Saccharides and Sweetning agents(Additives for Foods)

Additives for Foods and Drugs

Preservatives were analyzed using RSpak DS-613 (a column for reversed phase chromatography).

Sample :
1. Saccharin sodium
2. p-Hydroxybenzoic acid
3. Sorbic acid
4. Benzoic acid
5. Methyl paraben, Methyl p-hydroxybenzoate
6. Ethyl paraben, Ethyl p-hydroxybenzoate
7. Dehydroacetic acid
8. Propyl paraben, Propyl p-hydroxybenzoate

Column       : Shodex RSpak DS-613 (6.0mmID*150mm)
Eluent       : (0.05M KH2PO4 + 0.1% H3PO4) aq./CHSUB>3CN=60/40
Flow rate    : 1.0mL/min
Detector     : UV(210nm)
Column temp. : 22deg-C

Preservatives for foods were analyzed using RSpak DE-613 (a column for reversed phase chromatography).

Sample : p-Hydroxybenzoate
Me Methyl paraben, Methyl p-hydroxybenzoate
Et Ethyl paraben, Ethyl p-hydroxybenzoate
n-Pr Propyl paraben, Propyl p-hydroxybenzoate
n-Bu Butyl paraben, Butyl p-hydroxybenzoate

Column       : Shodex RSpak DE-613 (6.0mmID*150mm)
Eluent       : CH3CN/H2O=50/50
Flow rate    : 2.0mL/min
Detector     : UV(254nm)
Column temp. : 50deg-C

Preservatives were analyzed using RSpak RP18-613 (a column for reversed phase chromatography).

Sample :
1. Saccharin sodium
2. p-Hydroxybenzoic acid
3. Sorbic acid
4. Benzoic acid
5. Methyl paraben, Methyl p-hydroxybenzoate
6. Ethyl paraben, Ethyl p-hydroxybenzoate
7. Dehydroacetic acid
8. Propyl paraben, Propyl p-hydroxybenzoate

Column       : Shodex RSpak RP18-613 (6.0mmID*150mm)
Eluent       : (0.05M KH2PO4 + 0.1% H3PO4) aq./CH3CN=60/40
Flow rate    : 1.0mL/min
Detector     : UV(210nm)
Column temp. : 50deg-C

Food aditives, which have wide polarity range, can be analyzed using RSpak DE-413 (a column for reversed phase chromatography) by simple isocratic mode in a short time.

Sample : 10micro-L
1. Saccharin sodium 0.005%
2. p-Hydroxybenzoic acid 0.005%
3. Sorbic acid 0.02%
4. Benzoic acid 0.02%
5. Methyl paraben, Methyl p-hydroxybenzoate 0.01%
6. Dehydroacetic acid 0.01%
7. Ethyl paraben, Ethyl p-hydroxybenzoate 0.02%
8. Propyl paraben, Propyl p-hydroxybenzoate 0.01%

Column       : Shodex RSpak DE-413 (4.6mmID*150mm)
Eluent       : (0.05M KH2PO4 + 0.1% H3PO4) aq./CH3CN=35/65
Flow rate    : 1.0mL/min
Detector     : UV(210nm)
Column temp. : 40deg-C

Acesulfame K which is sweetening agent and some saccharides were analyzed using SUGAR SC1011 (a column for the analysis of saccharides).

Sample : 10micro-L
1. 0.005% Acesulfame K
2. 0.5% Sucrose
3. 0.5% Glucose
4. 0.5% Fructose

Column       : Shodex SUGAR SC1011 (8.0mmID*300mm)
Eluent       : 10mM CaSO4 aq.
Flow rate    : 1.0mL/min
Detector     : Shodex RI, UV(254nm)
Column temp. : 80deg-C

Sucralose which is sweetening agent and some saccharides were analyzed using SUGAR SP0810 (a column for the analysis of saccharides). Eluent is just water.

