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Figure 3 Optimizing retention with the Allure PFP Propyl gives
high sensitivity and low matrix interference when analyzing fluoro-
quinolones by LC/MS/MS.
2007.01
Foods, Flavors & Fragrances
Allure® PFP Propyl Columns (USP L43)
Excellent Columns for LC/MS and ELSD
5µm Column, 4.6mm cat. # price
150mm 9169565 $405
150mm (with Trident™ Inlet Fitting) 9169565-700 $420
ordering note
For guard cartridges for these columns, visit our website at
.
interactions, we evaluated a cyanopropyl phase
under the same conditions. This combination
produced similar retention, but less selectivity
(Figure 2). In contrast, an Allure® PFP Propyl col-
umn (pentafluorophenyl propyl phase), used
under the same conditions, enabled us to achieve
comparable retention capacities with the water
content of the mobile phase reduced to 40%
(Figure 2). In addition to greater retention capac-
ity than the other phases, the Allure® PFP Propyl
stationary phase has better selectivity – unlike
with the C18 and cyano phases, there are no coelu-
tions.
Another advantage to the Allure® PFP Propyl col-
umn’s high retention capacity for fluoro-
quinolones is in LC/MS analysis. Maximizing
retention causes the analytes to elute in mobile
phases having higher percentages of the organic
component. This can increase desolvation efficien-
cy in electrospray ionization (ESI), and can elimi-
nate unwanted adduct formation or charge com-
petition from matrix interferences that are less
retained by the column. The result is a potential
for increasing sensitivity, while using simple ana-
lytical conditions. A simple mobile phase gradient,
starting with a highly aqueous content and mov-
ing to a highly organic content, can be employed to
elute salts and low molecular weight sample
matrix interferences ahead of the compounds of
observed the same improved retention
when we assayed our fluoroquinolone test mix
through positive ESI LC/MS/MS on an Applied
Biosystems/MDS SCIEX API 3200 triple quadru-
pole LC/MS/MS mass spectrometer equipped with
a Shimadzu Prominence binary pump LC system
(Figure 3).
The Allure® PFP Propyl phase will retain polar
analytes much more effectively than a C18 phase.
When greater retention is needed to give the
desired selectivity, or when LC/MS analysis is
desired or required, simplify your method – use an
Allure® PFP Propyl column and a simple mobile
phase rather than a C18 column and an ion-pair-
ing technique.
Reference
1. United States Pharmacopoeia, 28th revision; National Formulary, 23rd
edition.
Sample:
Inj.: 5µL
Conc.: 50ng/mL
Sample diluent: mobile phase
Column: Allure ® PFP Propyl
Cat. #: 9169352
Dimensions: 50 x 2.1 mm
Particle size: 3µm
Pore size: 60Å
Conditions:
Instrument: Shimadzu Prominence HPLC
Mobile phase: A: 0.1% formic acid in water
B: 0.1% formic acid in acetonitrile
Time (min.) %B
0.00 10
10.00 90
10.10 10
15.00 10
Flow: 300µL/min.
Temp.: 30°C
Det.: Applied Biosystems API 3200
Triple Quadrupole LC/MS/MS
Mass Spectrometer
Ion mode: positive
Temp.: 600°C
Ion source: TurboIonSpray® , Electrospray 4000V
Precursor Fragment Declustering Collision
Compound Ion Ion Potential (V) Energy (V)
1. Norfloxacin 319.9 276.0 36.00 23.00
233.1 36.00 35.00
2. Ciprofloxacin 332.1 288.2 41.00 23.00
244.9 41.00 31.00
3. Levofloxacin 362.1 318.1 31.00 25.00
261.0 31.00 41.00
4. Lomefloxacin 351.9 265.1 41.00 29.00
308.0 41.00 23.00
5. Enrofloxacin 360.2 316.1 36.00 25.00
245.3 36.00 37.00
6. Sparfloxacin 393.1 349.4 36.00 25.00
292.2 36.00 29.00
1. norfloxacin
2. ciprofloxacin
3. levofloxacin
4. lomefloxacin
5. enrofloxacin
6. sparfloxacin
LC_PH0426
thank you
Instrument provided courtesy
of Applied Biosystems