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Table of Content

25 August 2016, Volume 37 Issue 16
Processing Technology
Optimized Preparation of New Anthocyanin Derivative Oxovitisin A and Its Antiproliferation Effect on Cancer Cells in vitro
WU Nao, WANG Jingyi, ZHANG Qian, GUO Ying, LI Shuyi, HE Yi, ZHU Zhenzhou, HE Jingren
2016, 37(16):  1-7.  doi:10.7506/spkx1002-6630-201616001
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A novel polyphenolic compound, named oxovitisin, with unique lactone pyranone ring and non-oxonium
structures was prepared. In this study, oxovitisin A was synthesized by two-step reactions in micro-oxygenation system
by using carboxypyranoanthocyanins (vitisin A) as a precursor, which was formed from pyruvic acid and anthocyanins.
The preparation conditions of oxovitisin A, namely temperature, pH, ethanol and reaction time, were optimized using
combination of single factor and orthogonal array experiments. High performance liquid chromatography-with diodearray
detection (HPLC-DAD) was used to monitor the reaction process and the main adduct was qualitatively and
quantitatively analyzed by HPLC-DAD combined with electron ionization tandem mass spectrometry (HPLC-DAD-EIMS/
MS). The sulforhodamine B (SRB) assay was employed to evaluate the anti-tumor effects of oxovitisin A, vitisin A,
methyl-pyranoanthocyanin and malvidin-3-O-glucoside against Caco-2 and MCF-7 cells in vitro. The results showed that
the optimal preparation conditions were found to be reaction at 45 ℃ for 21 d with 20% ethanol acidified to pH 3.6 using
1.0 mg/mL of vitisin A. The effects of four important variables on oxovitisin A yield followed the decreasing order: pH >
reaction temperature > reaction duration > ethanol concentration. The main reaction product was confirmed by HPLCMS/
MS to be oxovitisin A with a yield of 26.59% after 21 d. The SRB assay showed that oxovitisin A displayed obvious
inhibitory effect on the proliferation of Caco-2 and MCF-7 cells with an IC50 value of (238.8 ± 24.6) and (85.7 ± 9) μg/mL,
respectively, obviously stronger than that of vitisin A and methyl-pyranoanthocyanin.

Optimization of Preparation Process for Corn Resistant Starch with Psychrophilic Pullulanase and Analysis of Its Properties
KANG Huaibin, YOU Xiaoyan, XIAO Tiantian, ZHANG Xiaoyu, LIU Chaojian
2016, 37(16):  8-13.  doi:10.7506/spkx1002-6630-201616002
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In this research, corn resistant starch was produced by autoclaving treatment combined with psychrophilic
pullulanase hydrolysis, and starch slurry content, the amounts of heat α-amylase and psychrophilic pullulanase, and
psychrophilic pullulanase reaction time were optimized for the enhanced production of corn resistant starch through
orthogonal array experiments. Then, the crystal structure characterization and formation mechanisms of resistant starch
were investigated by using the scanning electron microscope (SEM), X-ray diffraction (XRD) and differential scanning
calorimeter (DSC). It was found that the optimum parameters for resistant starch preparation were as follows: corn
starch slurry concentration, 18%; thermostable alpha-amylase dosage, 7 U/g; psychrophilic-pullulanase dosage, 10 U/g; and
reaction time, 9 h. The yield of resistant starch was 16.84% under these conditions. The crystal structure of resistant starch
belonged to the V-type, which was different from that the A-type crystal structure of raw starch. Compared to raw starch,
the resistant starch showed higher phase onset temperature (117.07 ℃), peak temperature (140.69 ℃), end temperature
(153.03 ℃) and phase change enthalpy (1 858.12 J/g), respectively, resulting in better thermo stability.

Optimization of Hydrolysis Conditions of Corn Bran Hemicellulose by Supported Solid Acid Catalyst with Response Surface Methodology
ZHAO Wei, ZHOU Jinkun, WANG Dawei
2016, 37(16):  14-21.  doi:10.7506/spkx1002-6630-201616003
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This work reports on the optimization of hydrolysis conditions of corn bran hemicellulose by a supported solid
acid catalyst, SO42-/Fe2O3/γ-Al2O3. First of all, the one factor-at-a-time method was used to examine the effects of catalyst
dosage, hydrolysis temperature, time and material-to-solvent ratio on total sugar content and pentose yield. Subsequently,
optimization of the four reaction conditions was performed using response surface methodology based on Box-Behnken
experimental design. Results indicated that the optimal conditions for hydrolysis of corn bran hemicellulose were determined
as follows: catalyst amount, 12 mL/10 g corn bran; hydrolysis temperature, 120 ℃; hydrolysis time, 4 h; and solid-toliquid
ratio, 1:10. The maximum pentose yield of 22.45%, which was close to the predicted value, was obtained under these
conditions. Monosaccharide composition and contents of the hydrolysate, as analyzed by HPLC, showed that the relative
contents of three main monosaccharides were glucose 4.99%, xylose 34.78%, and arabinose 55.90%, respectively. Infrared
(IR) spectra of corn bran and its hydrolysis residues showed that the structure of xylose and arabinose changed, indicating
that hemicellulose is degraded.

Optimized Extraction Conditions and Physicochemical Characteristics of Dietary Fiber from Faba Bean Testa from Different Cultivars
LIU Hongkai, LI Fang, ZHANG Yahong, LIU Hong, KANG Yufan
2016, 37(16):  22-28.  doi:10.7506/spkx1002-6630-201616004
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Dietary fiber (DF) contents of faba bean testa from 20 different cultivars were determined and DF was extracted
by an enzyme-alkali method. The optimization of extraction conditions was investigated using single factor experiments
and orthogonal array design. For this purpose, the effects of four crucial parameters including cellulose dosage, pH, alkali
treatment time and temperature, on extraction efficiency were examined. Physicochemical characteristics of DF from
different faba bean cultivars were compared and analyzed. The results indicated that the DF contents of faba bean testa from 20
different cultivars were 78.7–87.12 g/100 g and the optimum extraction conditions were as follows: cellulose dosage, 30 U/g; pH,
12.0; and alkali treatment for 60 min at 60 ℃. Further, it was found that DF from the ‘Lincan 6’ had the highest water holding
capacity (7.67 g/g), the best Fe3+ and Cu2+ chelating ability (140.09 mg/g and 33.63 mg/g), whereas DF from ‘Fengdou 16’ had
the lowest bulk density (0.206 g/cm3), the largest swelling capacity (4.00 mL/g), the highest adsorption ability for unsaturated and
saturated fatty acids (4.48 g/g and 4.23 g/g), and the highest DPPH-scavenging ability (13.23 μmol/g).

Extraction Optimization and Functional Properties of Protein Isolates from Elaeagnus mollis Seed Kernels
FENG Xiaoxiao, SU Yingjie, WU Mengyue, WU Mengyang, YANG Jing, LI Meiping, ZHANG Shengwan
2016, 37(16):  29-35.  doi:10.7506/spkx1002-6630-201616005
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The chemical composition of Elaeagnus mollis seed kernels was preliminarily analyzed. Then, several common
methods for extracting plant proteins were compared to establish a more efficient method to extract protein isolates from
Elaeagnus mollis seed kernels. Alkaline extraction and acid precipitation was selected for higher extraction efficiency,
and the extraction parameters were optimized using one-factor-at-a-time method and orthogonal array design. Moreover,
a systematic investigation was carried out to evaluate functional properties of protein isolates. The results showed that
E. mollis seed kernels contained 50.87% crude fat, 36.27% protein, 2.91% starch, 5.50% water and 3.10% ash. The optimal
parameters for extraction were determined as follows: pH, 11.0; S/L ratio, 1:50 (g/mL); extraction temperature, 55 ℃;
extraction time, 2.0 h; and two extraction cycles. Under these conditions, the extraction yield of protein was 93.85% with a
purity of 84.81%. The water-holding capacity and oil-absorbing capacity of the proteins extracted ranged from 1.2 to 1.8 g/g
and 2.8 to 3.3 g/g, respectively. The foaming capacity and the emulsifying capacity could reach 190% and 25 m2/g under
optimum pH conditions.

Extraction, Identification and Screening for Antioxidant Activity of Total Flavonoids from Cichorium endivia L. Seeds
ZHAO Yue, LI Rong, JIANG Zitao
2016, 37(16):  36-42.  doi:10.7506/spkx1002-6630-201616006
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The ultrasonic-microwave synergistic extraction of total flavonoids from Cichorium endivia L. seeds was
optimized by response surface methodology. The optimum conditions were determined as follows: ratio of liquid to solid,
40:1(mL/g); ethanol concentration, 71%; extraction temperature, 65 ℃; microwave power, 400 W; ultrasonic power,
50 W; and extraction time, 6 min. Under these extraction conditions, the content of total flavonoids in C. endivia seeds was
93.23 mg/g. The optimum dynamic adsorption and desorption conditions for purification of the crude extract obtained with
AB-8 macroporous resin were established as follows: sample pH, 4; sample flow rate, 2 BV/h; and 70% alcohol as eluant
at a flow rate of 2 BV/h. HPLC analysis results showed that total flavonoids from C. endivia seeds were mainly composed
of chlorogenic acid and cynarin. Off-line DPPH-HPLC assays showed that antioxidant activity of cynarin was stronger than
that of chlorogenic acid, which could scavenge 64.11% and 61.65% DPPH free radical, respectively.

