Penghasilan Sebatian Bioaktif, Azadirachtin Mentah Daripada Kultur Kalus dan Sel Ampalan Azadirachta Indica., A .Juss (Mambu)

Kalus dan sel ampaian tumbuhan mambu (Azadirachta indica A. JUSS) telah berjaya di perolehi daripada eksplan daun mudanya setelah dikulturkan di dalam suatu medium as as Murashige & S koog (MS) yang mengandungi 1.86mg/L NAA dan 0.2mg/L BAP. Profit pertumbuhan kalus dan sel ampaian serta analisis ke atas kandungan azadirachtin mentahnya turut di lakukan. Azadirachtin mentah yang telah berjaya di hasilkan daripada kalus dan sel ampaian A.indica di dapati meningkat secara maksimum semasa sel berada pada akhir fasa logarisma atau bermulanya fasa pegun di mana pertumbuhan atau pembahagian sel mula menurun atau berhenti. Ini menunjukkan bahawa penghasilan azadirachtin mentah ini tidak mempunyai perkaitan rapat dengan pertumbuhan kultur sel tersebut ('non-growth associated'). Penghasilan azadirachtin mentah di dapati lebih tinggi bagi kultur kalus dan sel ampaian yang telah dieramkan di dalam keadaan terang iaitu peningkatan maksimum masing-masing sebanyak 72% dan 59%, berbanding dengan peningkatan maksimum bagi penghasilan azadirachtin yang dikulturkan di dalam keadaan gelap iaitu 58% dan 34% . B ioasai sitotoksisiti telah menunjukkan bahawa fraksi organik etanol (F001), klorofom (F003) dan metanol (F005) yang di perolehi daripada tisu daun A.indica, kalus dan sel ampaiannya berupaya menunjukkan kesan sitotoksisiti kepada pertumbuhan bijibenih Brassica chinensis. Fraksi etanol (F001) daripada tisu daun A.indica berupaya menunjukkan aktiviti perencatan yang paling ketara berbanding dengan fraksi lain samada daripada tisu daun, kalus atau sel ampaiannya, iaitu rawatan pada kepekatan 250ppm ekstrak berupaya merencat pemanjangan hipokotil sebanyak 73% berbandi ng dengan kawalan. Bioasai larvasidal ke atas larva serangga Tribolium castaneum telah menunjukkan bahawa fraksi-fraksi organik etanol (F001), klorofom (F003), dan metanol (F005)) yang di perolehi daripada tisu daun A.indica, kalus dan sel ampaiannya berupaya menyebabkan perencatan terhadap pertumbuhan larva tersebut, walaupun tiada sebarang mortaliti larva di perhatikan sepanjang bioasai ini di jalankan. Keadaan ini berkemungkinan di sebabkan oleh sebatian bioaktif yang bertanggungjawab memberikan kesan perencatan ini, berupaya mengaruhkan pelbagai kesan perencatan ('multiple effect') yang menyebabkan berlakunya gangguan kepada pertumbuhan larva T.castaneum tersebut. Fraksi metanal daripada tisu daun Aindica telah menunjukkan aktiviti perencatan yang paling ketara berbanding dengan fraksi lain sam ada daripada tisu daun, kalus atau sel ampaiannya terutamanya rawatan pada kepekatan 125ppm ekstrak, di mana telah menyebabkan perencatan sebanyak 50% berbanding dengan kawalan. Ujian penghindar serangga telah menunjukkan bahawa hanya serbuk daun A.indica sahaja yang berupaya menunjukkan pengaruh sebagai suatu agen penghindar serangga terutamanya terhadap serangga Tribolium castaneum ini, sementara serbuk kalus dan sel ampaiannya tidak langsung menunjukkan sebarang pengaruh. Pendedahan 4% serbuk daun Aindica kepada serangga T. castaneum berupaya menyebabkan 40% daripada serangga ini menghindari atau menjauhi serbuk daun tersebut

The prospects of the cultivation of Arthrospira platensis under outdoor conditions in Malaysia

