Untargeted metabolomics using UHPLC-Q-Orbitrap HRMS for identifying cytotoxic compounds on MCF-7 breast cancer cells from Annona muricata Linn leaf extracts as potential anticancer agents.

INTRODUCTION: The leaves of Annona muricata L., known as "soursop" or "sirsak" in Indonesia, are used traditionally for cancer treatment. However, the bioactive components remain largely unidentified. OBJECTIVE: This study used untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics to identify potential cytotoxic compounds in A. muricata leaf extracts on MCF-7 breast cancer cells in vitro. METHODS: A. muricata leaves were macerated with water, 99% ethanol, and aqueous mixtures containing 30%, 50%, and 80% ethanol. Cytotoxic activity of the extracts against MCF-7 breast cancer cells was determined using the MTT assay. Ultra-high-performance liquid chromatography-Q-Orbitrap high-resolution mass spectroscopy (UHPLC-Q-Orbitrap-HRMS) was used to characterize the metabolite composition of each extract. The correlations between metabolite profile and cytotoxic activities were evaluated using orthogonal partial least square discriminant analysis (OPLS-DA). The binding of these bioactive compounds to the tumorigenic alpha-estrogen receptor (3ERT) was then evaluated by in silico docking simulations. RESULTS: Ninety-nine percent ethanol extracts demonstrated the greatest potency for reducing MCF-7 cell viability (IC50 = 22 µg/ml). We detected 35 metabolites in ethanol extracts, including alkaloids, flavonoids, and acetogenins. OPLS-DA predicted that annoreticuin, squadiolin C, and xylopine, and six unknown acetogenin metabolites, might reduce MCF-7 cell viability. In silico analysis predicted that annoreticuin, squadiolin C, and xylopine bind to 3ERT with an affinity comparable to doxorubicin. CONCLUSION: Untargeted metabolomics and in silico modeling identified cytotoxic compounds on MCF-7 cells and binding affinity to 3ERT in A. muricata leaf extracts. The findings need to be further verified to prove the screening results.

UHPLC-Q-Orbitrap HR-MS-Based Metabolomics for Profiling the Sida rhombifolia Metabolites with Different Plant Organs and Cultivation Ages.

Plant organs and cultivation ages can result in different compositions and concentration levels of plant metabolites. The metabolite profile of plants can be determined using liquid chromatography. This study determined the metabolite profiles of leaves, stems, and roots of Sida rhombifolia at different cultivation ages at 3, 4, and 5 months post-planting (MPP) using liquid chromatography-mass spectrometry/mass spectrometry (LC/MS/MS). The results identified that 41 metabolites in S. rhombifolia extract for all plant organs and cultivation ages. We successfully identified approximately 36 (leaves), 22 (stems), and 18 (roots) compounds in all extract. Using principal component analysis (PCA) with peak area as the variable, we clustered all sample extracts based on plant organs and cultivation ages. As a result of PCA, S. rhombifolia extracts were grouped according to plant organs and cultivation ages. In conclusion, a clear difference in the composition and concentration levels of metabolites was observed in the leaves, stems, and roots of S. rhombifolia harvested at 3-, 4-, and 5-MPP.

Metabolomics Study Reveals Biomarker L-Proline as Potential Stress-Protectant Compound for High-Temperature Bioethanol Fermentation by Yeast Pichia kudriavzevii 1P4.

