200 mesh sieve.
Samples screened through the sieve were used for subsequent experi­
ments;(2) The antibiotics and milk powder were added into the centri­
fuge tube according to the sample quality ratio table, and then were
labeled; (3) the prepared samples were put on the vortex mixer and
shaken for 3 min for thorough mixture; (4) the samples were put into a
plastic culture dish with a diameter of 30 mm and a height of about 8
mm, and then were scraped flat; (5) labels of corresponding concen­
tration and antibiotic type were stuck outside the culture dish. The
preparation methods of the 3 types of residual antibiotics samples were
all the same [17].The samples were randomly divided into calibration
set and prediction set at a ratio of about 3:1 by using K–S (Kennard-­
Stone) algorithm for subsequent modeling. Table 1 shows the true values
distribution of calibration set and prediction set for preparing infant
milk powder samples containing different antibiotic concentrations.
In this experiment, the Gaia Sorter system produced by Dualix
Spectral Imaging Technology Co.,LTD. Fig. 1 shows the Schematic and
structural of hyperspectral imaging system.The hyperspectral imaging
device is mainly composed of three parts: imaging lens, imaging spec­
trometer and detector. The light source range of the spectral camera is
980nm~2500 nm, and the main acquisition parameters are set as fol­
lows: the exposure time of the imaging system is 0.1− 20 ms (15 ms in
this experiment), the spectral resolution is 10 nm, the forward speed of
the motor platform is 5 mm/s, and the return speed is 20 mm/s.
2.2. Spectral acquisition
After the parameters were set, the image should be focused to get a
clear spectral image. In this experiment, SpecView software was used to
acquire hyperspectral images of mixed infant milk powder samples.
During the process of image acquisition, the sample acquisition surface
should face the camera lens and the sample should be placed in a po­
sition not exceeding the spectral image acquisition area.
In order to avoid the uneven distribution of the light source in each
wavelength and the influence of dark current in the camera on the image
quality, the image should be calibrated after the acquisition of hyper­
spectral images. Before the calibration of spectral image, the calibration
reference shall be obtained. The main steps to obtain the reference are as
J. Hu et al. Vibrational Spectroscopy 114 (2021) 103244

Table 1
The true value distribution of calibration set and prediction set of infant milk powder with different antibiotic concentrations.
Type of Milk Powder Classification of Sample Set Amount Minimum Value (%) Maximum Value (%) Average Value (%) STDEV

Total 105 0. 007 5.245 1.448 1.724

Type A Calibration set 79 0. 007 5.245 1.436 1.720
Prediction set 26 0. 007 5.245 1.485 1.768
Total 105 0. 006 5.028 1.400 1.649
Type B Calibration set 79 0. 006 5.028 1.387 1.646
Prediction set 26 0. 006 5.028 1.436 1.689
Total 105 0. 006 5.001 1.378 1.658
Type C Calibration set 79 0. 006 5.001 1.366 1.656
Prediction set 26 0. 006 5.001 1.417 1.697

Note: Type A milk powder is the prepared sample of infant milk powder containing aureomycin residues; Type B milk powder is the prepared sample of baby milk
powder containing doxycycline residues; Type C milk powder is the prepared sample of infant milk powder containing oxytetracycline residues.

Fig. 1. Hyperspectral Imaging System: (a) Schematic Diagram; (b) structural Diagram.

follows: Firstly, the CCD camera lens were covered to collect an all-black
image; then the lens cover was moved to collect the white frame image
of the white reference plate, which then was used to correct the original
image of the sample. The calibration Formula (1) is shown below.
I λ − Hλ
Rλ =
Bλ − H λ
In the formula, Rλ represents calibrated data; Hλ represents all-black
data; Bλ represents all-white data; and Iλ represents original data. After
all spectral images had been corrected, subsequent analysis and pro­
cessing were carried out.
ducting spectral matrix decomposition [18–20]. The discriminant
analysis process is as follows: (1) the classified variables of the cali­
bration set are established; (2) PLS analysis of classified variables and
spectral data is conducted to establish a PLS model between classified
variables and spectral data; (3) the PLS model of the classified variables
(1) and spectral characteristics is established according to the calibration
set, then the value of the classified variables of the verification set are
calculated. The calculation method is shown in Formula (2).
∑
n
Y= βi λi + bi (2)
i=1

