Estimation of Twelve Bacopa Saponins in Bacopa monnieri Extracts and Formulations by High-Performance Liquid Chromatography

B.S.Murthy, Papolu; Raju, Valivarti Raghava; Ramakrisana, Tummala; Chakravarthy, M. S.R. Krishna; Kumar, Katta Vijay; Kannababu, Sukala; Subbaraju, Gottumukkala V.

doi:10.1248/cpb.54.907

Cited by 70 publications

(46 citation statements)

References 15 publications

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“…Extraction and high performance liquid chromatography (HPLC) analysis of bacoside A were carried out by following the method of Murthy et al (2006). One gram of powdered shoot material was extracted in 100 ml of 70% methanol by heat refluxing and filtered through 0.45 lM membrane filters.…”

mentioning

confidence: 99%

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots

Naik

Manohar

Nagella

et al. 2009

Plant Cell Tiss Organ Cult

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Brahmi (Bacopa monnieri) is an important medicinal plant mainly used for the treatment of neurological disorders and depression. Recent investigations revealed that bacoside A is major chemical component shown to be responsible for memory facilitating action of brahmi. The current investigation was carried out to assess the potential for increasing biomass and the concentration of bacoside A in the in vitro regenerated shoots by varying sucrose and pH levels of shoot regeneration medium. The leaf explants were cultured on the Murashige and Skoog (MS) medium supplemented with 2 mg l -1 kinetin (KN) and with varying concentrations of sucrose (0, 1, 2, 3, 4, 5 and 6% at pH 5.8) and pH (4.5, 5.0, 5.5, 6.0 and 6.5 with 2% sucrose) with the objective of verifying the effects of sucrose and pH level on shoot regeneration and to verify the accumulation of bacoside A in the regenerated shoots. The shoot biomass increased (150.50 ± 2.84 shoots per explant, fresh wt 6.31 ± 0.12 g and dry wt 250 ± 5.00 mg) on the medium supplemented with 2% sucrose and pH which was set at 4.5. The results of HPLC analysis indicate that increase in sucrose concentration (0, 1, 2, 3, 4, 5 and 6% at pH 5.8) lead to decrease in the bacoside A content (39.51, 22.43, 13.05, 12.17, 10.73, 9.56 and 8.93 mg g -1 dry wt, respectively) in regenerated shoots. These findings provide evidence that stressful condition of inadequate supply of carbon elevated synthesis of bacoside A in brahmi shoots. However, 2% sucrose is found suitable for biomass accumulation. Therefore, medium supplemented with 2% sucrose and pH set at 4.5 was found suitable for both biomass (6.31 ± 0.12 g fresh wt and 250 ± 5.00 mg dry wt) and bacoside A accumulation (13.09 mg g -1 dry wt).

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mentioning

confidence: 99%

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots

Naik

Manohar

Nagella

et al. 2009

Plant Cell Tiss Organ Cult

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“…Extraction and HPLC analysis of bacoside A were carried out by following the method of Murthy et al (2006) with some modifications. Thirty milligram of powdered plant material was extracted in 25 mL of 70% methanol by heat refluxing for 45 minutes and filtered through 0.45 µm membrane filters.…”

Section: Extraction and Hplc Analysismentioning

confidence: 99%

Influence of different media, medium strength and carbon sources on adventitious shoot cultures and production of bacoside A in <em>Bacopa monnieri</em>(L.)

et al. 2017

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Abstract:Bacopa monnieri (L.) Wettst. (Plantaginaceae), commonly known as Brahmi, is used in Ayurveda as a medicinal herb, found throughout the Indian subcontinent in wet, damp and marshy areas. The herb yields pharmaceutically active compounds called bacosides. The current investigation was carried out to assess the potential for increasing biomass and the concentration of bacoside A from the in vitro regenerated shoots in different media (MS, B5, NN and N6), the strength of the MS medium (0.25, 0.50, 0.75, 1.0, 1.50 and 2.0X) and carbon source [sucrose, glucose, fructose, maltose, sucrose + glucose (1:1), sucrose + maltose (1:1), glucose + fructose (1:1), glucose + maltose (1:1), fructose + sucrose (1:1) and fructose + maltose (1:1)]. All the culture media were supplemented with 2 mg/L Kinetin (Kin) at pH 5.8. High performance liquid chromatography (HPLC) was used to determine the bacoside A content from the regenerated shoots. Optimum number of adventitious shoots (70.75 shoots per explant), fresh weight (2.344 g), dry weight (0.166 g) and bacoside A content (13.052 mg/g DW) were obtained from the cultures grown in the MS medium. Bacoside A production was highest in the shoots grown in the glucose + fructose combination (15.588 mg/g DW) among the different carbon sources tested. These findings suggest that B. monnieri culture requires full strength MS medium and a moderate level of sucrose. The present study is useful to obtain optimum production of biomass and bacoside A from the in vitro regenerated shoots of B. monnieri.

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“…Extraction and high-performance liquid chromatography (HPLC) analysis of bacoside A were carried out following the method of Murthy et al (2006). One gram of powdered plant material was extracted in 100 mL of 70% methanol by heat refluxing and filtered through 0.45-µM membrane filters.…”

Section: Estimation Of Bacoside Amentioning

confidence: 99%

Genetic fidelity assessment of encapsulated in vitro tissues of Bacopa monnieri after 6 months of storage by using ISSR and RAPD markers

Muthiah¹,

Shunmugiah²,

Ramesh³

2013

Turk J Bot

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The present study has investigated the efficiency of regrowth of Bacopa monnieri (L.) Pennell after 6 months of storage along with its clonal fidelity checking. Shoot tips and nodal segments of in vitro grown plantlets were encapsulated and stored at different temperatures and the highest regeneration was achieved by storing shoot tips at 4 °C for a period of 6 months. The response of shoot tips was found to be much better when compared to nodal segments in the case of forming healthy shoots and roots. The genetic stability of regrown plantlets was analysed by using RAPD and ISSR markers. Twenty ISSR primers produced 130 clear and reproducible amplicons, of which 125 bands were monomorphic. A total of 115 bands were produced by 25 RAPD markers with 94% monomorphism. This corresponds to a very low genetic variation pertaining to the clonal nature of the regrown plantlets with that of the mother plant. By means of HPLC biochemical analysis, bacoside A content in the regrown plants was also found to be comparable with that of the mother plant. Therefore, the protocol derived can be efficiently utilised for storing the encapsulated shoot tips of B. monnieri for about 6 months.

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Estimation of Twelve Bacopa Saponins in Bacopa monnieri Extracts and Formulations by High-Performance Liquid Chromatography

Cited by 70 publications

References 15 publications

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots

Influence of different media, medium strength and carbon sources on adventitious shoot cultures and production of bacoside A in <em>Bacopa monnieri</em>(L.)

Genetic fidelity assessment of encapsulated in vitro tissues of Bacopa monnieri after 6 months of storage by using ISSR and RAPD markers

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