Tacrolimus Standard solution 30 µg/ml. USP Organic impurities Procedure-2

LC Conditions

Column

Luna 3 µm C18(2) 100 Å, LC Column 150 x 4.6 mm, Ea

Brand

Luna

Part No

00F-4251-E0

Phase Name

C18(2)

Elution Type

Gradient

Mobile Phase

A: Solution -A: - 6mM Phosphoric acid in milliQ water. (0.4ml of 85% Orthophosphoric acid in 1000ml of water, filtered through 0.22µm membrane nylon filter.) Solution- B: - A Mixture of Acetonitrile tert-butyl methyl ether in ratio of 810:190ml v/v. Mobile phase A: Solution A and Solution B in a ratio of 4:1 (400:100 ml v/v)

B: Solution -A: - 6mM Phosphoric acid in milliQ water. (0.4ml of 85% Orthophosphoric acid in 1000ml of water, filtered through 0.22µm membrane nylon filter.) Solution- B: - A Mixture of Acetonitrile tert-butyl methyl ether in ratio of 810:190ml v/v. Mobile phase B: Solution A and Solution B in a ratio of 1:4 (400:400 ml v/v)

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

72

28

2

30

72

28

3

53

15

85

4

54

72

28

5

60

72

28

Flow Rate

1.50 mL/min

Temperature

60°C

System

Waters Arc HPLC Waters HPLC System

Detection

UV-Vis Abs.-Diode Array (PDA) @ 220 nm (nanometers)

Detection Info