Verbena Officinalis

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Hermann Stuppner - One of the best experts on this subject based on the ideXlab platform.

  • towards eco friendly secondary plant metabolite quantitation ultra high performance supercritical fluid chromatography applied to common vervain Verbena Officinalis l
    IEEE Journal of Solid-state Circuits, 2020
    Co-Authors: Nora Gibitzeisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Christoph Seger, Hermann Stuppner
    Abstract:

    This report presents the first ultra high performance supercritical fluid chromatography diode array detector based assay for simultaneous determination of iridoid glucosides, flavonoid glucuronides, and phenylpropanoid glycosides in Verbena Officinalis (Verbenaceae) extracts. Separation of the key metabolites was achieved in less than 7 min on an Acquity UPC2 Torus Diol column using a mobile phase gradient comprising subcritical carbon dioxide and methanol with 0.15% phosphoric acid. Method validation for seven selected marker compounds (hastatoside, Verbenalin, apigenin-7-O-glucuronide, luteolin-7-O-glucuronide, apigenin-7-O-diglucuronide, verbascoside, and luteolin-7-O-diglucuronide) confirmed the assay to be sensitive, linear, precise, and accurate. Head-to-head comparison to an ultra high performance liquid chromatography comparator assay did prove the high orthogonality of the methods. Quantitative result equivalence was evaluated by Passing-Bablok-correlation and Bland-Altman-plot analysis. This cross-validation revealed, that one of the investigated marker compound peaks was contaminated in the ultra high performance liquid chromatography assay by a structurally related congener. Taken together, it was proven that the ultra high performance supercritical fluid chromatography instrument setup with its orthogonal selectivity is a true alternative to conventional reversed phase liquid chromatography in quantitative secondary metabolite analysis. For regulatory purposes, assay cross-validation with highly orthogonal methods seems a viable approach to avoid analyte overestimation due to coeluting, analytically indistinguishable contaminants.

  • Towards eco‐friendly secondary plant metabolite quantitation: Ultra high performance supercritical fluid chromatography applied to common vervain (Verbena Officinalis L.)
    Journal of separation science, 2019
    Co-Authors: Nora Gibitz-eisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Christoph Seger, Hermann Stuppner
    Abstract:

    This report presents the first ultra high performance supercritical fluid chromatography diode array detector based assay for simultaneous determination of iridoid glucosides, flavonoid glucuronides, and phenylpropanoid glycosides in Verbena Officinalis (Verbenaceae) extracts. Separation of the key metabolites was achieved in less than 7 min on an Acquity UPC2 Torus Diol column using a mobile phase gradient comprising subcritical carbon dioxide and methanol with 0.15% phosphoric acid. Method validation for seven selected marker compounds (hastatoside, Verbenalin, apigenin-7-O-glucuronide, luteolin-7-O-glucuronide, apigenin-7-O-diglucuronide, verbascoside, and luteolin-7-O-diglucuronide) confirmed the assay to be sensitive, linear, precise, and accurate. Head-to-head comparison to an ultra high performance liquid chromatography comparator assay did prove the high orthogonality of the methods. Quantitative result equivalence was evaluated by Passing-Bablok-correlation and Bland-Altman-plot analysis. This cross-validation revealed, that one of the investigated marker compound peaks was contaminated in the ultra high performance liquid chromatography assay by a structurally related congener. Taken together, it was proven that the ultra high performance supercritical fluid chromatography instrument setup with its orthogonal selectivity is a true alternative to conventional reversed phase liquid chromatography in quantitative secondary metabolite analysis. For regulatory purposes, assay cross-validation with highly orthogonal methods seems a viable approach to avoid analyte overestimation due to coeluting, analytically indistinguishable contaminants.

