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Application High Performance Liquid Chromatography

News Analysis of Omeprazole by "i-Series" for USP and JP

Methods
No.L494

Omeprazole, a drug that effectively suppresses the excessive n Speed Enhancement for USP Method
secretion of gastric acid, is often used for the treatment of The permissible ranges within which the analytical
gastric ulcer and duodenal ulcer, in addition to the treatment conditions may be modified are specified in the USP General
of reflux esophagitis. Acting as a Proton Pump Inhibitor (PPI), Chapters: <621> Chromatography. Changing these
omeprazole is included in the WHO Model List of Essential analytical conditions within range makes it possible to
Medicine, and considered an important component of basic shorten the analysis time. For details regarding changes that
medical care. can be used to allow fast USP-compliant analysis, please
This Application News introduces an example of analysis of refer to Application News L464.
omeprazole in accordance with the Japanese Pharmacopoeia Shortening analysis time can be accomplished in two ways,
(JP) and the United States Pharmacopeia (USP). Also 1) by shortening the column, and 2) by increasing the
presented here is an example of analysis that can be flowrate (linear velocity). To preserve the resolution of the
completed in a significantly shorter time than that described column, the column length and particle size may be modified
in the USP General Chapter 621 Chromatography. as long as the ratio of L (column length) to dp (column
The Nexera-i integrated UHPLC was used for the analysis by particle size) remains in the specified range (permissible
the procedure described in the USP. The Nexera-i supports range: -25 % to +50 %). We selected a column size of
the use of analytical conditions specified for both HPLC and 50 mmL. × 3.0 mm I.D., and 2 µm particle size. For further
UHPLC. In the case of compliance (HPLC conditions) with details, please see Table 2. The flowrate, proportional to the
the Japanese Pharmacopoeia, we conducted analysis using column cross-sectional area, and inversely proportional to the
the Prominence-i integrated HPLC. particle diameter (see text for permissible limits), was
determined as 0.85 mL/min.
The instrument used for the analysis was the Nexera-i high-
n The USP Method - Original Method speed integrated UHPLC, suitable for multi-sample
The analytical conditions specified in the USP monograph processing. The Nexera-i permits analysis using both HPLC
are shown in Table 1. The results of analysis of the system and UHPLC conditions, without requiring changes to
suitability test solution (0.1 mg/mL, acetonitrile-boric acid plumbing or flow cell type. This flexibility can allow legacy
HPLC methods to be quickly transferred to UHPLC speed
solution) specified in the omeprazole test method are and performance.
shown in the upper chromatogram of Fig. 1. The results Table 3 shows the analytical conditions using the higher
obtained sufficiently satisfy the threshold required with speed analysis, and the chromatogram obtained from
respect to both tailing factor and relative standard analysis of the system suitability test solution is shown in the
deviation (n = 6) specified in the monograph (Table 4). lower part of Fig. 1. The analysis time was reduced more
than 80 percent compared to that using the analytical
conditions of Table 1 (Fig. 1 upper).
Table 1 Analytical Conditions (USP Original Method) The results of the system suitability test are shown in Table 4.
Clearly, the threshold value has been satisfied even using
System : Nexera-i high-speed analysis conditions.
Column : Shim-pack GIST C8
(150 mmL. × 4.6 mm I.D., 5 µm) mAU
Mobile Phase : Acetonitrile/Phosphate (Na) Buffer (pH 7.6) = 1/3 (v/v) 200
Flowrate : 0.80 mL/min USP Original Method Omeprazole
Column Temp. : 40 ˚C
Injection Volume : 20 µL
Detection : UV 280 nm (Cell temp. 40 ˚C)
100

Table 2 Selection of Column for Speed Enhancement
Column Size L/dp Ratio 0
USP 150 mmL. × 4.6 mm I.D., 1 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
Original Method 5 µm 30000 (100 %)
USP 50 mmL. × 3.0 mm I.D., 0.83 mAU
90
Fast Method 2 µm 25000 (-17 %) USP Fast Method Omeprazole
80
70
60
Table 3 Analytical Conditions (USP Fast Method) 50
40
System : Nexera-i 30
Column : Shim-pack GIST C8 20
(50 mmL. × 3.0 mm I.D., 2 µm) 10
Mobile Phase : Acetonitrile/Phosphate (Na) Buffer (pH 7.6) = 1/3 (v/v) 0
Flowrate : 0.85 mL/min 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min
Column Temp. : 40 ˚C
Injection Volume : 8 µL Fig. 1 Chromatograms Conforming to USP Method
Detection : UV 280 nm (Cell temp. 40 ˚C)
(Upper: USP Original Method, Lower: USP Fast Method)