A HPLC Method for Determination of Ursolic Acid and Betulinic Acids from their Methanolic Extracts of Vitex Negundo Linn

Ursolic acid is a pentacyclic triterpenoid compound which naturally occurs in a large number of vegetarian foods, medicinal herbs, and other plants. A simple, specific, accurate and precise HPLC method has been developed and validated for estimation of ursolic acid in three herbal extracts. Ursolic acid was estimated on a Grace smart RP C18 (5 μm; 25cm X 4.6 mm i.d.) using methanol: water: orthophosphoric acid (35:65:0.05, v/v) as the mobile phase and the detection was carried out 210 nm. The retention time was found to be 7.15 min. The linearity obtained at this detection wavelength was 25–475 μg/mL for ursolic acid. The limit of detection and limit of quantitation was found to be 6 and 25µg/mL, respectively, for ursolic acid. The co-relation and co-efficient for the calibration curve of ursolic acid was found to be 0.999. The developed method was used for separation and estimation of ursolic acid from 3 different herbal extracts (Alstonia scholaris, Ocimum sanctum, Mentha piperata). The estimated value of ursolic acid from herbal extracts of Alstonia scholaris, Ocimum sanctum and Mentha piperata was 1.87% w/w, 0.278% w/w and 0.575% w/w, respectively.

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The HPLC determination of the content of magnoflorine in epimedium alpinum L

Journal of the Serbian Chemical Society ◽

10.2298/jsc0603251d ◽

2006 ◽

Vol 71 (3) ◽

pp. 251-255 ◽

Cited By ~ 5

Author(s):

Zvezdana Doslov-Kokorus ◽

Ivana Ivanovic ◽

Milena Simic ◽

Vlatka Vajs ◽

Nada Kovacevic

Keyword(s):

Plant Material ◽

Aerial Part ◽

Hplc Method ◽

Underground Part ◽

Aporphine Alkaloid ◽

European Species ◽

Methanolic Extracts ◽

First Time ◽

Dose Dependent

The aporphine alkaloid magnoflorine, known as a constituent of Asian Epimedium species, was isolated for the first time from the wild growing European species, Epimedium alpinum L. Identification was done by comparison with literature data. The HPLC method was applied for the determination of the content of the alkaloid in methanolic extracts of plant material. The underground part of E. alpinum contains 1-2 % of magnoflorine, while its content in methanolic extracts was 9.2-11.8 %. The aerial part of the plant contains less alkaloid (0.06-0.12 %), while the magnoflorine content in methanolic extracts of the aerial part was 0.4-0.8 %. Probably, the high content of magnoflorine in the methanolic extracts of underground parts of E. alpinum influences its dose-dependent cytotoxic activity (300 ?g/ml; 39 % survival of cells in an experimental K562 cell culture).

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Analysis of Flavonoids and Iridoids in Vitex Negundo by HPLC-PDA and Method Validation

Natural Product Communications ◽

10.1177/1934578x1300800914 ◽

2013 ◽

Vol 8 (9) ◽

pp. 1934578X1300800

Author(s):

Somendu K. Roy ◽

Khemraj Bairwa ◽

Jagdeep Grover ◽

Amit Srivastava ◽

Sanjay M. Jachak

Keyword(s):

Quality Control ◽

Simultaneous Determination ◽

Method Validation ◽

Hplc Method ◽

Vitex Negundo ◽

Good Recovery ◽

Bioactive Constituents ◽

Ich Guidelines ◽

Validation Parameters

The leaves of Vitex negundo have been reported to contain various bioactive constituents including iridoids and flavonoids. This is the first report on the simultaneous determination of iridoids and flavonoids by HPLC in three different samples of V. negundo leaves collected from three regions of India. Separation of iridoids and flavonoids was accomplished by HPLC and further elaborated for their quantification in V. negundo leaves using a C-18 column with detection at 254 and 330 nm, respectively. The developed HPLC method showed good linearity (r2≥0.999), high precision (RSD<5%) and a good recovery (99.3–103.0%) of the compounds. All the validation parameters of the developed HPLC were found to be within the permissible limits according to the ICH guidelines. The developed method was robust, accurate and reliable for the quality control of V. negundo leaves.