Optimization of Extrusion-Assisted Extraction of Yellow Pigment from Corn Gluten Powder
JIAO Yan, CHANG Ying, YU Shifeng, WANG Yan, BO Liying, ZHENG Xiqun
2016, 37(16):  43-47.  doi:10.7506/spkx1002-6630-201616007
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In order to improve the efficiency of pigment extraction from corn gluten powder, the effects and underlying
mechanism of extrusion pretreatment on the extraction rate of yellow pigment from corn gluten powder were studied, and
components of the pigment were analyzed. The results indicated that pigment yield increased obviously after extrusion. The
optimal extrusion conditions were found as follows: moisture content, 18%; extruder barrel temperature, 150 ℃; and screw
speed, 160 r/min. Under these conditions, the pigment yield reached (59.03 ± 1.86) mg/g, which was 149.70% higher than
that from non-extruded corn gluten powder. The microstructure observed by scanning electron microscope (SEM) showed
that the external morphology and internal structure of corn gluten powder were destroyed and significantly changed after
extrusion, leading to increased yield of yellow pigment. The high performance liquid chromatographic (HPLC) profile of the
pigment showed that the main components consisted of zeaxanthin, lutein and other carotenoids.

Optimization of Ultrasonic-Assisted Extraction of Pectin from Muskmelon Rind and Its Physical Characteristics
YANG Xiaoqing, ZHANG Yingnan, WANG Lili, LIU Jianzhi
2016, 37(16):  48-53.  doi:10.7506/spkx1002-6630-201616008
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This study was intended to optimize the process conditions for ultrasonic-assisted extraction of pectin from
muskmelon rind by the combined use of one-factor-at-a-time method and response surface methodology. For this purpose,
a quadratic regression model was developed indicating the effects of extraction parameters on pectin yield, and the main
effects and interactions of ultrasonic irradiation time, ultrasonic power and solid-to-solvent ratio on extraction efficiency
were examined. The results showed the model was highly statistically significant and had a good prediction performance.
The decreasing order of the effects of the studied variables on pectin yield was as follows: solid-to-solvent ratio >
ultrasonic irradiation time > ultrasonic power. The optimum extraction conditions that provided the maximum pectin yield
(1.76%) were obtained as 41 min; 300 W and 1:21 (g/mL) for ultrasonic irradiation time, power and solid-to-solvent ratio,
respectively. Physicochemical analyses and Fourier transform infrared spectroscopic (FTIR) studies revealed that the
extracted pectin had a galacturonic acid content of 76.24% and an esterification degree of 67.31% as a high-ester pectin
which was in line with the requirements of the national standard. By adding the pectin, the gel strength of gel products could
be enhanced by up to 140%, in turn resulting in improved physical characteristics.

Process Optimization for Fluidized Bed Drying of Paddy Rice Using Response Surface Analysis Based on Multiple Quality Indexes
ZHANG Yue, DING Chao, YANG Guofeng, LIU Qiang, ZHU Hongyan, LI Xingchao
2016, 37(16):  54-62.  doi:10.7506/spkx1002-6630-201616009
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In order to optimize the fluidized bed drying process of paddy rice, a three-factor-three-level Box-Behnken
response surface experimental design was used to analyze the effects of air temperature, moisture removal in each drying
pass (MREDP) and tempering time on drying efficiency, additional crack percentage (ACP), chalky grain percentage (CGP),
fatty acid value (FAV), hardness and adhesiveness of dried rice. The results showed that the drying efficiency, ACP, CGP
and hardness of paddy rice increased with increasing either drying temperature or MREDP, while FAV and adhesiveness
decreased. However, the drying efficiency, ACP, FAV and hardness decreased and adhesiveness increased as the tempering
time increased. Moreover, at low drying temperature, as the tempering time increased, ACP and CGP decreased but not
significantly. The highest membership degree comprehensive rating score of 0.75 was achieved under the optimum drying
conditions established: hot-air temperature, 45 ℃; MREDP, 2.50% (dry basis); and tempering time, 3 h. The proposed
predictive model proved to be reliable and effective since no significant difference was seen between the experimental data
and model prediction (P < 0.05).

Optimization of Preparation of Acetylated Polysaccharides from Longan (Dimocarpus longan) Pulp by Response Surface Methodology and Its Antioxidant Ability
WANG Jing, WU Nini, HUANG Jing, WEI Yiming, YANG Yanfang, LI Xuehua, LUO Haixian, QIN Anni, GAN Rizhi
2016, 37(16):  63-68.  doi:10.7506/spkx1002-6630-201616010
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In this study, the synthesis of acetylated polysaccharides extracted from longan (Dimocarpus longan) pulp was
investigated using acetic anhydride. The influences of reaction time, reaction temperature and polysaccharides-to-acetic
anhydride ratio on the degree of substitution (DS) of the synthesized derivatives were examined to select and optimize
important reaction conditions by response surface methodology. The antioxidant activity of acetylated polysaccharides
in vitro was determined. The optimized conditions that provided the maximum DS of 0.443 were determined as follows:
reaction time, 30 min; polysaccharides-to-acetic anhydride ratio, 10.21:1; and reaction temperature, 42 ℃. The acetylated
derivatives could scavenge hydroxyl free radical, and inhibit lipid peroxidation and red cell hemolysis induced by H2O2, with
50% inhibition concentrations (IC50) of 702.41, 646.04 μg/mL and 380.11 μg/mL, respectively. The antioxidant ability of the
synthesized acetylated derivatives was improved as compared to the native polysaccharides.

Macroporous Adsorption Resin for the Purification of Polyphenols from Aronia melanocarpa Berries
LI Bin, GAO Ningxuan, LIU Hui, MENG Xianjun, ZHANG Qi, JIAO Xinyao, LI Enhui1, CHOU Shurui, ZHANG Ye, CHEN Shifu
2016, 37(16):  69-74.  doi:10.7506/spkx1002-6630-201616011
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In this study, six different macroporous adsorption resins were evaluated for their use to purify polyphenols from
Aronia melanocarpa berries. Macroporous resin XAD-7 was selected as the most suitable one. The purification parameters
optimized by static adsorption and desorption tests were as follows: adsorption and desorption time, 4 and 2 h, respectively;
sample concentration, 3.6 mg/mL; sample pH, 4; and desorption solvent, 95% ethanol (pH 7). The best purification
parameters optimized by dynamic adsorption and desorption tests were as follows: sample loading amount, 560 mL; flow
rate, 2 mL/min; washing the column with 350 mL of water; and subsequent elution with an eluent volume of 300 mL at
2 mL/min flow rate. Under these conditions, the purity of polyphenols from A. melanocarpa was improved from 11.62% to
64.37%, indicating that macroporous resin XAD-7 has a better effect in purifying polyphenols from A. melancarpa.

Preparation of Reducing Sugar from Corn Bran by Steam Explosion Combined with Acid Hydrolysis: Process Optimization Using Response Surface Methodology and Analysis of Hydrolysis Degree
ZHANG Lili, NING Dongxue, KANG Lijun, KOU Fang, XIA Tiantian, CAO Longkui
2016, 37(16):  75-82.  doi:10.7506/spkx1002-6630-201616012
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In order to increase reducing sugar yield from corn bran as a by-product of the production of corn starch by
wet milling, the optimal conditions for preparation of reducing sugar from corn bran by steam explosion combined with
acid hydrolysis were investigated in this research. First of all, the one-factor-at-a-time method was employed to examine
the effects of sulphuric acid concentration, hydrolysis time, temperature and material-to-liquid ratio on reducing sugar
content. Subsequently, the optimization of process parameters for improved reducing sugar content was performed using
Box-Behnken design and response surface methodology. Hydrolysates were analyzed by ion chromatography. The results
showed that the maximum reducing sugar content of 54.61% was obtained under the following optimal conditions: sulphuric
acid concentration, 1.66%; hydrolysis time, 1.5 h; temperature, 120 ℃; and material-to-liquid ratio, 1:10 (g/mL), which
was increased by 9.58% as compared to that obtained without steam explosion pretreatment. Ion chromatography analysis
indicated that the hydrolysate obtained was mainly composed of D-glucose (19.34 mg/mL), D-xylose (16.01 mg/mL)
and L-arabinose (10.37 mg/mL). In comparison to the raw material, the fiber structure of the residue left after the preparation of
reducing sugar was more loose and more significantly cracked, showing more holes and cracks on its surface, revealing that the
cellulose, hemicellulose and lignin in corn bran can be well decomposed by successive steam explosion and acid hydrolysis. This
result is in accordance with the reducing sugar content in the degradation solution obtained after both treatments.

