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Blossmate ® Phenyl 2.1mm×100mm, 3.5 μm, 450Å Solve the problem of semaglutide residue

Blossmate Phenyl 2 Solve the problem of semaglutide residue

Test Report

Chromatography conditions:

Instruction information

Xevo TQ-S micro

Column 

Blossmate ® Phenyl 2.1mm×100mm,3.5 μm,450Å

Mobile Phase

A: 0.02% difluoroacetic acid in acetonitrile solution; B: 0.4% formic acid in water;

Column Temperature

65 ℃

Injection Volume

5μL

Flow Rate

See reference chromatographic conditions

Reference chromatographic conditions

Time (min)

Flow rate mL/min

A(%)

B(%)

0

0.2

40

60

6.0

0.2

90

10

6.5

0.8

90

10

7.0

0.8

90

10

8.0

0.8

95

5

12.0

0.8

50

50

14.0

0.8

80

20

14.4

0.8

40

60

14.5

0.2

40

60

15.0

0.2

40

60


Note

/

Chromatogram and Data: 


Mass Spectrometry Conditions:
Ion Source: ESI

Detection Mode: MRM
Drying Gas: Nitrogen, 400 ℃,

Flow Rate: 1000 L/Hr
Collision Gas: Argon Ion

Spray Voltage:1kV


Solution preparation

1. Preparation of Mobile Phase:
0.1% Formic Acid Aqueous Solution: Take 0.05 mL of formic acid, dilute with water to 100 mL, mix well, and set aside;
0.02% Trifluoroacetic Acid Acetonitrile Solution: Take 0.1 mL of trifluoroacetic acid, dilute with acetonitrile to 500 mL, mix well, and set aside.


2. Preparation of Solutions:
Methanol-Water Solution (50+50): Take 100 mL of methanol, add an equal volume of water, mix well, and set aside.


3. Preparation of Standard Solutions:
Dissolve the Somatropin standard substance in pure water to a concentration of 1000 mg/L; Dilute the 1000 mg/L Somatropin standard substance with 50% methanol-water to concentrations of 20 mg/L, 1 mg/L, 50 ng/L, 20 ng/L, 10 ng/L, and 5 ng/L; The 1 mg/L Somatropin standard dilution solution is used for mass spectrometry tuning after adding an appropriate amount of formic acid.


4. Sample Volume:


1) 20 mg/L Standard Injection Solution:


2) Blank injection 1:


3) Blank injection 2:


4) Blank injection 3:


5) Blank injection 4:


6) Blank injection 5:


7) 5 ng/L standard solution:


8) 10 ng/L standard solution:


9) 20 ng/L standard solution:


10) 50 ng/L standard solution:


Conclusion:

Using the Welch Blossmate® Phenyl 2.1mm×100mm, 3.5 μm, 450Å HPLC column, after injecting the 20 mg/L Somatropin standard injection solution in the mobile phase system, blank solution (50% methanol-water) was injected. The first blank injection showed approximately 2 ng/mL, and subsequent blank injections showed no significant residue.