Standardization, Safety And Antiobesity Studies Of Piper Sarmentosum Roxburgh Extracts
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Date
2016-03
Authors
Hamil, Mohd Shahrul Ridzuan
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
This study involves quality control, standardization, safety and biological
activities using Piper sarmentosum extracts. Extracts using ethanol, ethanol (50%) and
water were standardized and further evaluated using anti-obesity and antihyperlipidemic
assays. Physico-chemical, limit test, qualitative and quantitative plant
fingerprint was performed using spectroscopic and chromatographic techniques. The
contents of primary and secondary metabolites in the extracts were determined. The
extracts were analyzed for asarones content using HPLC and FT-IR. Chemometrics
(PCA and HCA) were applied in standardization. Standardized extracts were screened in
vitro for cytotoxicity against brine shrimp assay (BSLA), pancreatic lipase (PPL)
inhibition and anti-angiogenesis activities. The anti-obesity and anti-hyperlipidemic
assays were performed in vivo on high fat diet (HFD) induced rats. Distinctive
fingerprint of the plant materials and the presence of vitexin and naringenin in the
extracts were observed. Samples from Penang and Perak were complying with the
acceptable safety limit tests. HPLC shows varying amount of vitexin (0.03 ± 0.002 –
0.98 ± 0.01% wt/wt) in P. sarmentosum extracts. Naringenin was detected in ethanol
extracts from Perlis and Terengganu (0.06 ± 0.001% and 0.12 ± 0.0003% wt/wt),
respectively. Extracts from Penang (PSP) showed the highest total polysaccharides
(87.71 ± 0.26 – 111.10 ± 0.06 mg/g), proteins (19.32 ± 0.25 – 39.81 ± 0.40 mg/g),
glycosaponins (15.55 ± 1.60 – 203.73 ± 0.39 mg/g), phospholipids (0.00 ± 0.00 – 46.25 ± 0.92 mg/g), phenolics (19.88 ± 0.18 – 37.39 ± 0.54 mg/g) and flavonoids (5.65 ± 0.04
– 43.66 ± 0.11 mg/g), respectively thus, selected as the standardized extracts. A new
HPLC method for evaluation of two toxic markers, α- and β-asarone was developed and
validated. Ethanol and SFE extracts contained the highest amount of asarones followed
by ethanol (50%) and absent in water extracts. Hence, water extract was considered as
safe for human consumption. FT-IR combined with PCA and HCA revealed that ethanol
and SFE extracts were clustered together with asarones and correlated with the amount
of asarones in the extracts. Cytotoxicity screening using BSLA showed that the
standardized ethanol, ethanol (50%) and water extracts exhibited low cytotoxicity after
24 hours of treatment. In vitro PPL inhibition of P. sarmentosum showed mild to
moderate inhibition activity in ethanol (50%) (21.81 ± 1.25%) and water extracts (9.41 ±
1.41%), respectively. Anti-angiogenesis activity using rat aortic ring assay on ethanol
(50%) extract showed 26.50 ± 0.89% inhibition whereas water extract showed 43.40 ±
0.93% inhibition. The in vivo treatment with standardized ethanol (50%) and water
extracts for 45 days influenced the cholesterol metabolism in HFD rats. Doses of 500
mg/kg significantly lower the percentage of total body weight increase, body mass
index, lipid profiles and adipose tissue weight compared to HFD untreated rats. No sign
of fatty liver disease were observed at doses of 250 and 500 mg/kg. As conclusion,
information on the quality, safety and efficacy of P. sarmentosum extracts were
obtained. The standardized ethanol (50%) and water extracts shows positive results for
in vitro and in vivo obesity-related studies.
Description
Keywords
Piper sarmentosum extracts