Page 56 - BR20828_E Accucore 0713S

2
Figure 1: Separation of naproxen, fenoprofen, and ibuprofen, with theophylline as a t0 marker.
Injections 1, 750, 1500, and 2,200 are shown.
mAU
min
0.50 0.0 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00
1
750
1500
2200
Separation Conditions Part Number
Instrumentation: Thermo Scientific DionexTM UltiMateTM 3000 HPLC system
Column: Accucore XL C8 4 μm, 50 x 2.1 mm 74204-052130
Mobile phase: Acetonitrile / 20 mM ammonium formate, pH 3 (40:60 v/v)
Backpressure: 50 bar
Column temperature: 30 °C
Injection volume: 2 μL (partial loop)
Flow rate: 0.3 mL/min
UV detection: 233 nm ( data rate 20 Hz)
Results
A stable analytical HPLC method was developed to assess the robustness of the Accucore XL HPLC column.
Using isocratic conditions, full separation of the non-steroidal, anti-inflammatory drugs (NSAIDs) naproxen,
fenoprofen, and ibuprofen was performed. The Accucore XL C8 HPLC column was shown to be stable for the
entire experiment with no indication of deterioration in chromatography even after 2,200 injections, which equates
to 45,000 column volumes.
The reproducibility of the Accucore XL HPLC column for the chromatographic separation of the NSAIDs is
summarized in Table 1. It is evident that the data for all analytes is matched with excellent precision with the
% RSD retention factor of ≤0.80% and % RSD normalized efficiency ≤1.52%. A typical chromatogram of the
NSAIDs is seen in Figure 1.
Figures 2 and 3 are illustrations of the stability data. It is apparent there was a degree of variability within the
middle of the analysis, brought about by instrument related issues. However, there is no indication of instability
of the Accucore XL HPLC column, even after 2,200 injections, as both the retention and efficiency remained
consistent to the end of the analysis.
Table 1: Assessment of the stability on the Accucore XL C8 HPLC column assessed for 2,200 injections.
Efficiency was assessed by normalizing efficiency values to the mean of the efficiencies for the first
five samples injected.
Naproxen Fenoprofen Ibuprofen
k' N/N(n1-5) k' N/N(n1-5) k' N/N(n1-5)
Mean 2.26 0.986 4.80 0.996 7.99 1.002
% RSD 0.80 1.52 0.77 0.66 0.77 0.36