Page 94 - Application Handbook - Liquid Chromatography

LAAN-A-LC-E226

Application High Performance Liquid Chromatography

News Ultra-High Speed Analysis of Ibuprofen by Nexera

in Accordance with USP 621
No.L448

High throughput analysis has been advancing n High Speed Analysis of Impurities in Ibuprofen
dramatically in recent years with the increasing with UHPLC Column
necessity to improve productivity and operational
efficiency. HPLC has also been in the spotlight thanks to Ibuprofen is a type of non-steroidal anti-inflammatory drug
significant advances in ultra-high-speed analysis (NSAID) that is used as an antipyretic and analgesic.
technology, in particular ultra high performance LC and Monographs on ibuprofen-related substances using
micro-particle column packing material. The recently conventional columns associated with the USP column
1)
revised General Chapter 621 of the United States category L1 (C18) are listed in the USP-NF , in which the
Pharmacopoeia (USP 621) now permits a degree of analysis method and system suitability are specified for three
adjustment of HPLC and GC parameters, specifically substances; ibuprofen and its degradation products
aimed at satisfying the requirements of system 4-isobutyl acetophenone and valerophenone (internal
suitability. standard substance).
Here, using the Nexera ultra high performance liquid This paper presents our investigation into speeding up the
chromatograph and the Shim-pack VP-ODS analysis of ibuprofen-related substances listed in USP-NF in
conventional column, in addition to the Kinetex XB-C18 compliance with USP 621. For the analytical column, we
Series Core-Shell fast analysis column, we introduce selected the Kinetex XB-C18 (100 mmL. × 4.6 mm ID,
examples of high-speed analysis of ibuprofen-related 2.6 µm) high-speed analytical column, which is within the
substances in conformance with USP 621. acceptable range of column adjustment specified in USP 621.
Except for the column, all other conditions were the same as
n Allowable Adjustments to HPLC Parameters those listed in USP-NF. Fig. 1 shows the results of analysis of
a mixtur e of Ibupr ofen, Valer ophenone and
Table 1 shows the parameters which may be changed 4-Isobutylacetophenone using the Shim-pack VP-ODS and
according to USP 621, such as column length, particle Kinetex XB-C18 columns, respectively, and Table 2 shows the
size and flowrate, etc., in addition to the actual analytical conditions used for each. Use of the Kinetex XB-
permissible ranges within which these LC parameters C18 made it possible to reduce both the analysis time and
may be changed. After the allowable changes are solvent consumption to about 1/4 without compromising the
implemented, no re-validation is required since the separation. The results of the system suitability test are
changes are interpreted only as method adjustments. shown in Table 3 on the following page. The results of this
study clearly indicate that all of the system suitability
Table 1 Allowable Adjustments to HPLC Parameters According to USP 621 requirements have been met using the Kinetex XB-C18.
USP General Chapter <621>
Column Length ±70 % change allowed mAU
Column Internal Changes permitted as long as linear velocity is 300 Shim-pack VP-ODS (250 mmL) 2
Diameter the same 250 1
May be reduced to 50 % at maximum. 200
Particle Size
However, may not be increased. 150
Flowrate ±50 % change allowed 100
Column Temperature ±10 °C change allowed 50 3
Change is permitted if it satisfies the
Injection Volume 0
requirements for system suitability.
pH ±0.2 change allowed 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0
UV Wavelength ±3 nm change allowed mAU
300
Salt Concentration ±10 % change allowed 250 KInetex XB-C18 (100 mmL) 2
Mobile Phase The smaller of ±30 % or ±10 % of absolute 200 1
Composition volume to be selected.
150
100
Table 2 Analytical Conditions 50 3
System : Nexera Method Scouting 0
Column : (1) Shim-pack VP-ODS (250 mmL. × 4.6 mm I.D., 4.6 µm) 0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00
(2) Kinetex XB-C18 (100 mmL. × 4.6 mm I.D., 2.6 µm) ■Peaks
Mobile Phase : A: 1 % (wt / v)Chloroacetic Acid Water 1. Ibuprofen (12 mg/mL)
(pH 3.0 adjusted with ammonium hydroxide) 2. Valerophenone (0.35 mg/mL)
B: Acetonitrile 3. 4-Isobutylacetophenone (0.012 mg/mL)
A/B = 2/3 (v/v)
Flowrate : 2.0 mL/min Fig. 1 Comparison of the Shim-pack VP-ODS and Kinetex XB-C18
Column Temp. : 30 °C Series Columns -Chromatograms of a Standard Mixture of
Injection Vol. : (1) 5 µL Ibuprofen, 4-Isobutylacetophenone and Valerophenone
(2) 1 µL Upper: Shim-pack VP-ODS (250 mmL)
Detection : SPD-20AV at 254 nm Lower: Kinetex XB-C18 (100 mmL)