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2. Materials and methods2.1. Plant
2. Materials and methods
2.1. Plant materials
The carrots defined as No. 1 Orange-red variety grew in Gaoling
(Shaanxi, China) Planting base in autumn season. After harvesting,
the fresh carrots were transported to the laboratory by truck at
about 15 C, and stored in woven polypropylene bags in batches
of approximately 10 kg each. Storage temperature was 1 C and
the relative humidity was 90–95%. The storage time was less than
15 days before processing.
2.2. Preparation of carrot juices with different treatments
The raw material carrots were randomly separated into 4
batches. Washing process was conducted on a pilot plant scale with
commonly used as industrial equipment. Then selected good ones,
peeled and treated as follows (all treatments were selected for the
best in preliminary experiments):
(1) Fresh carrot juice: The carrots were sliced into 3 mm thick
and pressed into juice by HR2864 juice extractor (Philips
Electric Appliance Co., Holland) without any treatment used
as control.
(2) Blanching treatment: The carrots were placed in water bath
(Model HH-S; Jiangsu Zhenjiang Instrument Co. Ltd, Jiangsu,
China). Water blanching was carried out at 86 C for 10 min.
Then the carrots were sliced into pieces and pressed into
juice as above.
(3) Enzyme liquefaction treatment: The carrots were sliced into
pieces and treated by 1.5% (w/w) pectase (12,475 U mL1)
and 1.5% (w/w) cellulase (18,200 U g1). The optimal conditions
of pectase and cellulase treatments were 45 C, pH4.5,
120 min, and 50 C, pH5.0, 60 min, respectively. After that,
the pieces of carrots were pressed into juice.
(4) Pasteurisation treatment: The carrots were sliced into pieces
and pressed into juice, and then the juice was sterilized in
water bath at 100 C for 30 s.
Each treatment was replicated thrice and all the carrot juices
with different treatments were stored at 4 C and the storage time
was less than 15 days before analysis.
2.3. Extraction of the essential oil of carrot juice with different
treatments
The extraction of essential oil was carried out according to the
method with slight modifications (Diao, Hu, Zhang, & Xu, 2014;
Gao et al., 2011). The carrot juices with different treatments were
subjected to hydro distill for 6 h in a Clevenger-type apparatus. The
oily layer obtained on top of the aqueous distillate was separated
and dried with anhydrous sodium sulphate, then stored in a tightly
closed dark vial at 4 C until chemical composition analysis and
antimicrobial activity studies.
2.4. GC/MS analysis
The carrot juice essential oil with different treatments were
analysed by GC–MS, a gas chromatograph, Agilent Technologies
5973A using a HP 6890A Mass spectrometer with electron impact
ionisation (70 eV). A SE-54 capillary column (30 m 0.25 mm;
film thickness, 0.25 lm) was used. Oven temperature was programmed
to rise from 40 to 280 C at a flow rate of 4 C/min, sample
injection volume, 0.5 lL. The carrier gas was He with a flow
rate of 1 ml/min and a split ratio of 30:1, scan speed and mass
range were 0.5 s/dec and 45–450 m/z, respectively (Zaouali, Hnia,
Rim, & Mohamed, 2013).
The components were identified by comparing their mass spectra
with the data from the library of essential oil constituents, U.S.
National Bureau of Standards library, and Adams GC/MS libraries.
Use INCOS data system for retrieval and mass spectrum analysis.
Only the similarity of the components is greater than 80% would
be recorded.
2.5. Microbial strains
The antimicrobial activity of the carrot juice essential oil with
different treatments was tested against 8 selected food-related
microorganisms. Two gram-positive strains were Listeria monocytogenes
ATCC 19115 and Staphylococcus aureus ATCC 6538 P. Four
gram-negative bacteria were Salmonella typhimurium ATCC
19430, Salmonella enteritidis ATCC 13076, Shigella dysenteriae CMCC
(B) 51252 and Escherichia coli ATCC 25922. Aspergillus niger ATCC
02512 and Yeast were the two fungal microorganisms studied.
All the microorganisms were provided by the Microbiological Laboratory,
College of Life Science, Shaanxi Normal University, China.
2.6. Determination of antimicrobial activity of the carrot juice
essential oil with different treatments
The antimicrobial activity of the essential oil with different
treatments was evaluated using the standardised filter paper disk
diffusion method as described previously (Gao et al., 2011). Briefly,
100 ml of a suspension containing approximately 107 colonyforming
units (CFU)/ml of bacteria and 104 spore/ml of fungus
were spread on nutrient agar (NA) and potato dextrose agar
(PDA), respectively. Filter paper disks (6 mm in diameter) were
impregnated with 10 lL (4 mg/ml) of dilutions in dimethyl sulfoxide
(DMSO, Sigma) of the essential oil and placed onto the solid
medium (20 ml) plates. The diameter of inhibition zones (DIZ)
was measured after 24 h of incubation at 37 C for bacteria, after
4–5 d of incubation at 28 C for A. niger and after 24 h of incubation
at 25 C for Yeast. Ten microlitres of gentamicin (4 mg/ml) and
DMSO were used as positive and negative controls respectively.