After the original hyperspectral images of the experimental samples

In the formula, Y is the predicted classified value; n is the number of
were corrected, the characteristics of the hyperspectral images could be
spectral variables not involved in modeling; β is the energy spectrum
extracted. The spectra of the hyperspectral images were extracted by
intensity; λ is the regression coefficient; b is the intercept of the model.
using ENVI 4.5 software. Firstly, the file was imported and the hyper­
The classified predicted value of the PLS model can be obtained by
spectral image was opened. Then, the region of interest was selected.
adding the weighted sum of spectral variables and regression co­
Afterwards, ENVI software was used to calculate the average spectrum
efficients within the full spectrum, and the intercept.
of the region of interest for subsequent data modeling.
2.3.2. RF algorithm
RF is an integrated algorithm based on decision tree classification. Its
2.3. Algorithm introduction
basic idea is to select training samples to minimize the residual sum of
squares until a complete decision tree is formed, so that multiple deci­
2.3.1. PLS-DA algorithm
sion trees can be formed and the final prediction can be obtained by
PLS-DA is a multivariate method based on PLS, which combines
voting. The most important parameter of RF is the number of decision
principal component analysis (PCA) and correlation analysis to perform
trees, and the more the number is, the higher the prediction accuracy
linear regression on spectral data and classified variables. Meanwhile,
will be [21,22]. Compared with the neural network, RF not only reduces
the spectrum was decomposed and fitted by setting factor matrix. Then,
the computation but also improves the prediction accuracy of the model.
the factors could be substituted into the spectral data when the matrix
was decomposed, thus avoiding the impact on the data of only con­

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J. Hu et al.
2.3.3. LS-SVM algorithm
LS-SVM is a learning method based on kernel function. It is opti­
mized on the basis of support vector machine to reduce the amount of
computation so as to reduce the calculation time, thus improving the
efficiency of model establishment [23–25]. LS-SVM has good mathe­
matical properties and can solve practical problems such as small sam­
ple, local minimum point, nonlinear and high dimension. Its main kernel
functions include linear kernel function (Link_kernel) and radial basis
kernel function (RBF_kernel). The algorithm of RBF_kernel adopted in
this paper is shown in Formula (3).
⃦ ⃦2 /
K(xi , xj ) = exp(− ⃦xi − xj ⃦ 2σ2 ) (3)
In the formula, xi represents the sample point; xj represents the
center point of the kernel function; σ2 represents the kernel parameter.
Eq. (4) was calculated according to the slope of the fitting curve
between true value and predictive value established by hyperspectral
data and the variance of prediction error. The calculation result shows
that the limit of detection (LOD) had a confidence interval of 99.86 %
[26]. In Eq. (4),σ refers to the standard error of predictive concentration;
m refers to the slope of the fitting curve, and RMSEP is equal to the value
of σ in the model.
3σ
LOD =
m
2.4. Model evaluation criteria
The parameters of the evaluation model mainly include correlation
coefficient (rc and rp) of calibration set and prediction set and RMSE
(RMSEC and RMSEP) between those two sets. The higher the correlation
coefficient of the model, the closer its value is to 1, the better the pre­
dictive effect; the smaller the RMSEP, the higher the accuracy of the
Fig. 2. The flow chart of establishing detection model of antibiotic residues in
milk powder.
Vibrational Spectroscopy 114 (2021) 103244
model, and the closer the values of RMSEC and RMSEP are, the better
the robustness of the model. Fig. 2 shows the flow chart of establishing
the detection model of antibiotic residues in infant milk powder.
3. Experimental results and analysis
3.1. Hyperspectral response characteristics of pure infant milk powder
and aureomycin samples
3.1.1. Hyperspectral contrast analysis of different antibiotic samples
Hyperspectrum has abundant data and message. In this experiment,
the frequency range of hyperspectral is 957.49~2567.67 nm, with a
total of 288 band points. Fig. 3 shows the spectral comparison of the
three pure antibiotics, it can be seen that the reflectance of the three
antibiotics as a whole had a similar trend. As the wavelength increased,
the reflectance gradually decreased, but there were certain differences
in different wavelengths. Oxytetracycline has the highest reflectivity
among the three antibiotics at 957.79~1400 nm, 1700~1900 nm and
2050nm~2567.67 nm.
3.1.2. Hyperspectral analysis of infant milk powder containing antibiotic
residues
Figs. 4–6 show the spectral comparison of different contents of the
(4) three antibiotics in milk powder, among which the spectral reflection
intensities of the three pure antibiotics were all higher than that of the
milk powder. With the increase of spectral wavelength, the spectral
refraction intensities of the infant milk powder containing antibiotic
residues all presented a downward trend. Fig. 4 shows the samples of
mixed milk powder containing aureomycin residues, and the spectral
peaks were relatively obvious at 1154.54 nm, 1261.51 nm, 1368.48 nm,
1706.28 nm, 1908.96 nm, 2004.67 nm and 2252.39 nm. Fig. 5 shows the
spectral peaks of the mixed milk powder containing doxycycline resi­
dues at 1154.54 nm, 1261.51 nm, 1368.48 nm, 1706.28 nm, 1908.96
nm, 2004.67 nm and 2252.39 nm. Fig. 6 shows the spectral peaks of the
mixed milk powder containing oxytetracycline residues at 1154.54 nm,
1261.51 nm, 1368.48 nm, 1706.28 nm, 1908.96 nm, 2004.67 nm and
2252.39 nm. And with the increase of antibiotic concentration, the in­
tensity of absorption peak increased gradually. The reason for the
emergence of this peak might be that the concentration of antibiotics
mixed in infant milk powder increased, making the hyperspectral
reflection of samples more intense.
3.2. Establishment of a qualitative model for different antibiotic residues
in infant milk powder
In this experiment, 105 spectra were collected for each type of
sample, a total of 315 spectra, with the wavelength range of
Fig. 3. Spectral comparison of different types of antibiotics.
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Fig. 4. Hyperspectral comparison diagram of samples with different concen­ Fig. 7. The regression coefficient curve chart of the PLS-DA discriminant model
trations of aureomycin in infant milk powder. for antibiotic types in milk powder.