High-temperature ethanol fermentation (> 40 °C) can be applied as effective bioprocess technology to increase ethanol production. Thermotolerant yeast Pichia kudriavzevii 1P4 showed the ability to produce ethanol at optimum 37 °C. Thus, this study evaluated the ethanol productivity of isolate 1P4 at high-temperature ethanol fermentation (42 and 45 °C) and the identification of metabolite biomarkers using untargeted metabolomics with liquid chromatography-tandem mass spectrometry (LC-MS/MS). 1P4 showed tolerance to temperature stress up to 45 °C and thus relevant for high-temperature fermentation. As measured by gas chromatography (GC), bioethanol production of 1P4 at 30, 37, 42, and 45 °C was 5.8 g/l, 7.1 g/l, 5.1 g/l, and 2.8 g/l, respectively. The classification of biomarker compounds was based on orthogonal projection analysis to latent structure discriminant analysis (OPLS-DA), resulting in L-proline being a suspected biomarker compound for isolate 1P4 tolerance against high-temperature stress. Indeed, supplementation of L-proline on fermentation medium supported the growth of 1P4 at high temperatures (> 40 °C) than without L-proline. The bioethanol production with the addition of the L-proline resulted in the highest ethanol concentration (7.15 g/l) at 42 °C. Supplementation of L-proline as a stress-protective compound increased ethanol productivity at high-temperature fermentation of 42 and 45 °C by 36.35% and 83.33%, respectively, compared without the addition of L-proline. Preliminary interpretation of these results indicates that bioprocess engineering through supplementation of stress-protective compounds L-proline increases the fermentation efficiency of isolate 1P4 at higher temperatures (42 °C and 45 °C).

Antioxidant Activity and Metabolite Profiling of Xylocarpus granatum Extracts Using Gas Chromatography-Mass Spectrometry.

The potential application of Xylocarpus granatum, a mangrove species, as traditional medicine has been widely linked to its high secondary metabolite and antioxidant contents. However, few studies have been reported to identify and classify active metabolites responsible for such excellent biological activities. Therefore, the aim of this work was to determine the antioxidant activity, identify the metabolite profiles, and predict the metabolites acting as antioxidants in X. granatum extract using a gas chromatography-mass spectrometry (GC-MS)-based metabolomics approach. The seeds, stems, fruit peel, pulp, leaves, and twigs of X. granatum were macerated with ethanol. Each extract was analyzed with GC-MS, and the data were processed using mass spectrometry data-independent analysis (MS-DIAL) software to identify the metabolites. The IC50 value of plant parts of X. granatum ranged from 7.73 to 295 ppm. A total of 153 metabolites were identified and confirmed in the X. granatum extracts. Among the identified metabolites, epicatechin and epigallocatechin were the two most abundant in the stem extracts and are expected to have the greatest potential as antioxidants. Principal component analysis (PCA) succeeded in grouping all parts of the plant into three groups based on the composition of the metabolites: group 1 (stems, fruit peel, and twigs), group 2 (seeds and pulp), and group 3 (leaves).

Effects of two contrasting potting media on the leaf development index, photosynthetic rate, and metabolite profile of camphor (Dryobalanops aromatica) seedlings.

Camphor (Dryobalanops aromatica C. F. Gaertn.) is a vulnerable tropical tree species that has been exploited for its timber as well as its resin, which is used for medicinal uses. The use of camphor in Indonesia is limited owing to the decreasing size of the species' population in its native habitat. Therefore, replanting programs have been encouraged for this species owing to its adaptability to mineral soils and shallow peatlands. However, experimental evidence of the effect of different growing media on morphology, physiology, and biochemistry is very limited, which is needed to evaluate the replanting program's success. Therefore, this study aimed to determine the responses of camphor (D. aromatica) seedlings grown in two different types of potting media i.e. mineral and peat, for 8 weeks of planting. In particular, the types of bioactive compounds produced in camphor leaves and their levels were assessed by analyzing their metabolite profiles. Leaf growth was evaluated morphologically using the plastochron index, while photosynthetic rates were measured with LI-6800 Portable Photosynthesis System. Metabolites were identified by using liquid chromatography-tandem mass spectrometry. The percentage of LPI of 5 or more was lower in the peat medium at 8% than in the mineral medium at 12%. The photosynthetic rate of camphor seedlings was 1-9 µmol CO2 m⁻2 s⁻1, with a higher rate in the peat medium than in the mineral medium, suggesting that the peat medium was better for growth. Lastly, the metabolomic analysis in the leaf extract revealed the presence of 21 metabolites, which were dominated by flavonoid compounds.

Electrochemical Sensors Based on a Composite of Electrochemically Reduced Graphene Oxide and PEDOT:PSS for Hydrazine Detection.