Tests were performed in triplicate.
2.7. Determination of minimum inhibitory concentration (MIC) and
minimum bactericide concentration (MBC) with different treatments
MIC values and MBC values were determined for the microorganisms
that were sensitive to the essential oil in the filter paper
disk diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock solution of
essential oil with different treatments was prepared in DMSO.
Two fold serial dilutions of essential oil were prepared in sterile
2.1. Plant materials
The carrots defined as No. 1 Orange-red variety grew in Gaoling
(Shaanxi, China) Planting base in autumn season. After harvesting,
the fresh carrots were transported to the laboratory by truck at
about 15 C, and stored in woven polypropylene bags in batches
of approximately 10 kg each. Storage temperature was 1 C and
the relative humidity was 90–95%. The storage time was less than
15 days before processing.
2.2. Preparation of carrot juices with different treatments
The raw material carrots were randomly separated into 4
batches. Washing process was conducted on a pilot plant scale with
commonly used as industrial equipment. Then selected good ones,
peeled and treated as follows (all treatments were selected for the
best in preliminary experiments):
(1) Fresh carrot juice: The carrots were sliced into 3 mm thick
and pressed into juice by HR2864 juice extractor (Philips
Electric Appliance Co., Holland) without any treatment used
as control.
(2) Blanching treatment: The carrots were placed in water bath
(Model HH-S; Jiangsu Zhenjiang Instrument Co. Ltd, Jiangsu,
China). Water blanching was carried out at 86 C for 10 min.
Then the carrots were sliced into pieces and pressed into
juice as above.
(3) Enzyme liquefaction treatment: The carrots were sliced into
pieces and treated by 1.5% (w/w) pectase (12,475 U mL1)
and 1.5% (w/w) cellulase (18,200 U g1). The optimal conditions
of pectase and cellulase treatments were 45 C, pH4.5,
120 min, and 50 C, pH5.0, 60 min, respectively. After that,
the pieces of carrots were pressed into juice.
(4) Pasteurisation treatment: The carrots were sliced into pieces
and pressed into juice, and then the juice was sterilized in
water bath at 100 C for 30 s.
Each treatment was replicated thrice and all the carrot juices
with different treatments were stored at 4 C and the storage time
was less than 15 days before analysis.
2.3. Extraction of the essential oil of carrot juice with different
treatments
The extraction of essential oil was carried out according to the
method with slight modifications (Diao, Hu, Zhang, & Xu, 2014;
Gao et al., 2011). The carrot juices with different treatments were
subjected to hydro distill for 6 h in a Clevenger-type apparatus. The
oily layer obtained on top of the aqueous distillate was separated
and dried with anhydrous sodium sulphate, then stored in a tightly
closed dark vial at 4 C until chemical composition analysis and
antimicrobial activity studies.
2.4. GC/MS analysis
The carrot juice essential oil with different treatments were
analysed by GC–MS, a gas chromatograph, Agilent Technologies
5973A using a HP 6890A Mass spectrometer with electron impact
ionisation (70 eV). A SE-54 capillary column (30 m 0.25 mm;
film thickness, 0.25 lm) was used. Oven temperature was programmed
to rise from 40 to 280 C at a flow rate of 4 C/min, sample
injection volume, 0.5 lL. The carrier gas was He with a flow
rate of 1 ml/min and a split ratio of 30:1, scan speed and mass
range were 0.5 s/dec and 45–450 m/z, respectively (Zaouali, Hnia,
Rim, & Mohamed, 2013).
The components were identified by comparing their mass spectra
with the data from the library of essential oil constituents, U.S.
National Bureau of Standards library, and Adams GC/MS libraries.
Use INCOS data system for retrieval and mass spectrum analysis.
Only the similarity of the components is greater than 80% would
be recorded.
2.5. Microbial strains
The antimicrobial activity of the carrot juice essential oil with
different treatments was tested against 8 selected food-related
microorganisms. Two gram-positive strains were Listeria monocytogenes
ATCC 19115 and Staphylococcus aureus ATCC 6538 P. Four
gram-negative bacteria were Salmonella typhimurium ATCC
19430, Salmonella enteritidis ATCC 13076, Shigella dysenteriae CMCC
(B) 51252 and Escherichia coli ATCC 25922. Aspergillus niger ATCC
02512 and Yeast were the two fungal microorganisms studied.
All the microorganisms were provided by the Microbiological Laboratory,
College of Life Science, Shaanxi Normal University, China.
