Agilent Manuals

The document describes the sample preparation method using 0.1 N HCl for extracting beta-agonists from vitreous humor and retina extracts of pigs. However, under strong acidic conditions, the beta-agonists are protonated and not well retained on reversed phase SPE sorbents. To improve recovery, the pH of the acidic sample extract needs to be adjusted to basic conditions, which is time-consuming. High SPE recoveries can be achieved using Bond Elut Plexa PCX, which does not require pH adjustment and offers time savings. The polymeric sorbent combines reversed phase with strong cation-exchange functionalities and binds the basic analytes under acid conditions. Recoveries for many compounds can be improved to >90%. Additionally, Bond Elut Plexa PCX reduces ion suppression and minimizes binding of proteins and lipids due to its highly polar, hydroxylated polymer surface.

This document introduces the LC/MS solutions provided by Agilent Technologies for drug discovery and development, covering applications for compound identification and quantification, metabolite identification, organic impurities analysis, high throughput screening, and bioanalysis.

This application note describes a sensitive, precise, and accurate quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the measurement of beta-estradiol acetate in human plasma. The sample preparation method employed solid phase extraction on Agilent SampliQ cartridges. The processed samples were chromatographed on an Agilent ZORBAX Eclipse Plus C18 analytical column and analyzed by triple-quadrupole tandem mass spectrometry in the multiple reaction monitoring (MRM) mode using negative electrospray ionization. The principal advantage of the LC/MS/MS method described in this application note is the simultaneous achievement of high absolute recovery (~ 80%), high sensi- tivity (LLOQ of 100 fg injected on-column or 28.6 pg/mL plasma), high precision (≤ 5.9% CV) and accuracy (81.5-118.1%), as well as excellent linearity over the concentration range 0.1 to 10000 pg/μL (R2 ≥ 0.998) with a short run time of only 1.2 min.

This document describes a rapid analysis method for adenovirus type 5 particles, using a bio-monolith anion-exchange HPLC column to support the development of a high-titre manufacturing platform. The method incorporates regeneration conditions to extend the column's functional life and efficiently separates intact adenovirus type 5 particles from contaminating proteins and DNA. The method was used to analyze samples throughout a downstream process and the samples were detected at 260 nm and 280 nm using a photodiode array detector (PDA). The method was validated based on performance criteria including repeatability, intermediate precision, and linearity. The linear working range of analysis was established between 7.5 x 10^8 to at least 2.4 x 10^10 viral particles (3 x 10^10 to 9.6 x 10^11 viral particles/mL), with a correlation coefficient of 0.9992. The relative standard deviations (RSDs) for intra- and inter-day repeatability and precision for retention time and peak area were less than 1 and 2.5%, respectively.

The Agilent MassHunter Analytical Studio Reviewer software provides an LC/MS data browsing solution for rapid and accurate analysis of synthesis products. The software offers a simplified user interface for LC/MS single-quadrupole and time-of-flight (TOF) analyses, required for compound confirmation or purification of small molecules or proteins. Agilent Analytical Studio Reviewer allows for fast, flexible, and accurate review and reporting of LC/MS data to expedite the development process. The combination of these two software packages, along with Agilent's rugged LC and MS systems, provides a flexible and powerful solution for rapidly obtaining high-quality data.

This application note describes MassHunter Easy Access, a software package that provides a simplified user interface for LC/MS single-quadrupole and time-of-flight (TOF) analyses required for either compound confirmation or purification of small molecules or proteins.

This application note describes a rapid method for the simultaneous detection and quantification of 37 acylcarnitines and 12 amino acids in dried blood spots using the Agilent Triple Quadrupole LC/MS System in neutral loss scan, precursor ion scan and multiple reaction monitoring (MRM) data acquisition modes. With a run time of 1.6 minutes, the assay provides the ability to analyze 400-500 samples per day, and custom reports provide all the information needed to rapidly list the critical elements for the analysis.

This Application Note describes the design and performance of an HPLC-Chip with a hybrid TiO2 and C18 enrichment column for selective phosphopeptide enrichment and analysis.

This application note outlines a viable method for reliably detecting low-fem- togram levels of perfluorinated carboxylates (PFCA) and sulphonates (PFSA) without background component interference, with an inline contaminant trap LC modification. Good chromatographic resolution of all perfluorinated compounds was observed. A representative suite of PFCAs and PFSAs were analyzed herein and were all detected at on-column levels lower than 75 fg in drinking water matrices (S/N >3.) The most sensitive analytes PFHxS, PFDS and PFBS were detected at 2.6, 3.2 and 5 fg levels, respectively. Method detection limits for spiked pork liver matrix extract samples were below 600 fg on-column for the entire analyte suite. No detectable background conta- mination was observed in blank injections for any analyte in this study. Linearity for up to five orders of magnitude with R2 values above 0.996 for the entire suite were recorded.

The Agilent 7700s ICP-MS is a high-performance mass spectrometer designed for semiconductor applications, offering unmatched sensitivity, matrix tolerance, interference removal, and stability.

A rapid gradient and elevated temperature UHPLC method for flavonoids in citrus fruits