square method to discriminant analysis and establishes a regression

model between hyperspectral characteristics and the samples’ classified
variables [27]. The main idea is to replace the concentration vector with
the classified vector of the sample, and then establish the model by
quantitative PLS method. K–S algorithm was used to randomly divide
the 315 collected groups of hyperspectral data into calibration set and
prediction set according to the ratio of 3:1. It means that the numbers of
spectral data occupied by the two sets were 236 and 79 respectively.
Based on the divided calibration set and prediction set, the PLS-DA
model was built by using unscrambler8.0 software. In this experiment,
the number of optimal principal component factors in the PLS-DA model
was 19. Fig. 7 shows the regression coefficient curve of the PLS-DA
discriminant model for antibiotic types in milk powder. Regression co­
efficients were obtained through the model regression calculation,
namely the weighted value of spectral variables corresponding to
different wavelength points. By adding the weighted sum of regression
coefficients and spectral variables and the intercept (b=-11.64), the
class value of the PLS-DA model could be obtained for qualitative
Fig. 5. Hyperspectral comparison diagram of samples with different concen­
determination of residual antibiotic types in milk powder.
trations of doxycycline in infant milk powder.
Fig. 8 shows the PLS-DA model fitting diagram for the classified
variables of the prediction set (3 types of samples). The correlation co­
efficient of the prediction set (rp) was 0.9468 and RMSEP was 0.4875 %.
The intermediate value of the categorization vector was taken as the
threshold with an interval of 2, and the intermediate value was taken as
the threshold with corresponding thresholds T1 = 2.0 and T2 = 4.0
respectively.

Fig. 6. Hyperspectral comparison diagram of samples with different concen­

trations of oxytetracycline in infant milk powder.

957.49~2567.67 nm. The spectra of Type A, B and C samples were

marked as “1′′ , “3′′ and “5′′ respectively, and then the model was con­
structed for qualitative discrimination.

