Nuri Andarwulan

Improvement of pea (Pisum sativum) seed vigour response by fish protein hydrolysates in combination with acetyl salicylic acid

Abstract Acetyl salicylic acid (ASA) and fish protein hydrolysate (FPH) were used for stimulation of vigour of pea ( Pisum sativum ) seeds. Pea seeds were pre-hydrated in water which contained 50 μM of ASA, 2 ml l −1 of FPH (standardized mackerel hydrolysates) and 2 ml l −1 of a combination of ASA and FPH (ASA/FPH) for 24 h and followed by germination in potting soil for 5 and 10 days. After 5 and 10 days, the average plant height and weight increased in plants with FPH and ASA/FPH treatments, but decreased slightly in plants with ASA treatment. After 10 days, the average plant height with FPH and ASA/FPH treatments increased by 9% (23.3 mm) and 11% (28.8 mm) respectively, while the average fresh weight of seedling increased by 15% (13 mg) for both treatments, respectively. The average phenolics content and guaiacol peroxidase (GPX) activity of pea seedling increased following ASA and FPH treatment alone on day 5. Antioxidant activity in all pea seedlings was high on day 10 compared to day 5. On either day there were no difference in antioxidant activity between treatments. Glucose-6-phosphate dehydrogenase (G-6-PDH) activity and free proline content were higher on day 5 than on day 10. The average G-6-PDH activity in pea seedling treated with FPH and ASA/FPH was higher than ASA and control treatments. The plant response to ASA/FPH was better than FPH treatment alone. These results suggest that combination of 50 μM of ASA and FPH with the concentration of 2 ml l −1 (ASA/FPH) can enhance seed performance of peas according to some criteria of seedling vigour and thus have potential for use as a seed treatment.

Protective Role of Ternatin Anthocyanins and Quercetin Glycosides from Butterfly Pea (Clitoria ternatea Leguminosae) Blue Flower Petals against Lipopolysaccharide (LPS)-Induced Inflammation in Macrophage Cells.

Twelve phenolic metabolites (nine ternatin anthocyanins and three glycosylated quercetins) were identified from the blue flowers of Clitoria ternatea by high-performance liquid chromatography diode array detection and electrospray ionization/mass spectrometry (HPLC-DAD-ESI/MS(n)). Three anthocyanins not reported in this species before show fragmentation pattern of the ternatin class. Extracts were fractionated in fractions containing flavonols (F3) and ternatin anthocyanins (F4). In general, C. ternatea polyphenols showed anti-inflammatory properties in lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells with distinct molecular targets. Flavonols (F3) showed strong inhibition of COX-2 activity and partial ROS suppression. On the other hand, the ternatin anthocyanins (F4) inhibited nuclear NF-κB translocation, iNOS protein expression, and NO production through a non-ROS suppression mechanism. Accordingly, quercetin glycosides and ternatin anthocyanins from the blue flower petals of C. ternatea may be useful in developing drugs or nutraceuticals for protection against chronic inflammatory diseases by suppressing the excessive production of pro-inflammatory mediators from macrophage cells.

Stimulation of novel phenolic metabolite, epoxy‐pseudoisoeugenol‐(2‐methylbutyrate) (EPB), in transformed anise (pimpinella anisum L.) root cultures by fish protein hydrolysates

Abstract Acetyl salicylic acid (ASA) and fish protein hydrolysate (FPH) were investigated for stimulating valuable phenolics production in transformed anise root cultures (Pimpinella anisum L.) for food and nutraceutical applications. Anise root cultures were grown in Murashige‐Skoog (MS) hormone free medium. Total phenolics in anise root cultures decreased with ASA and ASA/FPH treatments and may possibly be diverted to lignification as indicated by increase in guaiacol peroxidase (GPX) activity. Epoxy‐pseudoisoeugenol‐(2‐methylbutyrate) (EPB) production in root cultures in response to FPH treatment increased 3 fold compared to control and 6 fold higher compared to ASA treatment on day 60. The optimum concentration of FPH for EPB stimulation was 2 ml/L of standardized mackerel hydrolysates and if the concentration was higher than 2 ml/L, FPH was inhibitory to growth of root cultures. Antioxidant activity in all treatments was high on day 60 compared to day 30. On either day, there was no difference in antioxidant activity between treatments. The activity of glucose‐6‐phosphate dehydrogenase (G‐6‐PDH) did not vary. Free proline content in response to control and ASA treatments was higher than in FPH and ASA/FPH treatments on day 30. These results clearly indicate that mackerel hydrolysate can be used to stimulate EPB in transformed anise root cultures. EPB is potentially a valuable phenolic metabolite to regulate nutraceutical type phytochemicals during seed germination.

