Capsanthin Analysis Service - Creative Proteomics

Service Details Q&A Case Study

What is Capsanthin?

Capsanthin is a natural red pigment found in chili peppers, which is the material basis for the red color of chili peppers' fruits, and is crucial to the appearance and quality of chili peppers' fruits, and plays a vital role in protecting chili peppers from environmental stresses and regulating the ripening process of the fruits. It is widely used in food processing, cosmetic and pharmaceutical industries because of its strong coloring ability and safety for human beings

Analysis of capsanthin is critical to understanding the physiology, biochemistry, and impact on human health of the pepper plant. Analyzing capsanthin provides you with valuable information on how the pepper plant adapts to different environmental conditions (e.g., light, temperature, and nutrient availability) and how it regulates its growth, development, and fruit ripening. In addition, it provides insights into the metabolic pathways involved in the body's response to oxidative stress and inflammation.

By analyzing and quantifying capsanthin and their metabolites and determining their patterns of change under specific chili pepper growth conditions, environmental factors, or human health conditions, you can lay the scientific groundwork for improving chili pepper plant resistance, increasing the nutritional and commercial value of chili peppers, and promoting human health.

Why Choose Us?

High Sensitivity: Detection limits as low as 0.1 ng/mL using advanced LC-MS/MS systems.
Comprehensive Coverage: Simultaneous analysis of capsanthin isomers, esters, and degradation products.
Rapid Turnaround: Optimized workflows ensure results within 10–20 business days.
Multi-Method Validation: Cross-validated results from HPLC, NMR, and MS platforms for unmatched accuracy.
Custom Solutions: Tailored protocols for complex matrices (e.g., oils, plant extracts, processed foods).

Capsanthin Analysis Services by Creative Proteomics

Quantitative Analysis of Capsanthin: Accurate determination of capsanthin concentration in different matrices.
Capsanthin Purity Testing: Identification of purity levels in food additives and nutraceuticals.
Metabolite Profiling: Comprehensive analysis of capsanthin-related metabolic pathways.
Structural Characterization: Advanced spectroscopic analysis for molecular structure elucidation.
Capsanthin Stability Studies: Investigation of capsanthin stability under different environmental conditions.

Detectable Capsanthin, Related Metabolites, and Metabolic Pathways

Compound Name	Metabolic Pathway	Function/Significance
Capsanthin	Carotenoid Biosynthesis	Primary red pigment in red peppers, antioxidant properties
Capsorubin	Carotenoid Biosynthesis	Red xanthophyll, contributes to pepper coloration
β-Cryptoxanthin	Carotenoid Metabolism	Precursor to vitamin A, antioxidant
Zeaxanthin	Xanthophyll Cycle	Enhances eye health, protects against oxidative stress
Violaxanthin	Xanthophyll Cycle	Involved in photoprotection in plants
Lutein	Xanthophyll Cycle	Antioxidant, important for vision and skin health
Neoxanthin	Carotenoid Biosynthesis	Intermediate in the biosynthesis of ABA (abscisic acid)
Antheraxanthin	Xanthophyll Cycle	Involved in non-photochemical quenching in plants
Phytoene	Early Carotenoid Biosynthesis	Precursor to all carotenoids
Phytofluene	Early Carotenoid Biosynthesis	Intermediate in lycopene biosynthesis
Lycopene	Carotenoid Biosynthesis	Precursor for β-carotene, strong antioxidant
β-Carotene	Carotenoid Metabolism	Precursor to vitamin A, essential for human nutrition
α-Carotene	Carotenoid Metabolism	Pro-vitamin A carotenoid, antioxidant
Abscisic Acid (ABA)	Carotenoid Degradation Pathway	Plant hormone derived from carotenoids, regulates stress responses

Methods Used for Capsanthin Analysis

Ultra-Performance Liquid Chromatography (UPLC-QTOF-MS) – High-resolution separation and mass spectrometry-based identification.

High-Performance Liquid Chromatography (HPLC-DAD) – UV-visible detection for precise quantification.

Gas Chromatography-Mass Spectrometry (GC-MS) – For volatile carotenoid derivatives.

1260 Infinity II LC System (Figure from Agilent)

Waters Xevo G2-XS QTOF (Figure from Waters)

Workflow for Capsanthin Analysis Service

Sample Requirements for Capsanthin Assay

Sample Type	Required Amount	Storage Conditions	Notes
Plant Tissues (e.g., Red Pepper, Chili, Paprika)	≥ 100 mg (dry) or ≥ 1 g (fresh)	-80°C (long-term) or -20°C (short-term)	Freeze-dried samples preferred to prevent degradation
Food Products (e.g., Sauces, Spices, Beverages)	≥ 5 g	4°C or -20°C	Avoid light exposure to prevent oxidation
Nutraceuticals (Capsules, Tablets, Powders)	≥ 10 capsules/tablets or ≥ 500 mg powder	Room temperature (sealed)	Provide ingredient list if available
Liquid Samples (Juices, Extracts, Fermentation Broths)	≥ 10 mL	4°C or -20°C	Store in amber vials to minimize light degradation
Purified Capsanthin Standards	≥ 1 mg	-20°C or -80°C	Dissolved in appropriate solvent (e.g., ethanol)

Applications of Capsanthin Analysis

Q: What are the common analytical methods for Capsanthin?

