Agilent Manuals

This document describes the analysis challenges of D-penicillamine enkephalin (DPDPE) and the rapid sensitive analysis of DPDPE in cerebrospinal fluid using an Agilent 1100 Series LC/MSD mass spectrometer. DPDPE is a synthetic enkephalin used for pain control, which has proven difficult to analyze in the past. The document also provides detailed information on the experimental method and sample preparation process.

This application note provides step-by-step procedures for creating and using a mass spectral library with NIST MS Search software.

Metabolomics is the comparative analysis of metabolites found in sets of similar biological samples. Since metabolites play vital roles in biological systems, metabolomics can be useful for finding and identifying biomarkers, or for obtaining a better understanding of the effects of drugs or diseases on both known and unexpected biological pathways. Successful metabolomic research requires effective metabolite extraction. For non-targeted metabolomics, extraction methods need to capture a broad range of cellular and biofluid metabolites, while excluding components such as proteins that are not intended for analysis. Extraction is made more challenging by the physico-chemical diversity of metabolites and by metabolite abundances that can vary by many orders of magnitude. Biphasic, liquid-liquid extraction is often used to extract metabolites. The nature of the organic and aqueous solvents, their volumes, solvent ratios, and aqueous solvent pH, however, must be considered carefully. They can significantly affect the total number of metabolites extracted and experimental reproducibility. This application note describes a method for liquid-liquid extraction of metabolites from erythrocytes. It demonstrates the importance of adjusting the aqueous/organic ratio to favor biphasic extraction.

A fast, sensitive, and validated technique for confirming the presence of amphetamine drugs in whole blood using the Agilent G6410A Triple Quadrupole Mass Spec-trometer (QQQ) is presented. Excellent linearity is demon- strated over the range of approximately 15 to 1,000 ng/mL. The amphetamine drugs analyzed in this work include amphetamine, methamphetamine, methyl-enedioxyamphetamine (MDA), and methylenedioxymeth-amphetamine (MDMA) in blood. The drugs have been prepared using an extractive alkylation technique. The sample preparation is then followed by reverse-phase LC/MS/MS using a 1.8-µm particle size C18 column for high chromatographic resolution with a high-speed sepa-ration. As a result, elution times for both analytes and internal standards are all less than or equal to 3.6 min- utes.

This application note describes the use of mass spectrometry to identify metabolites in rice and related bacteria, and to investigate the relationship between metabolites and susceptibility or resistance of rice to bacterial blight

A rapid, simple, highly sensitive procedure for the simultaneous analysis of 14 benzodiazepines and six metabolites in urine and blood, using the Agilent 6410 Triple Quadrupole Mass Spectrometer in electrospray mode, is described.

A quantitative analytical procedure for the determination of benzodiazepines and metabolites in hair has been developed and validated. The hair samples were washed, incubated, and any drugs present were quantified using mixed mode solid-phase extraction and liquid chromatography with tandem mass spectrometric detection (LC/MS/MS) in positive electrospray ionization mode.

A quantitative analytical procedure for the determination of cocaine, benzoylecgonine, cocaethylene, and norco- caine in hair has been developed and validated. The hair samples were washed, incubated, and any drugs present were quantified using mixed-mode solid-phase extraction and liquid chromatography with tandem mass spectro- metric detection in positive atmospheric pressure chemi- cal ionization mode. For confirmation, two transitions were monitored and one ion ratio was determined, which was within 20% of that of the known calibration stan- dards. The monitoring of the qualifying transition and requirement for its presence within a specific ratio to the primary ion limited the sensitivity of the assay, particu- larly for benzoylecgonine; however, the additional confi- dence in the final result as well as forensic defensibility were considered to be of greater importance. Even with simultaneous monitoring, the concentrations proposed by the United States federal guidelines for hair analysis were achieved. The limits of quantitation were 50 pg/mg; the limit of detection was 25 pg/mg. The intra-day precision Determination of Cocaine, Benzoylecgo- nine, Cocaethylene, and Norcocaine in Human Hair Using Solid-Phase Extraction and Liquid Chromatograp

This Application Note describes the rapid development of a MRM method for triple quadrupole mass spectrometry, the rapid development of a liquid chromatography method for high throughput conditions with a 1.8 µm RRHT column, the use of the developed LC/MS MRM method for rapid determina- tion of a metabolic stability profile of a pharmaceutical drug, and the relative quantification of the acquired data to generate the metabolic stability profile.

This Application Note is based on scientific poster MOP 253 presented at the ASMS conference of mass spectrometry, June 4, 2007, in Indianapolis, Indiana, USA. The note describes: - Rapid, computer-assisted identification of drug metabolites using a multi-algorithm approach. - High resolution chromatographic separation of drug metabolites from an in vitro experiment. - High mass accuracy measurement of drug metabolites by QTOF MS and MS/MS.

This Application Note describes: • The rapid development of an MRM triple quadrupole MS method for compound quantification. • The fast liquid chromatography separation on 1.8 µm RRHT columns for high throughput of DMPK samples. • The application of the developed LC/MS MRM method for the quantification of compounds in a mixture by a multi MRM method.

This article introduces the techniques for isolating metabolites from biological samples using liquid chromatography with sub-2-micron particle columns. The method improves the rate and resolution of metabolite separation without compromising the overall purity or recovery of the metabolites.

This article describes the optimization of selectivity for antibody separations using HPLC and LC/MS by increasing the percentage of n-propanol and acetonitrile in 0.1% trifluoroacetic acid using ZORBAX 300 Å SB-CN LC Columns.

This document describes the development of a rapid resolution liquid chromatography (RRLC) method for the analysis of notoginseng. The method has good resolution and can be used to quickly analyze complex components, thereby increasing sample throughput and lowering costs. The method can also be used to analyze compounds in different parts of the notoginseng plant, and different peak profiles and different concentrations of certain saponins are detected.

This application note describes how to use Agilent’s Tiling Microarray Technology to monitor genome-wide methylation patterns in CpG islands.