2.6. Determination of antimicrobial activity of the carrot juice
essential oil with different treatments
The antimicrobial activity of the essential oil with different
treatments was evaluated using the standardised filter paper disk
diffusion method as described previously (Gao et al., 2011). Briefly,
100 ml of a suspension containing approximately 107 colonyforming
units (CFU)/ml of bacteria and 104 spore/ml of fungus
were spread on nutrient agar (NA) and potato dextrose agar
(PDA), respectively. Filter paper disks (6 mm in diameter) were
impregnated with 10 lL (4 mg/ml) of dilutions in dimethyl sulfoxide
(DMSO, Sigma) of the essential oil and placed onto the solid
medium (20 ml) plates. The diameter of inhibition zones (DIZ)
was measured after 24 h of incubation at 37 C for bacteria, after
4–5 d of incubation at 28 C for A. niger and after 24 h of incubation
at 25 C for Yeast. Ten microlitres of gentamicin (4 mg/ml) and
DMSO were used as positive and negative controls respectively.
Tests were performed in triplicate.
2.7. Determination of minimum inhibitory concentration (MIC) and
minimum bactericide concentration (MBC) with different treatments
MIC values and MBC values were determined for the microorganisms
that were sensitive to the essential oil in the filter paper
disk diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock solution of
essential oil with different treatments was prepared in DMSO.
Two fold serial dilutions of essential oil were prepared in sterile
0/5000
2. Materials and methods2.1. Plant materialsThe carrots defined as No. 1 Orange-red variety grew in Gaoling(Shaanxi, China) Planting base in autumn season. After harvesting,the fresh carrots were transported to the laboratory by truck atabout 15 C, and stored in woven polypropylene bags in batchesof approximately 10 kg each. Storage temperature was 1 C andthe relative humidity was 90–95%. The storage time was less than15 days before processing.2.2. Preparation of carrot juices with different treatmentsThe raw material carrots were randomly separated into 4batches. Washing process was conducted on a pilot plant scale withcommonly used as industrial equipment. Then selected good ones,peeled and treated as follows (all treatments were selected for thebest in preliminary experiments):(1) Fresh carrot juice: The carrots were sliced into 3 mm thickand pressed into juice by HR2864 juice extractor (PhilipsElectric Appliance Co., Holland) without any treatment usedas control.(2) Blanching treatment: The carrots were placed in water bath(Model HH-S; Jiangsu Zhenjiang Instrument Co. Ltd, Jiangsu,China). Water blanching was carried out at 86 C for 10 min.Then the carrots were sliced into pieces and pressed intojuice as above.(3) Enzyme liquefaction treatment: The carrots were sliced intopieces and treated by 1.5% (w/w) pectase (12,475 U mL 1)and 1.5% (w/w) cellulase (18,200 U g 1). The optimal conditionsof pectase and cellulase treatments were 45 C, pH4.5,120 min, and 50 C, pH5.0, 60 min, respectively. After that,the pieces of carrots were pressed into juice.(4) Pasteurisation treatment: The carrots were sliced into piecesand pressed into juice, and then the juice was sterilized inwater bath at 100 C for 30 s.Each treatment was replicated thrice and all the carrot juiceswith different treatments were stored at 4 C and the storage timewas less than 15 days before analysis.2.3. Extraction of the essential oil of carrot juice with differenttreatmentsThe extraction of essential oil was carried out according to themethod with slight modifications (Diao, Hu, Zhang, & Xu, 2014;Gao et al., 2011). The carrot juices with different treatments weresubjected to hydro distill for 6 h in a Clevenger-type apparatus. Theoily layer obtained on top of the aqueous distillate was separatedand dried with anhydrous sodium sulphate, then stored in a tightlyclosed dark vial at 4 C until chemical composition analysis andantimicrobial activity studies.2.4. GC/MS analysisThe carrot juice essential oil with different treatments wereanalysed by GC–MS, a gas chromatograph, Agilent Technologies5973A using a HP 6890A Mass spectrometer with electron impactionisation (70 eV). A SE-54 capillary column (30 m 0.25 mm;film thickness, 0.25 lm) was used. Oven temperature was programmedto rise from 40 to 280 C at a flow rate of 4 C/min, sampleinjection volume, 0.5 lL. The carrier gas was He with a flowrate of 1 ml/min and a split ratio of 30:1, scan speed and massrange were 0.5 s/dec and 45–450 m/z, respectively (Zaouali, Hnia,Rim, & Mohamed, 2013).The components were identified by comparing their mass spectrawith the data from the library of essential oil constituents, U.S.National Bureau of Standards library, and Adams GC/MS libraries.Use INCOS data system for retrieval and mass spectrum analysis.Only the similarity of the components is greater than 80% wouldbe recorded.2.5. Microbial strainsThe antimicrobial activity of the carrot juice essential oil withdifferent treatments was tested against 8 selected food-relatedmicroorganisms. Two gram-positive strains were Listeria monocytogenesATCC 19115 and Staphylococcus aureus ATCC 6538 P. Fourgram-negative