Vitamins (WaterSoluble)
HPLC
UltraAqueousC18
Vitamins (FatSoluble)
HPLC
UltraC18
FOODS, FLAVORS, & FRAGRANCES
12
Phone: 800-356-1688 or 814-353-1300
Vitamins
Vitamins
Vitamins play vital roles in the human body, and
include a wide range of organic compounds. The
quantitative analysis of vitamins has become a
necessity in both themainstream food and dietary
supplement markets. However, because the body
needs vitamins in very small quantities, the analyst
oftenmust perform trace-level analyses, which can
be challenging. Vitamin assays are used to ensure
product quality and to verify nutritional label
claims. In addition, assays are important in devel-
opingmanufacturingand storageprocessesbecause
manyvitamins are light- and/or air-sensitive.
Vitamins can be classified broadly into two
groups—water-soluble and fat-soluble. Fat-soluble
vitamins includeA (retinol), E (alpha-tocopherol),
D, andK.They are quite hydrophobic andmust be
dissolved in an organic solvent. The
Ultra C18
HPLC column features a retentive, high-purity
packing that is ideal for separating a range of fat-
soluble vitamins.The fully end-capped silica elimi-
nates unwanted analyte-silanol interactions and
improves column-to-column reproducibility.
ordering
note
Please see pages 40–45 for HPLC columns featured in this catalog.
Peak List:
Conc.: (mg/mL)
1. solvent front
n/a
2. menadione (vitamin K
3
)
0.45
3. all-
trans
-retinol (vitamin A)
0.34
4. vitaminD
3
0.4
5. unknown
n/a
6. alpha tocopherol (vitamin E)
2.4
7. alpha tocopherol acetate
(vitamin E acetate)
2.4
8. unknown
n/a
9. phylloquinone (vitamin K
1
)
0.84
Sample:
Solvent:
diethyl ether
Column:
Ultra C18
Cat.#:
9174575
Dimensions:
250 x 4.6mm
Particle size:
5µm
Pore size:
100Å
Conditions:
Mobile phase: acetonitrile:methanol (90:10, v/v)
Time (min.)
Flow (mL/min.)
1.00
1.00
5.00
1.00
5.01
2.00
50.0
2.00
Temp.:
30°C
Det.:
UV@280nm
1
2
3
4
5
6
7
8
9
min.
0
10
20
30
40
LC_0144
min.
0
2
4
6
8
10 12 14
1
2
3
4
5 6
9
7
8
Peak List:
Conc.: (mg/mL)
1. thiamin (B
1
)
250
2. ascorbic acid (C)
1000
3. unknown
n/a
4. nicotinic acid (B
3
)
1000
5. unknown
n/a
6. pantothenic acid (B
5
)
1000
7. folic acid (B
9
)
500
8. riboflavin (B
2
)
250
9. methyl paraben
0.2
Sample:
Initial dilutions of B
1
and B
2
basifiedwith
ammonium hydroxide (to promote solubility)
Column:
Ultra Aqueous C18
Cat.#:
9178575
Dimensions:
250 x 4.6mm
Particle size:
5µm
Pore size:
100Å
Conditions:
Mobile Phase: A: 25mM potassium phosphate,
pH 2.00:methanol (95:5, v/v)
B: methanol:25mM potassium
phosphate, pH 3.5 (60:40, v/v)
Time (min.)
%B
0-6
0
6.01
25
6.01-11
25-100
11-16
100
Flow:
1.0mL/min.
Temp.:
27°C
Det.:
UV@254nm
LC_0141
restek
innovation
!
Monitorwater-soluble vitaminswithout
ion pairing reagents.
