Pseudoephedrine hydrochloride (I), brompheniramine maleate (II), and dextromethorphan hydrobromide (III) in a cough-cold sytup were separated and determined by ion-pair reversed-phase high-pressure liquid chromatography. The separation was carried out using a muBondapak C18 column (30 cm x 3.9 mm i.d.) and a mobile phase of acetonitrile-water-acetic acid (40:60:1) with 0.01 N 1-octanesulfonic acid sodium salt and 0.05 N potassium nitrate. Detection was accomplished using a UV detector at 265 nm for I and II; III was monitored at 280 nm. Concentration versus peak height plots in the ranges of 0.37-1.9 mg/ml for I, 0.025-0.126 mg/ml for II, and 0.125-0.625 mg/ml for III were linear. Ten consecutive injections of a mixture gave a percent relative standard deviation of less than 1% for all three components. Average recoveries from laboratory-prepared samples were 100.5% for I, 100.9% for II, and 100.1% for III. No precolumn cleanup was necessary, and the chromatogram was complete in 16 min.
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http://dx.doi.org/10.1002/jps.2600681137 | DOI Listing |
Anal Chem
January 2025
Synthetic Molecule Analytical Chemistry, Genentech Inc., South San Francisco, California 94080, United States.
Accurate characterization of therapeutic RNA, including purity and identity, is critical in drug discovery and development. Here, we utilize denaturing and non-denaturing chromatography for the analysis of ∼25 kDa divalent small interfering RNA (di-siRNA), which comprises a complex 2:1 triplex structure. Ion pair reversed-phase (IPRP) liquid chromatography (LC) experiments with UV absorbance and mass spectrometry (MS) showcase a single denaturing LC method for identity confirmation, impurity profiling, and sequencing with automated MS data interpretation.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Bioscience, Research and Early Development, Oncology, AstraZeneca, Cambridge, Cambridgeshire, UK.
A protocol for the preparation of tissue extracts for the targeted analysis ca. 150 polar metabolites, including those involved in central carbon metabolism, is described, using a reversed phase ion pair U(H)PLC-MS method. Data collection enabled in high-resolution mass spectrometry detection provides highly specific and sensitive acquisition of metabolic intermediates with wide range physicochemical properties and pathway coverage.
View Article and Find Full Text PDFAnal Chem
January 2025
State Key Laboratory of Genetic Engineering, School of Life Sciences, Human Phenome Institute, Zhangjiang Fudan International Innovation Center, Metabonomics and Systems Biology Laboratory at Shanghai International Centre for Molecular Phenomics, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Simultaneous analysis of multiple phosphorylated metabolites (phosphorylated metabolome) in biological samples is vital to reveal their physiological and pathophysiological functions, which is extremely challenging due to their low abundance in some biological matrices, high hydrophilicity, and poor chromatographic behavior. Here, we developed a new method with ion-pair reversed-phase ultrahigh-performance liquid chromatography and mass spectrometry using BEH C18 columns modified with hybrid surface technology. This method demonstrated good performances for various phosphorylated metabolites, including phosphorylated sugars and amino acids, nucleotides, NAD-based cofactors, and acyl-CoAs in a single run using standard LC systems.
View Article and Find Full Text PDFJ Chromatogr A
January 2025
Ionis Pharmaceuticals, Carlsbad, CA 92010, USA.
The increased effectiveness of small interfering RNAs (siRNAs) to induce gene silencing has brought a great therapeutic promise to many diseases. siRNAs are under highly active current research and development. Ligand conjugation and chemical modifications of the sense (SS) and antisense (AS) strands of the siRNA duplex improve stability and facilitate delivery, but significantly increase the complexity of the analytical requirements.
View Article and Find Full Text PDFJ Chromatogr A
January 2025
Department of Engineering and Chemical Sciences, Karlstad University SE-651 88 Karlstad, Sweden. Electronic address:
In indirect detection, sample components lacking detectable properties are detected by adding a detectable component to the eluent, a so-called probe that interacts with the analytes to be detected. This study focuses on modeling indirect detection in two principally different cases. In case (1), the analyte component has the same charge as the probe component, so the probe acts as a co-ion of the analyte.
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