Page 9 - Shimadzu’s Solutions for Impurities Analysis

Analysis of Ultra-Trace Impurities
Co-Sense for LC/MS System
2D-LC/LCMS-IT-TOF ®
Data

Sulfadimethoximne-based sulfa drug impurities with four 2ndD-LC Unk-1
similar structures were prepared as a model sample. The uV(x1,000)
1.50
UV chromatogram
main components of the impurities were adjusted at 500 1.25 At sample injection
1.00 At solvent injection
ug/ml. The structures are described below. The 0.75
0.50
measurement results are shown in Fig.1 to 3 and every 0.25
®
impurity was confirmed. The 2D-LC/LCMS-IT-TOF System 0.00 FDA Regulations on
-0.25 Genotoxic Impurities
makes it possible for researchers to eliminate troublesome 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min
LC/MS measurement under volatile mobile phase (x1,000,000)
1.00 1:TIC (1.00) 1:265.0757 (8.21) Mass chromatogram
conditions. Impurities can be confirmed in comparison 0.75
0.50
with the blank data, enabling effective identification of
0.25
impurities at CMC. 0.00
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5
pseudo main Inten.
components: H 3C Sulfamerazine (x1,000,000)
Sulfadimethoxine O
7.5 265.0757 [M+H] + Mass spectrum
NH 2 1+
N N NH 2 Error: 1.24 ppm 1
N 2
O 5.0 6 6
O + & & 1 1+
H 3C S 2
O NH S Composition Formula= C11H12N4O2S
O H 3C N NH 2.5
O Monoisotopic Mass = 264.068096 Da
[ M + H ] + = 265.075372 Da
Composition Formula=C12H14N4O4S Composition Formula=C11H12N4O2S 0.0
Monoisotopic Mass=310.073575 Da Monoisotopic Mass=264.068096 Da 100 200 300 400 500 600 700 800 900 m/z
[M+H] + =311.080851 Da [M+H] + =265.075372 Da
Fig 2. UV chromatogram, MS chromatogram and
Sulfadimidine Sulfamonomethoxine Sulfaquinoxaline MS spectrum of Unk-1
Pink in UV chromatogram: At sample injection
CH 3 Residual Solvents FDA Regulations on
N
NH 2 NH 2 NH 2 Black in UV chromatogram: At solvent injection (blank)
N N N
O O O Red arrow: Impurity
H 3C N NH S H 3C O NH S N NH S
O O O
Composition Formula=C12H14N4O2S Composition Formula=C11H12N4O3S Composition Formula=C14H12N4O2S
Monoisotopic Mass=278.083746 Da Monoisotopic Mass=280.06301 Da Monoisotopic Mass=300.068096 Da
+ + =281.070287 Da
[M+H] =279.091022 Da [M+H] [M+H] + =301.075372 Da
2ndD-LC Unk-2
uV(x1,000)
mAU(x10) MPa 1.00 UV chromatogram
3.75 19.0 At sample injection
270nm,4nm(1.00) Main 18.0 0.75
3.50 At solvent injection
17.0
3.25 16.0 0.50
3.00 15.0
2.75 14.0 0.25
2.50 13.0 0.00
2.25 12.0
2.00 11.0 -0.25
1.75 10.0
9.0
1.50 Loop#4 Loop#5 Loop#6 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min
Loop#1 Loop#2 Loop#3 8.0
1.25 7.0
1.00 (x100,000,000)
6.0
0.75 5.0 1.00 1:TIC (1.00) 1:279.0905 (4.76) Mass chromatogram
Pump pressure fluctuation
0.50 4.0 0.75
0.25 Unk-1 Unk-2 Unk-3 Unk-4 3.0 0.50
0.00 2.0 Inorganic Impurities FDA Regulations on
1.0 0.25
-0.25 PDA chromatogram (270nm)
-0.50 0.0 0.00
0.0 2.5 5.0 7.5 10.0 12.5 15.5 17.5 20.0 22.5 min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5
Fig.1 1st dimensional PDA chromatogram
Inten.
Blue line illustrates variation of system pressure and black (x10,000,000) &+
1.5 279.0905 [M+H] + 1+ Mass spectrum
line shows trace at UV270nm. Both conform to each other Error: 1.88 ppm 1 2
1.0 + & 1 1+ 6
and the elution peaks are fractionated inside the loop. Composition Formula = C12H14N4O2S
2
0.5
Monoisotopic Mass = 278.083746 Da
[ M + H ] + = 279.091022 Da
0.0
Analytical Conditions <LC 1stD>
100 200 300 400 500 600 700 800 900 m/z
Column : Shim-pack VP-ODS, 4.6mm i.d.×150mm, 5 μm
Mobile phase : 0.01mol/L Phosphate buffer (pH2.6) Fig. 3 UV chromatogram, MS chromatogram and
Methanol mixture MS spectrum of Unk-2
Mobile phase flow rate : 1 mL/min
Column temperature : 40ºC
Sample injection volume : 10 μL
PDA detection wavelength : 200-350 nm (monitoring 270 nm)
Analytical Conditions <LC 2ndD>
Column : Shim-pack XR-ODS 2.0mm i.d.×75mm, 2.2 μm
Mobile phase A : 0.1% Formic acid solution
Mobile phase B : Methanol
Mobile phase ratio : 10%B (0 min)-50%B (10 min)-10%B (10.01-20 min) Support from Shimadzu
Mobile phase flow rate : 0.3 mL/min
Column temperature : 40ºC
Sample injection volume : 10 μL (Loop capacity)
UV detection wavelength : 270 nm
<MS>
Ionization mode : ESI+
Nebulizer gas flow rate : 1.5 L/min
Drying gas pressure : 0.15 MPa
Applied voltage : 4.5 kV Ref: Technical Report (Improved Drug Impurity
CDL temperature : 200ºC
BH temperature : 200ºC ID Efficiency under CMC using 2-D LC/MS) C146-E149
Scan range : m/z 100-1000
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