ACE UHPLC and HPLC Columns
Analytes
0 9 10 11 12
900
800
700
600
500
400
300
200
100
VVVitamins in Fruit Juice by Fast LC-MS
182
Analytes
1
2
4
123456789
3
5
6
5
6
7
8
9
0 9 10 11 12
10
10
Vitamins – Water Soluble (Isocratic II)
Analytes
900
800
700
600
500
400
300
200
100
0.0 0.0
0.5 0.5
1.0 1.0
1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 min
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Application #AN2610
Conditions
Column: ACE Excel 3 C18-PFP
Dimensions: 100 x 2.1 mm
Part Number: EXL -1110-1002U
Mobile Phase: A: 15 mM formic acid, adjusted to
pH 3.8 with ammonia solution
B: MeOH
Gradient: Time (mins) %B
0.00 1
1.00 1
3.00 8
3.10 25
6.00 50
6.50 50
6.51 1
9.00 1
Flow Rate: 0.4 mL/min
Temperature: 30 °C
Detection: LCMS-8040 triple quad MS
ESI positive mode (ESI negative
for ascorbic and citric acid)
DL temperature: 250 °C
Heat block temperature: 400 °C
1. Thiamine (Vitamin B1)
(m/z 266.10 122.15)
2. Pyridoxine (Vitamin B6)
(m/z 170.20 152.15)
3. Nicotinic acid (Vitamin B3)
(m/z 124.00 78.00)
4. Pantothenic acid (Vitamin B5)
(m/z 220.30 90.05)
5. Cyanocobalamin (Vitamin B12)
(m/z 678.50 147.05)
6. Riboflavin (Vitamin B2)
(m/z 377.20 243.10)
7. Biotin (Vitamin B7)
(m/z 245.10 227.05)
8. Nicotinamide (Vitamin B3)
(m/z 123.20 80.05)
9. Ascorbic acid (Vitamin C)
(m/z 175.10 114.80)
10. Citric acid
(m/z 191.10 87.15)
3.0
3.5
4.0
Time - Minutes
1
2
3
4
7
8
9
10
123456789
1
1
2
2
3
3
4
4
5
5
6
6
7
7
8
8
9
9
10
10
10
1.5
2.0
2.5
4.5
5.0
5.5
6.0
6.5
0.0 0.0
0.5 0.5
1.0 1.0
1.5 1.5
2.0 2.0
2.5 2.5
3.0 3.0
3.5 3.5
4.0 4.0
4.5 4.5
5.0 5.0
5.5 5.5
6.0 6.0
6.5 6.5
min
Reproduced with permission of Shimadzu Europe GmbH
Standard
Juice sample
Application #AN2980
Conditions
Column: ACE 5 C8
Dimensions: 250 x 4.6 mm
Part Number: ACE-122-2546
Mobile Phase: 50 mM KH2PO4 pH 2.5 in
H2O/MeOH (97:3 v/v)
Flow Rate: 1 mL/min
Temperature: Ambient
Detection: UV, 205 nm
1. Pyridoxamine
2. Thiamine (Vitamin B1)
3. L -Ascorbic acid (Vitamin C)
4. Niacinamide (Vitamin B3)
5. Pyridoxal
6. Nicotinic acid
7. Pyridoxine
1 2 3 4 5 6 7 8
Time - Minutes
0
2
1
3
4 5
6
7
Vitamins – Water Soluble (Isocratic I)
Application #AN2990
Conditions
Column: ACE 5 C18
Dimensions: 250 x 4.6 mm
Part Number: ACE-121-2546
Mobile Phase: 50 mM KH2PO4 pH 3.0
in H2O/MeOH (97:3 v/v)
Flow Rate: 1 mL/min
Temperature: Ambient
Detection: UV, 205 nm
1. Pyridoxamine
2. Thiamine (Vitamin B1)
3. L -Ascorbic acid (Vitamin C)
4. Nicotinic acid
5. Pyridoxal
6. Impurity
7. Pyridoxine
1 2 3 4 5 6 7 8
Time - Minutes
0
2
1
3
4
5
6
7
Response - MilliVolts
Response - MilliVolts