bacteria were Salmonella typhimurium ATCC19430, Salmonella enteritidis ATCC 13076, Shigella dysenteriae CMCC(B) 51252 and Escherichia coli ATCC 25922. Aspergillus niger ATCC02512 and Yeast were the two fungal microorganisms studied.All the microorganisms were provided by the Microbiological Laboratory,College of Life Science, Shaanxi Normal University, China.2.6. Determination of antimicrobial activity of the carrot juiceessential oil with different treatmentsThe antimicrobial activity of the essential oil with differenttreatments was evaluated using the standardised filter paper diskdiffusion method as described previously (Gao et al., 2011). Briefly,100 ml of a suspension containing approximately 107 colonyformingunits (CFU)/ml of bacteria and 104 spore/ml of funguswere spread on nutrient agar (NA) and potato dextrose agar(PDA), respectively. Filter paper disks (6 mm in diameter) wereimpregnated with 10 lL (4 mg/ml) of dilutions in dimethyl sulfoxide(DMSO, Sigma) of the essential oil and placed onto the solidmedium (20 ml) plates. The diameter of inhibition zones (DIZ)was measured after 24 h of incubation at 37 C for bacteria, after4–5 d of incubation at 28 C for A. niger and after 24 h of incubationat 25 C for Yeast. Ten microlitres of gentamicin (4 mg/ml) andDMSO were used as positive and negative controls respectively.Tests were performed in triplicate.2.7. Determination of minimum inhibitory concentration (MIC) andminimum bactericide concentration (MBC) with different treatmentsMIC values and MBC values were determined for the microorganismsthat were sensitive to the essential oil in the filter paperdisk diffusion assay. The MIC values were determined by tube dilutionmethod as described by Gao et al. (2011). Stock solution ofessential oil with different treatments was prepared in DMSO.Two fold serial dilutions of essential oil were prepared in sterile
การแปล กรุณารอสักครู่..
2. Materials and methods
2.1. Plant Materials
The carrots defined as No. 1 Orange-Red Variety grew in Gaoling
(Shaanxi, China) Planting Base in Autumn Season. After harvesting,
the Fresh carrots were transported to the Laboratory by Truck at
About 15? C, and stored in polypropylene woven bags in batches
of approximately 10 kg each. Storage Temperature was 1? C and
the Relative humidity was 90-95%. Storage the time was less than
15 days before Processing.
2.2. Preparation of Carrot juices with different treatments
were randomly Separated Into The RAW Material carrots 4
batches. Washing Pilot Plant Process was conducted on a scale with
commonly used as Industrial Equipment. Then selected good Ones,
peeled and treated as follows (all treatments were selected for the
Best in Preliminary experiments):
(1) Fresh Carrot Juice: The carrots were sliced Into 3 mm Thick
and pressed Into Juice by HR2864 Juice Extractor (Philips
Electric Appliance. Co., Holland) Without any Treatment used
as Control.
(2) blanching Treatment: The carrots were Placed in Water bath
(Model HH-S; Jiangsu Zhenjiang Instrument Co. Ltd, Jiangsu,
China). Water blanching was carried out at 86? C for 10 min.
Then the carrots were sliced Into pieces and pressed Into
Juice as above.
(3) Enzyme liquefaction Treatment: The carrots were sliced Into
pieces and treated by 1.5% (W / W). Pectase (twelve thousand four hundred and seventy-five U mL? 1)
and 1.5% (W / W) cellulase (18,200 U G? 1). Optimal the conditions
of Pectase and cellulase treatments were 45? C, PH4.5,
120 min, and 50? C, PH5.0, 60 min, respectively. After that,
the pieces of carrots were pressed Into Juice.
(4) Pasteurisation Treatment: The carrots were sliced Into pieces
and pressed Into Juice, and then the Juice was sterilized in
Water bath at 100? C for 30 s.
Each Treatment was Replicated. thrice and all the Carrot juices
with different treatments were stored at 4? C and the Storage time
was less than 15 days before Analysis.
2.3. Carrot Juice extraction of the Essential Oil of with different
treatments
of Essential Oil The extraction was carried out according to the
method with slight modifications (Diao, Hu, Zhang, & Xu, 2,014th;
Gao et al., 2011). The Carrot juices with different treatments were
subjected to hydro distill for 6 H in a Clevenger-Type Apparatus. The
oily layer on top of the aqueous distillate was obtained Separated
and Dried with anhydrous Sodium sulphate, then stored in a tightly
Dark Closed Vial at 4? C until Analysis Chemical composition and
antimicrobial Activity Studies.
2.4. GC / MS Analysis
The Carrot Juice Essential Oil with different treatments were
analyzed by GC-MS, a gas chromatograph, Agilent Technologies
5973A using a HP 6890A Mass spectrometer with Electron Impact
ionisation (70 eV). A SE-54 capillary column (30 M? Twelve twenty-five a.m. mm;
Film thickness, twelve twenty-five LM) was used. Temperature oven was programmed
to rise from 40 to 280? C at a flow rate of 4? C / min, sample
volume Injection, 0.5 Ll. He was the Carrier gas with a flow
rate of 1 ml / min and a Split ratio of 30: 1, speed and mass Scan
Range were 0.5 s / dec and forty-five to four hundred fifty M / Z, respectively (Zaouali, Hnia,
Rim, &. mohamed, the 2013th).
