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Related Concept Videos

High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
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Clinically Relevant Drug Product Specifications: Methods of Establishment01:29

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Product specifications define the acceptable quality of a pharmaceutical product by ensuring identity, purity, potency, and strength. These specifications serve as benchmarks during development, manufacturing, and post-approval quality control. Clinically relevant specifications are particularly important because they directly relate to a drug's safety and efficacy in clinical use.Dissolution studies are critical biopharmaceutic tools that link in vitro behavior to in vivo performance. They...
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High-Performance Liquid Chromatography: Instrumentation00:57

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High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.
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Drug Dissolution: Requirements and Profile Comparison01:14

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The acceptance criteria for dissolution profile data are anchored in Q values, representing the percentage of drug dissolved within a specified period. This assessment unfolds in three stages:First Stage: The test passes if all six drug dosage units are equal to or greater than Q plus 5%; otherwise, the sample proceeds to the second stage.Second Stage: The average of twelve units must be equal to or greater than Q, with no unit falling below Q - 15% to pass; if not, it progresses to the final...
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High-Performance Liquid Chromatography: Elution Process01:05

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In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
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Rational drug product design integrates knowledge of the drug’s physicochemical properties, formulation components, manufacturing techniques, and intended route of administration. Each factor influences the drug’s performance, including how it is released, absorbed, and eliminated in the body.The physicochemical properties of a drug—such as solubility, stability, and particle size—affect its compatibility with excipients and the choice of dosage form. Excipients, though...
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Related Experiment Video

Updated: Feb 19, 2026

Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
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UHPLC method for multiproduct pharmaceutical analysis by Quality-by-Design.

Ludivine Ferey1, Adrien Raimbault1, Isabelle Rivals2

  • 1ChemBioPharm Team, ARNA Laboratory, INSERM U1212, CNRS UMR 5320, Bordeaux University, France.

Journal of Pharmaceutical and Biomedical Analysis
|November 8, 2017
PubMed
Summary
This summary is machine-generated.

A new Analytical Quality-by-Design (AQbD) method was developed for separating 16 active pharmaceutical ingredients (APIs) using ethanol-based mobile phases. This robust UHPLC method ensures reliable separation for pharmaceutical repositioning.

Keywords:
Design of experimentsGreen chemistryPharmaceutical analysisQuality-by-DesignRobustness

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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
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Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Sciences

Background:

  • Pharmaceutical repositioning involves exploring new therapeutic uses for existing drugs.
  • Diverse physicochemical properties of active pharmaceutical ingredients (APIs) pose challenges for simultaneous analysis.
  • Buccal per mucous (BPM®) administration offers an alternative drug delivery route.

Purpose of the Study:

  • To develop a specific and robust Ultra-High-Performance Liquid Chromatography (UHPLC) method for separating 16 diverse APIs.
  • To apply an Analytical Quality-by-Design (AQbD) methodology for method development and optimization.
  • To investigate the use of ethanol as a greener alternative to acetonitrile in mobile phases.

Main Methods:

  • Analytical Quality-by-Design (AQbD) methodology, including screening designs and response surface methodology.
  • Investigation of C18 and Xbridge BEH Shield RP18 columns with ethanol-based mobile phases.
  • Optimization of critical process parameters (CPPs) such as gradient slope, pH, and temperature.
  • Development of an original data treatment based on modeling time intervals between peaks.

Main Results:

  • A robust UHPLC method was successfully developed using ethanol as the mobile phase.
  • Optimal conditions were determined: 2.57%/min gradient slope, pH 4.85, and 33.7°C temperature.
  • Baseline separation of 16 APIs was achieved with resolution > 2.4.
  • Method robustness was experimentally validated.

Conclusions:

  • The developed AQbD methodology provides a robust and specific UHPLC method for analyzing multiple APIs.
  • Ethanol is a viable and greener alternative to acetonitrile for UHPLC mobile phases.
  • The method is suitable for supporting pharmaceutical repositioning studies and BPM® administration route development.