3.2.1. Establishment of PLS-DA model for infant milk powder

PLS-DA is a qualitative discriminant method that applies partial least Fig. 8. Fitting diagram of prediction set for PLS-DA model.

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Table 2
Classification results of RF discriminant model.
Samples Calibration set Prediction Set True Correct rate

315 236 79 79 100.00%

Fig. 9. OOB error rate of spectra.

There were three types of milk powder samples in total: Type A

(containing residual aureomycin),Type B (containing residual doxycy­
cline), and Type C (containing residual oxytetracycline). The judgment
results are as follows: the judgment result of Type A was correct, and the
misjudgment rate was 0%; the judgment result of Type B was correct,
and 1 sample was misjudged; there were 2 errors in Type C judgment
results; there were 3 misjudgments in 79 predicted samples. In conclu­
sion, the comprehensive misjudgment rate was 96.2 %.
3.2.2. Establishment of random forest discriminant model for infant milk
powder
Essentially, RF model is a classifier containing multiple decision
trees. These decision trees are formed by a random method, so they are
also called random decision trees and there is no correlation between the
trees in RF. The test data enter into the random forest to be classified by
each decision tree, and finally the category with the most classified re­
sults among all decision trees is taken as the final result. The method of
randomly selecting split attribute sets is adopted by RF to build decision
trees [28]. Firstly, K–S algorithm was used to divide 315 samples of
hyperspectral data into a calibration set and a prediction set at a ratio of
about 3:1. The parameter settings are as follows: Ntree was the number
of decision trees in RF, which was set to be 500; Mtry was the number of
attributes in the split attribute set, serving as the square root of the
√̅̅̅̅̅̅̅̅̅
number of spectral variables ( 288 (rounded down)), which was about
16. Next, the RF classification model was constructed. Fig. 9 shows the
relationship between out-of-bag (OOB) error rate and the number of
decision trees. When the number of decision trees was 27, the OOB error
Fig. 11. Selecting process diagram of wavelength variable based on
SPA algorithm.
rate reached the minimum value of 0.0085. Fig. 10 shows the variation
of contribution rate of variables. Variables near the wavelength range of
957.5~1064.5 nm and 2455~2567.7 nm could be better used to iden­
tify different types of antibiotics.
Table 2 shows the classification results of the RF discriminant model.
The accuracy of classification for different types of antibiotics was 100
%, showing good classification effect.
3.3. Quantitative model analysis of antibiotic residues in infant milk
powder
3.3.1. Effective wavelength selection of antibiotic residues in infant milk
powder
In order to simplify the model and reduce computation, SPA, UVE
and CARS were used to select the spectral wavelengths in order to find a
better wavelength selection method.
3.3.1.1. Selection of hyperspectral wavelength variables based on SPA
algorithm. SPA is a forward cyclic selection method, which uses vector
projection analysis to select the effective wavelength with minimum

Fig. 12. The results of the mixed samples of milk powder variables selection
Fig. 10. The variation diagram of variable contribution rate. by UVE.

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J. Hu et al. Vibrational Spectroscopy 114 (2021) 103244

uses the UVE algorithm to select 37 variables from the original spectrum
of 288 variables. The spectrum of Class C milk powder uses the UVE
algorithm in the original spectrum. 28 variables were selected out of 288
variables. The algorithm eliminated the redundant information of the
original spectrum and optimized the model.

