Page 16 - Shimadzu Journal vol.5 Issue1

Selection 5 Clinical research
High-sensitivity and simultaneous analysis of Psychoactive drugs using LC-MS/MS with full-automat-
ed pretreatment system
LC-MS/MS has become a preferred method for the routine analysis for forensic toxicology. LC-MS/MS allows for the simultaneous
analysis of multiple compounds in a single run, thus enabling a fast and high throughput analysis. In recent years that it seems the
number of incident and accident is increasing caused by dosed with psychotropic drugs and the number of drug testing with
LC-MS/MS is also increasing to investigate the cause of death. However, manual sample preparation often involves several
complicated manual steps which can introduce error into the results. In this study, we investigated the processing capability to
analyze serum, whole blood and urine spiked sixty psychotropic drugs by LC-MS/MS with automated sample preparation unit.

Selection 6 Clinical research
Strategies for Multiple-Target Screening using LC-MS/MS with Merged Spectrum Database for
Forensic Toxicology
In Forensic Toxicology, LC/MS/MS has become a preferred method for the routine quantitative and qualitative analysis of drugs of
abuse. LC/MS/MS allows for the simultaneous analysis of multiple compounds in a single run, thus enabling a fast and high
throughput analysis. In this study, we developed Multiple-Target Screening (MTS) method for forensic toxicology to reduce false
positive and negative using MS/MS spectral library database. MTS method consists of multiple reaction monitoring (MRM) and
product ion scans at three collision energies to confirm the compound identification based on mass spectral library searching. The
mass spectral library was created using certified reference materials from over 1,200 compounds for forensic toxicology.

Selection 7 Clinical research
Analytical method development for widely targeted perfluoroalkyl acids (PFAAs) and their precursors
in plasma using multi-gradient eluent system by LC-MS/MS
As perfluoroalkyl acids (PFAAs) had been widely used because of their excellent surfactant. While the product and usage of
perfluorooctane sulfonic acid (PFOS), its salts and perfluorooctane sulfonyl fluoride (PFOS-F) were restricted by the Stockholm
Convention in 2009, many kinds of precursors which cause PFAAs by decomposition keep being used because they are not
restricted.
LC-MS/MS analysis methods for PFAAs have been developed, where a high selectivity analysis is available, but it was very difficult
to implement the simultaneous analysis of PFAAs and their precursors since the wideness of their chemical property in terms of
hydrophilic / hydrophobic.
This time, we developed the analytical method for widely targeted PFAAs and their precursors in plasma using multi-gradient
eluent system by LC-MS/MS.

Selection 8 Clinical research
Determination of Unbound Urinary Amino Acids Incorporated with Creatinine Normalization by
LC-MS/MS Method with CLAM-2000 Online Sample Pre-treatment
Free or unbound amino acids are important metabolites in human blood and urine [1]. The profile of unbound amino acids in urine
are the reference indication of metabolic imbalances and amino acid transport disorders as well as dietary protein adequacy and
assimilation. Creatinine produced by muscle metabolism is excreted in the urine, which can be used to normalize the metabolite
levels to compensate the large variation due to different intakes of water and fluid food [2-4]. The aim of this study is to develop a
reliable LC-MS/MS method for quantitation of 22 free amino acids and creatinine in urine samples. A derivatization-free LC-MS/MS
amino acid method [5] with stable isotope labelled IS was employed. An on-line sample pre-treatment module CLAM-2000
coupled with LC-MS/MS makes the analysis fully-automated, which enables from adding internal standards, sample and solvent
mixing, shaking for protein-crash and filtration to transferring the final sample solution to LC-MS/MS for analysis.

Selection 9 Clinical research
A Novel Platform of On-line Sample Pre-treatment and LC/MS/MS Analysis for Screening and
Quantitation of Illicit Drugs in Urine
In recent years, LC/MS/MS methods are adopted in analyses of illicit and prescription drugs in toxicological samples such as urine
and serum. Sample pre-treatment is always a critical step in the whole analysis procedure and on-line sample pre-treatment is
desired not only for improving analysis throughput, but also minimizing human errors. The CLAM-2000 module is designed for
on-line sample pre-treatment in high throughput LC/MS/MS analysis of drugs and metabolites in biological samples such as
plasma/serum and urine. Many sample preparation process can be performed automatically such as dispensing solvents,
sample-reagent mixing by vortexing, sample filtering by vacuum filtration, and sample derivatisation with heating. Internal
standard and reagent for derivatization or other purposes can be added to a sample before or after protein crash. We describe
development of an automated sample pre-treatment using a Shimadzu CLAM-2000 module coupled with Shimadzu LCMS-8040
TQ system. It involves IS addition, protein precipitation, filtration and transferring the final solution to LC/MS/MS for analysis. This
new platform was applied and evaluated for quantitation of 18 illicit drugs with 14 isotope-labelled internal standards (IS).