Sample : 1% each, 10micro-L
1. Sucrose
2. Glucose
3. Fructose
4. meso-Erythritol
5. Sucralose

Column       : Shodex SUGAR SP0810 (8.0mmID*300mm)
Eluent       : H2O
Flow rate    : 1.0mL/min
Detector     : Shodex RI
Column temp. : 80deg-C

Food Yellow No.4, one of food aditives was analyzed using a reversed phase column, Asahipak ODP-50 4D. Tartazine is the another name of Food Yellow No.4.

Sample : Food Yellow No.4

Column       : Shodex Asahipak ODP-50 4D (4.6mmID*150mm)
Eluent       : 0.01M (NH4)2HPO4 aq./CH3OH=95/5
Flow rate    : 0.6mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Methylxantines were analyzed using RSpak DS-613 (a column for reversed phase chromatography).

Sample : 0.02% each, 6micro-L
1. Xanthine
2. Theobromine
3. Theophyline
4. Caffeine

Column       : Shodex RSpak DS-613 (6.0mmID*150mm)
Eluent       : H2O/CH3CN=88/12
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Theobromine, theophylline, caffeine were separated by RSpak DE-413 (4.6mmID x 150mm) and DE-213 (2.0mmID x 150mm). As DE-213 is a semi-micro type column, it can be also used for LC/MS.

Sample :
1. Theobromine
2. Theophylline
3. Caffeine

              (Left)
Column       : Shodex RSpak DE-413(4.6mmID*150mm)   
Eluent       : H2O/CH3CN=75/25
Flow rate    : 0.5mL/min
Detector     : UV(254nm)
Column temp. : 30deg-C

 (Right)
: Shodex RSpak DE-213(2.0mmID*150mm)
: H2O/CH3CN=75/25
: 0.1mL/min
: UV(254nm)
: 30deg-C

RSpak DE series is stable in wide eluent pH range.

Sample : 10micro-L
1. Theobromine
2. Theophylline
3. Caffeine

Column       : Shodex RSpak DE-413 (4.6mmID*150mm) 
Eluent       : 10mM Sodium phosphate buffer/CH3CN=82/18
Flow rate    : 0.6mL/min 
Detector     : UV(254nm) 
Column temp. : 40deg-C

Synthetic food colorants Red 2, Red 102, as well as azorubine and patent blue V are commonly added to red wine to deepen its red color. In this application, these 4 synthetics food colorants were analyzed at high temperature with the polymer-based reversed phase chromatography cplumn ET-RP1 3D. The high-temperature analysis was completed in about 10 min, compared to a usual analysis time of over 40 min.

Sample : 45μg/mL each, 20micro-L
1. Food Red No.2, Amaranth
2. Food Red No.102, New coccine
3. Azorubine
4. Patent blue V

Column       : Shodex ET-RP1 3D (3.0mmID*150mm)
Eluent       : 6mM Tetrabutylammonium bromide in (25mM Ammonium acetate aq./CH3CN=70/30)
Flow rate    : 0.5mL/min
Detector     : UV(254nm)
Column temp. : 70deg-C

Asahipak NH2P-50 4E was used to analyze betaine. Betaine (Trimethylglycine, or (Carboxylmethyl)-Trimethylammonium) is a dipole ion which bears cationic and anionic charges in its structure, found in sugar beets (Beta vulgaris), shrimps and crabs. Betaine is often used as a food additive and a moisturizer in cosmetics.

Sample : 20micro-L
1. Betaine (Trimethylglycine,
Carboxymethyl)trimethylammonium) 0.1%

Column       : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm)
Eluent       : H2O/CH3CN=25/75
Flow rate    : 1.0mL/min
Detector     : Shodex RI
Column temp. : 40deg-C

Polymer-based-amino column, Asahipak NH2P-50 4E was used to separate stevioside and rebaudioside A, that were extracted from stevia.

