MUAL- Phenolics & Flavonoids
1
Phenolics & Flavonoids RESULTS What is being measured: Global profiling of phenolic compounds in plants. Phenolic metabolites biosynthesized by plants are broadly categorized as simple phenolic acids, flavonoids, tannins and lignins. The phenolic acids and flavonoids together constitute the phenylpropanoids, and is the focus of this analysis. The flavonoids are further categorized into six broad groups based on the chemical structure: flavanones, flavones, flavonols, flavan-3-ols, anthocyanins, and proanthocyanidins. These compounds perform a myriad of functions in plants by serving as signaling molecules, structural polymers, attractants and deterrents, regulators of phytohormones, and stress mitigators. Flavonoids are also of greater importance in human health due to their antioxidant and anti-cancerous properties. More than 5000 phenolic compounds have been characterized from plants. How it is done: Phenolics and flavonoids are extracted from samples using aqueous- methanol and are analyzed using liquid chromatograph coupled to a high resolution mass spectrometer (LC-MS/MS). Compound identification is achieved through the matching accurate mass and fragmentation pattern of the precursors with in-house mass spectral libraries. Absolute quantitation is possible when the authentic standards are available. Relative quantitation is determined by normalization with isotope labeled internal standard. MUAL has a large repository (>1500) of authentic flavonoid and phenylpropanoid standards for confident identification and quantitation. Figure 5. Intelligent, automated product ion- dependent MS n mass spectrometric method for deciphering the glycosylation pattern in flavonoids on the Orbitrap ID-X Tribrid MS instrument. Figure 2. Phenolic compounds in different berries as influenced by cultivation practices (organic vs conventional). INSTRUMENTATION 111 Biosystems Research Complex (BRC) 105 Collings Street, Cl`emson SC 29634 Ph: (864) 656-5060 Website: https://www.clemson.edu/cafls/mual/ CONTACT US • Combines quadrupole, ion trap and Orbitrap mass analysis in Tribrid architecture • Ultrahigh resolution up to 500,000 FWHM • Sub ppm mass accuracy • Multiple dissociation techniques—CID, HCD, ETD • ID-X capability for small molecule identification • Coupled to HESI and nano-ESI interfaces • UltiMate 3000 RS UHPLC & UltiMate 3000 RSLCnano Thermo Orbitrap Fusion™ Tribrid™ Mass Spectrometer Nishanth Tharayil Facility Director [email protected] Elizabeth Leonard Research Associate [email protected] Khang Huynh Postdoctoral Fellow [email protected] Jisun Lee Research Associate [email protected] Figure 1. Scheme of the biosynthesis of polyphenolics in plants. Figure 4. In-house spectral trees used for the accurate identification of unknown flavonoids. Figure 3. Approaches adopted by MUAL to establish identities of unknown compounds using accurate mass MSn libraries.
Transcript of MUAL- Phenolics & Flavonoids
Phenolics & Flavonoids
RESULTS
What is being measured: Global profiling of phenolic compounds in plants. Phenolic
metabolites biosynthesized by plants are broadly categorized as simple phenolic acids,
flavonoids, tannins and lignins. The phenolic acids and flavonoids together constitute
the phenylpropanoids, and is the focus of this analysis. The flavonoids are further
categorized into six broad groups based on the chemical structure: flavanones,
flavones, flavonols, flavan-3-ols, anthocyanins, and proanthocyanidins. These
compounds perform a myriad of functions in plants by serving as signaling molecules,
structural polymers, attractants and deterrents, regulators of phytohormones, and
stress mitigators. Flavonoids are also of greater importance in human health due to
their antioxidant and anti-cancerous properties. More than 5000 phenolic compounds
have been characterized from plants.
How it is done: Phenolics and flavonoids are extracted from samples using aqueous-
methanol and are analyzed using liquid chromatograph coupled to a high resolution
mass spectrometer (LC-MS/MS). Compound identification is achieved through the
matching accurate mass and fragmentation pattern of the precursors with in-house
mass spectral libraries. Absolute quantitation is possible when the authentic standards
are available. Relative quantitation is determined by normalization with isotope labeled
internal standard. MUAL has a large repository (>1500) of authentic flavonoid and
phenylpropanoid standards for confident identification and quantitation.
Figure 5. Intelligent, automated product ion-
dependent MSn mass spectrometric method for
deciphering the glycosylation pattern in flavonoids
on the Orbitrap ID-X Tribrid MS instrument.
Figure 2. Phenolic compounds in different berries
as influenced by cultivation practices (organic vs
conventional).
INSTRUMENTATION
111 Biosystems Research Complex (BRC)
105 Collings Street, Cl`emson SC 29634
Ph: (864) 656-5060
Website: https://www.clemson.edu/cafls/mual/
CONTACT US
• Combines quadrupole, ion trap and Orbitrap mass analysis in Tribrid architecture
• Ultrahigh resolution up to 500,000 FWHM
• Sub ppm mass accuracy
• Multiple dissociation techniques—CID, HCD, ETD
• ID-X capability for small molecule identification
• Coupled to HESI and nano-ESI interfaces
• UltiMate 3000 RS UHPLC & UltiMate 3000 RSLCnano
Thermo Orbitrap Fusion™ Tribrid™ Mass Spectrometer
Nishanth Tharayil
Facility Director
Elizabeth Leonard
Research Associate
Khang Huynh
Postdoctoral Fellow
Jisun Lee Research Associate [email protected]
Figure 1. Scheme of the biosynthesis
of polyphenolics in plants.
Figure 4. In-house spectral trees used for the
accurate identification of unknown flavonoids.Figure 3.
Approaches
adopted by
MUAL to
establish
identities of
unknown
compounds
using accurate
mass MSn
libraries.
