Application Notes: LC

This Application Note describes the method transfer of a USP HPLC method for amlodipine besylate tablets to a UHPLC method using the Agilent 1290 Infinity II LC.

11-nor-9-Carboxy-THC, also known as THCA or carboxy-THC, is the main secondary metabolite of THC (the active component of marijuana) formed in the human body [1]. THCA is excreted in urine in the form of glucuronide conjugates. THCA is not psychoactive but has a long half-life of up to several days or even weeks in very heavy users, thus determination of THCA in urine plays an important role in confirmation of marijuana consumption. The Substance Abuse and Mental Health Services Administration (SAMHSA) has set the THCA cutoff concentration of confirmatory testing at 15 ng/mL. Typical sample preparation methods for THCA in urine include liquid-liquid extraction (LLE) and solid phase extraction (SPE). This application utilizes a novel sample preparation technique, QuEChERS to effectively quantitate THCA levels in human urine.

Using a targeted solid phase extraction (SPE) method and GC–ECD analysis, chlorinated pesticides are extracted from poultry fat resulting in a method that decreases the amount of labor and reagents when compared to the USDA-FSIS CHC2 method.

A simple, automatable cleanup method is developed for the analysis of acrylamide from coffee, yielding absolute recoveries that exceed 92% when analyzed by LC–MS-MS.

Using a targeted solid phase extraction (SPE) method and GC–ECD analysis, chlorinated pesticides are extracted from poultry fat resulting in a method that decreases the amount of labor and reagents when compared to the USDA-FSIS CHC2 method.

A simple, automatable cleanup method is developed for the analysis of acrylamide from coffee, yielding absolute recoveries that exceed 92% when analyzed by LC–MS-MS.

Cannabinoids are successfully extracted from a complex brownie matrix using the QuEChERS sample preparation technique, significantly reducing the presence of matrix interferences. The resulting clean samples are analyzed by GC–MS using a specialized deactivated GC column to provide heightened peaks for the cannabinoid compounds.

EPA Method 625 is greatly simplified by using solid phase extraction (SPE) as an alternative to traditional liquid-liquid extraction (LLE). Using this SPE technique, paired with a sensitive GC–MS analysis, increases throughput and data quality, while decreasing manual labor and solvent usage.

Cannabinoids are successfully extracted from a complex brownie matrix using the QuEChERS sample preparation technique, significantly reducing the presence of matrix interferences. The resulting clean samples are analyzed by GC–MS using a specialized deactivated GC column to provide heightened peaks for the cannabinoid compounds.

EPA Method 625 is greatly simplified by using solid phase extraction (SPE) as an alternative to traditional liquid-liquid extraction (LLE). Using this SPE technique, paired with a sensitive GC–MS analysis, increases throughput and data quality, while decreasing manual labor and solvent usage.

Phospholipid removal proves to yield superior results as compared to a traditional protein precipitation step when analyzing a complex plasma matrix via LC–MS-MS. The results displayed a decrease in ion suppression, increased analyte sensitivity, and an improvement in column lifetime.

Enhancing the extraction of vitamins A and E in serum using simplified liquid extraction (SLE) significantly improves the LC–MS-MS detection of target compounds, all while preserving a simple and robust method.

Phospholipid removal proves to yield superior results as compared to a traditional protein precipitation step when analyzing a complex plasma matrix via LC–MS-MS. The results displayed a decrease in ion suppression, increased analyte sensitivity, and an improvement in column lifetime.

Enhancing the extraction of vitamins A and E in serum using simplified liquid extraction (SLE) significantly improves the LC–MS-MS detection of target compounds, all while preserving a simple and robust method.

Using oral fluid as the matrix, a viable and simple solid phase extraction method for a wide range of drugs is developed. Several oral fluid collection devices were evaluated to determine the effectiveness of the cleanup procedure, ultimately confirming that the method is both robust and widely applicable.

Improving and optimizing EPA Method 593 using solid phase extraction (SPE) enhances sensitivity and maximizes efficiency while screening by LC–MS-MS for different female birth control hormones present in drinking and waste water.

Using oral fluid as the matrix, a viable and simple solid phase extraction method for a wide range of drugs is developed. Several oral fluid collection devices were evaluated to determine the effectiveness of the cleanup procedure, ultimately confirming that the method is both robust and widely applicable.

An ultra-sensitive sample preparation technique using solid phase extraction (SPE) microelution 96-well plates effectively concentrates peptides and allows for cleanup of small volume.

A suite of unknown drug compounds from whole blood are analyzed by LC–MS-MS, requiring the need for an effective yet nonspecific cleanup technique. The resulting method demonstrates a simple and fast sample preparation procedure that is suitable for screening many compounds.

A suite of unknown drug compounds from whole blood are analyzed by LC–MS-MS, requiring the need for an effective yet nonspecific cleanup technique. The resulting method demonstrates a simple and fast sample preparation procedure that is suitable for screening many compounds.

An ultra-sensitive sample preparation technique using solid phase extraction (SPE) microelution 96-well plates effectively concentrates peptides and allows for cleanup of small volume.

Intact and reduced IgGs have been successfully retained and separated using a non-porous column. Potential applications of Presto columns for a single-injection separation of free drug, antibody, and antibody-drug complexes (ADCs) is also discussed.

This Application Note demonstrates the separation and quantification of beta-blockers, listed as prohibited substances by the World Anti-Doping Agency, in urine using the Agilent 1260 Infinity Analytical SFC System.

HPLC column stability is a critical factor. This note describes how the Agilent Poroshell HPH-C18 column performed brilliantly in an elevated pH mobile phases such as ammonium bicarbonate buffer.

This Application Note demonstrates the separation and quantification of beta-blockers, listed as prohibited substances by the World Anti-Doping Agency, in urine using the Agilent 1260 Infinity Analytical SFC System.

Characterization of glycosylation is a major quality parameter in the production of biotherapeutics. This note demonstrates the benefits of using a new, small particle TSKgel Amide-80 HILIC column which improves peak capacity and sensitivity for UHPLC and LC-MS analysis of labelled glycans.

In new drug development, the number of diverse chiral compounds is increasing and sensitive chiral methods are often needed quickly. Many new CSPs are available on the market making it challenging to select the most important ones for the initial screening stages and expedite method development. The focus of this study is to evaluate high selectivity CSPs and to suggest the best screening method with a limited number of high success rate chiral columns.

‘Old’ HPLC methods with long run times are being altered or surpassed by newer UHPLC or core-shell methods in order to save time and cost. In this application note we show how with the use of 3 simple equations transfer of older methods can be easily achieved onto newer core-shell particles. We show the example of a pharmaceutical drug and its impurities being reduced from a 30minute run time down to less than 10minutes. Using the calculations correctly means that no loss of resolution is seen even with the decrease in retention time.