Simultaneous Quantification of Betulinic Acid, Lupeol, and β-Sitosterol in Madhuca longifolia Methanolic Extract of Bark by Liquid Chromatography-Tandem Mass Spectrometric Method.
Journal
Journal of AOAC International
ISSN: 1944-7922
Titre abrégé: J AOAC Int
Pays: England
ID NLM: 9215446
Informations de publication
Date de publication:
21 May 2021
21 May 2021
Historique:
received:
05
07
2020
revised:
19
08
2020
accepted:
03
09
2020
pubmed:
23
2
2021
medline:
29
6
2021
entrez:
22
2
2021
Statut:
ppublish
Résumé
Liquid chromatography with tandem mass spectrometry is used widely used for the quantitative analysis of phytoconstituents present in medicinal plants to assess the quality of extract used for different investigations. A sensitive, precise, and accurate liquid chromatographic method with tandem mass spectrometric detection was developed for simultaneous quantification of lupeol, betulinic acid, and β-sitosterol in the methanolic extract of Madhuca longifolia bark. The three compounds were eluted with a stationary phase Gemini C18 column (50 × 2.0 mm, 3 μm id) and the temperature of the column was maintained by a column oven at 40 ± 0.3°C; mobile phase A (water and 0.1% formic acid) and mobile phase B [acetonitrile-methanol (50+50, v/v) and 0.1% formic acid] were used in a gradient mode and the flow rate was 0.4 mL/min. With these conditions, the retention time for betulinic acid, lupeol, and β-sitosterol was found to be 1.25, 3.08, and 3.53 minutes, respectively. The total run time was 5.0 min. Detection and quantitation of all three phytoconstituents were carried out by the mass spectrometer, a triple quadrupole equipped with atmospheric pressure chemical ionization, and multiple reaction monitoring using the predominantly positive ion mode and obtained much higher and more stable response nebulizer gas flow at 3.0 L/min. Linear responses were exhibited for all three phytoconstituents with a dynamic linear range of 10-100 μg/mL with the values of the regression coefficient more than 0.995 for betulinic acid, lupeol, and β-sitosterol. The values of percentage RSD for intraday and interday precision were found to be within the accepted limits for analytical methods (<15%). Selectivity, linearity, LOD, LOQ, accuracy, and precision were evaluated for all three phytoconstituents as per International Conference on Harmonization guidelines. The proposed method is accurate and sensitive and can be used for the routine quantification of betulinic acid, lupeol, and β-sitosterol from the herbal extract and its poly-herbal formulations.
Sections du résumé
BACKGROUND
BACKGROUND
Liquid chromatography with tandem mass spectrometry is used widely used for the quantitative analysis of phytoconstituents present in medicinal plants to assess the quality of extract used for different investigations.
OBJECTIVE
OBJECTIVE
A sensitive, precise, and accurate liquid chromatographic method with tandem mass spectrometric detection was developed for simultaneous quantification of lupeol, betulinic acid, and β-sitosterol in the methanolic extract of Madhuca longifolia bark.
METHOD
METHODS
The three compounds were eluted with a stationary phase Gemini C18 column (50 × 2.0 mm, 3 μm id) and the temperature of the column was maintained by a column oven at 40 ± 0.3°C; mobile phase A (water and 0.1% formic acid) and mobile phase B [acetonitrile-methanol (50+50, v/v) and 0.1% formic acid] were used in a gradient mode and the flow rate was 0.4 mL/min.
RESULTS
RESULTS
With these conditions, the retention time for betulinic acid, lupeol, and β-sitosterol was found to be 1.25, 3.08, and 3.53 minutes, respectively. The total run time was 5.0 min. Detection and quantitation of all three phytoconstituents were carried out by the mass spectrometer, a triple quadrupole equipped with atmospheric pressure chemical ionization, and multiple reaction monitoring using the predominantly positive ion mode and obtained much higher and more stable response nebulizer gas flow at 3.0 L/min. Linear responses were exhibited for all three phytoconstituents with a dynamic linear range of 10-100 μg/mL with the values of the regression coefficient more than 0.995 for betulinic acid, lupeol, and β-sitosterol. The values of percentage RSD for intraday and interday precision were found to be within the accepted limits for analytical methods (<15%). Selectivity, linearity, LOD, LOQ, accuracy, and precision were evaluated for all three phytoconstituents as per International Conference on Harmonization guidelines.
CONCLUSIONS
CONCLUSIONS
The proposed method is accurate and sensitive and can be used for the routine quantification of betulinic acid, lupeol, and β-sitosterol from the herbal extract and its poly-herbal formulations.
Identifiants
pubmed: 33615381
pii: 5909643
doi: 10.1093/jaoacint/qsaa128
doi:
Substances chimiques
Pentacyclic Triterpenes
0
Plant Extracts
0
Sitosterols
0
gamma-sitosterol
5LI01C78DD
lupeol
O268W13H3O
Methanol
Y4S76JWI15
Betulinic Acid
4G6A18707N
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
498-505Informations de copyright
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