Agilent Manuals

The Agilent 1290 Infinity UHPLC is a new liquid chromatograph that can be used for environmental applications. The two important innovations of this instrument are 1.8-µm columns and fast gradient. These two innovations can improve the analysis efficiency and reliability.

This Application Bulletin describes a protocol for the Promega P450-Glo kit using the Agilent Bravo Platform for the measurement of Cytochrome P450 activity. The protocol involves a two-step reaction, in which the P450 enzyme metabolizes a luminogenic substrate (step 1) into a luciferin product in the presence of an NADPH regenerating system. The addition of a luciferin detection reagent (step 2) stops the enzyme reaction and converts the luciferin product into a luminescent signal that is directly proportional to the enzyme activity.

This application note provides detailed information about the Agilent BioCel system configuration for DNA purification and sequencing applications. The system consists of the Agilent Vertical Pipetting Station high-precision high-speed pipetter, which with its versatile shelf configuration options is ideally suited for complex pipetting sequences while simultaneously achieving very high throughput. The system can process 200 prepared DNA plates in four hours, eliminates sample contamination and ensures the highest possible quality of sequencing preparations.

The document describes the application of the Agilent Bravo Automated Liquid Handling Platform in quantitative polymerase chain reaction (QPCR) assays. It has been shown that the Bravo platform can effectively automate the setup of various QPCR assays and has similar slopes and good linearity compared to manual setup. Dilution samples set up by both methods showed similar slopes and good linearity, and the target genes amplified by both methods also had similar Ct values. When examining cDNA from fetal and adult lung tissues, differential expression was observed in the B2M gene, while expression of the GUSB and TBP genes remained similar. The Bravo Automated Liquid Handling Platform is useful for scientists who want to speed up setup while maintaining reliability in QPCR studies.

The present study developed a miRNA microarray analysis method suitable for tissue pathology applications. This method uses the miRNA Complete Labeling and Hyb Kit to extract miRNAs from FFPE samples, and then miRNAs are analyzed using a miRNA microarray. The results showed that this method can effectively extract miRNAs from FFPE samples and accurately analyze miRNA expression. This method provides a feasible method for miRNA analysis using FFPE samples.

An Application Kit for the Screening of Samples for Analytes of Forensic and Toxicological Interest using LC/QQQ MS/MS with a Dynamic MRM Transition Database

Extended ionization capability of thermal gradient focusing ESI in high-throughput in-vitro ADME assays

This article introduces a method for the high-pressure and high-flow analysis of polyaromatic hydrocarbons using the Agilent 1290 Infinity LC and Eclipse PAH columns.

This Application Note describes the development of a rapid resolution LC method for the separation of the Chinese herbal prescription Zuo Jin Pill into its six major con- stituents: magnoflorine, jatrorrhizine, coptisine, palmatine, berberine and evodiamine. The method uses an Agilent 1200 Series Rapid Resolution Liquid Chromatography (RRLC) system coupled with an Agilent 1200 Series Diode Array Detector (DAD). The separation was carried out on an Agilent ZORBAX Eclipse Plus C18 column (4.6 × 150 mm, 1.8 µm). The linear gradient was eluted with the mobile phase of an acetoni- trile – acetate buffer. All calibration curves showed good linearity (r2 >0.9998) within the test range 1.2-1200 µg/ml. The results demonstrate that this analytical method is simple, sensitive and reliable for rapidly analyzing six major bioactive compounds in ZJPs. It is useful for the comprehensive evaluation of the quality of this traditional Chinese Medicine (TCM).

This application note describes the method development options for separation of vitamin D2 and D3 by UHPLC. Partial separation of vitamin D2 and D3 was achieved by an isocratic LC method using a C18 column and methanol/acetonitrile mobile phase. Several optimization parameters were examined to increase resolution (Rs) including column selectivity and column length. A 2.1 mm × 150 mm Agilent ZORBAX StableBond-C18 column provided the best resolution. Further fine tuning included faster flow rates and mobile phase strength resulting in short analysis times of 3 to 1.5 minutes with Rs of 1.6 to 1.4. The performance of a 3.0 mm internal diameter (id) column was also compared to that of the 2.1 mm id column demonstrating higher sample loading before any loss of resolution in the closely spaced peaks.

A precolumn backflush system based on capillary columns using midpoint pressure control is described. Midpoint backflush is made possible with a Capillary Flow Technology (CFT) purged union controlled by an AUX EPC channel on the Agilent 7890A GC system. The key application discussed is prefractionation of crude oil that provides a high resolution separation of the C4 to C12 cut. A general backflush method using Polywax 500 is presented to illustrate the backflush concept.

This article describes a method for the analysis of phthalate content in childcare items and toys using GC/MS

This document introduces the method of SampliQ OPT solid phase extraction sorbent to clean up flavonoids in Ginkgo biloba tablets by HPLC-DAD.

The effects of extra-column volume on 50 and 150 mm columns in 2.1 and 4.6 mm internal diameters were made using a range of capillary tubing volumes installed between the autosampler and column. The results showed that, in using short columns, increases in tubing volume will reduce the column separation efficiency, while increases in tubing volume will have little effect on the separation efficiency of the long column, which is consistent with the results of other scholars.

This application note describes an approach for automating the biodiesel standard and sample preparation prior to GC analysis using the enhanced capabilities of the Agilent 7693A Automated Liquid Sampler (ALS).