There is no virtual report on the commercial cultivation of Arthrospira in Malaysia beyond the laboratory scale probably because of the high costs of production and the lower yield which are highly interconnected with the algal cultivation techniques. One way to alleviate the production cost is through outdoor mass cultivation under natural conditions using all available resources. Therefore, the present study was conducted to investigate the prospects of the production of Arthrospira platensis under Malaysian tropical climate using enhanced cultivation techniques to reach a maximum yield. In this study, the growth and yield of A. platensis were investigated under three different cultivation conditions: laboratory (control), outdoor shaded (greenhouse, T1), and outdoor non-shaded (field, T2). The algal growth was measured through optical density, biomass dry weight, and chlorophyll a content. The algal yield was determined by calculating its productivity and specific growth rate. The A. platensis cultivation under outdoor non-shaded conditions achieved significantly higher growth (p < 0.05) with 1.62 ± 0.038 ABS of maximum optical density, 0.88 ± 0.020 g L-1 of maximum biomass dry weight, 8.77 ± 0.219 mg L-1 of maximum chlorophyll a content, 0.091 ± 0.0022 g L-1 d-1 of productivity and 0.220 ± 0.0017 µ d-1 of specific growth rate over a cultivation period of eight days. The present finding showed that the Malaysian climate is suitable for a satisfactory A. platensis productivity with proper cultivation techniques such as the pre-adaptation of the algal culture, inoculation in the late evening, continuous agitation and compensation of the evaporated culture medium

1 H NMR-based metabolomics approach in investigating the chemical profile, antioxidant and anti-inflammatory activities of Gynura procumbens and Cleome gynandra

Gynura procumbens and Cleome gynandra are two herbs commonly used in Malaysia to treat various ailments and are also consumed as salads (ulam) and vegetables. The present study aims to evaluate the relationship between the chemical compositions of both herbs and their antioxidant and anti-inflammatory properties using nuclear magnetic resonance (NMR) metabolomics approach, which is being reported for the first time. Different ethanolic extracts of both herbs were tested for DPPH scavenging and inhibition of nitric oxide (NO) via RAW 264.7 macrophage cell induction. Principal component analysis (PCA) revealed a good separation between the extracts and the corresponding metabolites identified via 1H NMR spectroscopy. The 100% ethanolic extract from both herbs and 20% ethanolic extract of C. gynandra were found to have the best antioxidant and anti-inflammatory activities. Kaempferol, quercetin, caffeoylquinic, dicaffeoylquinic acids, gallic acid, mallic acid, citric acid, phenylalanine, and choline are among the metabolites that contributed to bioactivities. The partial least square (PLS) model for both herbs have an overall acceptable goodness of fit and predictive power, which further strengthens the validity of this study. The present study provides a preliminary reference for the selection of optimum extract and will shed some light on the potential use of G. procumbens and C. gynandra as a phytomedicinal preparation

Comparative analysis of metabolites and antioxidant potentials from different plant parts of Curcuma aeruginosa roxb

A comparative analysis of metabolites from different parts of Curcuma aeruginosa, i.e. leaves, stems, adventitious roots and rhizomes was performed by GC-MS/MS coupled with multivariate statistical analysis. The GC-MS/MS analysis confirmed the occurrence of 26 metabolites belonged to terpenoids in almost all the samples. The Principal Component Analysis (PCA) indicated that there was a clear distinction between rhizomes and other plant parts, i.e. stems, leaves, and adventitious roots that could be explained by relatively higher contents of terpenoids including curzerene, alpha-farnesen, furanocoumarin, velleral, germacrone cineole, borneol, beta- and gamma- elemene and methenolone. The results of Hierarchical Clustering Analyses (HCA) corresponded with the PCA results where many terpenoids found abundantly high in rhizome were clustered together. This was supported by the Pearson correlation analysis that showed a significantly good relationship between those terpenoids. The adventitious roots demonstrated the strongest antioxidant activity as compared to the other plant parts which could be attributed to its highest Total Phenolic Contents (TPC). Total phenolic contents of all the plant parts were positively correlated with their antioxidant activities which indicate that phenolic compounds may play a role in the overall antioxidant activities of the plants. The results of the study highlighted the potential of this underexploited Curcuma species which could serve as a new source of important phytochemicals and natural antioxidant that could be incorporated in functional foods and nutraceuticals. In addition, chemical and biological evidence shown in the present work has rationalised the different uses of various plant parts of C. aeruginosa

GC-MS-based metabolite profiling of Cosmos caudatus leaves possessing alpha-glucosidase inhibitory activity

Cosmos caudatus, which is known as “Ulam Raja,” is an herbal plant used in Malaysia to enhance vitality. This study focused on the evaluation of the α-glucosidase inhibitory activity of different ethanolic extracts of C. caudatus. Six series of samples extracted with water, 20%, 40%, 60%, 80%, and 100% ethanol (EtOH) were employed. Gas chromatography-mass spectrometry (GC-MS) and orthogonal partial least-squares (OPLS) analysis was used to correlate bioactivity of different extracts to different metabolite profiles of C. caudatus. The obtained OPLS scores indicated a distinct and remarkable separation into 6 clusters, which were indicative of the 6 different ethanol concentrations. GC-MS can be integrated with multivariate data analysis to identify compounds that inhibit α-glucosidase activity. In addition, catechin, α-linolenic acid, α-D-glucopyranoside, and vitamin E compounds were identified and indicate the potential α-glucosidase inhibitory activity of this herb