Optimization of Coating Formulations for Low-Sugar Preserved Peony Using Response Surface Methodology
ZHAI Fangfang, ZHU Wenxue*, MA Yitong, CHEN Zixian
2016, 37(16):  83-87.  doi:10.7506/spkx1002-6630-201616013
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With the aim to develop the optimal coating formulation to be used in low-sugar candied peony for improved
qualities, the optimization of coating formulations was investigated using single-factor experiments and response surface
methodology with Box-Behnken design. The optimal combination of ingredients found were apple pectin powder 5 g/L,
peony red pigment 4 g/L, and xylitol 28.77 g/100 mL. With this optimized coating formulation, the total color value (ΔE) of
low-sugar preserved peony was 9.01 with a shrinkage rate of 14.59%. Compared with the control group, the experimental
group with the coating was significantly improved. This study can provide a reference for the development of peony foods.

Ultra High Pressure Treatment for Improvement of Viscosity, Dispersion and Stability of Low Fat Mayonnaise
ZHANG Xing, PAN Jian, WANG Xi, ZHANG Enguang
2016, 37(16):  88-93.  doi:10.7506/spkx1002-6630-201616014
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Purpose: To improve the viscosity, dispersion and stability of low fat mayonnaise by ultra high pressure (UHP)
treatment. Methods: The optimal conditions for UHP treatment were investigated by single factor and orthogonal array
design experiments taking viscosity, median diameter and centrifugal demulsification rate as responses. Results: The amount
of oil added in mayonnaise was reduced to 50% by the application of UHP treatment at 400 MPa with a pressure-holding
duration of 10 min. Moreover, this treatment resulted in an over 6 times increase in viscosity, an increase in dispersibility of
14.5% and an elevation in stability of 90.7% as well as better sensory characteristics of mayonnaise compared to untreated
control. Conclusions: On the basis of ensuring the safety of mayonnaise, the application of UHP treatment can reduce the
amount of oil added to 50% and simultaneously improve its viscosity, dispersion, stability and sensory characteristics,
thereby having broad application prospect.

Component Analysis
Correlation Analysis of Volatile Components and Sensory Properties in Fresh Waxy Corn
WU Jianping, NIU Liying, LI Dajing, LIU Chunquan, XIAO Lixia
2016, 37(16):  94-99.  doi:10.7506/spkx1002-6630-201616015
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Two waxy corn cultivars, namely ‘Jingtianzihuanuo No. 2’ and ‘Suyunuo No. 11’, were harvested at four time
points from day 14 to 33 after pollination and pulped freshly or after blanching. Volatile components of corn juice were
analyzed using solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) and sensory properties
were evaluated using quantitative analysis. Results showed that the major volatile components of the fresh samples were
aliphatic chain aldehydes, alcohols and ketones, with hexanal being the most dominant compound. At the same time, grassy
and green bean flavor were clearly perceived. The contents of aldehydes, alcohols and ketones were decreased after blanching
and dimethyl sulfide was detected as the most abundant compound. Moreover, the sensory properties of the blanched samples
were described as cooked immature corn or popcorn flavor. The fresh corn juice was unacceptable (1.57–2.00), while
blanched samples were acceptable (2.86–3.90). Correlation analysis showed that the high contents of hexanal and hexanol
of the fresh samples may contribute to the unacceptable odor while the typical corn flavor of the blanched samples may be
related to the reduced contents of aldehydes and alcohols and the formation of dimethyl sulfide. Factor analysis showed that
the earlier harvested corn was more obviously tender. The most tender corn was the blanched sample of ‘Jingtianzihuanuo
No. 2’ harvested at 14 d after pollination.

Interactions of Silver Carp Meat Proteins with Characteristic Fishy Volatile Compounds Analyzed Using HS-SPME-GC-MS
LIU Lin, CHONG Yunqing, DING Yuting, GU Saiqi, ZHOU Xuxia
2016, 37(16):  100-105.  doi:10.7506/spkx1002-6630-201616016
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In this work, the binding abilities of four characteristic fishy volatile compounds (heptanal, octanal, nonanal,
and 1-octen-3-ol) to silver carp meat proteins (myofibrillar proteins, myosin, and actin) were studied by headspace solid
phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). Results indicated
that myosin, one of the major myofibrillar proteins, was the primary binding receptor of the volatiles. Based on the studies
of thermodynamic models (Scatchard equation and Klotz plot), the Gibb’s free energy (ΔG) of binding of myosin to the
four fishy volatiles studied were all below 0, indicating that the interactions are spontaneous. Compared with other linerchain
aldehydes (heptanal, octanal and nonanal), 1-octen-3-ol had more binding sites and higher binding constant to myosin
protein. Moreover, the binding affinity of myosin to linear-chain aldehydes decreased with the chain length of the aldehydes.
These results could provide theoretical basis for improving the existing rinsing method, aiming to achieve effective
deodorization of surimi.

Analysis of Volatile Compounds in 11 Kinds of Health Wines
ZHANG Yanan, SUN Baoguo, SUN Jinyuan, SUN Xiaotao, HUANG Mingquan, LI Hehe
2016, 37(16):  106-111.  doi:10.7506/spkx1002-6630-201616017
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Volatile compounds of 11 kinds of health wines were analyzed by silica gel column chromatography (SGCC) and
gas chromatography-mass spectrometry (GC-MS). The identification and quantification were carried out by comparing with
standards and searching the NIST 11 Library. The results showed a total of 37 compounds were identified in 11 wine samples,
consisting of 6 alcohols, 12 hydrocarbons, 9 esters, 4 aldehydes, 2 acids, 2 ketones, eugenol and propyl lactate. Among these
compounds, n-propanol, 2-methyl-1-propanol, 3-methyl-1-butanol, ethyl lactate, triethyl citrate, 5-hydroxymethyfurfural
(5-HMF) and acetic acid were found to be prominent components. Methyl lactate was used as an internal standard
to quantify propyl lactate. In these samples, the concentrations of propyl lactate ranged from 0.02 to 0.74 mg/L.
The regression equation for propyl lactate demonstrated good linearity (R2 = 0.995). The limits of detection (LOD) was
0.005 mg/L and the limit of quantitation (LOQ) was 0.01 mg/L. Pareto analysis and one-way analysis of variance (ANOVA)
were carried out to qualitatively analyze the important volatile compounds and differences in the percentage compositions of
alcohols, hydrocarbons, esters and aldehydes in Chinese Jinjiu, Yangchun Zibujiu, Tuopai Goji, Yedao Haiwang, Zhuyeqing
and Zhangyu Sanbian wines by MATLAB 7.0. The ANOVA results showed that a significant difference in hydrocarbons
was detected. The important volatile compounds included 3-methyl-1-butanol, 1-propanol, 2-methyl-1-propanol, 2-butanol,
5-HMF, eugenol, ethyl lactate, ethyl hexanoate, isoamyl acetate, phenethyl acetate, ethyl caprate, triethyl citrate, furfural,
cinnamyl aldehyde, acetic acid, styrene, tetradecane, pentadecane, cetane, 1,4-dimethyl benzene, and diphenylamine.

Analysis of Oligosaccharides Released from Weak Acid Hydrolysis of Abalone Gonad Polysaccharide Using PMP-HPLC-MSn
LU Jiaojiao, AI Chunqing, WEN Chengrong, XU Xin, ZHANG Bao, SONG Shuang
2016, 37(16):  112-116.  doi:10.7506/spkx1002-6630-201616018
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In order to investigate oligosaccharides released from abalone gonad sulfated polysaccharide (AGSP) through
weak acid hydrolysis, hydrolysates obtained at different acid concentrations were labeled with 1-phenyl-3-methyl-5-
pyrazolone (PMP), and then analyzed by high performance liquid chromatograph-mass spectrometry (HPLC-MSn). Two
disaccharides (DS1 and DS2) and one tetrasaccharide (TS) were found in the AGSP hydrolysates. The two disaccharides
were composed of a hexuronic acid linked to a hexose through a glucosidic bond. DS2 was deduced as β-GlcA(1→2)-Man,
and TS was proposed as a dimer of DS2. In hydrolysis reaction with 0.1–2.0 mol/L trifluoroacetic acid at 100 ℃ for 1 h, the
yields of the three oligosaccharides all increased with increasing acid concentration.