Sample : 0.05% each, 20micro-L
1. Stevioside
2. Rebaudioside A

Column       : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm)
Eluent       : H2O/CH3CN=25/75
Flow rate    : 1.0mL/min
Detector     : UV (210nm)
Column temp. : 30deg-C

Aromatic Compounds

Benzoates were analyzed using RSpak DE-413 (a column for reversed phase chromatography).

Sample :
1. Methyl benzoate
2. Butyl benzoate
3. Hexyl benzoate

Column       : Shodex RSpak DE-413 (4.6mmID*150mm)
Eluent       : (left): CH3CN/H2O=65/35, (center): CH3CN/H2O=80/20, (right): THF/H2O=50/50
Flow rate    : 0.6mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Alkylbenzenes were analyzed using RSpak DS-613 (a column for reversed phase chromatography).

Sample :
R=H Benzene
Me Methylbenzene
Et Ethylbenzene
n-Pr n-propylbenzene
n-Bu n-Butylbenzene

Column       : Shodex RSpak DS-613 (6.0mmID*150mm)
Eluent       : THF/H2O=55/45
Flow rate    : 0.8mL/min
Detector     : UV(254nm)
Column temp. : 50deg-C

Alkylbenzenes were analyzed using RSpak DS-613 (a column for reversed phase chromatography).

Sample :
1. m-Cresol
2. 2,4-Xylenol
3. Benzene
4. Toluene
5. Ethylbenzene
6. n-Propylbenzene

Column       : Shodex RSpak DS-613 (6.0mmID*150mm)
Eluent       : CH3CN/THF/H2O=40/30/30
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Xylenols were analyzed using RSpak RP18-413 (a column for reversed phase chromatography).

Sample : 200micro-g/mL each, 20micro-L
1. 3,4-Xylenol
2. 3,5-Xylenol
3. 2,3-Xylenol
4. 2,6-Xylenol

Column       : Shodex RSpak RP18-413 (4.6mmID*150mm)
Eluent       : THF/H2O=35/65
Flow rate    : 0.5mL/min
Detector     : UV(254nm)
Column temp. : 50deg-C

Alkylbenzenes and naphtalene were analyzed using RSpak DE-413 (a column for reversed phase chromatography).

Sample : 1. Benzene, 2.Toluene, 3.Ethylbenzene, 4.n-Propylbenzene, 5.Naphthalene

Columns      : Shodex RSpak DE-413 (4.6mmID*150mm), ODS column (other manufacturer)
Eluent       : CH3CN/H2O=65/35
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 30deg-C

Polycyclic aromatic hydrocarbons were analyzed using RSpak DE-413 (a column for reversed phase chromatography).

Sample :
1. Benzene
2. Naphthalene
3. Anthracene

Column       : Shodex RSpak DE-413 (4.6mmID*150mm)
Eluent       : CH3CN/H2O=65/35
Flow rate    : 1.0mL/min
Detector     : UV(254nm)

Phenols were analyzed using RSpak DE-413 (a column for reversed phase chromatography).

Sample : 5micro-L

No. UV(220nm) Fluorescence(Ex.225nm Em.300nm)
1. Phenol 0.5ppm 50ppm
2. 4-tert-Butylphenol 1ppm 100ppm
3. .4-Octylphenol 1ppm 100ppm
4. Nonylphenol 1ppm 100ppm 
Column       : Shodex RSpak DE-413 (4.6mmID*150mm)
Eluent       : CH3CN/H2O=70/30
Flow rate    : 1.0mL/min
Detector     : UV(220nm), Fluorescence(Ex. 225nm, Em. 300nm)
Column temp. : 40deg-C

Chlorobenzenes were analyzed using RSpak DE-613 (a column for reversed phase chromatography).

Sample :
1. o-Chlorophenol
2. Benzene
3. Chlorobenzene
4. o-Dichlorobenzene
5. 1,2,4-Trichlorobenzene

Column       : Shodex RSpak DE-613 (6.0mmID*150mm)
Eluent       : CH3CN/H2O=70/30
Flow rate    : 0.8mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Iodobenzene diacetate was analyzed using GPC K-801 (a column for organic SEC (GPC)). The separation mode is SEC and the elution order is iodobenzene diacetate, iodobenzene acetate and iodobenzene.