Chemical profiling of Curcuma aeruginosa Roxb. rhizome using different techniques of solvent extraction

Objective: To investigate the possible phytochemical constituents of Curcuma aeruginosa Roxb. (C. aeruginosa) rhizome using two different techniques of direct solvent extraction. Methods: Two different techniques of direct solvent extractions, i.e. methyl tert-butyl ether (MTBE) extraction and two-phase methanol/chloroform (M/C) system, were used in this study. The analysis of the phytochemical constituents in MTBE and M/C extracts was performed using gas chromatography-mass spectrometry/mass spectrometry. The mass spectra of the compounds was matched with the NIST 08 mass spectral library. Results: The present study revealed that the extraction using two-phase M/C have resulted in higher metabolite coverage compared to the extraction with MTBE. Direct solvent extraction using MTBE revealed the presence of 27 compounds; whereas, M/C allowed the extraction of 18 and 36 compounds in polar (methanol) and nonpolar (chloroform) fractions respectively. The major compounds detected in the MTBE extract that based on the peak area percentage were methenolone (16.64%), cycloisolongifolene, 8,9-dehydro-9-formyl- (15.93%), labd-13-en-15-oic acid,8,12-epoxy-12-hydroxy-γ-lactone (10.77%), propiolic acid, 3-(1-hydroxy)-2 isopropyl-1,5-methylcyclohexyl) (7.84%), 4-oxo-β-isodamascol (5.17%), velleral (3.11%) and Z-α-farnesene (2.00%). The most prevailing major compounds identified in the polar fraction of the M/C extraction were α-D glucopyranoside, 1,3,4,6 tetrakis-O-(TMS) (trimethylsilyl)-β-D-fructofuranosyl 2,3,4,6-tetrakis-O-(TMS)- (38.08%), d-glucose, 2,3,4,5,6-pentakis-O-(TMS)-, O-methyloxime (14.61%), D-fructose, 1,3,4,5,6-pentakis-O-(TMS)-, O-methyloxime (5.28%), isocitric acid (TMS) (3.06%), oxalic acid, bis (TMS) ester (2.96%), hexadecanoic acid, TMS ester (2.16%), citric acid, ethyl ester, tri-TMS (1.91%) and butanedioic acid, [(TMS) oxy]-, bis (TMS) ester (1.14%); whereas in the nonpolar extract, among the major compounds detected were cycloisolongifolene, 8, 9-dehydro −9-formyl (15.70%), propiolic acid, 3-(1-hydroxy-2-isopropyl-5-methylcyclohexyl) (11.09%), stearic acid, TMS ester (2.78%), hexadecanoic acid, TMS ester (2.33%), oleic acid, TMS ester (1.62%), curzerene (1.56%); Z-α-farnesene (1.52%), germacrone (1.41%) and β-elemene (1.33%). Conclusions: It was evident from the results that C. aeruginosa rhizome extracted using two different techniques of solvent extractions (MTBE and M/C) contained various chemical classes of compounds including terpenoids, sterols, organic acids, fatty acids and sugars. Different methods of extraction have led to different compounds extraction for C. aeruginosa rhizome. The results also indicated that the plant was a source of phytochemical importance

UHPLC-ESI-Orbitrap-MS Analysis of Biologically Active Extracts from Gynura procumbens (Lour.) Merr. and Cleome gynandra L. Leaves

This study aimed to determine the total phenolic content, DPPH scavenging, α-glucosidase, and nitric oxide (NO) inhibition of Gynura procumbens and Cleome gynandra extracts obtained with five different ethanolic concentrations. The findings showed that the 100% ethanolic extract of G. procumbens had the highest phenolic content and the lowest IC50 values for DPPH scavenging and NO inhibition activity compared to the properties of the other extracts. For C. gynandra, the 20% and 100% ethanolic extracts had comparably high total phenolic contents, and the latter possessed the lowest IC50 value in the NO inhibition assay. In addition, the 20% ethanolic extract of C. gynandra had the lowest IC50 value in the DPPH scavenging assay. However, none of the extracts from either herb had the ability to inhibit α-glucosidase enzyme. Pearson correlation analysis indicated a strong relationship between the phenolic content and DPPH scavenging activity in both herb extracts. A moderately strong relationship was also observed between the phenolic content and NO inhibition in G. procumbens extracts and not in C. gynandra extracts. The UHPLC-ESI-Orbitrap-MS revealed major phenolics from the groups of hydroxycinnamic acids, hydroxybenzoic acids, and flavonoid derivatives from both herbs, which could be the key contributors to their bioactivities. Among the identified metabolites, 24 metabolites were tentatively assigned for the first time from both species of studied herbs. These two herbs could be recommended as prospective natural products with valuable medicinal properties