Analysis of Functional Components and Volatile Components from Loquat Peel Before and After Hot-Air Drying
ZHANG Qiao, GU Xinzhe, WU Yongjin, TU Kang
2016, 37(16):  117-122.  doi:10.7506/spkx1002-6630-201616019
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In this research study, we determined the effect of hot-air drying on three functional components (polyphenols,
flavonoids and triterpene acids) and volatile components of loquat peel. Fresh samples of loquat peel were hot-air dried 60 ℃
to a moisture of 9%–10% to investigate changes in the contents of functional components before and after the drying
process. The identification and quantitation of volatile components by using headspace solid phase microextraction coupled
with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Results illustrated that the contents of polyphenols and
flavonoids in dried loquat peel were significantly lower than in fresh loquat peel (P < 0.05), which were (37.94 ± 0.84) and
(19.29±0.76) mg/g vs. (63.66 ± 1.68) and (56.78 ± 1.44) mg/g, respectively. On the other hand, no significant change in
triterpene acids was noted. Totally, 35 volatile compounds were detected in fresh loquat peel, mainly aldehydes, esters,
ketones, which were dominated by hexanal (19.79%) and trans-2-hexenal (26.59%). In comparison with fresh loquat
peel, hot-air drying resulted in the formation of new volatiles as indicated by increasing the number of volatiles identified
to 73, which mainly included aldehydes, esters and ketones. Twenty-one volatile compounds were common to both
samples. Nonanal (31.47%) was the most dominant volatile of the dried sample, followed by β-cyclocitral, β-ionone,
dihydroactinidiolide and decanal, which altogether contributed to the distinct aroma of the dried loquat peel.

Determination of Flavonoid in Ziziphus jujuba Mill. cv. Jinsixiaozao Fruits by DAD-HPLC and LC-MS
SHI Renli, ZHAI Longfei, YU Wenlong, LIANG Na, GONG Zhengsi, WANG Xianghong
2016, 37(16):  123-127.  doi:10.7506/spkx1002-6630-201616020
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In this study, the flavonoids in Ziziphus jujuba Mill. cv. Jinsixiaozao fruits were analyzed by using high
performance liquid chromatography with diode array detector (DAD-HPLC) and liquid chromatography-mass spectrometry
(LC-MS). The chromatographic separation was achieved on a Hypersil BDS-C18 column (250 mm × 4.6 mm, 5 μm) using a
mobile phase consisting of methanol and 0.1% formic acid aqueous solution at 0.8 mL/min flow rate by gradient elution. The
column temperature was set at 30 ℃, and the injection volume was 10 μL. Dual-wavelength detection was carried out at 270
and 370 nm. Then LC-MS was employed to determine the flavonoids. Results demonstrated that Jinsixiaozao jujube contained
four flavonoids, myricetin, rutin, quercetin, and isorhamnetin and their contents were 40.90, 8.40, 32.60 and 12.56 μg/g,
respectively. In the range of 5–125 μg/mL, the method exhibited a good linear relationship and repeatability. The limits of
detection (LODs) were in the range of 1.23–1.73 ng/L at a signal-to-noise ratio of 3. The relative standard deviation (RSD)
values for precision were 1.10%–4.31%. The recoveries from spiked samples were 93.50%–106.70% (RSD = 0.68%–2.90%).
Conclusively, this method had the advantages of simple pretreatment, high recovery rate and good precision, and it could
provide a convenient and precise method for the analysis of flavonoids in Ziziphus jujuba Mill. cv. Jinsixiaoza fruits.

Identification and Analysis of Characteristic Flavor Components of Tricholoma matsutake Cookies
TAO Hongling, YANG Wenjian, PEI Fei, ZHAO Liyan, AN Xinxin, SU Kaimei, HU Qiuhui
2016, 37(16):  128-134.  doi:10.7506/spkx1002-6630-201616021
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White bean paste and Tricholoma matsutake powder, both of which are rich in functional active factors and
protein, were selected for production of cookies instead of traditional low gluten flour for flavor enrichment and nutritional
complementation. As an extension of our previous work where we selected the raw materials and established the optimized
manufacturing process, electronic nose and headspace solid-phase microextraction gas chromatography-mass spectrometry
(HS-SPME-GC-MS) were applied to investigate and compare the volatile components of T. matsutake powder, common
cookies and T. matsutake cookies. The results showed that there were 42, 12 and 25 volatile components detected in
T. matsutake powder, common cookies and T. matsutake cookies, respectively. Compared to common cookies, T. matsutake
cookies were richer in flavor components and contained the characteristic volatile components, pyrazines heterocyclic
compounds including 2-methyl pyrazine (2.98%), 2,5-dimethyl pyrazine (3.72%) and 3-ethyl-2,5-dimethyl pyrazine (1.30%).
This bakery product with distinctive characteristic is more acceptable for consumers in sensory evaluation.

Analysis of Fruit Quality Traits and Volatiles in Red and Pink Cherry and Large-Fruited Tomato Accessions
ZHAO Jiantao, ZHANG Jing, ZHANG Yating, ZOU Zhirong
2016, 37(16):  135-141.  doi:10.7506/spkx1002-6630-201616022
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The contents of ascorbic acid, soluble solid, lycopene and beta-carotenoid were measured in 8 red and pink
cherry and large-fruited tomato accessions using routine analytical procedures. As demonstrated by gas chromatographymass
spectrometry (GC-MS) analysis, the main sugars in red and pink cherry tomatoes were fructose, glucose and sucrose,
and the main organic acids were malic acid, butanedioic acid and citric acid. Twenty-five volatiles were co-detected in 8
tomato accessions, consisting mainly of alcohols, aldehydes, ketones and esters. Cluster analysis led to four groups: red and
pink cherry tomatoes as well as red large-fruited and pink large-fruited tomatoes, showing a closer distance to each other.
The principal component analysis of the main nutritional traits and the 25 detected volatiles showed that the first three
components accounted for over 82.293% of the total cumulative variance.

Analysis of the Volatile Compositions of Water Extracts from Walnut Diaphragm
LI Ping, JIA Hongjie, JIN Yu, JI Yali, FAN Tingwen, LI Meiping, ZHANG Shengwan
2016, 37(16):  142-148.  doi:10.7506/spkx1002-6630-201616023
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The volatile compounds of water extracts from walnut diaphragm were extracted by using four different polar
solid-phase micro-extraction (SPME) fibers, separated by either of two chromatographic columns, HP-5 (with weak polarity)
and RTX-WAX (with strong polarity), and analyzed by using head-space solid-phase micro-extraction (HS-SPME) and gas
chromatography-mass spectrometry (GC-MS). Further, the effects of different chromatographic columns and extraction
fibers on the analysis of volatile components were examined. Results showed that a total of 115 volatile compounds with
known structure from four walnut diaphragm extracts were separated on different chromatographic columns and identified
by GC-MS, consisting of 21 alcohols, 18 esters, 15 ketones, 14 aromatic compounds, 13 hydrocarbons, 12 aldehydes,
6 acids, 5 phenolic compounds, 2 ethers and 9 other compounds. The main volatile components were acetic acid, ethyl
acetate, 2,6-dibutyl-4-methylphenol, menthol, cedrol, 2,3-butanediol, 1-hexanol, phenethylalcohol, 3-butyl-4-hydroxyanisole,
hexanal, and azulene.

Effects of Boron Fertilizer on Volatiles of “Jinpeng No.1” Tomato Fruit
XU Weinan, ZHANG Xin, ZHANG Jing, DING Ming, ZHEN Ai, CHANG Xiaoxiao, BAI Yongjuan, HU Xiaohui
2016, 37(16):  149-155.  doi:10.7506/spkx1002-6630-201616024
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Boron fertilizer applications by foliar spraying (L) and nutrient solution application (W) at different levels of
boron (1.9, 3.8, and 5.7 mg/L H3BO3), respectively, were performed to investigate the effects of boron fertilizer on volatiles
of tomato (cv. “Jinpeng No.1”) fruit using boron-free nutrient solution treatment as control (CK). The composition and
content of aroma compounds were measured using solid phase microextraction coupled to gas chromatography-mass
spectrometry (SPME-GC-MS). The results showed that a total of 79 volatiles were detected in seven treatment groups of
tomato fruits, with aldehydes, esters, alcohols, hydrocarbons and ketiones being the predominant compounds. The decreasing
order of total volatile component contents was as follows: L1 > W2 > W1 > L3 > W3 > L2 > CK. Meanwhile, 14 of these
were characteristic aroma components and their total contents were in the descending order: W2 > L1 > L3 > W1 > W3 >
L2 > CK. Different application methods for the same concentration of boron and different boron levels applied in the same
way had different effects on the characteristic compounds of tomato. All 14 characteristic aroma components and 3 types
of aroma components (‘fruity aroma’, ‘green aroma’ and ‘floral aroma’) were detected in the treatment group of 3.8 mg/L
H3BO3 nutrient solution, with fruity aroma being the most predominant as well as 6-methyl-5-hepten-2-one being the biggest
contributor to the aroma, suggesting the presence of a wide range of aroma compounds in the tomato fruits. Therefore,
the application of 3.8 mg/L H3BO3 nutrient solution can result in the formation of a wide variety of aroma compounds
responsible for the strong fruity aroma in tomato.

