5989_5672.qxd

Fast LC/MS/MS Analysis of Group 4
Pharmaceuticals from EPA-1694 with
RRHD HILIC Plus
Abstract
The analysis of the Group 4 compounds in EPA-1694 is sped up using an Agilent 1290 Infinity UHPLC and an Agilent ZORBAX RRHD HILIC Plus column. Excellent peak shape is found for all compounds, while the flow rate and sample throughput are increased by four times the original rate.
Introduction
Pharmaceuticals and personal care products (PPCPs) are an important group of conta-minants targeted by environmental laboratories. Several methods address these ana-lytes, including EPA-1694. In this work, the Group 4 compounds from this method(cimetidine, albuterol, ranitidine and metformin) will be addressed, and the methodwill be improved upon through the use of UHPLC. Previous Agilent application noteshave addressed the implementation of UHPLC with the compounds found in Groups1–3 of EPA-1694, which use an Agilent ZORBAX RRHD Eclipse Plus C18 column (referto publication numbers 5990-4409EN and 5990-4605EN).
Advancements in liquid chromatography have lead to significantly improved samplethroughput, which is advantageous to many environmental laboratories. AgilentTechnologies’ 1290 Infinity UHPLC and Agilent ZORBAX Rapid Resolution High Definition(RRHD) columns are manufactured to withstand pressures up to 1200 bar, thus allow-ing the use of faster flow rates and higher throughput.
The newly released Agilent ZORBAX RRHD HILIC Plus is a 1.8 μm column that is stable to 1200 bar. The non-bonded silica is based on the silica used to manufactureEclipse Plus columns to ensure excellent peak shape. HILIC columns are ideal for theretention of small, polar analytes, such as those found in Group 4 of EPA-1694. Experimental
An Agilent 1290 Infinity UHPLC with an Agilent 6410 Triple Quadrupole Mass Spectrometer, and an Agilent ZORBAX A: 10mM Ammonium Acetate pH 6.7 in H2O; B: CH3CN RRHD HILIC Plus 2.1 mm × 100 mm, 1.8 μm column (p/n 959758-901) were used in this experiment. 0.1 μL injection of sample (0.1 mg/mL each in CH3CN/H2O) dMRM, ESI positive mode, cycle time 35 ms, drying gas: 9 L/min, 300 °C; nebulizer pressure: 40 psig; capillary voltage: 4000 0.1 μL injection of 0.1 mg/mL each in acetonitrile/water (3:1): cimetidine, albuterol, ranitidine and metformin dMRM, ESI positive mode, cycle time 35 ms, drying gas: 9 L/min, 300 °C; nebulizer pressure: 40 psig; capillary voltage: 4000; see Table 1 for MRM transition MassHunter versions B.03.01, B.02.00 and B.03.01 were used for data acquisition, qualitative and quantitative analysesrespectively An Agilent RRHD HILIC Plus 2.1 mm × 100 mm, 1.8 μm column isused to rapidly analyze the Group 4 pharmaceuticals from EPA-1694, see Experimental section for detailed method parameters. MRM Transitions for the Pharmaceuticals in Group 4 of EPA-1694 Precursor
Fragmentor
Collision
Conclusions
Compound
The Agilent 1290 Infinity UHPLC coupled with a ZORBAX RRHD HILIC Plus column successfully analyzes the Group 4 pharma- ceutical compounds in EPA-1694. The flow rate and analysis is sped up by a factor of 4, while maintaining good peak shape for For More Information
Results and Discussion
For more information on our products and services, visit ourWeb site at www.agilent.com/chem.
A ZORBAX RRHD HILIC Plus column is used to quickly screenthe Group 4 analytes found in EPA-1694, shown in Figure 1.
This analysis is typically run at 0.25 mL/min, however this 1200bar stable RRHD HILIC column is capable of running 1 mL/min,which reduces run time by 75% while generating a maximumpressure of 960 bar. Excellent peak shape is found for all four compounds: cimetidine, albuterol, ranitidine and metformin. Agilent shall not be liable for errors contained herein or for incidental or consequentialdamages in connection with the furnishing, performance, or use of this material.
Information, descriptions, and specifications in this publication are subject to changewithout notice.
Agilent Technologies, Inc., 2011Printed in the USAJune 28, 20115990-8433EN

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