  • Development and validation of a rapid ultra-high performance liquid chromatography diode array detector method for Verbena Officinalis L.
    Journal of pharmaceutical and biomedical analysis, 2018
    Co-Authors: Nora Gibitz-eisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Hermann Stuppner
    Abstract:

    Abstract This study presents a fast and validated ultra-high performance liquid chromatography diode array detector (UHPLC-DAD) method for the simultaneous determination of the major compounds in Verbena Officinalis L. (Verbenaceae), a medicinal plant listed in the European, British, and, Chinese Pharmacopoeias. In order to get reference substances, nine compounds, belonging to iridoids, flavonoids, and phenylpropanoid glycosides, were isolated from the herb extract. Two of them, namely cistanoside D and leucosceptoside A, were found in this plant species for the first time. Chromatographic separation was achieved in less than 7 min on a Kinetex  1.7 u XB-C18 (50 × 2.10 mm) column by using a solvent gradient of water-acetonitrile modified with 0.1% formic acid. Method validation confirmed the assays sensitivity, linearity (R2 ≥ 0.997), precision (intraday precision ≤ 1.71%; interday precision ≤ 1.46%) and accuracy (recovery rates between 93.9% and 108.8%) for the quantitative analysis of the eight selected marker compounds. Identity and peak purity of the analytes was confirmed by coupling the UHPLC instrument to a quadrupole time-of-flight mass spectrometer via an electrospray ionization interface (ESI-QTOF-MS) operating in the negative ionization mode. Finally, the applicability of the developed UHPLC-DAD method was successfully proven for the sensitive quantitation of the major compounds in Verbena herb extracts, thereby providing a reliable tool for its rapid quality control.

  • effects of differential extraction of Verbena Officinalis on rat models of inflammation cicatrization and gastric damage
    Planta Medica, 2007
    Co-Authors: Ester Speroni, A. Müller, Rinaldo Cervellati, Stefano Costa, M C Guerra, Aneli Utan, Paolo Govoni, A Berger, Hermann Stuppner
    Abstract:

    Verbena Officinalis L. is used in folk medicine for the treatment of inflammatory disorders, skin burns, abrasions, and gastric diseases. Extracts obtained with different solvents (methanol, VoME; enriched flavonoids, VoEF; supercritical CO2, VoCO2) were evaluated for anti-inflammatory, gastroprotective and cicatrizing activities. Additionally, the antioxidant capacity was determined in vitro. In order to confirm the activities investigated, histological observations were performed. All extracts induce a remarkable anti-inflammatory activity. The gastric damage is significantly reduced by all extracts administered, whereby the most pronounced protection is observed for the VoCO2 and VoEF extracts. Finally, a wound healing effect is obtained particularly by the CO2 extract, suggesting the presence of some lipophilic active principles. Histological evidence confirms the results evaluated with the animal procedures. The results obtained after oral administration of V. Officinalis extracts are also in agreement with the antioxidant capacity evaluated in vitro, confirming the relationship between pharmacological activities and antiradical efficacy.

  • Analysis of the Aerial Parts of Verbena Officinalis L. by Micellar Electrokinetic Capillary Chromatography
    Chromatographia, 2004
    Co-Authors: A. Müller, Markus Ganzera, Hermann Stuppner
    Abstract:

    A micellar electrokinetic capillary chromatography (MECC) method was developed for the qualitative and quantitative determination of five marker compounds (iridoid glycosides, flavonoids and phenylethanoids) in Verbena Officinalis. Optimum separation was achieved using a 50 mM sodium borate solution (pH 9.3), containing 50 mM sodium dodecylsulfate (SDS) as surfactant, at an applied voltage of 25 kV and a temperature of 30 °C, respectively. Because of their different absorption maxima, the compounds were detected either at 205 or 235 nm. Calibration data confirmed linearity of the detector response within the concentration range injected (R2 from 0.997 to 0.999), and revealed detection limits ranging from 5.0 μg mL−1 (verbascoside) to 13.6 μg mL−1 (hastatoside). The five markers were readily assignable in several samples of Verbena.

Halina Ekiert - One of the best experts on this subject based on the ideXlab platform.

  • In Vitro Cultures of Some Medicinal Plant Species (Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and Scutellaria baicalensis) as a Rich Potential Source of Antioxidants-Evaluation by CUPRAC and QUENCHER-CUPRAC Assays.
    Plants (Basel Switzerland), 2021
    Co-Authors: Michał Dziurka, Halina Ekiert, Paweł Kubica, Inga Kwiecień, Jolanta Biesaga-kościelniak, Shaimaa A. M. Abdelmohsen, Fatemah Farraj Alharbi, Diaa O. El-ansary, Hosam O. Elansary, Agnieszka Szopa
    Abstract:

    Comparative estimations of the antioxidant activity of methanolic extracts from biomasses of different types of in vitro cultures of Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and S. baicalensis and also from plant raw materials were performed. The antioxidant measurements were based on the modern assays-cupric ion reducing antioxidant capacity (CUPRAC) and quick, easy, new, cheap, and reproducible CUPRAC (QUENCHER-CUPRAC). The total extractable antioxidants (CUPRAC assay) ranged from 10.4 to 49.7 mmol (100 g)-1 of dry weight (DW) expressed as Trolox equivalent antioxidant capacity (TEAC), and the global antioxidant response (QUENCHER-CUPRAC assay) ranged from 16.0 to 79.1 mmol (100 g)-1 DW for in vitro cultures, whereas for plant raw materials the total extractable antioxidants ranged from 20.9 to 69.5 mmol (100 g)-1 DW, and the global antioxidant response ranged from 67.2 to 97.8 mmol (100 g)-1 DW. Finally, the in vitro cultures could be regarded as an antioxidant-rich alternative resource for the pharmaceutical, health food and cosmetics industries.

  • in vitro cultures of some medicinal plant species cistus incanus Verbena Officinalis scutellaria lateriflora and scutellaria baicalensis as a rich potential source of antioxidants evaluation by cuprac and quencher cuprac assays
    Plants (Basel Switzerland), 2021
    Co-Authors: Michał Dziurka, Pawel Kubica, Halina Ekiert, Inga Kwiecień, Shaimaa A. M. Abdelmohsen, Fatemah Farraj Alharbi, Hosam O. Elansary, Jolanta Biesagakościelniak, Diaa O Elansary, Agnieszka Szopa
    Abstract:

    Comparative estimations of the antioxidant activity of methanolic extracts from biomasses of different types of in vitro cultures of Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and S. baicalensis and also from plant raw materials were performed. The antioxidant measurements were based on the modern assays-cupric ion reducing antioxidant capacity (CUPRAC) and quick, easy, new, cheap, and reproducible CUPRAC (QUENCHER-CUPRAC). The total extractable antioxidants (CUPRAC assay) ranged from 10.4 to 49.7 mmol (100 g)-1 of dry weight (DW) expressed as Trolox equivalent antioxidant capacity (TEAC), and the global antioxidant response (QUENCHER-CUPRAC assay) ranged from 16.0 to 79.1 mmol (100 g)-1 DW for in vitro cultures, whereas for plant raw materials the total extractable antioxidants ranged from 20.9 to 69.5 mmol (100 g)-1 DW, and the global antioxidant response ranged from 67.2 to 97.8 mmol (100 g)-1 DW. Finally, the in vitro cultures could be regarded as an antioxidant-rich alternative resource for the pharmaceutical, health food and cosmetics industries.

  • Accumulation of volatile constituents in agar and bioreactor shoot cultures of Verbena Officinalis L.
    Plant Cell Tissue and Organ Culture (PCTOC), 2021
    Co-Authors: Adam Kokotkiewicz, Bozena Zabiegala, Pawel Kubica, Agnieszka Szopa, Adam Bucinski, Halina Ekiert, Maria Luczkiewicz
    Abstract:

    In vitro shoot cultures of common vervain ( Verbena Officinalis L., Verbenaceae) were established and evaluated for essential oil content. The shoots were maintained as stationary microshoot culture, grown on Schenk–Hildebrandt medium supplemented with 9.84 µM of 2-isopentenyladenine and 1.0 µM of thidiazuron. The shoots were further adapted to grow in two temporary immersion bioreactors (RITA and Plantform), using Schenk–Hildebrandt medium supplemented with 4.92 µM indole-3-butyric acid. The best growth, in terms of dry weight production, was recorded for agar culture (0.73 g l^−1 day^−1), followed by 35 days (day) Plantform (0.53 g l^−1 day^−1), 35 days RITA (0.42 g l^−1 day^−1) and 56 days Plantform culture (0.29 g l^−1 day^−1). The volatile fractions were obtained by simultaneous hydrodistillation-extraction of the plant materials using Likens–Nickerson apparatus. Aerial parts of V. Officinalis intact plant and 56 d Plantform culture had the highest essential oil content (8.1 g kg^−1) whereas the stationary agar culture excelled in essential oil productivity (5.0 mg l^−1 day^−1). The volatile oils isolated from in vitro shoots contained 1-octen-3-ol, benzyl alcohol, phenethyl alcohol, coumaran and hexadecenoic acid as major constituents. 1-Octen-3-ol was the predominant (> 50%) constituent of 35 day shoot cultures maintained in temporary immersion bioreactors whereas coumaran (19%) and hexadecanoid acid (21%) prevailed in stationary microshoot culture. The volatile fraction obtained from aerial parts of V. Officinalis intact plant was composed mainly of hexadecanoic acid (> 20%), linalool (> 8%), anethole (> 5%) and carvone (> 3%). Microshoot cultures of Verbena Officinalis , grown in temporary immersion bioreactors, were investigated in terms of essential oil content and composition of volatile fraction