Transfer of Metal Elements in the Distillation Process of Chinese Luzhou-Flavor Liquor
LI Yongjiao, ZHANG Suyi, HUO Danqun, ZENG Na, ZHOU Tao, LI Delin, YANG Yan, WANG Ming
2016, 37(16):  156-161.  doi:10.7506/spkx1002-6630-201616025
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A total of 17 common metal elements including Na, K, As, Pb, Cd, Sn, Ti, Mg, Fe, Cu, Mn, Zn, Ca, Al, Ni, Cr
and Ba were measured in fermented grains, distillery spent wash and liquor samples using inductively coupled plasma
mass spectrometry (ICP-MS) coupled with microwave digestion before and after distillation. SPSS was applied to analyze
the origin of related elements in liquor samples by t-test. The results suggested that more than 94% of the overall metal
elements remained in the wine lees after distillation, while less than 6% metal ions were transferred into either spent wash
or the resultant liquor. Heavy metal ions such as Pb, Mn and Cr showed lower transfer ratio than Na, K, Ca and Mg, which
significantly reduced the possibility of transfer of those hazardous components from the polluted raw materials to liquor. It
was also found that Ca, Cu, K, Mg, Na, Zn and Ba in liquor samples were mainly from wine lees and the equipment used in
the distillation process would introduce Fe, Ni, Al and Sn, and even Pb and Cr to the liquor.

Comparison of Analytical Methods for the Quantitation of Xylan in Sugarcane Bagasse
MIAO Lu, ZHOU Yuheng, ZHANG Hourui, HAO Zaibin
2016, 37(16):  162-167.  doi:10.7506/spkx1002-6630-201616026
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This study aimed to establish an accurate method for quantitatively determining xylan for the purpose
of broadening its range of application. A comparative evaluation of two sample pretreatment methods including
hydroformylation and hydrolytic saccharification respectively coupled with various analytical techniques such as gas
chromatography (GC), high performance liquid chromatography (HPLC), colorimetry and titrimetrywas was carried out for
the determination of xylan in the same batch of sugarcane bagasse. The results showed that the hydroformylation method
could only detect the pentose but it could not detect the hexose, which was unable to be converted into furfural. On the
other hand, by the combined use of hydrolytic saccharification with 3,5-dinitrosalicylic acid (DNS) colorimetry, HPLC with
Ca2+ column and pre-column derivatization HPLC, respectively, the results obtained were basically identical and authentic, that
is, both pentose and hexose could be detected using these methods. Single HPLC with Ca2+ column or pre-column derivatization
HPLC could not separate effectively some of the constituent monosaccharides. However, using a combination of the two HPLC
methods the four monosaccharides consisting of xylan could be accurately detected including xylose, arabinose, galactose and
glucose. This combination of HPLC methods is suitable for monosaccharide composition analysis of xylan in most plants.

Analysis of Aroma Compounds of Duyun Maojian Tea
JIANG Chenkai, LI Yingxiang, HUANG Yahui, ZHANG Lijuan, YANG Jiagan, XU Xingguo, LIU Qing, CHEN Yingyu, LI Dan, ZENG Zhen
2016, 37(16):  168-172.  doi:10.7506/spkx1002-6630-201616027
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This study aimed to explore the effects of tea varieties and environmental factors on the quality of Duyun
Maojian tea, and to compare the aroma composition of Duyun Maojian tea made from local tea varieties and the ‘Fuding
Dabai’ variety in Qiandongnan prefecture, Guizhou province. Aroma components were determined by gas chromatographymass
spectrometry (GC-MS), and their relative contents were calculated by peak area normalization. A total of 50 aroma
components were detected from all samples with hydrocarbon compounds and alcohols being the most predominant.
There were distinct differences in aroma composition among Duyun Maojian tea made from different tea varieties grown
in the same area and from ‘Fuding Dabai’ grown in different areas. The contents of β-linalool, 2-ethenyl-1,1-dimethyl-3-
methylene-cyclohexane, cis-β-ocimene, and dimethyl sulfide in Duyun Maojian tea were significantly higher than in other
famous green teas.

SPME-GC-MS Analysis of Aroma Components in Black Tea Prepared with Main Tea Cultivars in Sichuan
LUO Xueping, LI Lixia, MA Chaolong, ZHAO Xianming
2016, 37(16):  173-178.  doi:10.7506/spkx1002-6630-201616028
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The aroma components of black teas prepared with 5 main tea varieties grown in Sichuan, including
Mingshanbaihao 131, Fudingdabaicha, Fuxuan No. 9, Zaobaijian and Sichuan medium- and small-leafed population cultivar,
were analyzed by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). A total of 148
aroma components including alcohols, aldehydes, esters, ketones, hydrocarbon and acids were detected in the black teas,
and the major aroma compounds were alcoholic compounds with relative contents of 45.97%–63.78%, mainly geraniol,
linalool and its oxides, phenylethyl alcohol, 3,7-dimethyl-1,5,7-octatrien-3-ol, benzyl alcohol, nerolidol, methyl salicylate,
ethyl caprate, cis-3-hexenyl caproate, benzene acetaldehyde, citral and others, which were the important components that
formed the sweet floral and fruity aroma in Sichuan black tea. The aroma of the black teas prepared with five tea varieties
belonged to the middle type, which contained abundant amounts of linalyl alcohol and geraniol. The terpene index of black
teas prepared with Mingshanbaihao 131, Fudingdabaicha, and Sichuan medium- and small-leafed population cultivar was
0.57, 0.60 and 0.65, respectively, suggesting that their flavors were rich in high aroma, which was consistent with the results
of sensory evaluation. Therefore, Mingshanbaihao 131, Fudingdabaicha, and Sichuan medium- and small-leafed population
cultivar are suitable to produce high aroma or floral aroma type black tea.

Comparison of Key Flavor Substances of Fermented Grass Carp with Lactobacillus plantarum Based on Two Growth Media
MING Tinghong, SU Xiurong, ZHOU Jun, LI Ye, ZHANG Chundan, JI Lu, SUN Tingting, HUANG Zhongbai, HE Shan, QIU Dihong
2016, 37(16):  179-186.  doi:10.7506/spkx1002-6630-201616029
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The goal of this study was to examine and compare the changes and differences in volatile substances of
fermented grass carp with Lactobacillus plantarum based on two different growth media at different times and consequently
to provide a reference for the research of flavor compounds of fermented grass carp. Electronic nose and headspace solidphase
microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) were used to detect and analyze the
changes of volatile substances. The relative odor activity value (ROAV) combined with odor threshold was applied to
confirm the key odor compounds of fermented grass carp. Electronic nose could sensitively detect the changes in volatile
substances of fermented grass carp during the fermentation process, and the LDA results revealed that there were obvious
differences among these samples. GC-MS analysis indicated that a total of 90 and 70 volatile components were found to
be present respectively in fermented grass carp with L. plantarum in MRS broth medium and 12% skimmed milk medium,
consisting mainly of hydrocarbons, aldehydes, alcohols, ketones, esters, amines and aromatic compounds. The principal
flavor components common to both samples were D-limonene, 1-octene-3-ol, heptanal, octanal, decanal and acetoin.
The unique flavor components of fermented grass carp produced using MRS broth medium were 2,3-butanedione and
(E)-2-nonenal, while the unique flavor components of fermented grass carp produced using 12% skimmed milk medium
were hexanal, nonanal and butyric acid ethyl ester.

Evaluation of Uncertainty for Zinc Determination in Oyster by Microwave Digestion-ICP-OES
SUN Lingling, SONG Jinming, YU Ying, SUN Xuan, WEN Tingyu, LIN Qiang
2016, 37(16):  187-192.  doi:10.7506/spkx1002-6630-201616030
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The sources of uncertainty were analyzed for the determination of zinc in oyster by microwave digestioninductively
coupled plasma optical emission spectrometry (ICP-OES). According to JJF 1059—1999, the sub-uncertainty
was calculated and used to obtain combined uncertainty and expanded uncertainty of the measurement results. The results
showed that the uncertainty was mainly derived from fitting to the standard curve, and repeated experiments of samples,
preparation of the standard solution, and sample weighing and volume had a nonneligible effect on it with relative standard
uncertainty values of 0.016, 0.004 7, 0.001 4, 0.002 2 and 0.000 8, respectively. In this study, the combined standard
uncertainty of the measurement results was 13.29 mg/kg, and the expanded uncertainty was 26.58 mg/kg (k = 2). The final
result of zinc content in oyster was (781.98 ± 26.58) mg/kg (k = 2, P = 95%).

