Page 100 - Application Handbook - Liquid Chromatography

LAAN-A-LC-E255

Application High Performance Liquid Chromatography

News High-Speed Analysis of Impurities of Pramipexole

Dihydrochloride by Prominence-i
No.L478

High-resolution, high-sensitivity chromatograms can be
obtained using a column with small-size particle
packing material. However, as the particle size mV
decreases, the pressure applied to the column increases. 2500
The Kinetex 5 µm core-shell column, in which the
applied pressure is equivalent to that using a 5 µm 2000 N: 28500 Pramipexole
particle column, permits analysis at very high resolution.
Therefore, high-resolution, high-sensitivity 1500
chromatograms can be obtained even when using a
general-purpose LC system. 1000 N: 17500
Here, we introduce an example in which we used the 500 (a)
Kinetex 5 µm core-shell column in the Prominence-i (b)
integrated high-performance liquid chromatograph, 0
with the aim of speeding up the analysis of a drug-
related substance in accordance with criteria 0.0 5.0 10.0 15.0 min
requirements of the European Pharmacopoeia (EP). mV
10
9
n Analysis of Impurities in Pramipexole Dihydrochloride 8 Pramipexole
Using the Kinetex 5 µm Core-Shell Column
Pramipexole dihydrochloride, known as a therapeutic 7
agent for Parkinson's disease, is used as a drug to 6
replenish the neurotransmitter dopamine. (a) RS : 3.341
Here, as specified in the EP, a standard solution of 5
pramipexole dihydrochloride (1.5 mg/mL) was analyzed 4 E A B C
using the analytical conditions of Table 1. 1)
Chromatograms (a) and (b) of Fig. 1 correspond to the 3
measurement results using the Kinetex 5 µm core-shell 2
column and the Shim-pack VP-ODS column, (b) B C
respectively. The lower part of the figure shows an 1 E A
enlarged view of the chromatograms from 3 to 10 0
minutes. Use of the Kinetex 5 µm core-shell column
increased sensitivity for the principal component, and 3.0 5.0 7.5 min
the circled peak of Fig. 1 demonstrates that peak
separation is achieved. In addition, the elution time of
the principal component occurs about two minutes
earlier than when using the Shim-pack VP-ODS. Fig. 1 Comparison of Chromatograms of Pramipexole Dihydrochloride
Using (a) Kinetex 5 µm Core-shell Column and (b) Shim-pack
VP-ODS
Table 1 Analytical Conditions
n Compatibility with Third-Party LC Systems
Column (1) : Kinetex 5µ C18 100A (150 mm L × 4.6 mm I.D., 5 µm)
Column (2) : Shim-pack VP-ODS (150 mm L × 4.6 mm I.D., 5 µm) The Prominence-i is compatible with LC systems of
Flowrate : 1.5 mL/min other companies. Here, we conducted analysis of a
Mobile Phase : A) 67 mmol/L Phosphate (Potassium) Buffer (pH 3.0)
Containing 21 mmol/L 1-Octanesulfonic Acid standard solution of pramipexole dihydrochloride
Sodium Salt (1.5 mg/mL) using the analytical conditions shown in
B) Solution A / Acetonitrile (1/1)
Time program : B Conc. 40 % (0 min) → 80 % (15 min) Table 1 (using Column 1) in a similar manner as used in
→ 40 % (15.1 - 20 min) the previous section. The chromatogram in the upper
Column Temp. : 40 ˚C
Injection Volume : 5 µL window of Fig. 2 was acquired using the Prominence-i,
Detection : LC-2030C at 264 nm and those in the lower window are enlarged views of
Flow Cell : Conventional Cell for Integrated
chromatograms acquired with (a) a third-party LC
system, and (b) the Prominence-i. According to the
results using the third-party system and the
Prominence-i, there is clear compatibility of
chromatogram separation.