Page 30 - Pharmaceutical Solution for Pharma Analysis

LAAN-A-LC-E291

Application Supercritical Fluid Chromatography

News
Analysis and Evaluation of Chiral Drugs in Biological
Samples Using the Nexera UC-MS/MS System
No. L517

As introduced in Application News No. L495, the Table 1 Analytical Conditions
optimization for chiral separation using supercritical Column : CHIRALPAK®, IC-3
fluid chromatography (SFC) starts from employing (100 mm L. × 3.0 mm I.D., 3 Pm)
Mobile phase : A) Super critical fluid of CO2
column scouting to find the column and mobile phase B) Modifier: Methanol
appropriate to separation. This article introduces an A/B = 5/1 (v/v for omeprazole, isocratic)
example of the selectivity and sensitivity of drug level = 4/1 (v/v for rabeprazole, isocratic)
monitoring in a biological sample and the evaluation Flow rate : 3 mL/min
Column temp. : 40 qC
results of the analysis method, as an application to the Injection volume : 3 μL
pharmacokinetics research of chiral separation using BPR pressure : 10 MPa
SFC/MS/MS, after having selected an appropriate BPR temp. : 50 qC
Detector
: LCMS- 8050 (ESI, MRM mode)
column. Make-up : Methanol
Y. Watabe, T. Hattori, T. Iida Make-up flow rate : 0.1 mL/min
MRM : (+) m/z 346.1 > 198.1 (for omeprazole)
(+) m/z 359.9 > 150.1 (for rabeprazole)
„ Analysis of Omeprazole in a Plasma Sample
The applicability of human plasma matrix to SFC was Calibration curve was created based on human plasma
evaluated taking an example of enantiomeric drug samples that contained 1, 2, 10, 2 and 100 μg/L of
omeprazole, well-known as a proton pump inhibitor. standard omeplazole to confirm the linearity of loaded
Fig. 1 shows the chemical structure of omeprazole. Fig. amounts.
2 shows the pretreatment procedure employed for the Fig. 3 and Fig. 4 show the MRM chromatograms for
blood plasma sample. Table 1 lists the analytical 2 μg/L and 20 μg/L respectively. Among the optically
conditions. CHIRALPAK® IC-3 from Daicel Company, separated peaks, (A) is the fast-eluting isomer and (B) is
2
which exhibited good separation when utilized in the slow-eluting isomer. The linearity (r ) obtained after
Application News No. L495 was used as the column. correcting by 1/(concentration squared) was favorable
Detection was performed using the LCMS-8050 triple at 0.99996 for omeprazole (A) and 0.99998 for
quadrupole mass spectrometer. omeprazole (B).

(× 1,000)
1:346.10 > 198.10 (+)
2.5
(B)
Omeprazole 2.0
(A)

1.5
Omeprazole Structure

1.0
20 μL of plasma
0.5
250 μL of acetonitrile
0
0 1.0 2.0 3.0 4.0 5.0
Filtrate

250 μL of acetonitrile/ Omeprazole Added to Human Plasma (2 μg/L)
28 % aqueous ammonia= 1000/1 (v/v)

Inject 3 μL

Plasma Sample Pretreatment Procedure