  • Verbena Officinalis (Common Vervain) - A Review on the Investigations of This Medicinally Important Plant Species.
    Planta medica, 2020
    Co-Authors: Paweł Kubica, Agnieszka Szopa, Maria Luczkiewicz, Jakub Dominiak, Halina Ekiert
    Abstract:

    Verbena Officinalis (common vervain) is a medicinal plant species widely distributed in the world and commonly used in folk medicine of different countries, including traditional Chinese medicine. Monographs on “Verbenae herba” have been included in the European Pharmacopoeia since 2008, and in the Chinese Pharmacopoeia since 1995. This work presents botanical characteristics of this species. It reviews the current knowledge of its chemical composition, which is a rich source mostly of iridoids, phenylpropanoid glycosides, phenolic acids, flavonoids, terpenoids, and essential oil. A large part of this article summarizes traditional medicinal uses and professional pharmacological in vitro and in vivo studies that prove new important applications, e.g., antioxidant, antimicrobial, anti-inflammatory, neuroprotective anticancer, analgesic, or anticonvulsant of Verbena herb extracts and individual metabolites. Moreover, emphasis is put on the use of V. Officinalis in the food and cosmetics industries, especially due to its antioxidant, antibacterial, and anti-inflammatory properties, and the presence of essential oil with an attractive fragrance composition. This paper also presents the state of biotechnological studies of this species.

  • production of verbascoside and phenolic acids in biomass of Verbena Officinalis l vervain cultured under different in vitro conditions
    Natural Product Research, 2017
    Co-Authors: Pawel Kubica, Agnieszka Szopa, Halina Ekiert
    Abstract:

    AbstractMethanolic extracts from the biomass of Verbena Officinalis cultured under continuous artificial light and in darkness on 12 variants of the Murashige and Skoog medium containing different concentrations (0.5–3.0 mg/L) of plant growth regulators: 6-benzyladenine, kinetin, 1-naphthaleneacetic acid and indole-3-butyric acid, were analysed for the amounts of verbascoside and phenolic acids, before and after acid hydrolysis, using the HPLC-DAD method. The amounts of verbascoside were very high (max. 2454.12 mg/100 g DW – light, and 2135.59 mg/100 g DW – darkness). The total amounts of phenolic acids reached a maximum of 46.02 mg/100 g DW (free phenolic acids) and 141.05 mg/100 g DW (bound compounds). The main metabolites were: ferulic, o-coumaric and caffeic acids. The maximum amount of verbascoside was 3.28 times higher than in extracts from the herb of the parent plant. The cultures could be proposed as a potential biotechnological source for selected biologically active compounds.

Agnieszka Szopa - One of the best experts on this subject based on the ideXlab platform.

  • In Vitro Cultures of Some Medicinal Plant Species (Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and Scutellaria baicalensis) as a Rich Potential Source of Antioxidants-Evaluation by CUPRAC and QUENCHER-CUPRAC Assays.
    Plants (Basel Switzerland), 2021
    Co-Authors: Michał Dziurka, Halina Ekiert, Paweł Kubica, Inga Kwiecień, Jolanta Biesaga-kościelniak, Shaimaa A. M. Abdelmohsen, Fatemah Farraj Alharbi, Diaa O. El-ansary, Hosam O. Elansary, Agnieszka Szopa
    Abstract:

    Comparative estimations of the antioxidant activity of methanolic extracts from biomasses of different types of in vitro cultures of Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and S. baicalensis and also from plant raw materials were performed. The antioxidant measurements were based on the modern assays-cupric ion reducing antioxidant capacity (CUPRAC) and quick, easy, new, cheap, and reproducible CUPRAC (QUENCHER-CUPRAC). The total extractable antioxidants (CUPRAC assay) ranged from 10.4 to 49.7 mmol (100 g)-1 of dry weight (DW) expressed as Trolox equivalent antioxidant capacity (TEAC), and the global antioxidant response (QUENCHER-CUPRAC assay) ranged from 16.0 to 79.1 mmol (100 g)-1 DW for in vitro cultures, whereas for plant raw materials the total extractable antioxidants ranged from 20.9 to 69.5 mmol (100 g)-1 DW, and the global antioxidant response ranged from 67.2 to 97.8 mmol (100 g)-1 DW. Finally, the in vitro cultures could be regarded as an antioxidant-rich alternative resource for the pharmaceutical, health food and cosmetics industries.

  • in vitro cultures of some medicinal plant species cistus incanus Verbena Officinalis scutellaria lateriflora and scutellaria baicalensis as a rich potential source of antioxidants evaluation by cuprac and quencher cuprac assays
    Plants (Basel Switzerland), 2021
    Co-Authors: Michał Dziurka, Pawel Kubica, Halina Ekiert, Inga Kwiecień, Shaimaa A. M. Abdelmohsen, Fatemah Farraj Alharbi, Hosam O. Elansary, Jolanta Biesagakościelniak, Diaa O Elansary, Agnieszka Szopa
    Abstract:

    Comparative estimations of the antioxidant activity of methanolic extracts from biomasses of different types of in vitro cultures of Cistus × incanus, Verbena Officinalis, Scutellaria lateriflora, and S. baicalensis and also from plant raw materials were performed. The antioxidant measurements were based on the modern assays-cupric ion reducing antioxidant capacity (CUPRAC) and quick, easy, new, cheap, and reproducible CUPRAC (QUENCHER-CUPRAC). The total extractable antioxidants (CUPRAC assay) ranged from 10.4 to 49.7 mmol (100 g)-1 of dry weight (DW) expressed as Trolox equivalent antioxidant capacity (TEAC), and the global antioxidant response (QUENCHER-CUPRAC assay) ranged from 16.0 to 79.1 mmol (100 g)-1 DW for in vitro cultures, whereas for plant raw materials the total extractable antioxidants ranged from 20.9 to 69.5 mmol (100 g)-1 DW, and the global antioxidant response ranged from 67.2 to 97.8 mmol (100 g)-1 DW. Finally, the in vitro cultures could be regarded as an antioxidant-rich alternative resource for the pharmaceutical, health food and cosmetics industries.

  • Accumulation of volatile constituents in agar and bioreactor shoot cultures of Verbena Officinalis L.
    Plant Cell Tissue and Organ Culture (PCTOC), 2021
    Co-Authors: Adam Kokotkiewicz, Bozena Zabiegala, Pawel Kubica, Agnieszka Szopa, Adam Bucinski, Halina Ekiert, Maria Luczkiewicz
    Abstract:

    In vitro shoot cultures of common vervain ( Verbena Officinalis L., Verbenaceae) were established and evaluated for essential oil content. The shoots were maintained as stationary microshoot culture, grown on Schenk–Hildebrandt medium supplemented with 9.84 µM of 2-isopentenyladenine and 1.0 µM of thidiazuron. The shoots were further adapted to grow in two temporary immersion bioreactors (RITA and Plantform), using Schenk–Hildebrandt medium supplemented with 4.92 µM indole-3-butyric acid. The best growth, in terms of dry weight production, was recorded for agar culture (0.73 g l^−1 day^−1), followed by 35 days (day) Plantform (0.53 g l^−1 day^−1), 35 days RITA (0.42 g l^−1 day^−1) and 56 days Plantform culture (0.29 g l^−1 day^−1). The volatile fractions were obtained by simultaneous hydrodistillation-extraction of the plant materials using Likens–Nickerson apparatus. Aerial parts of V. Officinalis intact plant and 56 d Plantform culture had the highest essential oil content (8.1 g kg^−1) whereas the stationary agar culture excelled in essential oil productivity (5.0 mg l^−1 day^−1). The volatile oils isolated from in vitro shoots contained 1-octen-3-ol, benzyl alcohol, phenethyl alcohol, coumaran and hexadecenoic acid as major constituents. 1-Octen-3-ol was the predominant (> 50%) constituent of 35 day shoot cultures maintained in temporary immersion bioreactors whereas coumaran (19%) and hexadecanoid acid (21%) prevailed in stationary microshoot culture. The volatile fraction obtained from aerial parts of V. Officinalis intact plant was composed mainly of hexadecanoic acid (> 20%), linalool (> 8%), anethole (> 5%) and carvone (> 3%). Microshoot cultures of Verbena Officinalis , grown in temporary immersion bioreactors, were investigated in terms of essential oil content and composition of volatile fraction