The components were identified by comparing their mass Spectra
with the Data from the Library of Essential Oil constituents, US
National Bureau of Standards Library, Adams and GC / MS libraries.
Use INCOS Data Retrieval System for Spectrum Analysis and mass.
the only similarity is Greater than 80% of the components would
be Recorded.
2.5. Microbial strains
The antimicrobial Activity of the Essential Oil Carrot Juice with
different treatments was tested against 8 selected Food-related
microorganisms. Gram-positive Listeria monocytogenes Two strains were
Staphylococcus aureus ATCC 19115 and ATCC 6538 P. Four
Gram-negative bacteria Salmonella typhimurium were ATCC
19430, Salmonella Enteritidis ATCC 13076, Shigella dysenteriae CMCC
(B) and Escherichia coli ATCC 25922. Aspergillus niger 51,252th ATCC
02512th. Yeast and fungal microorganisms studied were the Two.
All the microorganisms were provided by the Microbiological Laboratory,
College of Life Science, Shaanxi Normal University, China.
2.6. Activity of the determination of antimicrobial Carrot Juice
Essential Oil with different treatments
of the Activity The antimicrobial Essential Oil with different
treatments was evaluated using the standardized Filter Paper Disk
Diffusion method as described previously (Gao et al., in 2011). Briefly,
100 ml of a suspension containing approximately 107 Colonyforming
Units (CFU) / ml of bacteria and Spore 104 / ml of Fungus
were spread on nutrient agar (NA) and potato dextrose agar
(PDA), respectively. Filter Paper Disks (6 mm in Diameter) were
impregnated with 10 Ll (4 mg / ml) in dimethyl sulfoxide of dilutions
(DMSO, Sigma) of the Essential Oil and Placed onto the Solid
Medium (20 ml) plates. Diameter of the inhibition zones (DIZ)
was measured after 24 H of Incubation at 37? C for bacteria, after
4-5 D of Incubation at 28? C for A. niger and after 24 H of Incubation
at 25? C for Yeast. Ten Microlitres of gentamicin (4 mg / ml) and
DMSO were used as positive and negative respectively Controls.
Tests were performed in Triplicate.
2.7. Minimum inhibitory concentration determination of (MIC) and
Minimum bactericide concentration (MBC) with different treatments
MIC values and MBC values were determined for the microorganisms
that were sensitive to the Essential Oil in the Filter Paper
Disk Diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock Solution of
Essential Oil with different treatments was prepared in DMSO.
Two Fold Serial dilutions of Essential Oil were prepared in sterile.
2.1. Plant Materials
The carrots defined as No. 1 Orange-Red Variety grew in Gaoling
(Shaanxi, China) Planting Base in Autumn Season. After harvesting,
the Fresh carrots were transported to the Laboratory by Truck at
About 15? C, and stored in polypropylene woven bags in batches
of approximately 10 kg each. Storage Temperature was 1? C and
the Relative humidity was 90-95%. Storage the time was less than
15 days before Processing.
2.2. Preparation of Carrot juices with different treatments
were randomly Separated Into The RAW Material carrots 4
batches. Washing Pilot Plant Process was conducted on a scale with
commonly used as Industrial Equipment. Then selected good Ones,
peeled and treated as follows (all treatments were selected for the
Best in Preliminary experiments):
(1) Fresh Carrot Juice: The carrots were sliced Into 3 mm Thick
and pressed Into Juice by HR2864 Juice Extractor (Philips
Electric Appliance. Co., Holland) Without any Treatment used
as Control.
(2) blanching Treatment: The carrots were Placed in Water bath
(Model HH-S; Jiangsu Zhenjiang Instrument Co. Ltd, Jiangsu,
China). Water blanching was carried out at 86? C for 10 min.
Then the carrots were sliced Into pieces and pressed Into
Juice as above.
(3) Enzyme liquefaction Treatment: The carrots were sliced Into
pieces and treated by 1.5% (W / W). Pectase (twelve thousand four hundred and seventy-five U mL? 1)
and 1.5% (W / W) cellulase (18,200 U G? 1). Optimal the conditions
of Pectase and cellulase treatments were 45? C, PH4.5,
120 min, and 50? C, PH5.0, 60 min, respectively. After that,
the pieces of carrots were pressed Into Juice.
(4) Pasteurisation Treatment: The carrots were sliced Into pieces
and pressed Into Juice, and then the Juice was sterilized in
Water bath at 100? C for 30 s.
Each Treatment was Replicated. thrice and all the Carrot juices
with different treatments were stored at 4? C and the Storage time
was less than 15 days before Analysis.
2.3. Carrot Juice extraction of the Essential Oil of with different
treatments
of Essential Oil The extraction was carried out according to the
method with slight modifications (Diao, Hu, Zhang, & Xu, 2,014th;
Gao et al., 2011). The Carrot juices with different treatments were
subjected to hydro distill for 6 H in a Clevenger-Type Apparatus. The
oily layer on top of the aqueous distillate was obtained Separated
and Dried with anhydrous Sodium sulphate, then stored in a tightly
Dark Closed Vial at 4? C until Analysis Chemical composition and
antimicrobial Activity Studies.