Capsanthin analysis primarily relies on chromatography and spectroscopy techniques:

HPLC-PDA: High-performance liquid chromatography with a photodiode array detector enables simultaneous detection of capsanthin, zeaxanthin, lutein, and other carotenoids. It offers a short analysis time (<18 minutes) and high sensitivity (LOD as low as 0.02 mg/L).
LC-MS/MS: Used for the qualitative and quantitative analysis of trace amounts of capsanthin and its degradation products (e.g., norisoprenoids) in complex samples, supporting isomer differentiation.
Spectrophotometry and NMR: Suitable for rapid screening and structural identification, though requiring high-purity reference standards.

Q: What are the key steps in sample preparation?

Drying method: Freeze-drying or low-temperature drying is preferred to minimize the loss of heat-sensitive compounds (e.g., vitamin C).
Solvent selection: Acetone or n-hexane is commonly used for extraction, while a saponification step helps remove chlorophyll interference.
Ultrasound/microwave-assisted extraction: Enhances extraction efficiency while reducing solvent consumption, making it suitable for industrial applications.

Q: How to ensure the stability of Capsanthin during analysis?

Light and low-temperature storage: Capsanthin is light-sensitive, requiring amber sample vials and temperature control below 4°C.
Antioxidant addition: Compounds like vitamin E or BHT help prevent oxidative degradation.
Rapid analysis: Reducing the time between extraction and detection prevents degradation due to prolonged exposure to high temperatures.

Q: How to distinguish Capsanthin from other carotenoids (e.g., β-carotene)?

Chromatographic separation: HPLC gradient optimization (e.g., acetone-water mobile phase) allows baseline separation (resolution R > 1.5) of capsanthin and β-carotene.
Spectral characteristics: Capsanthin exhibits a maximum absorption peak at 450 nm, while β-cryptoxanthin peaks at 470 nm.
Mass spectrometry identification: Molecular ion peak differences aid iden
tion (Capsanthin: m/z 584.9; Zeaxanthin: m/z 568.9).

Q: What are the special requirements for Capsanthin analysis in processed foods (e.g., chili sauce)?

Matrix interference removal: Saponification helps eliminate oils, while solid-phase extraction (SPE) is used for sample purification.
Degradation product detection: LC-MS/MS can identify degradation compounds like apocarotenals and xanthophyll epoxides.
Capsaicin co-existence: Optimized chromatographic conditions prevent the overlapping of capsaicin and capsanthin peaks.

Q: What should be considered when selecting a standard for Capsanthin quantification?

Purity verification: Standards should be ≥95% pure, validated via HPLC and NMR.
Calibration curve range: The linear range should match the sample concentration (e.g., 0.1–50 mg/L), with R² > 0.998.
Stability testing: Regularly verify concentration changes in standard solutions over storage periods.

Q: How can the bioactivity of Capsanthin be reflected through analytical data?

Antioxidant activity correlation: Combine DPPH/ABTS free radical scavenging assays with HPLC quantification to evaluate capsanthin's synergy with vitamin C/E.
Metabolic pathway studies: Metabolomics techniques help track capsanthin distribution in lipoproteins and its clearance rate, which is faster than lycopene.

Case. Simultaneous extraction of edible oil from avocado and capsanthin from red bell pepper using supercritical carbon dioxide as solvent

Background:

Vegetable extracts containing bioactive compounds and supercritical CO2 were analyzed.

The study aimed to obtain capsanthin-rich avocado oil using scCO₂ extraction and examine the effect of avocado oil as a co-solvent for capsanthin extraction from red bell pepper.

Samples:

Avocado (variety Haas) from Jaguacy Brasil and red bell pepper from the local market in Campinas/SP, Brazil were used. The pulps were freeze-dried after processing.

Technical methods procedure:

The moisture content was determined by gravimetric method for fresh samples.

The freeze-dried samples was determined by Karl Fischer method.

Particle size was measured by ASAE method after crushing/grinding and sieving.

Bed density and porosity were calculated.

Extraction experiments were conducted in a fixed bed extractor at 50 °C and 400 bar with a 1.5 L/min CO₂ flow rate. Extraction curves were constructed by collecting samples at different times and measuring extract and solvent masses.

Lipid content of avocado was determined by Soxhlet method.

Capsanthin content in raw material was measured by exhaustive extraction with acetone followed by isolation, identification and quantification using open-column chromatography and UHPLC-PDA.

Results:

The moisture contents of avocado and red bell pepper decreased after freeze-drying.

Avocado had a high lipid content (68%), and red bell pepper had a certain capsanthin level (625 ± 87 g/g in freeze-dried).

The extraction curves showed the oil solubility in scCO₂ and the extraction kinetics of both materials.

The overall extraction yield of avocado oil was 57% with 84.3% recovery. Capsanthin extraction yield was low (15%) without co-solvent.

In simultaneous extraction, as the proportion of red bell pepper to avocado decreased, capsanthin recovery increased (from 15% to 32%, 35%, and 56% with increasing avocado concentration).

Capsanthin concentration in the extract decreased with more avocado oil as co-solvent due to dilution.

Chromatogram of the freeze-dried red pepper extract.

Reference

Barros H D F Q, et al. "Simultaneous extraction of edible oil from avocado and capsanthin from red bell pepper using supercritical carbon dioxide as solvent." Journal of Supercritical Fluids 107 (2016): 315 - 320.

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