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HPLC-fingerprints and antioxidant constituents of Phyla nodiflora.

Lin FJ, Yen FL, Chen PC, Wang MC, Lin CN, Lee CW, Ko HH - ScientificWorldJournal (2014)

Bottom Line: In the present study, ten compounds, namely, 3,7,4',5'-tetrahydroxy-3'-methoxyflavone (1), nodifloretin (2), 4'-hydroxywogonin (3), onopordin (4), cirsiliol (5), 5,7,8,4'-tetrahydroxy-3'-methoxyflavone (6), eupafolin (7), hispidulin (8), larycitrin (9), and β-sitosterol were isolated from the methanolic extract of the aerial part of P. nodiflora (PNM) and their structures were identified by 1D-NMR comparing their spectra with the literature.The antioxidant activities of these compounds were evaluated by free radical scavenging activity and tyrosinase inhibitory effect in cell-free systems.Compounds 4, 5, and 7 showed strong antioxidant activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan.

ABSTRACT
Phyla nodiflora is a creeping perennial herb, widely distributed in the most tropical and subtropical regions. It has been used as a folk medicine, herbal beverage, or folk cosmetic. For these usages, the development of a chemical quality control method of this plant is necessary. In the present study, ten compounds, namely, 3,7,4',5'-tetrahydroxy-3'-methoxyflavone (1), nodifloretin (2), 4'-hydroxywogonin (3), onopordin (4), cirsiliol (5), 5,7,8,4'-tetrahydroxy-3'-methoxyflavone (6), eupafolin (7), hispidulin (8), larycitrin (9), and β-sitosterol were isolated from the methanolic extract of the aerial part of P. nodiflora (PNM) and their structures were identified by 1D-NMR comparing their spectra with the literature. The antioxidant activities of these compounds were evaluated by free radical scavenging activity and tyrosinase inhibitory effect in cell-free systems. Compounds 4, 5, and 7 showed strong antioxidant activity. To control the quality of P. nodiflora, a simple and reliable method of high-performance liquid chromatography combined with ultraviolet detector (HPLC-UV) was established for both the fingerprint analysis and the quantitative determination of two selected active compounds, onopordin (4) and eupafolin (7). Statistical analysis of the obtained data demonstrated that our method achieved the desired linearity, precision, and accuracy. The results indicated that the developed method can be used as a quality evaluation method for PNM.

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(A) The chromatogram of mixture standard compounds: nodifloretin (2), onopordin (4), and eupafolin (7). (B) Chromatographic fingerprint of PNM (2010-06-PNM) by HPLC-UV at 326 nm.
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fig3: (A) The chromatogram of mixture standard compounds: nodifloretin (2), onopordin (4), and eupafolin (7). (B) Chromatographic fingerprint of PNM (2010-06-PNM) by HPLC-UV at 326 nm.

Mentions: The wavelength for the detection of PNM and the active constituents in this plant was selected by the UV-Vis (200–400 nm) detector. Due to a full-scan experiment of PNM, its UV spectrum showed maximum adsorption at the wavelengths of 210, 273, and 326 nm. The chromatographs monitored at 210 and 273 nm revealed more peaks than at 326 nm within 15–20 min. However, due to the fact that some peaks were missing between 40 and 70 min and due to the presence of serious baseline noise, some peaks seen at 210 and 273 nm were not well separated. Meanwhile, signals response for flavones and flavonols is commonly referred to as band I (usually 300–380 nm) and band II (usually 240–280 nm). In order to detect more common peaks while achieving suitable detection of PNM and active constituents, 326 nm was selected as the most appropriate detection wavelength. The HPLC chromatograms of standards and PNM are shown in Figure 3.


HPLC-fingerprints and antioxidant constituents of Phyla nodiflora.

Lin FJ, Yen FL, Chen PC, Wang MC, Lin CN, Lee CW, Ko HH - ScientificWorldJournal (2014)

(A) The chromatogram of mixture standard compounds: nodifloretin (2), onopordin (4), and eupafolin (7). (B) Chromatographic fingerprint of PNM (2010-06-PNM) by HPLC-UV at 326 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4129175&req=5

fig3: (A) The chromatogram of mixture standard compounds: nodifloretin (2), onopordin (4), and eupafolin (7). (B) Chromatographic fingerprint of PNM (2010-06-PNM) by HPLC-UV at 326 nm.
Mentions: The wavelength for the detection of PNM and the active constituents in this plant was selected by the UV-Vis (200–400 nm) detector. Due to a full-scan experiment of PNM, its UV spectrum showed maximum adsorption at the wavelengths of 210, 273, and 326 nm. The chromatographs monitored at 210 and 273 nm revealed more peaks than at 326 nm within 15–20 min. However, due to the fact that some peaks were missing between 40 and 70 min and due to the presence of serious baseline noise, some peaks seen at 210 and 273 nm were not well separated. Meanwhile, signals response for flavones and flavonols is commonly referred to as band I (usually 300–380 nm) and band II (usually 240–280 nm). In order to detect more common peaks while achieving suitable detection of PNM and active constituents, 326 nm was selected as the most appropriate detection wavelength. The HPLC chromatograms of standards and PNM are shown in Figure 3.

Bottom Line: In the present study, ten compounds, namely, 3,7,4',5'-tetrahydroxy-3'-methoxyflavone (1), nodifloretin (2), 4'-hydroxywogonin (3), onopordin (4), cirsiliol (5), 5,7,8,4'-tetrahydroxy-3'-methoxyflavone (6), eupafolin (7), hispidulin (8), larycitrin (9), and β-sitosterol were isolated from the methanolic extract of the aerial part of P. nodiflora (PNM) and their structures were identified by 1D-NMR comparing their spectra with the literature.The antioxidant activities of these compounds were evaluated by free radical scavenging activity and tyrosinase inhibitory effect in cell-free systems.Compounds 4, 5, and 7 showed strong antioxidant activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan.

ABSTRACT
Phyla nodiflora is a creeping perennial herb, widely distributed in the most tropical and subtropical regions. It has been used as a folk medicine, herbal beverage, or folk cosmetic. For these usages, the development of a chemical quality control method of this plant is necessary. In the present study, ten compounds, namely, 3,7,4',5'-tetrahydroxy-3'-methoxyflavone (1), nodifloretin (2), 4'-hydroxywogonin (3), onopordin (4), cirsiliol (5), 5,7,8,4'-tetrahydroxy-3'-methoxyflavone (6), eupafolin (7), hispidulin (8), larycitrin (9), and β-sitosterol were isolated from the methanolic extract of the aerial part of P. nodiflora (PNM) and their structures were identified by 1D-NMR comparing their spectra with the literature. The antioxidant activities of these compounds were evaluated by free radical scavenging activity and tyrosinase inhibitory effect in cell-free systems. Compounds 4, 5, and 7 showed strong antioxidant activity. To control the quality of P. nodiflora, a simple and reliable method of high-performance liquid chromatography combined with ultraviolet detector (HPLC-UV) was established for both the fingerprint analysis and the quantitative determination of two selected active compounds, onopordin (4) and eupafolin (7). Statistical analysis of the obtained data demonstrated that our method achieved the desired linearity, precision, and accuracy. The results indicated that the developed method can be used as a quality evaluation method for PNM.

Show MeSH