Page 101 - Application Handbook - Liquid Chromatography

Application No.L478
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Fig. 3 shows the chromatogram obtained from analysis of a
pramipexole dihydrochloride standard solution (1.5 mg/mL)
mV
2000 using the analytical conditions of Table 3, in which the
permissible limits of Table 2 are satisfied. The lower
Pramipexole chromatogram presents an expanded view of the region from
1500 1 to 3 minutes. To conduct the accelerated analysis, the
optional "Low-volume Tubing Kit" was used, and for the
column, the Kinetex 2.6 µm core-shell column for high-speed
1000 analysis was used. The elution time of the principal
component occurred about three minutes earlier than when
using the Kinetex 5 µm core-shell column.
500
In addition, it is clear from the results of the system suitability
test shown in Table 4, the system suitability requirement is
0 fully satisfied in all respects, including conforming to the EP
conditions as well as from the standpoint of satisfying the
0.0 2.5 5.0 7.5 min
conditions associated with high-speed analysis.
mV
Table 3 Analytical Conditions - High Speed Analysis
Pramipexole Mobile Phase : A) 67 mmol/L Phosphate (Potassium) Buffer (pH 3.0)
5 Column : Kinetex 2.6µ C18 100A (75 mm L × 4.6 mm I.D., 2.6 µm)
: 2.0 mL/min
Flowrate
4 Containing 21 mmol/L 1-Octanesulfonic Acid
Sodium Salt
B) Solution A / Acetonitrile (1/1)
Time program : B Conc. 40 % (0 min) → 80 % (5.63 min)
→ 40 % (5.64 - 8.63 min)
3 Ontime injection: 230 µL
Column Temp. : 40 ˚C
B Injection Volume : 2 µL
2 E A C Detection : LC-2030C at 264 nm
Flow Cell
: High-speed High-sensitivity Cell
(a) Table 4 Results of System Suitability Test According to EP
1
System Reference EP-Equivalent High-Speed
(b) Suitability Item Value Condition Condition Judgment
Resolution
0
- Impurity A and ≧ 6.0 8.2 9.7 PASS
Pramipexole
0.0 2.5 5.0 7.5 min
mV
Fig. 2 Chromatograms of Pramipexole Dihydrochloride 1500
Upper: Prominence-i
Lower: Enlarged Views of Data Using (a) Third-Party 1000 Pramipexole
System and (b) Prominence-i
500
n High-Speed Analysis of Impurities of Pramipexole
Dihydrochloride 0
We investigated the speed-up of analysis of pramipexole
hydrochloride-related substances in accordance with the 0.0 1.0 2.0 3.0 min
EP. Table 2 shows the permissible ranges within which the mV
HPLC parameters may be changed as stipulated in the EP. 2) 3
Pramipexole
2
Table 2 Permissible Ranges of HPLC Conditions Specified in EP
Item Permissible Range 1 B
Mobile Phase (Minor substances) Select ±30 % or absolute quantity ±2 %, E A C
whichever is larger
(Other substances) Absolute quantity ±10 % change is permitted 0
pH ±0.2 % change permissible
Salt Concentration ±10 % change permissible
UV Wavelength No change permitted 1.0 1.5 2.0 2.5 3.0 min
Column Inner Diameter ±25 % change permissible
Particle Size Decrease up to 50 % permissible,
increase not permitted Fig. 3 Chromatograms of Pramipexole Dihydrochloride – High-Speed Analysis
Flowrate ±50 % change permissible (with Low-volume Tubing Kit)
Column Temperature ±10 % change permissible (Max 60 °C)
Injection Volume Change permissible as long as system [References]
suitability requirement is satisfied 1) E. Pharmacopoeia 01/2012: 2416 “Pramipexole Dihydrochloride”
2) E. Pharmacopoeia 01/2008: 20246 General Chapters
2. 2. 46. Chromatographic separation technique

First Edition: Jan. 2015

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