2D NMR Spectroscopy Services

Advanced two-dimensional NMR techniques for complete structure elucidation of complex molecules.

2D NMR spectroscopy provides correlations between different nuclei, revealing connectivity and spatial relationships that are impossible to determine from 1D spectra alone. Essential for natural products, complex synthetic compounds, peptides, and structural biology.

What is 2D NMR?

Two-dimensional NMR spectroscopy displays data in a two-dimensional plot, revealing correlations between different nuclei. While 1D NMR (H-1 or C-13) provides a list of chemical shifts, 2D NMR shows which atoms are connected or spatially close, providing crucial information for:

  • Structure elucidation - Determine complete molecular structure
  • Connectivity analysis - Identify which atoms are bonded together
  • Stereochemistry - Determine 3D spatial arrangements
  • Complex molecule analysis - Resolve overlapping signals in 1D spectra

2D NMR Techniques Available

COSY (Correlation Spectroscopy)

What it shows: 1H-1H correlations through bonds

COSY reveals which protons are coupled to each other (typically 2-3 bonds apart), showing the connectivity pattern in your molecule.

What You Learn

  • Which protons are adjacent or nearby
  • Spin system identification
  • Structural connectivity patterns
  • Assignment of overlapping signals

Best Used For

  • Determining proton connectivity
  • Resolving complex multiplets
  • Small to medium-sized molecules
  • Initial structure determination

HSQC (Heteronuclear Single Quantum Coherence)

What it shows: Direct 1H-13C correlations (1-bond)

HSQC is one of the most useful 2D experiments, showing which protons are directly attached to which carbons. This is typically the first heteronuclear 2D experiment performed.

What You Learn

  • Direct C-H connectivity
  • Carbon and proton assignments simultaneously
  • Distinguishes CH, CH₂, and CH₃ groups
  • Cleaner than HMQC (similar technique)

Best Used For

  • Complete NMR assignment
  • Natural product structure elucidation
  • Complex organic molecules
  • Pharmaceutical compound verification

HMBC (Heteronuclear Multiple Bond Correlation)

What it shows: Long-range 1H-13C correlations (2-4 bonds)

HMBC reveals which protons are near which carbons, even when they're not directly bonded. This is crucial for determining the carbon skeleton and quaternary carbon positions.

What You Learn

  • Long-range C-H correlations
  • Quaternary carbon positions
  • Carbonyl carbon connections
  • Complete carbon skeleton

Best Used For

  • Identifying quaternary carbons
  • Connecting molecular fragments
  • Natural product structure determination
  • Verifying proposed structures

NOESY (Nuclear Overhauser Effect Spectroscopy)

What it shows: Spatial proximity between protons (through space, not bonds)

NOESY reveals which protons are physically close in 3D space (typically <5 Å), regardless of bonding. This is essential for determining stereochemistry and conformation.

What You Learn

  • 3D spatial relationships
  • Stereochemistry (cis/trans, relative configuration)
  • Molecular conformation
  • Proximity of functional groups

Best Used For

  • Determining stereochemistry
  • Conformational analysis
  • Resolving structural ambiguities

TOCSY (Total Correlation Spectroscopy)

What it shows: All protons within a spin system

TOCSY reveals complete spin systems, showing all protons that are connected through a chain of couplings. Particularly useful for complex molecules like sugars and peptides.

What You Learn

  • Complete spin system identification
  • All coupled protons at once
  • Amino acid residues in peptides
  • Sugar ring assignments

Best Used For

  • Carbohydrate structure
  • Complex natural products
  • Overlapping signal resolution

When is 2D NMR Necessary?

2D NMR becomes essential when 1D NMR alone cannot solve your structural problem:

Classic Scenarios for 2D NMR

  • Complex natural products - Multiple rings, many functional groups, overlapping signals
  • Unknown compound identification - Need complete structure determination
  • Quaternary carbons - HMBC required to locate carbons with no attached protons
  • Stereochemistry questions - NOESY needed to determine relative configuration
  • Overlapping 1D signals - 2D spreads signals into second dimension
  • Publication requirements - Many journals require 2D NMR for structure proof
  • Structural confirmation - Verify proposed structure is correct

Typical 2D NMR Workflow

Most structure elucidation projects follow this sequence:

Step Experiment Information Gained
1 H-1 NMR Proton count and chemical shifts
2 C-13 NMR + DEPT Carbon count and types (CH₃, CH₂, CH, C)
3 HSQC Which H's are on which C's
4 COSY Which H's are coupled to which H's
5 HMBC Long-range C-H correlations, quaternary carbons
6 (if needed) NOESY Stereochemistry and 3D structure

Not all experiments are needed for every compound. We can recommend the optimal suite of experiments based on your structure and goals.

Sample Requirements for 2D NMR

Sample Amount

  • Minimum: 10-20 mg
  • Recommended: 20-100 mg for best results
  • 2D NMR is less sensitive than 1D
  • More material = faster acquisition, better data
  • Contact us if sample is limited

Sample Concentration

  • Higher concentration shortens experiment time
  • Need enough for good signal-to-noise
  • Typical volume: 0.5-0.7 mL in NMR tube

Sample Preparation Tips

  • Purity: Higher purity gives cleaner 2D spectra
  • Solubility: Must be completely soluble - particles interfere with shimming
  • Solvent: Deuterated solvents (CDCl₃, DMSO-d₆, D₂O, etc.)
  • Dry sample: Remove residual H₂O when possible
  • Reference:solvent peak

Turnaround Time & Pricing

Standard Turnaround: 3-7 business days (depends on complexity and number of experiments)

Rush Service: Contact us to discuss accelerated timelines

Factors Affecting Turnaround

  • Number of experiments - Basic package (COSY, HSQC) vs. complete suite
  • Sample concentration - Lower concentration requires longer acquisition
  • Data quality requirements - Publication-quality needs more signal averaging
  • Instrument availability - Some experiments require significant instrument time

For pricing information, please visit our Pricing page or contact us. Pricing depends on which experiments are needed and sample complexity.

What You'll Receive

Data Files

  • Raw 2D data (Bruker format)
  • Processed 2D spectra
  • Publication-quality images (PDF, PNG)
  • 1D spectra (H-1, C-13 if acquired)
  • All acquisition parameters

Optional Services

  • Consultation - Discuss results with our chemists

Need 2D NMR for Structure Elucidation?

Contact us to discuss which 2D experiments are right for your compound

Submit a Sample Request a Quote

Phone: (858) 793-6057 | Email: NuMegaLab@NuMegaLabs.com

H-1 NMR

Start with 1D proton NMR

C-13 NMR

Carbon skeleton determination

DEPT/APT

Distinguish CH₃, CH₂, CH

Quantitative NMR

Accurate purity analysis