Accumulation patterns of lipophilic organic contaminants in surface sediments and in economic important mussel and fish species from Jakarta Bay, Indonesia

Non-target screening analyses were conducted in order to identify a wide range of organic contaminants in sediment and animal tissue samples from Jakarta Bay. High concentrations of di-iso-propylnaphthalenes (DIPNs), linear alkylbenzenes (LABs) and polycyclic aromatic hydrocarbons (PAHs) were detected in all samples, whereas phenylmethoxynaphthalene (PMN), DDT and DDT metabolites (DDX) were detected at lower concentrations. In order to evaluate the uptake and accumulation by economic important mussel (Perna viridis) and fish species, contaminant patterns of DIPNs, LABs and PAHs in different compartments were compared. Different patterns of these contaminant groups were found in sediment and animal tissue samples, suggesting compound-specific accumulation and metabolism processes. Significantly higher concentrations of these three contaminant groups in mussel tissue as compared to fish tissue from Jakarta Bay were found. Because P. viridis is an important aquaculture species in Asia, this result is relevant for food safety.

Mobilization of primary metabolites and phenolics during natural fermentation in seeds of Pangium edule Reinw.

Abstract Fermented seeds of the tropical tree Pangium edule Reinw. are a speciality in Indonesia and have been used as spices. The fermentation process of the seeds is a natural spontaneous process, which occurs 40 days following seed maturity and treatment. This study reports some biochemical changes, especially primary metabolites, and antioxidant activity associated with mobilization of lipids and phenolics during seed fermentation. The lipid content increased slightly (46.07–50.95% db) although the dominant fatty acid composition did not change. The dominant fatty acids were oleic acid (C 18:1 n-9) and linoleic acid (C 18:2 n-6). During fermentation, the decrease in fatty acid content in lipid coincided with the increasing acid value, which indicated that free fatty acids increased in seeds during fermentation. The dominant tocol in the seed, γ-tocotrienol, increased (69.8–123.3 μg g −1 freeze-dried seed) during fermentation. In general, overall protein content and amino acid composition did not change but non-soluble protein increased while soluble protein decreased. The changes in carbohydrate fraction showed that total crude carbobydrate, neutral detergent fibre (NDF, as cellulose, hemicellulose, and lignin) decreased, but reducing sugar increased and starch content did not change. Enzyme assays showed that microorganisms may be involved in the fermentation process. β-glucosidase, an enzyme that can cleave glycosidic bonds of conjugated phenolics and guaiacol peroxidase (GPX) activities increased. The total phenolics content in seeds increased substantially corresponding to the increase in β-glucosidase but antioxidant activity of phenolic extracts did not change.

HPLC-based metabolomics to identify cytotoxic compounds from Plectranthus amboinicus (Lour.) Spreng against human breast cancer MCF-7Cells

The objective of this study was to identify the active compounds in Plectranthus amboinicus (Lour.) Spreng which play a role to inhibit viability of breast cancer MCF-7 cells using HPLC-based metabolomics approach. Five fractions of the plant extract were observed including ethanol, hexane, chloroform, ethyl acetate and water fraction. There were 45 HPLC chromatograms resulted from 5 fractions with 3 replications and 3 wavelengths detection. The chromatograms were compared to the data of IC50 from MTT assay of each fraction against human breast cancer MCF-7 cells using metabolomics. The OPLS analysis result promptly pointed towards a chloroform fraction at retention time of 40.16-41.28min that has the greatest contribution to the cytotoxic activity. The data of mass spectra indicated that an abietane diterpene namely 7-acetoxy-6-hydroxyroyleanone was the main compound that contributed to the cytotoxic activity. This metabolomics application method can be used as a quick preliminary guideline to uncover the most dominant compound related to the bioactivity.