Safety Detection
Development of a Double-Antibody Sandwich Enzyme Linked Immunosorbent Assay for Detection of Staphylococcal Enterotoxin I (SEI)
ZHU Anni, TANG Junni, ZHAO Yanying, TANG Cheng, CHEN Juan, LIU Ji
2016, 37(16):  193-198.  doi:10.7506/spkx1002-6630-201616031
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Objective: To establish a simple and sensitive double-antibody sandwich enzyme linked immunosorbent
assay (DAS-ELISA) for the detection of a newly identified Staphylococcal aureus enterotoxin, SEI. Methods: Different
combinations of coating antibody, polyclonal antibody and goat anti-rabbit IgG/HRP were tested through square matrix
titration. The experimental conditions were optimized such as buffer, blocking time, antigen incubation time, goat antirabbit
IgG/HRP incubation time, and chromogenic time of 3,3’,5,5’-tetramethylbenzidine (TMB). Further, the developed
method was analyzed and evaluated by sensitivity, intra/inter-batch coefficients of variation and recovery of spiked samples.
Results: The optimum experimental conditions were determined as follows: anti-SEI monoclonal antibody concentration,
2.89 mg/L; dilution ratio of polyclonal antibody, 1:2 000; and dilution ratio of goat anti-rabbit IgG/HRP, 1:6 000,
respectively. Moreover, 1 × PBS (pH 7.4) buffer solution was the optimal coating buffer, and the optimal blocking time,
antigen incubation time, goat anti-rabbit IgG/HRP incubation time, and TMB chromogenic time were 60, 90, 30, and
15 min, respectively. The equation between SEI concentration and optical density at 450 nm (OD450 nm) was fitted as follows:
y = 0.040 9 x + 0.042 9 (R2 = 0.993 3). The sensitivity of the developed method was 0.5 μg/L, with intra-batch coefficient of
variation < 10% and inter-batch variation < 15%. The recoveries for spiked saline, minced yak meat, steamed rice and UHT
milk were all above 90%, except for pasteurized milk. Conclusion: This study has established a DAS-ELISA method for
detecting newly identified staphylococcal enterotoxin I (SEI).

Determination of the Bioavailability of Lead in Rice by in vitro Simulate Digestion/Caco-2 Cell Model
FANG Yong, XIA Ji, LI Hongmei, CHEN Yue, PEI Fei, KONG Lingyan, YANG Peiling, HU Qiuhui
2016, 37(16):  199-204.  doi:10.7506/spkx1002-6630-201616032
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A method to determine both the bioaccessibility and bioavailability of lead in rice grains using in vitro digestion/
Caco-2 cell model was established. The results indicated that the bioaccessibility of lead in uncooked rice and cooked rice
in simulated gastric digestion were 61.34%–70.59% and 39.69%–47.48%, as well as 24.39%–41.79% and 13.57%–15.13%
in simulated intestinal digestion, respectively. The lead bioaccessibility of indica rice was higher than that of japonica rice,
and uncooked rice was higher than cooked rice. A Caco-2 cell model was established to evaluate the bioavailability of lead
in rice. The bioavailability of tetraethyl lead (TTEL) and inorganic lead spiked in cooked rice were 3.32% and 7.0%, while
those in spiked uncooked rice were 2.65% and 5.7%, respectively. In vitro digestion/Caco-2 cell model can be used as an
effective method to evaluate the bioavailability of lead in rice.

Optimization of Characteristic Wavelength Variables of Near Infrared Spectroscopy for Detecting Edible Oil Acid Value
WANG Liqi, LIU Yanan, ZHANG Qing, CUI Yue, GE Huifang, YU Dianyu
2016, 37(16):  205-210.  doi:10.7506/spkx1002-6630-201616033
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With the goal of achieving rapid detection of soybean oil acid value by using near-infrared (NIR) spectroscopy,
this study optimized the selection of the characteristic wavelength variables by combined use of interval partial least square
(iPLS), genetic algorithm (GA) and successive projection algorithm (SPA). A total of 100 soybean oil samples with different
acid values were collected, their NIR transmittance spectra in the range of 4 000–12 000 cm-1 were acquired. Firstly, the
characteristic wavebands of 4 540–5 346 cm-1and 6 807–7 004 cm-1 were extracted from the original spectra by iPLS, with a
determination coefficient (R2) and root mean square error of prediction (RMSEP) of 0.978 9 and 0.064 3, respectively. Then,
the characteristic wavelength variables closely related to oil acid value were selected by GA and SPA from the previously
selected bands, respectively. It was shown that the PLS calibration model established using 12 variables consisting of the top
6 characteristic wavelengths from optimal selection results of each of the two algorithms was optimum, with R2 and RMSEP
of 0.985 9 and 0.045 1, respectively. The research indicated that selection of the characteristic wavelength variables by
iPLS-GA-SPA in NIR analysis for oil acid value could effectively remove redundant information, and decrease the
complexity of the model. This paper can offer important reference for rapid and non-destructive detection of oil acid value.

Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA
ZHANG Shuai, QI Yingying, ZHANG Hongxing, WANG Yiwen, XIE Yuanhong, LIU Hui
2016, 37(16):  211-215.  doi:10.7506/spkx1002-6630-201616034
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This study aimed to prepare antibodies specific for Salmonella and to use them establish an enzyme linked
immunosorbent assay (ELISA) system for the rapid detection of six Salmonella serotypes in chilled fresh meat. In this
study, BALB/c mice and rabbits were immunized with mixed antigens from six pathogenic strains of Salmonella for the
production of monoclonal antibodies using hybridoma technology for cell fusion. The results showed that a hybridoma
cell line (6E7 CGMCC 10313) which could stably secret monoclonal antibody (McAb, M23) specific for Salmonella was
successfully screened and the polyclonal antibody (P23) was also obtained at the same time. The titer of the prepared highly
pure monoclonal and polyclonal antibodies was 1:1.28 × 106 and 1:8.0 × 105, respectively. P23 was adsorbed on 96-well
microtiter plates as coating antibody and M23 was labeled with horseradish peroxidase for the establishment of sandwich
ELISA. The limit of detection (LOD) for Salmonella in artificially contaminated meat samples was 800 CFU/g. There was
no cross-reactivity with Shigella, Enterobacter sakazakii, E. coli O157:H7, Staphylococcus aureus, Listeria monocytogenes
and other Salmonella.

Simultaneous Analysis of Nine Phytohormones in Porphyra haitanensis by High Performance Liquid Chromatography Coupled with Triple Quadrupole Tandem Mass Spectrometry
ZHAO Jiali, XU Panpan, CHEN Juanjuan, LUO Qijun, YANG Rui, CHEN Haimin, YAN Xiaojun
2016, 37(16):  216-222.  doi:10.7506/spkx1002-6630-201616035
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A quantitative method based on high performance liquid chromatography-tandem mass spectrometry
(HPLC-MS) has been established for simultaneous analysis of nine phytohormones in Porphyra haitanensis. The optimized
sample preparation procedure involved extraction with methonal-H2O-formic acid (15:4:1, V/V) and re-dissolution in
methonal-H2O-acetic acid (90:10:0.05, V/V) containing 10 mmol/L ammonium acetate. The chromatography was performed
using a Thermo Hypersil Gold C18 column (100 mm × 2.1 mm, 3 μm) as the stationary phase, and the mobile phase consisted
of aqueous solution containing 10 mmol/L ammonium acetate-methanol. Nine phytohormones (indole-3-acetic acid, N6-(2-
isopentyl) adenosine, N6-(2-isopentenyl)adenosine, trans-zeatin riboside, zeatin, abscisic acid, salicylic acid, gibberellin A3,
and brassinolide) were quantified using selective reaction mode. As results, good linear relationships between concentrations
and the corresponding peak areas of nine standards were achieved with correlation coefficients (R2) higher than 0.991. The
recoveries of eight standards except gibberellin A3 were higher than 71%. The limits of detection (LODs) of phytohormones
by this method were all lower than 3 μg/L and the limits of quantification (LOQs) were in the range from 0.45 to 5 μg/L.
Using the established method, the concentrations of nine phytohormones in different varieties of Porphyra haitanensis and at
different cultivation periods were successfully investigated and correlated with its morphology.

Analysis of Bacterial Diversity in O3-BAC Drinking Water Treatment Process by Using Illumina MiSeq High Throughput Sequencing Technology
GE Yingliang, YU Shuili, SHI Wenxin, YANG Fan, QI Qing
2016, 37(16):  223-228.  doi:10.7506/spkx1002-6630-201616036
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Illumina MiSeq high-throughput sequencing technology was used to study the bacterial diversity and abundance
of the effluent water from each unit process in O3-BAC water treatment. A total of 196 809 16S rDNA sequences were
acquired, which were classified as 522 genera in 38 phyla. The results showed that there was high bacterial community
diversity in the processing unit effluent. Pre-ozonation and ozone oxidation treatment of water influenced most significantly
bacterial diversity and abundance. Ozone could kill some bacteria which were hard to be killed by chlorine disinfection in
a reasonable ozone concentration range. Flocculation and sand filtration could recover the bacterial diversity in water, as
demonstrated number of bacterial species. In biological activated carbon filter treatment, bacterial diversity was increased
and their abundance distribution was more uniform, which would propose higher requirements for later disinfection technics.

 

Combined Use of ELISA and UPLC-MS/MS for Detection of Zearalenone from Grains
LI Yan, PEI Shichun, WANG Yan, GAO Jianwei, ZHEN Yuping, MA Mingxin, ZHANG Pin, XIE Tiezhi
2016, 37(16):  229-234.  doi:10.7506/spkx1002-6630-201616037
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An enzyme-linked immunosorbent assay (ELISA) coupled to ultra performance liquid chromatography-tandem
mass spectrometry (UPLC-MS/MS) method was developed for the determination of zearalenone in corn samples. Samples
were extracted with acetonitrile-water-acetic acid (70:29:1, V/V) and then the extract was determined by ELISA and UPLCMS/
MS. In the study, special attention was given to the stability of the system and the matrix effects frequently observed in
the process. The recovery of UPLC-MS/MS with and without sample purification was evaluated as well. Proper approaches
for dealing with these problems were obtained. This analysis method proved easy, fast and suitable for the determination of
zearalenone in bulk grain samples.