  • Verbena Officinalis (Common Vervain) - A Review on the Investigations of This Medicinally Important Plant Species.
    Planta medica, 2020
    Co-Authors: Paweł Kubica, Agnieszka Szopa, Maria Luczkiewicz, Jakub Dominiak, Halina Ekiert
    Abstract:

    Verbena Officinalis (common vervain) is a medicinal plant species widely distributed in the world and commonly used in folk medicine of different countries, including traditional Chinese medicine. Monographs on “Verbenae herba” have been included in the European Pharmacopoeia since 2008, and in the Chinese Pharmacopoeia since 1995. This work presents botanical characteristics of this species. It reviews the current knowledge of its chemical composition, which is a rich source mostly of iridoids, phenylpropanoid glycosides, phenolic acids, flavonoids, terpenoids, and essential oil. A large part of this article summarizes traditional medicinal uses and professional pharmacological in vitro and in vivo studies that prove new important applications, e.g., antioxidant, antimicrobial, anti-inflammatory, neuroprotective anticancer, analgesic, or anticonvulsant of Verbena herb extracts and individual metabolites. Moreover, emphasis is put on the use of V. Officinalis in the food and cosmetics industries, especially due to its antioxidant, antibacterial, and anti-inflammatory properties, and the presence of essential oil with an attractive fragrance composition. This paper also presents the state of biotechnological studies of this species.

  • production of verbascoside and phenolic acids in biomass of Verbena Officinalis l vervain cultured under different in vitro conditions
    Natural Product Research, 2017
    Co-Authors: Pawel Kubica, Agnieszka Szopa, Halina Ekiert
    Abstract:

    AbstractMethanolic extracts from the biomass of Verbena Officinalis cultured under continuous artificial light and in darkness on 12 variants of the Murashige and Skoog medium containing different concentrations (0.5–3.0 mg/L) of plant growth regulators: 6-benzyladenine, kinetin, 1-naphthaleneacetic acid and indole-3-butyric acid, were analysed for the amounts of verbascoside and phenolic acids, before and after acid hydrolysis, using the HPLC-DAD method. The amounts of verbascoside were very high (max. 2454.12 mg/100 g DW – light, and 2135.59 mg/100 g DW – darkness). The total amounts of phenolic acids reached a maximum of 46.02 mg/100 g DW (free phenolic acids) and 141.05 mg/100 g DW (bound compounds). The main metabolites were: ferulic, o-coumaric and caffeic acids. The maximum amount of verbascoside was 3.28 times higher than in extracts from the herb of the parent plant. The cultures could be proposed as a potential biotechnological source for selected biologically active compounds.

Hugh D. Wilson - One of the best experts on this subject based on the ideXlab platform.

Miriam Eichberger - One of the best experts on this subject based on the ideXlab platform.

  • towards eco friendly secondary plant metabolite quantitation ultra high performance supercritical fluid chromatography applied to common vervain Verbena Officinalis l
    IEEE Journal of Solid-state Circuits, 2020
    Co-Authors: Nora Gibitzeisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Christoph Seger, Hermann Stuppner
    Abstract:

    This report presents the first ultra high performance supercritical fluid chromatography diode array detector based assay for simultaneous determination of iridoid glucosides, flavonoid glucuronides, and phenylpropanoid glycosides in Verbena Officinalis (Verbenaceae) extracts. Separation of the key metabolites was achieved in less than 7 min on an Acquity UPC2 Torus Diol column using a mobile phase gradient comprising subcritical carbon dioxide and methanol with 0.15% phosphoric acid. Method validation for seven selected marker compounds (hastatoside, Verbenalin, apigenin-7-O-glucuronide, luteolin-7-O-glucuronide, apigenin-7-O-diglucuronide, verbascoside, and luteolin-7-O-diglucuronide) confirmed the assay to be sensitive, linear, precise, and accurate. Head-to-head comparison to an ultra high performance liquid chromatography comparator assay did prove the high orthogonality of the methods. Quantitative result equivalence was evaluated by Passing-Bablok-correlation and Bland-Altman-plot analysis. This cross-validation revealed, that one of the investigated marker compound peaks was contaminated in the ultra high performance liquid chromatography assay by a structurally related congener. Taken together, it was proven that the ultra high performance supercritical fluid chromatography instrument setup with its orthogonal selectivity is a true alternative to conventional reversed phase liquid chromatography in quantitative secondary metabolite analysis. For regulatory purposes, assay cross-validation with highly orthogonal methods seems a viable approach to avoid analyte overestimation due to coeluting, analytically indistinguishable contaminants.

  • Towards eco‐friendly secondary plant metabolite quantitation: Ultra high performance supercritical fluid chromatography applied to common vervain (Verbena Officinalis L.)
    Journal of separation science, 2019
    Co-Authors: Nora Gibitz-eisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Christoph Seger, Hermann Stuppner
    Abstract:

    This report presents the first ultra high performance supercritical fluid chromatography diode array detector based assay for simultaneous determination of iridoid glucosides, flavonoid glucuronides, and phenylpropanoid glycosides in Verbena Officinalis (Verbenaceae) extracts. Separation of the key metabolites was achieved in less than 7 min on an Acquity UPC2 Torus Diol column using a mobile phase gradient comprising subcritical carbon dioxide and methanol with 0.15% phosphoric acid. Method validation for seven selected marker compounds (hastatoside, Verbenalin, apigenin-7-O-glucuronide, luteolin-7-O-glucuronide, apigenin-7-O-diglucuronide, verbascoside, and luteolin-7-O-diglucuronide) confirmed the assay to be sensitive, linear, precise, and accurate. Head-to-head comparison to an ultra high performance liquid chromatography comparator assay did prove the high orthogonality of the methods. Quantitative result equivalence was evaluated by Passing-Bablok-correlation and Bland-Altman-plot analysis. This cross-validation revealed, that one of the investigated marker compound peaks was contaminated in the ultra high performance liquid chromatography assay by a structurally related congener. Taken together, it was proven that the ultra high performance supercritical fluid chromatography instrument setup with its orthogonal selectivity is a true alternative to conventional reversed phase liquid chromatography in quantitative secondary metabolite analysis. For regulatory purposes, assay cross-validation with highly orthogonal methods seems a viable approach to avoid analyte overestimation due to coeluting, analytically indistinguishable contaminants.

  • Development and validation of a rapid ultra-high performance liquid chromatography diode array detector method for Verbena Officinalis L.
    Journal of pharmaceutical and biomedical analysis, 2018
    Co-Authors: Nora Gibitz-eisath, Miriam Eichberger, Regina Gruber, Sonja Sturm, Hermann Stuppner
    Abstract:

    Abstract This study presents a fast and validated ultra-high performance liquid chromatography diode array detector (UHPLC-DAD) method for the simultaneous determination of the major compounds in Verbena Officinalis L. (Verbenaceae), a medicinal plant listed in the European, British, and, Chinese Pharmacopoeias. In order to get reference substances, nine compounds, belonging to iridoids, flavonoids, and phenylpropanoid glycosides, were isolated from the herb extract. Two of them, namely cistanoside D and leucosceptoside A, were found in this plant species for the first time. Chromatographic separation was achieved in less than 7 min on a Kinetex  1.7 u XB-C18 (50 × 2.10 mm) column by using a solvent gradient of water-acetonitrile modified with 0.1% formic acid. Method validation confirmed the assays sensitivity, linearity (R2 ≥ 0.997), precision (intraday precision ≤ 1.71%; interday precision ≤ 1.46%) and accuracy (recovery rates between 93.9% and 108.8%) for the quantitative analysis of the eight selected marker compounds. Identity and peak purity of the analytes was confirmed by coupling the UHPLC instrument to a quadrupole time-of-flight mass spectrometer via an electrospray ionization interface (ESI-QTOF-MS) operating in the negative ionization mode. Finally, the applicability of the developed UHPLC-DAD method was successfully proven for the sensitive quantitation of the major compounds in Verbena herb extracts, thereby providing a reliable tool for its rapid quality control.

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