In this study, hydrazine sensors were developed from a composite of electrochemically reduced graphene oxide (ErGO) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS), deposited onto a glassy carbon electrode (GCE). The structural properties, electrochemical characterization, and surface morphologies of this hydrazine sensor were characterized by Raman spectroscopy, Fourier transform infrared (FTIR) spectroscopy, electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). In addition, the proposed hydrazine sensor also demonstrates good electrochemical and analytical performance when investigated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and amperometry techniques under optimal parameters. Using these investigated parameters, DPV and amperometry were chosen as techniques for hydrazine measurements and showed a linear range of concentration in the range of 0.2-100 µM. The obtained limits of detection and limits of quantitation for hydrazine measurements were 0.01 and 0.03 µM, respectively. In addition, the proposed sensor demonstrated good reproducibility and stability in hydrazine measurements in eight consecutive days. This fabricated hydrazine sensor also exhibited good selectivity against interference from Mg2+, K+, Zn2+, Fe2+, Na+, NO2 -, CH3COO-, SO4 2-, Cl-, ascorbic acid, chlorophenol, and triclosan and combined interferences, as well as it depicted %RSD values of less than 5%. In conclusion, this proposed sensor based on GCE modified with ErGO/PEDOT:PSS displays exceptional electrochemical performance for use in hydrazine measurements and have the potential to be employed in practical applications.

Drug candidates and potential targets of Curculigo spp. compounds for treating diabetes mellitus based on network pharmacology, molecular docking and molecular dynamics simulation.

Curculigo spp. is a herb that is commonly used in Indonesia to treat diabetes mellitus (DM) . The main active components of Curculigo spp. were identified through our previous metabolomic study and online database platform. However, the biological mechanisms underlying Curculigo spp. activity in treating DM remain unclear. Therefore, in this study, a network pharmacology was used to explore the active compounds of Curculigo spp. and their potential molecular mechanisms for treating DM. Oral bioavailability and drug-likeness from the compounds of Curculigo spp. were screened using Lipinski's rule of five, BBB, HIA + and Caco-2 permeability criteria. A network of compound-target-disease-pathway was then constructed using Cytoscape. The highest degree compounds and targets were then confirmed by molecular docking and molecular dynamics (MD) simulations. The human body can absorb 33 compounds derived from Curculigo spp. In addition, 58 nodes and 62 edges generated a network analysis with the DM target. The highest degree of the compound-target-disease pathway was for orcinol glucoside, AKR1B1, autoimmune diabetes, bile acid and bile salt metabolism. Furthermore, the computational docking method on Curculigo spp. compounds with the highest degree revealed that orcinol glucoside interacted with PTPN1 through a hydrogen bond and resulted in a binding energy of -7.2 kcal mol-1. Through hydrogen bonds, orcinol glucoside in PTPN1 regulates multiple signaling pathways via the adherens junction pathway, which may play a therapeutic role in DM (type 2 diabetes: obesity). In addition, MD simulation confirmed that orcinol glucoside, is suitable for DM treatment by interacting with PTPN1.Communicated by Ramaswamy H. Sarma.

Natural Antioxidant Activities of Plants in Preventing Cataractogenesis.

A cataract is a condition that causes 17 million people to experience blindness and is the most significant cause of vision loss, around 47.9%. The formation of cataracts is linked to both the production of reactive oxygen species (ROS) and the reduction of endogenous antioxidants. ROS are highly reactive molecules produced by oxygen. Examples of ROS include peroxides, super-oxides, and hydroxyl radicals. ROS are produced in cellular responses to xenobiotics and bacterial invasion and during mitochondrial oxidative metabolism. Excessive ROS can trigger oxidative stress that initiates the progression of eye lens opacities. ROS and other free radicals are highly reactive molecules because their outer orbitals have one or more unpaired electrons and can be neutralized by electron-donating compounds, such as antioxidants. Examples of natural antioxidant compounds are vitamin C, vitamin E, and beta-carotene. Numerous studies have demonstrated that plants contain numerous antioxidant compounds that can be used as cataract preventatives or inhibitors. Natural antioxidant extracts for cataract therapy may be investigated further in light of these findings, which show that consuming a sufficient amount of antioxidant-rich plants is an excellent approach to cataract prevention. Several other natural compounds also prevent cataracts by inhibiting aldose reductase and preventing apoptosis of the eye lens.