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Simultaneous Determination of Oleanolic Acid and Ursolic Acid in Leaves of Paulownia by HPLC

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.781-784.787 ◽

2013 ◽

Vol 781-784 ◽

pp. 787-791 ◽

Cited By ~ 1

Author(s):

Ya Li Xing ◽

Liang Wu Bi ◽

Zhen Dong Zhao ◽

Tian Juan Xia

Keyword(s):

Phosphoric Acid ◽

Simultaneous Determination ◽

Ursolic Acid ◽

Oleanolic Acid ◽

Flow Rate ◽

Mobile Phase ◽

Leaf Extract ◽

Hplc Method ◽

Simultaneous Quantification

A quick and accurate HPLC method has been developed for the simultaneous quantification of two bioactive triterpenes, ursolic acid and oleanolic acid in Paulownia leaves. The samples were analyzed on a Shim-pack ODS-CLC (M) (4.6 mm × 250 mm, 5 μm) column kept at 21 °C, using the methanol and aqueous phase containing 0.05%phosphoric acid with the volumetric ratio of 91.7:8.3 as the mobile phase at a flow rate of 0.6 mL/ min, and the detection wavelength was set at 210 nm. The method was validated and applied to the simultaneous quantification of the two triterpenes in Paulownia leaf extract. The standard curves were established in the range of 0.44 ~ 8.75 μg for oleanolic acid and 0.92 ~ 18.37 μg for ursolic acid. The contents of oleanolic acid and ursolic acid in leaves of Paulownia were determinated using the HPLC method and the contents were 3.87 mg/g and 13.61 mg/g, respectively.

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A validated high performance liquid chromatography method for determination of three bioactive compounds p-hydroxy benzoic acid, negundoside and agnuside in Vitex species

Macedonian Journal of Chemistry and Chemical Engineering ◽

10.20450/mjcce.2015.500 ◽

2015 ◽

Vol 34 (2) ◽

pp. 321 ◽

Cited By ~ 1

Author(s):

Tushar Dhanani ◽

Sonal Shah ◽

Satyanshu Kumar

Keyword(s):

Benzoic Acid ◽

High Performance ◽

Limit Of Detection ◽

Solid Phase ◽

Hplc Method ◽

Vitex Negundo ◽

Limit Of Quantification ◽

Chromatography Method ◽

Vitex Species

A validated rapid and simple isocratic HPLC-PDA method was developed for identification and quantification of p- hydroxy benzoic acid and two iridoids negundoside and agnuside in the extracts of two Vitex species, Vitex negundo and Vitex trifolia. The separation of the three compounds was achieved on a RP-18 (250 mm X 4 mm, 5µm) column at 25o C using acetonitrile (15%) and 0.05% trifluoroacetic acid (TFA) in water (85%). Limit of detection (LOD) were 1, 2.5 and 2.5 μg/ml for p- hydroxy benzoic acid, negundoside and agnuside respectively. Similarly, limit of quantification (LOQ) were 2.5, 5 and 5 μg/ml for p- hydroxy benzoic acid, negundoside and agnuside respectively. Good linearity (r2 > 0.999) was observed for all the three compounds in wide concentration range. Using the developed HPLC method, the three compounds were identified and quantified in leaves and bark extracts of Vitex negundo and Vitex trifolia. The novelty of the developed HPLC method is that it does not require complex sample processing such as use of solid phase extraction as well as use of buffer in mobile phase. This is the first report of a validated HPLC method for the simultaneous determination of p- hydroxy benzoic acid, negundoside and agnuside in Vitex species.

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