Quality of Vegetable Oil Prior to Fortification Is an Important Criteria to Achieve a Health Impact

Unbranded palm cooking oil has been fortified for several years and can be found in the market with different oxidation levels. This study aimed to investigate the stability and shelf life of unbranded, bulk, vitamin A-fortified palm oils with the most commonly observed oxidation levels in Indonesia. Three types of cooking oils were tested: (i) cooking oil with a peroxide value (PV) below 2 mEq O2/kg (PO1); (ii) cooking oil with a PV around 4 mEq O2/kg (PO2); and (iii) cooking oil with a PV around 9 mEq O2/kg (PO3). The oil shelf life was determined by using accelerated shelf life testing (ASLT), where the product was stored at 60, 75 and 90 °C, and then PV, free fatty acid and vitamin A concentration in the oil samples were measured. The results showed that PO1 had a shelf life of between 2–3 months, while PO2’s shelf life was a few weeks and PO3’s only a few days. Even given those varying shelf lives, the vitamin A loss in the oils was still acceptable, at around 10%. However, the short shelf life of highly oxidized cooking oil, such as PO3, might negatively impact health, due to the potential increase of free radicals of the lipid peroxidation in the oil. Based on the results, the Indonesian government should prohibit the sale of highly-oxidized cooking oil. In addition, government authorities should promote and endorse the fortification of only cooking oil with low peroxide levels to ensure that fortification is not associated with any health issues associated with high oxidation levels of the cooking oil.

Effect of tocopherols, tocotrienols, β-carotene, and chlorophyll on the photo-oxidative stability of red palm oil

Effect of tocols, β-carotene, and chlorophyll on photo-oxidative stability of red palm oil (RPO) were studied. Model systems of triacylglycerols+tocols, triacylglycerols+β-carotene, triacylglycerols +tocols+β-carotene, and triacylglycerols+tocols+β-carotene+chlorophyll were exposed to fluorescent light at intensities of 5,000, 10,000, and 15,000 lux for 7 h at 30±2°C. Changes in concentrations of tocopherols, tocotrienols, β-carotene, chlorophyll, and peroxide values were evaluated every hour. Light intensity accelerated degradation of tocols in the triacylglycerols+tocols system and β-carotene in the triacylglycerols+β-carotene system. Gamma-tocotrienol showed the highest degradation rate and β-carotene was the most sensitive compound to changes in light intensity, indicated by the lowest light intensity coefficient (zi) value. The presence of tocols and β-carotene together showed protective effects for the photo-oxidative stability of RPO. The presence of chlorophyll increased the rate of photo-oxidation at high light intensities. Interactions between tocols and β-carotene contributed to the photo-oxidative stability of RPO.

The colour degradation of anthocyanin-rich extract from butterfly pea (Clitoria ternatea L.) petal in various solvents at pH 7

Abstract A spectroscopic study was conducted to evaluate the colour degradation mechanism of anthocyanin-rich extract from butterfly pea petal. The extract was diluted in four different solvent systems, which were buffer solution pH 7 (AQ7) and the mixture of organic solvent with buffer solution pH 7 (4:1 v/v). The organic cosolvent involved were methanol (ME7), ethanol (ET7) and acetone (AC7). The samples were stored in containers with 0% and 50% headspace, and their colour intensity, total anthocyanin and hypsochromic shift were evaluated periodically. The rank of colour and anthocyanin degradation from the biggest was AQ7 > ME7 > ET7 > AC7. The longest hypsochromic shift was AQ7 > ME7 > ET7, while in AC7 the shift was absent. There was evidence that the volume of package headspace provoked colour stability. The colour degradation in AC7 was proposed to occur through hydrophobic interaction unfolding, and in AQ7 was through the deacylation, while in ME7 and ET7 was due to both mechanisms.

Metabolomic approach for understanding phenolic compounds and melanoidin roles on antioxidant activity of Indonesia robusta and arabica coffee extracts

Our experiments investigated roles of phenolic compounds and melanoidins on antioxidant activity of Indonesia robusta and arabica coffee extracts. The 2,2,-diphenyl-1-picrylhydrazyl (DPPH) assay and a ferric reducing antioxidant power (FRAP) method were used to determine the antioxidant activity. An increase in the roasting degree (green, light, medium, and dark) reduced phenolic compounds and the antioxidant activity of coffee extracts, but enhanced melanoidin content. Principle component analysis (PCA) demonstrated that phenolic compounds showed stronger effects on antioxidant activity of coffee extracts in comparison with melanoidins. This finding was supported by the results of metabolomic fingerprint by partial least square (PLS), which describes the correlation of functional groups of coffee extracts on antioxidant activity. Based on the PLS analysis, hydroxyl groups (O–H) were observed to show a positive correlation, but carbonyl (C=O) and amine (N–H) groups were attributed to a negative correlation on antioxidant activity of coffee extracts.