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Rapid Bioassay-Guided Isolation of Antibacterial Clerodane Type Diterpenoid from Dodonaea viscosa (L.) Jaeq.

Khurram M, Lawton LA, Edwards C, Iriti M, Hameed A, Khan MA, Khan FA, Rahman SU - Int J Mol Sci (2015)

Bottom Line: Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery.This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16), 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC) = 64-128 µg/mL) against test bacteria.To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Shaheed Benazir Bhutto University, Sheringal, Dir Upper 18000, Pakistan. drmkhurram@sbbu.edu.pk.

ABSTRACT
Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery. This study describes the application of automated preparative-HPLC combined with a rapid off-line bacterial bioassay, using reduction of a tetrazolium salt as an indicator of bacterial metabolism. This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16), 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC) = 64-128 µg/mL) against test bacteria. To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa.

No MeSH data available.


Related in: MedlinePlus

Preparative separation of n-hexane extract of D. viscosa using generic gradient and volume-based fractionation (a), percentage inhibition (b) of S. aureus (NCIMB 6571), percentage inhibition (c) of E. coli (NCIMB 8797), where NEG represented the negative control of 10% (v/v) dimethyl sulfoxide in phosphate buffer saline (DMSO/PBS) and the positive controls (POS) of clarithromycin (25 µg/mL) and ciprofloxacin (50 µg/mL), respectively (n = 2 ± SD). Fraction 42 (arrow) contained high purity, bioactive compound. XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) Bioassay results are depicted in Figure S1.
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ijms-16-20290-f001: Preparative separation of n-hexane extract of D. viscosa using generic gradient and volume-based fractionation (a), percentage inhibition (b) of S. aureus (NCIMB 6571), percentage inhibition (c) of E. coli (NCIMB 8797), where NEG represented the negative control of 10% (v/v) dimethyl sulfoxide in phosphate buffer saline (DMSO/PBS) and the positive controls (POS) of clarithromycin (25 µg/mL) and ciprofloxacin (50 µg/mL), respectively (n = 2 ± SD). Fraction 42 (arrow) contained high purity, bioactive compound. XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) Bioassay results are depicted in Figure S1.

Mentions: The use of solvents such as methanol and water along with a volume-based fractionation strategy was necessary as the active component(s) may not contain a chromophore. For the extract, a total of 52 primary fractions were collected and each fraction was evaluated against the two test bacteria using the XTT assay. No bioactivity was detected in the fractions from the ethyl acetate extract of D. viscosa, despite the activity against both bacteria reported in disk diffusion assay (Table 1), and they were not investigated further. The butanol extract of D. viscosa yielded three fractions (18, 19 and 20) with moderate antibacterial activity, i.e., 50%–75% reduction in bacterial growth. In contrast, six fractions from the hexane extract inhibited S. aureus, with this antibacterial activity resulting in higher than 75% reduction in bacterial activity (Figure 1).


Rapid Bioassay-Guided Isolation of Antibacterial Clerodane Type Diterpenoid from Dodonaea viscosa (L.) Jaeq.

Khurram M, Lawton LA, Edwards C, Iriti M, Hameed A, Khan MA, Khan FA, Rahman SU - Int J Mol Sci (2015)

Preparative separation of n-hexane extract of D. viscosa using generic gradient and volume-based fractionation (a), percentage inhibition (b) of S. aureus (NCIMB 6571), percentage inhibition (c) of E. coli (NCIMB 8797), where NEG represented the negative control of 10% (v/v) dimethyl sulfoxide in phosphate buffer saline (DMSO/PBS) and the positive controls (POS) of clarithromycin (25 µg/mL) and ciprofloxacin (50 µg/mL), respectively (n = 2 ± SD). Fraction 42 (arrow) contained high purity, bioactive compound. XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) Bioassay results are depicted in Figure S1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4613204&req=5

ijms-16-20290-f001: Preparative separation of n-hexane extract of D. viscosa using generic gradient and volume-based fractionation (a), percentage inhibition (b) of S. aureus (NCIMB 6571), percentage inhibition (c) of E. coli (NCIMB 8797), where NEG represented the negative control of 10% (v/v) dimethyl sulfoxide in phosphate buffer saline (DMSO/PBS) and the positive controls (POS) of clarithromycin (25 µg/mL) and ciprofloxacin (50 µg/mL), respectively (n = 2 ± SD). Fraction 42 (arrow) contained high purity, bioactive compound. XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) Bioassay results are depicted in Figure S1.
Mentions: The use of solvents such as methanol and water along with a volume-based fractionation strategy was necessary as the active component(s) may not contain a chromophore. For the extract, a total of 52 primary fractions were collected and each fraction was evaluated against the two test bacteria using the XTT assay. No bioactivity was detected in the fractions from the ethyl acetate extract of D. viscosa, despite the activity against both bacteria reported in disk diffusion assay (Table 1), and they were not investigated further. The butanol extract of D. viscosa yielded three fractions (18, 19 and 20) with moderate antibacterial activity, i.e., 50%–75% reduction in bacterial growth. In contrast, six fractions from the hexane extract inhibited S. aureus, with this antibacterial activity resulting in higher than 75% reduction in bacterial activity (Figure 1).

Bottom Line: Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery.This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16), 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC) = 64-128 µg/mL) against test bacteria.To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Shaheed Benazir Bhutto University, Sheringal, Dir Upper 18000, Pakistan. drmkhurram@sbbu.edu.pk.

ABSTRACT
Plant extracts are complex matrices and, although crude extracts are widely in use, purified compounds are pivotal in drug discovery. This study describes the application of automated preparative-HPLC combined with a rapid off-line bacterial bioassay, using reduction of a tetrazolium salt as an indicator of bacterial metabolism. This approach enabled the identification of fractions from Dodonaea viscosa that were active against Staphylococcus aureus and Escherichia coli, which, ultimately, resulted in the identification of a clerodane type diterpenoid, 6β-hydroxy-15,16-epoxy-5β, 8β, 9β, 10α-cleroda-3, 13(16), 14-trien-18-oic acid, showing bacteriostatic activity (minimum inhibitory concentration (MIC) = 64-128 µg/mL) against test bacteria. To the best of our knowledge, this is the first report on antibacterial activity of this metabolite from D. viscosa.

No MeSH data available.


Related in: MedlinePlus