Discovery of Functional SNPs via Genome-Wide Exploration of Malaysian Pigmented Rice Varieties

Recently, rice breeding program has shown increased interests on the pigmented rice varieties due to their benefits to human health. However, the genetic variation of pigmented rice varieties is still scarce and remains unexplored. Hence, we performed genome-wide SNP analysis from the genome resequencing of four Malaysian pigmented rice varieties, representing two black and two red rice varieties. The genome of four pigmented varieties was mapped against Nipponbare reference genome sequences, and 1.9 million SNPs were discovered. Of these, 622 SNPs with polymorphic sites were identified in 258 protein-coding genes related to metabolism, stress response, and transporter. Comparative analysis of 622 SNPs with polymorphic sites against six rice SNP datasets from the Ensembl Plants variation database was performed, and 70 SNPs were identified as novel SNPs. Analysis of SNPs in the flavonoid biosynthetic genes revealed 40 nonsynonymous SNPs, which has potential as molecular markers for rice seed colour identification. The highlighted SNPs in this study show effort in producing valuable genomic resources for application in the rice breeding program, towards the genetic improvement of new and improved pigmented rice varieties

Volatile compounds of esssential oil from different stages of Michelia alba (cempaka putih) flower development

Seven developmental stages of Michelia alba (cempaka putih) flowers namely Stage 5 to 11 (S5–S11) were investigated for their volatile compounds. The essential oil was isolated by Simultaneous Distillation Extraction (SDE) technique and the oil obtained was subjected to Gas Chromatography-Mass Spectrometry (GC-MS) analysis. In total, 78 compounds representing 93–98% of the overall M. alba volatiles were identified. Thirty-three of these compounds belonged to isoprenoids group which comprised 30–50% of the total volatile compounds detected throughout S5–S11, whereas the remaining belonged to fatty acid derivatives, benzenoid, phenylpropanoid and other hydrocarbon compounds. The major compounds which represented more than 10% of the essential oil at each stage were dihydrocarveol (S5–S8), linalool (S9–S11), butanoic acid-2-methyl, methyl ester (S9) and cyclohexane, 1-ethenyl-1-methyl-2,4-bis (1- methylethenyl) (S6–S7). In this study, variations in the compounds of essential oil as well as their level in percentage within the flower development stages were observed. Dihydrocarveol was the most abundant compounds detected in S5–S8 (44–65%), while linalool was the most abundant compound detected in S9–S11, which accounted for 59–89% of the total essential oil obtained. Based on the profile of both compounds, it might suggest that dihydrocarveol was one of the compounds that contributed significantly during bud development through S5–S8 in which the bud became yellowish and started to swell until the petal just began to open, whereas linalool might contribute significantly to the characteristic fragrance through S9–S11 in which during these phases, the aroma of M. alba fragrance was very much intense

Effect of storage time on metabolite profile and alpha-glucosidase inhibitory activity of Cosmos caudatus leaves - GCMS based metabolomics approach

Cosmos caudatus, which is a commonly consumed vegetable in Malaysia, is locally known as “Ulam Raja”. It is a local Malaysian herb traditionally used as a food and medicinal herb to treat several maladies. Its bioactive or nutritional constituents consist of a wide range ofmetabolites, including glucosinolates, phenolics, amino acids, organic acids, and sugars. However, many of these metabolites are not stable and easily degraded or modified during storage. In order to investigate the metabolomics changes occurring during post-harvest storage, C. caudatus samples were subjected to seven different storage times (0 hours, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, and 12 hours) at room temperature. As the model experiment, the metabolites identified by gas chromatography-mass spectrometry (GC-MS) were correlated with a-glucosidase inhibitory activity analyzed with multivariate data analysis (MVDA) to find out the variation among samples and metabolites contributing to the activity. Orthogonal partial least squares (OPLS) analysis was applied to investigate the metabolomics changes. A profound chemical alteration, both in primary and secondary metabolites, was observed. The a-tocopherol, catechin, cyclohexen-1-carboxylic acid, benzoic acid, myo-inositol, stigmasterol, and lycopene compoundswere found to be the discriminatingmetabolites at early storage;however, sugars such as sucrose, a-D-galactopyranose, and turanose were detected, which was attributed to the discriminating metabolites for late storage. The result shows that the MVDA method is a promising technique to identify biomarker compounds relative to storage at different time