Collecting wood core samples from Macassar ebony (Diospyros celebica Bakh.) for multi-purpose analysis using pickering punch.

Sample collection activities for a study of population genetics across the natural distribution of targeted tree species require a lot of resources, mainly if repeated field visits are necessary. Conventionally, population genetic studies use good sample material like leaves. In addition, cambium or small pieces of fresh wood can be used to replace leaf samples. Currently, restrictions from the permit regulation have caused only a limited number of samples that can be collected. Therefore, efficient use of samples must be designed to maximize their uses for research. Due to the small amount of successfully sampled materials, hence there are limitations to extend their uses for other analyses and are often sufficient only for genetic analysis. Therefore, innovation in sampling methods using pickering punch (https://www.agroisolab.com/pickering-punch) to collect ebony wood cores in this study is required to cover multi-analyses not only limited to genetics but also for other analyses such as isotopes, near-infrared spectroscopy (NIRs), anatomy, and chemical compounds.•Pickering punch is recommended for efficient wood core sample collection from ebony standing trees.•323 wood core samples were successfully collected from 16 natural populations across Celebes (Sulawesi).•Multi-analyses studies on sampled wood cores are possible for ebony wood identification (e.g., species and origin/provenance).

Metabolite Fingerprinting Based on 1H-NMR Spectroscopy and Liquid Chromatography for the Authentication of Herbal Products.

Herbal medicines (HMs) are regarded as one of the traditional medicines in health care to prevent and treat some diseases. Some herbal components such as turmeric and ginger are used as HMs, therefore the identification and confirmation of herbal use are very necessary. In addition, the adulteration practice, mainly motivated to gain economical profits, may occur by substituting the high price of HMs with lower-priced ones or by addition of certain chemical constituents known as Bahan Kimia Obat (chemical drug ingredients) in Indonesia. Some analytical methods based on spectroscopic and chromatographic methods are developed for the authenticity and confirmation of the HMs used. Some approaches are explored during HMs authentication including single-component analysis, fingerprinting profiles, and metabolomics studies. The absence of reference standards for certain chemical markers has led to exploring the fingerprinting approach as a tool for the authentication of HMs. During fingerprinting-based spectroscopic and chromatographic methods, the data obtained were big, therefore the use of chemometrics is a must. This review highlights the application of fingerprinting profiles using variables of spectral and chromatogram data for authentication in HMs. Indeed, some chemometrics techniques, mainly pattern recognition either unsupervised or supervised, were applied for this purpose.

Metabolite profiling of Andrographis paniculata leaves and stem extract using UHPLC-Orbitrap-MS/MS.

Andrographis paniculata is known as the king of bitter and it has been widely used as a medicinal plant. The properties of A. paniculata are generally determined by the metabolite composition, which may be influenced by several factors, one of which is the part of the plant extracted. The objectives of this research are to identify putatively the metabolite composition of the stem and the leaves extracts using LC-MS/MS and classify them using PCA. The stem and the leaves samples were separated and extracted using sonication with 70% ethanol. A total of 31 metabolite compounds has been putatively identified. All compounds were identified in the stem and the leaves extracts, which only differed in their intensity. These metabolite compounds were divided into diterpene lactones, flavonoids, and phenolic acid groups. By using the peak intensities of the 18 compounds identified, the leaves and stem extracts were grouped using PCA.

Metabolite Fingerprinting Using 1H-NMR Spectroscopy and Chemometrics for Classification of Three Curcuma Species from Different Origins.

