• Application Note

On-Line Protocol for Basic Drugs in Cell Culture Lysate Using LC-MS/MS

On-Line Protocol for Basic Drugs in Cell Culture Lysate Using LC-MS/MS
  • Waters Corporation
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This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief highlights the analysis of basic drugs in cell culture lysate using Oasis HLB on-line protocol.

Introduction

Basic drugs has been analyzed in cell culture lysate using LC-MS/MS in this application brief.

Experimental

HPLC Method

Column:

XTerra MS C18, 2.1 x 30 mm, 3.5 μm (p/n: 186000398)

Mobile phase A:

Acetonitrile + 0.5% Formic Acid

Mobile phase B:

Water + 0.5% Formic Acid

Flow rate:

1.0 mL

LC1:

Alliance 2690–0.4 mL/min

LC2:

Waters 515–4.0 mL/min

Loading mobile phase:

100 % water

Eluting mobile phase:

1 minute gradient 5% ACN to 95% ACN

Eluting mobile Phase additive:

0.5 % Formic acid

Extraction column temp.:

40 °C

Switching valve:

Rheodyne LabPro 10 ports, 2 position

MS:

Waters Micromass Quattro Ultima

Source:

Electrospray positive

Sorce temp.:

150 °C

Desolvation gas:

600 L/hr

Gas cell:

1.5e-3 mbar

Cone voltage:

20 volts

Collision energy:

20

Gradient

Compound 2 name: Propranolol

Coefficient of Determination: 0.998650

Calibration curve: -1.52393e-5 * x2 + 0.00390583 * x + 0.000102086

Response type: Internal Std (Ref 1), Area * (IS Conc. / IS Area)

Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x2, Axis trans: None

Compound 3 name: Naloxone

Coefficient of Determination: 0.995263

Calibration curve: -2.47393e-5 * x2 + 0.00892751 * x + -5.00665e-5

Response type: Internal Std (Ref 1), Area * (IS Conc. / IS Area)

Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x2, Axis trans: None

On-Line Protocol

Oasis HLB 2.1 x 20 mm, 25 μm Extraction Column Part Number 186000706 

Results and Discussion

CID Mass Spectra

50/50 MEOH/Acetonitrile Cell Lysing

Featured Products

WA31764.112, June 2003

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