3.3.1.3. Effective wavelength variable selection based on CARS algorithm.

Fig. 13. The process of spectra variables selection by CARS.
redundancy and minimum multicollinearity. This method starts with a
wavelength. In each cycle, its projection on the unselected wavelength is
calculated and the wavelength with the maximum projection vector is
introduced into the wavelength combination [29]. Fig. 11 shows the
spectrogram after selecting the spectral variables of mixed infant milk
powder samples based on SPA algorithm (taking Type A milk powder as
an example). The spectrum of Type A milk powder selected 26 variables
out of 288 variables in the original spectrum using SPA algorithm; the
spectrum of Type B milk powder selected 37 variables out of 288 vari­
ables in the original spectrum using SPA algorithm; the spectrum of Type
C milk powder selected 28 variables out of 288 variables in the original
spectrum using SPA algorithm. This algorithm eliminated redundant
information and optimized the model to some extent.
3.3.1.2. Effective wavelength variable selection based on UVE algorithm.
The Uninformed Variable Elimination Method (UVE) is a wavelength
selecting method based on the partial least squares (PLS) regression
coefficient. The basic idea is to use the regression coefficient as a mea­
sure of the importance of wavelength. Fig. 12 shows the result of UVE
variable selecting of the spectrum of the mixed sample of infant milk
powder. The black spectrum line on the left side of the variable dividing
line represents the wavelength stability curve, and the right side is the
wavelength dividing line. The two parallel lines parallel to the X axis
represent the threshold of UVE variable selecting. The upper and lower
thresholds are ±12.1154, and the stability is at Variables other than the
upper and lower thresholds can be used as input variables, and the
variables whose stability is within the upper and lower thresholds will
be eliminated and not included in the built model. The spectrum of Class
A milk powder uses the UVE algorithm to select 26 variables from the
original spectrum of 288 variables. The spectrum of Class B milk powder
The Competitive Adaptive Reweighted Sampling (CARS) algorithm fol­
lows the principle of "survival of the fittest", using Monte Carlo sampling
(MCS) to randomly select 80 % of the sample set as the calibration set,
and then press the selected calibration set. The spectral array and con­
centration array of a part of the samples are proportionally extracted to
establish the PLS model respectively. And use exponential decay func­
tion (EDP) and adaptive reweight sampling (APS) to filter variables,
eliminate redundant information, and identify variables with high ab­
solute values of regression coefficients. Fig. 13 is a process diagram of
variable selecting of mixed samples of infant milk powder by CARS al­
gorithm. It can be seen from Fig. 13(a) that the number of reserved
wavelengths gradually decreases as the number of samples increases. In
the beginning, as the sampling runs increase, the number of reserved
wavelengths decreases faster, then decreases more slowly, and finally
does not change, which embodies the CARS wavelength variables se­
lection is the process from rough selection to fine selection. Fig. 13(b)
shows the trend diagram of the RMSECV value changing with the in­
crease of sampling times during the process of selecting wavelength
variables. When the sampling times are from 1 to 26, the RMSECV value
gradually becomes smaller. When the sampling time is 26, RMSECV get
the minimum value. After the number of samples exceeds 26, the
RMSECV value gradually increases. The above process shows that when
the number of sampling times is less than 26, the CARS algorithm filters
out spectral information that is not related to chlortetracycline antibi­
otics, and the important information related to chlortetracycline anti­
biotics is selected out after the number of sampling times 14 is greater
than 26. Fig. 13(c) shows the trend graph of the wavelength variable
regression coefficient changing with the increase of sampling times
during the wavelength selecting process. The position shown by "*" in
the Fig. is the sampling times when the RMSECV value takes the mini­
mum value. It can be seen from Fig. 13(a) that when the number of
samples is 26, RMSECV takes the minimum value, and the number of
wavelength variables corresponding to this time is 23.
3.3.2. Establishment of LS-SVM model for mixed infant milk powder
LS-SVM models mainly have two functions: RBF kernel function and
Lin linear kernel function. The established LS-SVM models of mixed
infant milk powder samples were compared and evaluated. In this paper,
the RBF kernel function which is better than Lin linear kernel function
was adopted. Table 3 compares the performance results of LS-SVM
models established after wavelength selecting based on RBF kernel
function. By comparing the spectral results of the three samples pro­
cessed by different waveband selecting methods, it was found that