Curcuma longa, Curcuma xanthorrhiza, and Curcuma manga have been widely used for herbal or traditional medicine purposes. It was reported that turmeric plants provided several biological activities such as antioxidant, anti-inflammatory, hepatoprotector, cardioprotector, and anticancer activities. Authentication of the Curcuma species is important to ensure its authenticity and to avoid adulteration practices. Plants from different origins will have different metabolite compositions because metabolites are affected by soil nutrition, climate, temperature, and humidity. 1H-NMR spectroscopy, principal component analysis (PCA), and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) were used for authentication of C. longa, C. xanthorrhiza, and C. manga from seven different origins in Indonesia. From the 1H-NMR analysis it was obtained that 14 metabolites were responsible for generating classification model such as curcumin, demethoxycurcumin, alanine, methionine, threonine, lysine, alpha-glucose, beta-glucose, sucrose, alpha-fructose, beta-fructose, fumaric acid, tyrosine, and formate. Both PCA and OPLS-DA model demonstrated goodness of fit (R2 value more than 0.8) and good predictivity (Q2 value more than 0.45). All OPLS-DA models were validated by assessing the permutation test results with high value of original R2 and Q2. It can be concluded that metabolite fingerprinting using 1H-NMR spectroscopy and chemometrics provide a powerful tool for authentication of herbal and medicinal plants.

Influence of Combined NPK and Manure on Improving Growth, Photosynthetic Characteristic and Yield of Justicia gendarussa Burm. F.

<b>Background and Objective:</b> <i>Justicia gendarussa </i>(family <i>Acanthaceae</i>) is regarded as a medicinal plant valued for its beneficial pharmacological activities. A polybag experiment has been conducted at Bogor (Indonesia) to determine the effects on growth, photosynthetic and yield of various NPK ratios and manure fertilizer application in <i>J. gendarussa</i>. <b>Materials and Methods:</b> Six treatments (control: 50 g/plant manure, NPK (1.1 g/plant N:0.7 g/plant P₂O₅ and 0.6 g/plant K), 50+50%/plant (manure+NPK), 10+50%/plant (manure+NPK) and 50+10%/plant (manure+NPK) were performed in a randomized complete block design. Data of individual plants were extracted from various treatments: plant height, number of leaves, number of branches, chlorophyll content, photosynthetic parameters, yield and sugar content were collected. <b>Results:</b> The plant treated with 10+50%/plant (manure+NPK) demonstrated a statistically significant increase in plant height, number of leaves and number of branches with the value of 59.00 cm, 85.00 per plant and 9.50 per plant, respectively. Chlorophyll a (0.538 mg g¹ FW), chlorophyll b (0.192 mg g¹ FW) and total chlorophyll (0.730 mg g¹ FW) were significantly improved by the treatment of NPK fertilizer. The application of manure and NPK at (50+50%) and (50+10%) was significantly increased stomatal conductance and transpiration rate, respectively. Leaves yield (9 g/plant DW) was improved by the application of 10+50%/plant (manure+NPK) fertilizer. The highest sugar content was obtained from <i>J. gendarussa</i> growth with 50+50%/plant (manure+NPK) followed by 10+50% plant (manure+NPK) fertilizer. <b>Conclusion:</b> The <i>J. gendarussa</i> plants could benefit from combined manure and NPK fertilizer application in growth, stomatal conductance, intercellular CO₂ concentration, transpiration rate, leaves yield and sugar content.

Investigation of Yacon Leaves (Smallanthus sonchifolius) for α-Glucosidase Inhibitors Using Metabolomics and In Silico Approach.

Yacon (Smallanthus sonchifolius (Poepp.) H. Robinson) leaves is traditionally consumed as herbal tea in many countries including Indonesia. This plant's antidiabetic properties have been extensively researched, but studies on the responsible active compound identification are scarce. Information on the active compounds is critical for the consistency of Yacon herbal tea quality. The aim of this study was to identify α-glucosidase inhibitors in Indonesian Yacon leaves grown in two different locations using FTIR- and LC-MS/MS-based metabolomics in combination with in silico technique. Yacon leaves ethanol (50 and 95%) and water extracts were tested for α-glucosidase inhibitory activity, with the 95% ethanol extract being the most active. Geographical origins were found to have no major impact on the activity. In parallel, chemical profile of Yacon leaves extract was determined using FTIR and LC-MS/MS. Orthogonal Projection to Latent Structure (OPLS) was used to analyze both sets of data. OPLS analysis of FTIR data showed that compounds associated to α-glucosidase inhibitor activity included those with functional groups -OH, stretched CH, carbonyl, and alkene. It was consistent with the result of OPLS analysis of LC-MS/MS data, which revealed that based on their VIP and Y-related coefficient value, nystose, 1-kestose, luteolin-3'-7-di-O-glucoside, and 1,3-O-dicaffeoilquinic acid isomers, strongly linked to Yacon's α-glucosidase inhibitor activity. In silico study supported these findings, revealing that the four compounds were potent α-glucosidase inhibitors with docking score in the range of - 100.216 to - 115.657 kcal/mol, which are similar to acarbose (- 115.774 kcal/mol) as a reference drug.

