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2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...

2D NMR: Overview of Heteronuclear Correlation Techniques

2D NMR: Overview of Heteronuclear Correlation Techniques

Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other axis.

Applications Of NMR In Biology

Applications Of NMR In Biology

Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
The...

¹H NMR: Interpreting Distorted and Overlapping Signals

¹H NMR: Interpreting Distorted and Overlapping Signals

Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are slanted or...

NMR Spectroscopy: Chemical Shift Overview

NMR Spectroscopy: Chemical Shift Overview

The position of the absorption signal of a sample is reported relative to the position of the signal of tetramethylsilane (TMS), which is added as an internal reference while recording spectra. The difference between the absorption frequencies of the sample and TMS (in Hz) is divided by the spectrometer operating frequency (in MHz) to obtain a dimensionless quantity called the chemical shift. It is reported on the δ (delta) scale and expressed in parts per million.
For instance, the proton...

Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule

Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule

In the AX proton spin system, proton A can sense the two spin states of a coupled proton X, resulting in a doublet NMR signal with two peaks of equal (1:1) intensity. When proton A is coupled to two equivalent protons (AX2 spin system), the spin states of each X can be aligned with or against the external field, creating three possible scenarios. This results in a 1:2:1 triplet signal, where the central peak corresponds to the chemical shift of A and is twice as large or intense as the others.

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Related Experiment Video

Updated: Jun 27, 2026

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

Published on: April 17, 2018

Quantitative ¹H NMR in Pharmaceutical and Biomedical Analyses: Methodologies and Applications.

Shangxiao An¹, Weiyi Zheng², Qi Tang²

¹School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.

Molecules (Basel, Switzerland)

|June 26, 2026

Summary

Quantitative 1H NMR (qNMR) offers structural and quantitative analysis without standards. This versatile tool is crucial for pharmaceutical drug analysis and biomedical research, with future advancements promising enhanced capabilities.

Keywords:

biomedical analysis drug analysis metabolomics quality control quantitative 1H NMR

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Last Updated: Jun 27, 2026

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

Published on: April 17, 2018

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

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A Strategy for Sensitive, Large Scale Quantitative Metabolomics

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Published on: May 27, 2014

Area of Science:

Analytical Chemistry
Spectroscopy
Biomedical Science

Background:

Quantitative 1H NMR (qNMR) is an analytical technique.
It provides structural and quantitative data without analyte-specific standards.
qNMR has diverse applications in pharmaceutical and biomedical fields.

Purpose of the Study:

To review the methodological fundamentals of qNMR.
To summarize its applications in pharmaceutical and biomedical analysis.
To discuss emerging technologies and future prospects of qNMR.

Main Methods:

Review of existing literature on qNMR methodology.
Analysis of qNMR applications in drug content determination, purity assessment, metabolomics, and cellular process monitoring.
Discussion of advancements like hyperpolarization, AI, and deep learning.

Main Results:

qNMR is effective for analyzing small molecules, polysaccharides, polymers, and herbal medicines.
It is a powerful tool for metabolomics, cellular process monitoring, and metabolite quantification in biofluids and tissues.
Emerging technologies promise to significantly improve qNMR's sensitivity, resolution, and automation.

Conclusions:

qNMR is a versatile and powerful analytical tool for both pharmaceutical and biomedical applications.
Ongoing integration into regulatory frameworks suggests expanding use in quality control and research.
Future technological advancements will further enhance its capabilities and applications.