2.4. GC / MS Analysis
The Carrot Juice Essential Oil with different treatments were
analyzed by GC-MS, a gas chromatograph, Agilent Technologies
5973A using a HP 6890A Mass spectrometer with Electron Impact
ionisation (70 eV). A SE-54 capillary column (30 M? Twelve twenty-five a.m. mm;
Film thickness, twelve twenty-five LM) was used. Temperature oven was programmed
to rise from 40 to 280? C at a flow rate of 4? C / min, sample
volume Injection, 0.5 Ll. He was the Carrier gas with a flow
rate of 1 ml / min and a Split ratio of 30: 1, speed and mass Scan
Range were 0.5 s / dec and forty-five to four hundred fifty M / Z, respectively (Zaouali, Hnia,
Rim, &. mohamed, the 2013th).
The components were identified by comparing their mass Spectra
with the Data from the Library of Essential Oil constituents, US
National Bureau of Standards Library, Adams and GC / MS libraries.
Use INCOS Data Retrieval System for Spectrum Analysis and mass.
the only similarity is Greater than 80% of the components would
be Recorded.
2.5. Microbial strains
The antimicrobial Activity of the Essential Oil Carrot Juice with
different treatments was tested against 8 selected Food-related
microorganisms. Gram-positive Listeria monocytogenes Two strains were
Staphylococcus aureus ATCC 19115 and ATCC 6538 P. Four
Gram-negative bacteria Salmonella typhimurium were ATCC
19430, Salmonella Enteritidis ATCC 13076, Shigella dysenteriae CMCC
(B) and Escherichia coli ATCC 25922. Aspergillus niger 51,252th ATCC
02512th. Yeast and fungal microorganisms studied were the Two.
All the microorganisms were provided by the Microbiological Laboratory,
College of Life Science, Shaanxi Normal University, China.
2.6. Activity of the determination of antimicrobial Carrot Juice
Essential Oil with different treatments
of the Activity The antimicrobial Essential Oil with different
treatments was evaluated using the standardized Filter Paper Disk
Diffusion method as described previously (Gao et al., in 2011). Briefly,
100 ml of a suspension containing approximately 107 Colonyforming
Units (CFU) / ml of bacteria and Spore 104 / ml of Fungus
were spread on nutrient agar (NA) and potato dextrose agar
(PDA), respectively. Filter Paper Disks (6 mm in Diameter) were
impregnated with 10 Ll (4 mg / ml) in dimethyl sulfoxide of dilutions
(DMSO, Sigma) of the Essential Oil and Placed onto the Solid
Medium (20 ml) plates. Diameter of the inhibition zones (DIZ)
was measured after 24 H of Incubation at 37? C for bacteria, after
4-5 D of Incubation at 28? C for A. niger and after 24 H of Incubation
at 25? C for Yeast. Ten Microlitres of gentamicin (4 mg / ml) and
DMSO were used as positive and negative respectively Controls.
Tests were performed in Triplicate.
2.7. Minimum inhibitory concentration determination of (MIC) and
Minimum bactericide concentration (MBC) with different treatments
MIC values and MBC values were determined for the microorganisms
that were sensitive to the Essential Oil in the Filter Paper
Disk Diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock Solution of
Essential Oil with different treatments was prepared in DMSO.
Two Fold Serial dilutions of Essential Oil were prepared in sterile.
การแปล กรุณารอสักครู่..
2. Materials and methods
2.1. Plant materials
The carrots defined as No. 1 Orange-red variety grew in Gaoling
(Shaanxi,, China) Planting base in autumn season. After harvesting
the, fresh carrots were transported to the laboratory by truck at
about 15, C. And stored in woven polypropylene bags in batches
of approximately 10 kg each. Storage temperature was 1 C and
.The relative humidity was 90 - 95%. The storage time was less than
15 days before processing.
2.2. Preparation of carrot. Juices with different treatments
The raw material carrots were randomly separated into 4
batches. Washing process was conducted. On a pilot plant scale with
commonly used as industrial equipment. Then selected, good ones
.Peeled and treated as follows (all treatments were selected for the
best in preliminary experiments)
(1) Fresh carrot. Juice: The carrots were sliced into 3 mm thick
and pressed into juice by HR2864 juice extractor (Philips
Electric Appliance. Co, Holland) without any treatment used
as control.
(2) Blanching treatment: The carrots were placed in water bath
(Model. HH-S;Jiangsu Zhenjiang Instrument Co. Ltd Jiangsu
China,,). Water blanching was carried out at 86 C for 10 min.
Then the carrots. Were sliced into pieces and pressed into
juice as above.
(3) Enzyme liquefaction treatment: The carrots were sliced into
pieces. And treated by 1.5% (w / W), pectase (12 475 U mL 1)
and 1.5% (w / W), cellulase (18 200 U g 1). The optimal conditions
.Of pectase and cellulase treatments were 45 C pH4.5
120 min,,,,, and 50 C pH5.0, 60 min respectively. After that
the,, Pieces of carrots were pressed into juice.
