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Versatile Solutions to Current Demands for Automated Sample Cleanup, High Throughput, and High-Resolution Chromatography on a Single LC-MS Platform
Overview
Purpose:
Use two case studies to demonstrate the versatility of an LC-MS platform for
research: high-throughput, online sample cleanup capabilities in a multi-channel
UHPLC t
l d t
hi h l ti
t
(HRAM) h b id
Methods
Sample Preparation
Case Study #1: Four stand
sys em coup e o a g -reso u on, accura e-mass
y r
quadrupole-Orbitrap™ mass spectrometer (MS).
Methods:
Case Study #1:
efavirenz, and zidovudine)
crashed plasma, water-dilut
acetonitrile-water neat mix.
1000 ng/mL. Warfarin was a
positive- or negative-ionizin
An LC/MS accurate-mass screen of antiretrovirals (ARVs) in biological matrix for
research was examined using online sample cleanup. In a high-throughput workflow
configuration, standards in several biological matrices were injected across two LC
channels (each with distinct sample extraction columns) in a cross-sequential manner
to one at-source guard column.
Case Study #2: A 100 ng/m
UHPLC columns, and mobil
lab.
Liquid Chromatography
Case Study #2:
One LC/MS research method for analysis of four major sulfatide molecular species in
an organic standard was optimized for demonstration of two discrete high-throughput
data acquisition styles. Four LC channels, each with distinct HPLC columns, were used
d i j ti
d i b th
h l
d
ti l
t
The multi-channel LC used i
Transcend™ II LX-4 configu
UltiMate™ 3000 Binary Rapi
interface module), and a CT
Zwingen, Switzerland). In b
an n ec ons were ma e n o a cross-c anne an a cross-sequen a manner o
the HRAM-MS.
Results:
Case Study #1:
waste, except during the dat
flow rates and, therefore, fa
Case Study #1: In a high-thr
with distinct TurboFlow sam
C18 guard column The LC
A 100 s LC/MS method for ARVs in human biological matrices with online sample
cleanup and chromatographic resolution were acquired within a data window of 30 s
using a multi-channel LC system. Four standard curve matrices were investigated. All
four curves gave linearity between R
2
= 0.96 and 0.99.
Case Study #2:
.
detector bypass position allo
TurboFlow technology.
1
Mul
across two channels of the
MX software version 2.1. Fi
of a neat standard with thre
Through the two discrete high-throughput acquisition styles examined
,
the percent
RSDs of each analyte across multiple injections of the organic standard were
maintained at less than 5.8%.
I t d ti
Case Study #2: The sulfatid
injections (10 µL) of the nea
50 mm, 1.7 µm) heated to 6
sequential manner and were
n ro uc on
Investigative demands in both clinical research and pharmaceutical laboratories
necessitate the use of simpler and faster LC/MS technologies. Market response to
these demands has produced high-throughput technologies that can perform online
l
l
f ll
d b
t l
l i Th h t
hi
ARVs LC Method Details
TABLE 1. LC method detai
samp e c eanup o owe y mass spec ra ana ys s. ese c roma ograp c
capabilities, however, can be limiting and their results inconsistent.
Here we present two case studies highlighting the versatility of a single multichannel
LC-MS platform as both a research and a pharmaceutical production tool. A multi-
channel LC provides the ability to make staggered injections through multiple LC
LC Method Length
Data Window
TurboFlow Columns
channels while utilizing a single MS, thus increasing sample throughput (Figure 1). This
platform also has the capacity to perform online sample cleanup utilizing Thermo
Scientific™ TurboFlow
TM
technology along with either baseline-resolved UHPLC
chromatography or resolution based solely on accurate mass, all in a rugged high-
throughput workflow.
Analytical Column
Loading Mobile Phase
Eluting Mobile Phase
Extraction Column Wash
FIGURE 1. Illustrative staggered chromatograph
Injection Volume
TABLE 2. LC Method Deta
Sulfatide LC Method Detai
LC Method Length
Data Window
Analytical Columns
Loading Mobile Phase
Eluting Mobile Phase
Injection Volume