Sample :
1. Iodobenzene diacetate
2. Iodobenzene acetate
3. Iodobenzene

Column       : Shodex GPC K-801 (8.0mmID*300mm)
Eluent       : CHCl3
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 40deg-C

Anilines were analyzed using Asahipak ODP-50 6D (a column for reversed phase chromatography).

Sample :
1. m-Aminophenylboronic acid
2. m-Aminophenol
3. o-Aminophenol
4. Aniline
5. m-Aminonitrobenzene

Column       : Shodex Asahipak ODP-50 6D (6.0mmID*150mm)
Eluent       : CH3CN/H2O=29/71
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 30deg-C

Perylene was analyzed using ODSpak F-411 (a column for reversed phase chromatography) and a chemiluminecsence detector (CLD).

Sample : 4.5fmol Perylene in CH3OH

Column       : Shodex ODSpak F-411 (4.6mmID*150mm)
Eluent       : 0.1mM Imidazol buffer(pH7.0,adjusted by nitrate)/CH3CN=1/5
Reagent      : [1mM *TCPO in Ethyl acetate]/[0.15M H2O2(30% aq.) in CH3CN]=25/75
Flow rate    : (Eluent); 0.5mL/min, (Reagent); 2.0mL/min 
Detector     : CLD
Column temp. : 30deg-C

*TCPO : Bis(2,4,6-trichlorophenyl) oxalate

Phenones were analyzed using Asahipak ODP-50 4D ( a column for reversed phase chromatography ), with alkaline eluent.

Sample :
1. Acetophenone
2. Butyrophenone
3. Hexanophenone
4. Heptanophenone
5. Octanophenone

Column       : Shodex Asahipak ODP-50 4D (4.6mmID*150mm)
Eluent       : 0.01M NaOH(pH12.0)/CH3CN=35/65
Flow rate    : 0.6mL/min
Detector     : UV(254nm)
Column temp. : 30deg-C

Isomers of Phenylenediamine analyzed using RSpak JJ-50 4D (a mutimode column) is shown.

Sample : 100ppm each, 20micro-L
1. p-Phenylenediamine
2. m-Phenylenediamine
3. o-Phenylenediamine

Column       : Shodex RSpak JJ-50 4D (4.6mmID*150mm)
Eluent       : 25mM CH3COONH4 buffer(pH9.2)/CH3CN=70/30
Flow rate    : 0.4mL/min
Detector     : UV(254nm)
Column temp. : 30deg-C

ODP2 HP is polymer based new reversed phase column. The theoretical plate number has improved nearly double comparing with current polymer based column ODP-50.

Sample : 5micro-L
1. Uracil 30mg/L
2. Theobromine 75mg/L
3. Caffeine 130mg/L
4. Phenol 300mg/L
5. Methyl benzoate 350mg/L
6. Toluene 1000mg/L
7. Naphthalene 150mg/L

Columns : Shodex ODP2 HP-4D (4.6mmID*150mm) : Shodex Asahipak ODP-50 4D (4.6mmID*150mm)
Eluent : H2O/CH3CN=55/45 : H2O/CH3CN=35/65
Flow rate : 0.6mL/min : 0.6mL/min
Detector : UV(254nm) : UV(254nm)
Column temp. : 40deg-C : 40deg-C

Imidazoles were analyzed using ODP2 HP-4E.

Sample : 0.1% each, 10micro-L
1. Imidazole
2. 2-Methylimidazole
3. 4-Methylimidazole

Column       : Shodex ODP2 HP-4E (4.6mmID*250mm)
Eluent       : 10mM Na2HPO4 aq./CH3CN=90/10
Flow rate    : 0.8mL/min
Detector     : UV(220nm)
Column temp. : 40deg-C