(4) Pasteurisation treatment: The carrots were sliced into pieces
and pressed. Into juice and then, the juice was sterilized in
water bath at 100 C for 30 s.
Each treatment was replicated thrice and. All the carrot juices
.With different treatments were stored at 4 C and the storage time
was less than 15 days before analysis.
2.3. Extraction. Of the essential oil of carrot juice with different
The treatments extraction of essential oil was carried out according. To the
method with slight modifications (Diao Hu Zhang,,,, & Xu 2014;
Gao et al, 2011). The carrot juices with different. Treatments were
.Subjected to Hydro distill for 6 h in a Clevenger-type apparatus. The
oily layer obtained on top of the aqueous distillate. Was separated
and dried with anhydrous, sodium sulphate then stored in a tightly
closed dark vial at 4 C until chemical. Composition analysis and
antimicrobial activity studies.
2.4. GC / MS analysis
The carrot juice essential oil with different. Treatments were
.Analysed by GC - MS a chromatograph Agilent, gas, Technologies
5973A using a HP 6890A Mass spectrometer with electron impact
ionisation. (70 eV). A SE-54 capillary column (30 m 0.25 mm;
film thickness 0.25 LM), was used. Oven temperature was programmed
to. Rise from 40 to 280 C at a flow rate of 4 C / min sample
injection, volume 0.5, lL. The carrier gas was He with a flow
.Rate of 1 ml / min and a split ratio of 30: 1 scan speed, and mass
range were 0.5 s / DEC and 45 - 450 m / Z respectively (Zaouali,,, Hnia
Rim,,, & Mohamed 2013).
The components were identified by comparing their mass spectra
with the data from the library. Of essential, oil constituents U.S.
National Bureau of Standards library and Adams, GC / MS libraries.
.Use INCOS data system for retrieval and mass spectrum analysis.
Only the similarity of the components is greater than 80% would
be. Recorded.
2.5. Microbial strains
The antimicrobial activity of the carrot juice essential oil with
different treatments. Was tested against 8 selected food-related
microorganisms. Two Gram-positive strains were Listeria monocytogenes
.ATCC 19115 and Staphylococcus aureus ATCC 6538 P. Four
Gram-negative bacteria were Salmonella typhimurium ATCC
19430 Salmonella,, Enteritidis, ATCC 13076 Shigella dysenteriae CMCC
(B) 51252 and Escherichia coli ATCC 25922. Aspergillus Niger ATCC
02512. And Yeast were the two fungal microorganisms studied.
All the microorganisms were provided by the, Microbiological Laboratory
.College of Life Science Shaanxi, Normal, University China.
2.6. Determination of antimicrobial activity of the carrot juice
essential. Oil with different treatments
The antimicrobial activity of the essential oil with different
treatments was evaluated using. The standardised filter paper disk
diffusion method as described previously (Gao et al, 2011), Briefly
.100 ml of a suspension containing approximately 107 colonyforming
units (CFU) / ml of bacteria and 104 spore / ml of fungus
were. Spread on Nutrient Agar (NA) and potato dextrose agar
(PDA), respectively. Filter paper disks (6 mm in diameter) were
impregnated. With 10 lL (4 mg / ml) of dilutions in dimethyl sulfoxide
(DMSO Sigma), of the essential oil and placed onto the solid
medium. (20 ml) plates.The diameter of inhibition zones (DIZ)
was measured after 24 h of incubation at 37 C, for bacteria after
4 - 5 d of incubation. At 28 C for A. Niger and after 24 h of incubation
at 25 C for Yeast. Ten microlitres of gentamicin (4 mg / ml) and
DMSO. Were used as positive and negative controls respectively.
Tests were performed in triplicate.
2.7.Determination of minimum inhibitory concentration (MIC) and
minimum bactericide concentration (MBC) with different treatments
MIC. Values and MBC values were determined for the microorganisms
that were sensitive to the essential oil in the filter paper
disk. Diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock solution. Of
.Essential oil with different treatments was prepared in DMSO.
Two fold serial dilutions of essential oil were prepared. In sterile.
2.1. Plant materials
The carrots defined as No. 1 Orange-red variety grew in Gaoling
(Shaanxi,, China) Planting base in autumn season. After harvesting
the, fresh carrots were transported to the laboratory by truck at
about 15, C. And stored in woven polypropylene bags in batches
of approximately 10 kg each. Storage temperature was 1 C and
.The relative humidity was 90 - 95%. The storage time was less than
15 days before processing.
2.2. Preparation of carrot. Juices with different treatments
The raw material carrots were randomly separated into 4
batches. Washing process was conducted. On a pilot plant scale with
commonly used as industrial equipment. Then selected, good ones
.Peeled and treated as follows (all treatments were selected for the
best in preliminary experiments)
(1) Fresh carrot. Juice: The carrots were sliced into 3 mm thick
and pressed into juice by HR2864 juice extractor (Philips
Electric Appliance. Co, Holland) without any treatment used
as control.