Detection of Genetically Modified Soybean Event DAS-44406-6 by Real-Time PCR Method and Digital PCR Method
YU Xiaofan, GAO Hongwei, SUN Min, XIAO Xizhi, LI Ronggui
2016, 37(16):  235-241.  doi:10.7506/spkx1002-6630-201616038
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Purpose: To determine the flanking sequence between exogenous and endogenous fragments using rapid
amplification of cDNA ends (5’-RACE) and consequently to examine genetically modified soybean (GMS) DAS-44406-
6. Methods: Specific primers and probes were designed based on the flanking sequences of exogenous fragments of DAS-
44406-6. The specificity of the developed method was validated by using it to detect a variety of other GM samples and non-
GM samples. DAS-44406-6 was used to prepare 6 content gradients to test the sensitivity of the method. At last, digital PCR
was applied to measure nucleic acid molecules for absolute quantification. Conclusions: A real-time PCR method has been
established for the identification of GMS soybean DAS-44406-6 with high specificity. The limit of detection (LODs) of the
real-time PCR method at template DNA concentration of 100 ng/reaction and 0.01% GM soybean content was 16.6 copies
of genomic DNA from DAS-44406-6. As for the LOD of the digital PCR method, the relative standard deviation (RSD) was
0.7% at template DNA concentration of 0.5 ng/reaction. The highly specific method combining real-time PCR and digital
PCR could meet the requirements for the detection of GMS DAS-44406-6.

Simultaneous Determination of Fluoroquinolone Residues in Honey and Royal Jelly by QuEChERS and High Performance Liquid Chromatography-Tandem Mass Spectrometry
ZHANG Zhongyin, ZHAO Liuwei, CAO Weirui, ZHOU Jinhui
2016, 37(16):  242-248.  doi:10.7506/spkx1002-6630-201616039
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A multi-residue analytical method for the determination of 14 fluoroquinolone residues in honey and royal
jelly using QuEChERS extraction coupled with high performance liquid chromatography-tandem mass spectrometry
(HPLC-MS/MS) was developed. Samples were extracted with acetonitrile containing 1% acetic acid, cleaned up by
dispersive solid phase extraction with C18-EC and PSA as absorbent. Recoveries from samples spiked at three levels were
between 80.4% and 110.2% with relative standard deviations (RSDs) lower than 16.3%. The limits of detection (LODs) and
limits of quantification (LOQs) for honey were 0.45-2.58 ng/g and 1.51-8.59 ng/g, respectively. For royal jelly, the LODs
and LOQs were 0.51-3.22 ng/g and 1.69-10.75 ng/g, respectively. The established method could meet the requirement for
the determination of fluoroquinolone residues in honey and royal jelly samples.

Determination of Four Nitrofurans in Water and Sediment Using Ultra Performance Liquid Chromatography
WANG Qiang, WANG Xufeng, ZHAO Donghao, HUANG Ke, LI Zhiguang, LI Yongxian, YANG Hongliang, LI Liudong
2016, 37(16):  249-253.  doi:10.7506/spkx1002-6630-201616040
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An ultra performance liquid chromatography (UPLC) method was developed for the simultaneous determination
of 4 nitrofuran antibiotics (furazolidone, furaltadone, nitrofurazone and nitrofurantion) in water and sediment. Water
samples were filtrated and cleaned up using mixed-mode cation exchange solid-phase extraction column (MCX). Sediment
samples were extracted with acetonitrile-0.1% formic acid solution (8:2, V/V), and then cleaned up also with MCX solidphase
extraction column. The analytes were separated on a BEH C18 column (100 mm × 2.1 mm, 1.7 μm) utilizing gradient
elution with a mixture of acetonitrile and 0.1% formic acid solution as the mobile phase. The method showed good linearity
in the range of 10.0–200 μg/L with correlation coefficients (R2) all above 0.999. The recoveries were 81.5%–103.2% and
73.3%–91.9% with relative standard deviations (RSDs, n = 5) of 2.0%–5.8% and 3.4%–9.6% for 4 nitrofurans in water and
sediment, respectively. The limits of detection (LODs) were 0.03 μg/L and 0.6 μg/kg and the limits of quantitation (LOQ)
were 0.1 μg/L and 2.0 μg/kg, respectively. The method can be applied to the determination of nitrofuran antibiotics residues
in water and sediment.

Determination of Polybrominated Diphenyl Ethers Residues in Water by Gas Chromatography-Mass Spectrometry
YANG Fang, ZHANG Xianchen, LU Junwen, YAN Aiguo, CHEN Lisi, ZHANG Pengjie, ZHENG Leiqing
2016, 37(16):  254-257.  doi:10.7506/spkx1002-6630-201616041
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An accurate, efficient and fast method for the determination of residues of polybrominated diphenyl ethers
in water using gas chromatography-mass spectrometry (GC-MS) was developed. The sample was extracted using
dichloromethane, concentrated under nitrogen blow, dissolved in hexane, and then detected by GC-MS. The calibration
curves of all 19 polybrominated diphenyl ethers presented good linearity with correlation coefficients (R2) higher than 0.99.
The relative standard deviations (RSD, n = 6) for precision were between 1.5% and 5.6%, and the limits of detection (LOD)
were in the range from 0.1 to 1.0 μg/L. The recoveries of PBDE-3 and PBDE-209 were from 84.3% to 93.2%, with RSD
(n = 6) between 3.6% and 10.1%. These results displayed that all the evaluated figures of merit of the method could meet the
analytical requirements. Thus, the developed method could be applied in the routine analysis of polybrominated diphenyl
ethers residues in water.

Determination of Ethylenediamine Tetraacetic Acid Disodium Salt Content in Canned Pickled Vegetables by High Performance Liquid Chromatography
MA Chiyuan, GUO Ping, CHEN Lin, WANG Ling, LU Fang, LUO Yan
2016, 37(16):  258-262.  doi:10.7506/spkx1002-6630-201616042
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A method for the determination of ethylenediamine tetraacetic acid disodium salt content in canned pickled
vegetables was developed using high performance liquid chromatography (HPLC). The target analyte was extracted with
pure water. The extract was then derivatized with FeCl3, cleaned up by PXA mixed anion solid phase extraction, detected
with HPLC detector and quantified by the external standard method. Under the optimal conditions, the calibration curve
of ethylenediamine tetraacetic acid disodium salt derivatives was linear in the range of 1.0–100.0 μg/mL with correlation
coefficient of 0.999 95. The limit of detection (LOD) was 0.09 mg/kg and the limit of quantitation (LOQ) was 0.31 mg/kg.
The average recoveries of spiked samples ranged from 99.09%–103.95%, with relative standard deviations (RSDs) between
0.4% and 3.3% at spiked levels of 0.03–0.3 g/kg. The method is accurate, repeatable and suitable for the determination of
ethylenediamine tetraacetic acid disodium salt in canned pickled vegetables.

Determination of Chloramphenicol in Propolis-Derived Health Foods by HPLC-MS/MS
YANG Li, LIAO Xiayun, YANG Weiting, XU Yangbiao, LIU Xianghong, LIU Jialing, LU Yixiang, GUO Wenwen, LIN Xiaojuan
2016, 37(16):  263-267.  doi:10.7506/spkx1002-6630-201616043
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A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established
to detect chloramphenicol residue in propolis-derived health foods. Samples were dissolved in 0.5% Ca(OH)2 solution,
followed by the removal of mucinous impurities after treatment with hydrochloric acid. Then chloramphenicol was back
extracted with ethyl acetate. The extract was separated on an Inertsil C8-3 column (2.1 mm × 100 mm, 2 μm) with gradient
elution using a mobile phase consisting of methanol and water. A tandem quadrupole mass spectrometer equipped with
electrospray ionization (ESI) source in positive ion mode was employed for the quantitative analysis of the analyte using
multiple reaction monitoring (MRM). The calibration curve for chloramphenicol displayed good linearity in the range of
0.1–2.0 ng/mL. The limit of detection (LOD) of the proposed method was 0.4 ng/mL. The absolute recoveries were 58.3%–
74.5% at spiked concentration levels of 0.1, 0.2, and 1.0 μg/kg with relative standard deviations (RSDs) of 1.34%–5.11%,
and the relative recoveries were 96.8%–114.0% with RSDs of 1.33%–5.36%. The method exhibited high sensitivity and
good reproducibility, and could be suitable for the determination of trace amounts of chloramphenicol in propolis-derived
health foods.

