Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

3.1K
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.
In HPLC, two phases play a critical role in the separation process:
3.1K
High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

2.9K
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.
2.9K
High-Performance Liquid Chromatography: Types of Detectors01:15

High-Performance Liquid Chromatography: Types of Detectors

2.3K
The role of the detectors in High-Performance Liquid Chromatography (HPLC) is to analyze the solutes as they exit from the chromatographic column. The detector recognizes the solute's property and generates corresponding electrical signals, which are converted into a readable graph of the detector's response versus elution time called a chromatogram at the computer. There are several types of HPLC detectors, each with its own advantages and limitations, depending on the analyte...
2.3K
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

2.1K
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
2.1K
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

2.1K
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...
2.1K
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

3.0K
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
3.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Redox-mediator enhanced electrochemiluminescence under non-aqueous conditions.

Chemical science·2026
Same author

Redox-Mediated Signal Enhancement of a Spatially Resolved Electrochemiluminescence Immunoassay.

ACS sensors·2026
Same author

Cost-effective routine pharmaceutical testing using radial flow stream splitting HPLC columns: Quantitative analysis and performance metrics in the analysis of over-the-counter drugs.

Journal of pharmaceutical and biomedical analysis·2026
Same author

Autocatalytic Electrochemiluminescence.

Angewandte Chemie (International ed. in English)·2025
Same author

Effectiveness of a washing protocol for the removal of gunshot residue from forensically relevant Lucilia sericata larvae.

Forensic science international·2025
Same author

Rethinking the Excited-State Redox Properties of Iron(III) Complexes for LMCT Photoredox Catalysis.

Journal of the American Chemical Society·2025

Related Experiment Video

Updated: May 6, 2026

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

11.3K

Comprehensive sample analysis using high performance liquid chromatography with multi-detection.

Sercan Pravadali1, Danielle N Bassanese, Xavier A Conlan

  • 1Australian Centre for Research on Separation Sciences (ACROSS), School of Science and Health, University of Western Sydney (Parramatta), NSW 1797, Australia.

Analytica Chimica Acta
|November 13, 2013
PubMed
Summary
This summary is machine-generated.

This study highlights limitations in analyzing complex tobacco leaf extracts using uni-dimensional liquid chromatography. Even with multiple detectors, identifying specific compounds proved challenging, suggesting multidimensional chromatography is essential.

Keywords:
AlkaloidsChemiluminescenceDPPHDetectionHigh performance liquid chromatographyTobacco

More Related Videos

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow
09:04

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Published on: April 18, 2019

14.1K
Tuning a Parallel Segmented Flow Column and Enabling Multiplexed Detection
08:01

Tuning a Parallel Segmented Flow Column and Enabling Multiplexed Detection

Published on: December 15, 2015

7.3K

Related Experiment Videos

Last Updated: May 6, 2026

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
10:17

High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

Published on: April 23, 2019

11.3K
Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow
09:04

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Published on: April 18, 2019

14.1K
Tuning a Parallel Segmented Flow Column and Enabling Multiplexed Detection
08:01

Tuning a Parallel Segmented Flow Column and Enabling Multiplexed Detection

Published on: December 15, 2015

7.3K

Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Spectrometry

Background:

  • Tobacco leaf extracts present a highly complex matrix for chemical analysis.
  • Uni-dimensional separation techniques often struggle to resolve all key chemical entities.
  • Advanced detection methods are needed to reduce data complexity and gain molecule-specific information.

Purpose of the Study:

  • To assess the separation capability of optimized uni-dimensional liquid chromatography for tobacco leaf extract analysis.
  • To evaluate the effectiveness of multiple detection modes in simplifying complex sample data.
  • To illustrate the limitations of current separation techniques and advocate for multidimensional approaches.

Main Methods:

  • Optimized uni-dimensional liquid chromatography (UDLC) was employed.
  • Multiple detection techniques were utilized: UV-visible absorbance, chemiluminescence (acidic potassium permanganate, manganese(IV), tris(2,2'-bipyridine)ruthenium(III)), mass spectrometry (MS), and DPPH radical scavenging.
  • Data complexity reduction and molecule-specific information acquisition were attempted.

Main Results:

  • A significant amount of chemical data was generated from the tobacco leaf extract.
  • Limitations were observed in assigning detector responses to specific compounds, even with complementary detection.
  • Mass spectrometry detected thirty-three compounds; 12 out of 32 compounds showed a peak height ratio (PHR) > 0.33 on at least one detector.

Conclusions:

  • Uni-dimensional chromatography, even with optimized conditions and multiple detectors, has inherent limitations for comprehensive analysis of complex matrices like tobacco leaf extracts.
  • The study serves as a case study demonstrating these limitations.
  • Multidimensional chromatography is crucial for developing robust and comprehensive chemical detection systems for such samples.