Agilent Manuals

Simultaneous assessment of drug metabolic stability and identification of metabolites using HPLC-Chip/ion trap mass spectrometry can effectively identify metabolites, save time and improve work efficiency

In this study, an established HPLC method for USP Pheno-barbital was sequentially and quickly improved to a high throughput method using Rapid Resolution (RR) and Rapid Resolution High Throughput (RRHT) techniques. New RRHT column technology is an especially powerful tool for improving lab productivity by reducing HPLC analysis time dramatically. Starting with a proven rugged method, conversion to a high throughput method can be achieved simply by replacing the original analytical-sized column with either a RR or RRHT column. The reasons for this direct conversion include the similar selectivity of smaller 1.8- and 3.5-µm particles compared to larger 5-µm particles, and an engineered particle size distribution that reduces unacceptably high system pressures that may be experienced with other sub-two micron packings.

This is an application note on a drug called pindolol. It is used to lower blood pressure and alleviate depression. The note describes a method for conducting HPLC separation using pindolol and USP method. It also describes a method for conducting HPLC separation using sub-2-micron support particles (ZORBAX SB-CN (L10) columns).

Automated software-guided identification of new buspirone metabolites using capillary LC coupled to ion trap and TOF mass spectrometry

The present application note describes a method for the analysis of small RNAs using the Agilent 2100 bioanalyzer. The method involves the use of the Total RNA 6000 Nano Kit to extract small RNAs, and then the use of the bioanalyzer to separate, detect, and quantify the RNAs. The results showed that the method is highly sensitive and accurate, and is suitable for the rapid and accurate analysis of small RNAs.

This article describes the analysis of a complex natural product extract from ginseng. By using LC/MS technology, it is possible to differentiate between different species of ginseng plants and authenticate ginseng products.

Determination of Toxic Elements in Traditional Chinese Medicine Using Inductively Coupled Plasma Mass Spectrometry

This Application Note describes a method for studying noncovalent protein interactions using the HPLC-Chip/MS system coupled with an ESI-TOF mass spectrometer. The application note studies apomyoglobin and myoglobin protein complexes. The results show that ESI/MS is an efficient and accurate method for studying noncovalent protein interactions.

Fast and Ultra-fast Analysis with the Agilent 1200 Series Rapid Resolution LC System Compared to a Conventional Agilent 1100 Series LC System Using Sub 2-µm Particle Columns

This application note describes the application of the Low RNA Input Fluorescent Linear Amplification kit for the use of poly (A)+ RNA or total RNA samples. It shows that micrograms of cRNA targets can be generated within 6 hours, using as little as 10 ng of poly (A)+ RNA, or 50 ng of total RNA sample input. The resulting cRNA targets were of high quality and per- formed reproducibly and robustly on Agilent's oligonu- cleotide microarrays. Furthermore, the log ratios of array results from poly (A)+ RNA sample input correlated with that from total RNA input. The easy-to-use Low RNA Input Fluorescent Linear Amplification kit can be applied to a wide range of samples for gene expression analysis.

The search for biomarkers in proteomics research continues to be a critical bottleneck in the development of new drugs as well as in the study of disease. This paper discusses the limitations of traditional protein analysis methods and proposes the use of specific interaction media for the selective removal of high-abundant proteins.

The document introduces the Agilent Technologies' Multiple Affinity Removal System, which combines the specificity of antibody-antigen recognition with the efficiency of standard liquid chromatography instrumentation. This system effectively removes multiple proteins from human serum, including albumin, transferrin, IgG, IgA, haptoglobin, and antitrypsin, in a single step with high specificity and reproducibility. The document also demonstrates several methods to optimize column binding capacities, such as coupling two identical columns to increase capacity, using a custom column with dimensions of 10x100mm to enhance capacity, and achieving optimal binding capacity by injecting samples at a low flow rate.

This paper introduces a new multidimensional separation technique to reduce the complexity of complex serum samples. The technique uses a high-capacity multiple affinity removal column to specifically remove six high-abundance proteins (albumin, IgG, transferrin, haptoglobin, IgA, and anti-trypsin), and then uses a macro-porous reverse-phase C18 (mRP-C18) column to fractionate the depleted serum. Under the optimized mRP-HPLC condition, protein recoveries for depleted serum samples were greater than 99%. Reproducibility among patient samples was excellent based on the UV trace overlays. The mRP-C18 fractionated serum samples were further analyzed by 1D-PAGE and 2D-LC-MS/MS, the results showed that the technique can successfully reduce the complexity of serum samples and improve the detection sensitivity of low-abundance proteins.

This study describes a method based on Agilent Multiple Affinity Removal System for removing high-abundance proteins from monkey plasma, which can increase the sensitivity of detecting low-abundance proteins.

A protocol for the isolation of high-purity RNA from gram-positive and gram-negative bacteria was developed for use with the Agilent Total RNA Isolation Mini Kit.