(2) Blanching treatment: The carrots were placed in water bath
(Model. HH-S;Jiangsu Zhenjiang Instrument Co. Ltd Jiangsu
China,,). Water blanching was carried out at 86 C for 10 min.
Then the carrots. Were sliced into pieces and pressed into
juice as above.
(3) Enzyme liquefaction treatment: The carrots were sliced into
pieces. And treated by 1.5% (w / W), pectase (12 475 U mL 1)
and 1.5% (w / W), cellulase (18 200 U g 1). The optimal conditions
.Of pectase and cellulase treatments were 45 C pH4.5
120 min,,,,, and 50 C pH5.0, 60 min respectively. After that
the,, Pieces of carrots were pressed into juice.
(4) Pasteurisation treatment: The carrots were sliced into pieces
and pressed. Into juice and then, the juice was sterilized in
water bath at 100 C for 30 s.
Each treatment was replicated thrice and. All the carrot juices
.With different treatments were stored at 4 C and the storage time
was less than 15 days before analysis.
2.3. Extraction. Of the essential oil of carrot juice with different
The treatments extraction of essential oil was carried out according. To the
method with slight modifications (Diao Hu Zhang,,,, & Xu 2014;
Gao et al, 2011). The carrot juices with different. Treatments were
.Subjected to Hydro distill for 6 h in a Clevenger-type apparatus. The
oily layer obtained on top of the aqueous distillate. Was separated
and dried with anhydrous, sodium sulphate then stored in a tightly
closed dark vial at 4 C until chemical. Composition analysis and
antimicrobial activity studies.
2.4. GC / MS analysis
The carrot juice essential oil with different. Treatments were
.Analysed by GC - MS a chromatograph Agilent, gas, Technologies
5973A using a HP 6890A Mass spectrometer with electron impact
ionisation. (70 eV). A SE-54 capillary column (30 m 0.25 mm;
film thickness 0.25 LM), was used. Oven temperature was programmed
to. Rise from 40 to 280 C at a flow rate of 4 C / min sample
injection, volume 0.5, lL. The carrier gas was He with a flow
.Rate of 1 ml / min and a split ratio of 30: 1 scan speed, and mass
range were 0.5 s / DEC and 45 - 450 m / Z respectively (Zaouali,,, Hnia
Rim,,, & Mohamed 2013).
The components were identified by comparing their mass spectra
with the data from the library. Of essential, oil constituents U.S.
National Bureau of Standards library and Adams, GC / MS libraries.
.Use INCOS data system for retrieval and mass spectrum analysis.
Only the similarity of the components is greater than 80% would
be. Recorded.
2.5. Microbial strains
The antimicrobial activity of the carrot juice essential oil with
different treatments. Was tested against 8 selected food-related
microorganisms. Two Gram-positive strains were Listeria monocytogenes
.ATCC 19115 and Staphylococcus aureus ATCC 6538 P. Four
Gram-negative bacteria were Salmonella typhimurium ATCC
19430 Salmonella,, Enteritidis, ATCC 13076 Shigella dysenteriae CMCC
(B) 51252 and Escherichia coli ATCC 25922. Aspergillus Niger ATCC
02512. And Yeast were the two fungal microorganisms studied.
All the microorganisms were provided by the, Microbiological Laboratory
.College of Life Science Shaanxi, Normal, University China.
2.6. Determination of antimicrobial activity of the carrot juice
essential. Oil with different treatments
The antimicrobial activity of the essential oil with different
treatments was evaluated using. The standardised filter paper disk
diffusion method as described previously (Gao et al, 2011), Briefly
.100 ml of a suspension containing approximately 107 colonyforming
units (CFU) / ml of bacteria and 104 spore / ml of fungus
were. Spread on Nutrient Agar (NA) and potato dextrose agar
(PDA), respectively. Filter paper disks (6 mm in diameter) were
impregnated. With 10 lL (4 mg / ml) of dilutions in dimethyl sulfoxide
(DMSO Sigma), of the essential oil and placed onto the solid
medium. (20 ml) plates.The diameter of inhibition zones (DIZ)
was measured after 24 h of incubation at 37 C, for bacteria after
4 - 5 d of incubation. At 28 C for A. Niger and after 24 h of incubation
at 25 C for Yeast. Ten microlitres of gentamicin (4 mg / ml) and
DMSO. Were used as positive and negative controls respectively.
Tests were performed in triplicate.
2.7.Determination of minimum inhibitory concentration (MIC) and
minimum bactericide concentration (MBC) with different treatments
MIC. Values and MBC values were determined for the microorganisms
that were sensitive to the essential oil in the filter paper
disk. Diffusion assay. The MIC values were determined by tube dilution
method as described by Gao et al. (2011). Stock solution. Of
.Essential oil with different treatments was prepared in DMSO.
Two fold serial dilutions of essential oil were prepared. In sterile.
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