Influence of Added Water on Conductivity of Fresh Raw Cow Milk
LIN Biying, YAN Yadong, GUO Wenchuan
2016, 37(16):  268-271.  doi:10.7506/spkx1002-6630-201616044
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Adding water to fresh raw milk is a common method in producing adulterated milk. The goal of the present
study was to establish a method for rapidly detecting the added water in milk. For this purpose, milk samples with different
proportions of added water ranging from 0%–14.02% were prepared by adding tap water to fresh raw cow milk. The
conductivities of these milk samples were measured at six temperatures from 10 to 35 ℃. The influence of added water
content and temperature on conductivity was comprehensively analyzed to establish and verify the corresponding model.
The results showed that the conductivity of cow milk deceased linearly with increasing amount of added water but increased
linearly with increasing temperature. The linear equation in two variables, with a coefficient of determination of 0.998 1,
could be used to describe the relationship between conductivity and the amount of added water and temperature. The
absolute error between calculated measured values for the amount of added water in cow milk was −2.15%–2.03%, with an
average of −0.06%, indicating that the amount of added water could be calculated precisely using the model.

Packaging & Storage
Effects of Inducer Treatment on Physiological Quality, Active Oxygen and Energy Metabolism of Postharvest Wax Apples during Cold Storage
LI Wenwen, WU Guangbin, CHEN Fahe
2016, 37(16):  272-279.  doi:10.7506/spkx1002-6630-201616045
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To investigate the effects of 0.01% colchicine treatment on physiological quality, active oxygen and
energy metabolism of postharvest wax apples during cold storage, and further to understand better the role of colchicine on
senescence of the fruits.Methods: Wax apples were dipped with 0, 0.01% colchicine for 15 minutes at room temperature
and then stored at 4 ℃. The physiological quality, active oxygen metabolism and energy metabolism of wax apples were
determined during storage. Results: Colchicine treatment could retard the increase of cottony softening extent and had a
significant effect from the 4th day of storage onwards (P < 0.05), the fruits treated with colchicine had a lower percentage
weight loss during the whole storage (P < 0.05). Colchicine treatment could also postponed the decrease of superoxide
dismutase (SOD) activity during the whole storage, maintained lower catalase (CAT) activity, and significantly increased
peroxidase (POD) activity during the later stage of storage (P < 0.05). At the same time, colchicine treatment could maintain
higher activity of succinate dehydrogenase (SDH) and cytochrome oxidase (CCO), as compared with the control fruits. The
SDH activity was significantly higher at 4, 8, and 12 day (P < 0.05) while the activity of CCO was significantly higher at
2, 6 and 8 day (P < 0.05). Also colchicine treatment could slow down the decrease of ATP content and energy charge at
later stage. Conclusions: Colchicine at 0.01% could retard the increase of cottony softening extent of wax apples. Cottony
softening extent and energy charge had a significantly negative correlation with each other (r = −0.793, P < 0.05). Energy
deficit could lead to the senescence of the fruits. Energy charge had a significantly negative correlation with H2O2 content
(r = −0.766, P < 0.05), but demonstrated a significantly positive correlation with SOD activity (r = 0.802, P < 0.05). Energy
metabolism could influence active oxygen metabolism. The role of colchicine in delaying wax apple senescence was mainly
related to its effects on energy metabolism.

Safety Detection
Effect of 1-Methylcyclopropene Treatment on Postharvest Quality of Apple Fruit Stored at Ambient Temperature
WANG Yunxiang, ZHANG Yanan, QU Guiqin, ZHAI Baiqiang, LUO Yunbo, FU Daqi
2016, 37(16):  280-285.  doi:10.7506/spkx1002-6630-201616046
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“Fuji” and “Golden Delicious” apples were used to investigate the effect of 1-methylcyclopropene (1-MCP)
treatment on fruit quality during a short-term storage at room temperature. The results showed that, 1 μL/L 1-MCP treatment
had no significant influences on weight loss ratio, firmness and soluble solid content in both apple cultivars. However,
the titratable acid contents in treated fruits were significantly higher than those of control after 7 days of storage. 1-MCP
treatment retarded the increase in the pH value of “Fuji” apples after 14 days, while for “Golden Delicious” apples, the delay
occurred after 7 days. 1-MCP treatment had no significant effect on vitamin C (VC) content in “Fuji” apples, but for “Golden
Delicious” apples VC contents were significantly higher in 1-MCP treated apples than in the control ones. These results
indicated that 1-MCP treatment had different effects on the fruit quality of different apple varieties.

Packaging & Storage
Principal Component Analysis to Evaluate the Effect of Different Packaging Films on the Quality of Agaricus bisporus
XIE Liyuan, PENG Weihong, TANG Jie, HUANG Zhongqian, TAN Wei, GAN Bingcheng
2016, 37(16):  286-291.  doi:10.7506/spkx1002-6630-201616047
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In this paper, the effect of different packaging films on quality of Agaricus bisporus were analyzed by principal
component analysis (PCA) method, and an assessment model was established to screen the best packaging material. The
results showed that 9 traits were consolidated into 2 principal components, which accounted for 86.224% of the total
variance. The PCA showed that the order of the quality of A. bisporus under different packaging films was HDPE > LDPE >
OPP > LLDPE > EVA > control check (CK). Thus, all these packaging films were conductive to mushroom preservation,
and HDPE was the best one. These findings can provide scientific and intuitive evidence for quality evaluation.

Effect of Perforation Mediated Nano-Packaging on Postharvest Quality of Volvariella volvacea during Storage
YU Kelin, FANG Donglu, CHEN Meixiang, HU Qiuhui, ZHAO Liyan
2016, 37(16):  292-298.  doi:10.7506/spkx1002-6630-201616048
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The effect of perforation-mediated packaging on the quality of Volvariella volvacea during storage was studied.
For this purpose, samples of V. volvacea were subjected to perforation-mediated nano-polyethylene (PE) or common
PE packaging for storage at (15 ± 1) ℃ and relative humidity of (85 ± 5) % for 6 days. Meanwhile, the sample without
packaging was served as control. Weight loss rate, browning degree, respiration rate, malonaldehyde (MDA) content, total
soluble sugar and protein contents, and catalase (CAT), polyphenol oxidase (PPO) and peroxidase (POD) activities of
V. volvacea were measured daily over the storage period. Results showed that the two perforation-mediated packaging
methods could extend the shelf life of V. volvacea by 2 d as compared with the control. In addition, the mushrooms treated
with nano-packaging showed the best quality. After 6 days of storage, weight loss rate, browning degree, respiration rate,
MDA content, and PPO and POD activities of V. volvacea treated with nano-packaging were 4.47%, 3.29, 988.26 mg/(kg·h),
8.17 mmol/g, 246.67 U/g, and 8.02 U/g, respectively, which were lower than those of common PE packaging, while CAT
activity (43.20 U/mg) was higher than that of common PE packaging. The contents of total soluble sugar and protein of
nano-packaging were 13.15 and 6.21 mg/g, which were increased by 13.85% and 6.5% as compared with those of common
packaging, respectively. Perforation-mediated nano-packaging can effectively inhibit the sensory quality deterioration of
V. volvacea, maintain its nutritional value at a higher level, improve its preservation quality and extend its preservation time.

Comparative Changes in Respiration Rate and Quality Parameters of ‘Xiangshui’ and ‘Garber’ Pears during Postharvest Ripening
MA Chaoling, BI Yang, ZHANG Zhong, ZHANG Wenli, WEI Yongbo
2016, 37(16):  299-303.  doi:10.7506/spkx1002-6630-201616049
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‘Xiangshui’ pear is a traditional cultivar of Pyrus ussuriensis Maxim, and ‘Garber’ pear is a hybrid of
P. communis × P. pyiflia. Both cultivars become more delicious after ripening and freeze-thwa. However, no report is
available on the physiological and quality of ‘Xiangshui’ and ‘Garber’ pears during postharvest ripening. In this study,
‘Xiangshui’ and ‘Garber’ pears harvested from the same orchard and under the commercial maturity stage were stored for
ripening at room temperature ((20 ± 3) ℃), relative humidity (RH) of 85%–90%, and the changes in physiological and
quality parameters were determined. The results showed that respiration rates of ‘Xiangshui’ and ‘Garber’ pears belonged to
typical climacteric fruits, reached the maximum after 5 and 15 days of storage, respectively. Moreover, the content of total
soluble solids of ‘Garber’ pears was higher than that of ‘Xiangshui’ pears, and reached the maximum after 15 days and 10
days of storage, respectively. The content of titratable acid was decreased gradually for both cultivars during storage, with
‘Xiangshui’ pears having a significantly higher level than ‘Garber’ pears. Among color parameters investigated, a* value
increased most significantly during storage and was higher in ‘Xiangshui’ pears than in ‘Garber’ pears. The content of stone
cell in two cultivars kept stable, while the firmness decreased and weight loss increased during storage; levels of all three
parameters in ‘Xiangshui’ pears were higher than in ‘Garber’ pears. It is suggested that both ‘Xiangshui’ and ‘Garber’ pears
belong to climacteric fruits, and their color, taste and texture become better during after-ripening.