Table 3
Performance comparison of LS-SVM models based on variable selection methods in different wavelengths.
Type Treatment Method No. of variable γ σ2 rp RMSEP
4
Full spectrum 288 5.7209 × 10 573.5550 0.9990 0.0008
SPA 26 7.2264 × 105 6.0030 × 103 0.9983 0.0010
Type A
UVE 95 2.8098 × 103 129.1420 0.9989 0.0008
CARS 23 9.7373×103 66.0053 0.9990 0.0008
Full spectrum 288 1.2456 × 103 6.452 × 103 0.9995 0.0006
SPA 37 1.1468×107 472.1220 0.9992 0.0006
Type B
UVE 113 3622.2995 295.9600 0.9976 0.0013
CARS 23 2.7910×105 655.5651 0.9996 0.0005
Full spectrum 288 1.0462 × 105 5362.3224 0.9995 0.0005
SPA 28 5.0517 × 105 288.0593 0.9996 0.0005
Type C
UVE 113 2.0917 × 105 1807.8376 0.9996 0.0005
CARS 23 2.5144×105 519.8977 0.9997 0.0004

7
J. Hu et al.
Fig. 14. Predicted antibiotic residues contents of LS-SVM models in
three types.
samples processed by CARS could be better modeled. After being
selected by CARS on wavelengths, only 7% of the spectral data of Type
A, B and C milk powder samples were collected. In the prediction set of
the three types, the correlation coefficient rp and RMSEP were 0.9990
and 0.08 %, 0.9996 and 0.05 %, 0.9997 and 0.04 % respectively, indi­
cating that the established CARS-LS-SVM model had a good effect.
Therefore, the method of CARS combined with LS-SVM could be used to
quantitatively determine antibiotic residues in infant milk powder.
Fig. 14 shows the predicted values and true values of the three
antibiotic residues in the mixed samples of milk powder. The predictive
correlation coefficients (rp) of the CARS-LS-SVM models for the three
samples were greater than 0.9990, and the detection accuracy and sta­
bility of the model were relatively ideal. Therefore, CARS-LS-SVM model
was more suitable for the detection of antibiotic residues in infant milk
powder. The LOD of aureomycin, doxycycline, and oxytetracycline were
2.44 × 10− 3, 1.51 × 10− 3, 1.2 × 10-3, respectively.
4. Conclusion
This paper verified the feasibility of qualitative and quantitative
determination of antibiotics in infant milk powder by using hyper­
spectral detection technology. PLS-DA and RF qualitative discrimination
models were established respectively, and the accuracy of the RF qual­
itative discrimination model was up to 100 %. In the establishment of
the quantitative model, wavelength selection was conducted on the
original spectrum using the UVE, CARS and SPA algorithm, then the LS-
SVM model was established. The LOD of aureomycin, doxycycline, and
oxytetracycline were 2.44 × 10− 3, 1.51 × 10− 3, 1.2 × 10-3, respectively.
It was found that the predictive correlation coefficients of the estab­
lished CARS-LS-SVM model for the three types of antibiotics were all
greater than 0.9990, indicating that this model had better detection
effect and best predictive effect. This experiment verified that hyper­
spectral detection technology could be applied in the detection of other
types of antibiotics in food, which was of great significance to ensure
food safety.
Author statement
Jun Hu: Conceptualization, Methodology, Software, Writing-
Reviewing and Editing. Zhen Xu : Data curation, Software, Writing-
Original draft preparation. Maopeng Li: Visualization, Investigation.
Yong He: Supervision. Yande Liu: Validation, Funding acquisition,
Project administration.
8
Vibrational Spectroscopy 114 (2021) 103244
Declaration of Competing Interest
The authors report no declarations of interest.
Acknowledgement
The authors gratefully acknowledged the financial support of Na­
tional 863 Program (SS2012AA101306), “the 12th Five-Year Plan”,
Jiangxi Advantageous Science and Technology Innovation Team Con­
struction Plan (20153BCB24002), Collaborative Innovation Center
Project of Intelligent Management Technology and Equipment for
Southern Mountain Orchards (G.J.G.Z. [2014] No.60),National Natural
Science Foundation of China (2002017018).Science and Technology
Research Youth Project of Jiangxi Education Department (GJJ190348);
Innovation Fund Project for Doctoral Students of Jiangxi Province
(YC2019-B106).
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Jun Hu, male, born in 1992 in Xiaogan, Hubei Province,doctor; research direction: THz
spectroscopy applications in agricultural product quality and safety testing.
Yande Liu, female, born in 1967 in Ji’an, Jiangxi Province, doctor, professor; research
direction: spectroscopic diagnostic technology in agricultural product quality and safety.