Untargeted Metabolomics Analysis Using FTIR and UHPLC-Q-Orbitrap HRMS of Two Curculigo Species and Evaluation of their Antioxidant and α-Glucosidase Inhibitory Activities.

Curculigo orchioides and C. latifolia have been used as traditional medicines such as antidiabetic and anticancer. This study measured the total phenolics and flavonoid contents as well as analyzed the functional groups and chemical compounds using Fourier-transform infrared (FTIR) spectra and UHPLC-Q-Orbitrap-HRMS profiling for the discrimination of plant parts, geographical origin, and compounds that presumably have a significant contribution as antioxidant and α-glucosidase inhibitors on both plants. The total phenolics and flavonoids contents in Curculigo species varied from 142.09 to 452.47 mg gallic acid equivalent (GAE/g) and from 0.82 to 5.44 mg quercetin equivalent (QE/g), respectively. The lowest IC50 for antioxidant and α-glucosidase inhibitory activities is presented by C. latifolia from a higher altitude region. Principal component analysis (PCA) from FTIR and UHPLC-Q-Orbitrap-HRMS data could discriminate the plant parts and geographical origin. Partial least squares (PLS) analysis has identified several functional groups, such as O-H, C-H, C=O, C-C, C-O, and chemical compounds, unknown-185 and unknown-85, that are most likely to contribute to the antioxidant and α-glucosidase inhibitory activities.

HPLC fingerprint and simultaneous quantitative analysis of phyllanthin and hypophyllanthin for identification and authentication of Phyllanthus niruri from related species

Abstract A precise and accurate method for the identification and authentication of Phyllanthus niruri L. from P. debilis Klein ex Willd. and P. urinaria L., Phyllanthaceae, was developed using high-performance liquid chromatography. Chromatographic fingerprint analysis was combined with simultaneous quantification of phyllanthin and hypophyllanthin for the developed method. Phyllanthin and hypophyllanthin were successfully separated and quantified under this proposed method. The highest amount of phyllanthin and hypophyllanthin was found in P. niruri compared to P. debilis and P. urinaria. Fingerprint chromatogram of the three Phyllanthus species showed distinct profiles that these may be used to identify and authenticate each Phyllanthus species, which improved by marker compounds present in each species. The combination of chromatographic fingerprint analysis and discriminant analysis was successfully discriminated all three species, including P. niruri adulterated with P. debilis or P. urinaria. The method can be used for the identification and authentication of P. niruri from related species, such as P. debilis and P. urinaria.

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.

Discrimination of red and white rice bran from Indonesia using HPLC fingerprint analysis combined with chemometrics.

HPLC fingerprint analysis combined with chemometrics was developed to discriminate between the red and the white rice bran grown in Indonesia. The major component in rice bran is γ-oryzanol which consisted of 4 main compounds, namely cycloartenol ferulate, cyclobranol ferulate, campesterol ferulate and ß-sitosterol ferulate. Separation of these four compounds along with other compounds was performed using C18 and methanol-acetonitrile with gradient elution system. By using these intensity variations, principal component and discriminant analysis were performed to discriminate the two samples. Discriminant analysis was successfully discriminated the red from the white rice bran with predictive ability of the model showed a satisfactory classification for the test samples. The results of this study indicated that the developed method was suitable as quality control method for rice bran in terms of identification and discrimination of the red and the white rice bran.

Fourier transform infrared spectroscopy combined with chemometrics for discrimination of Curcuma longa, Curcuma xanthorrhiza and Zingiber cassumunar.

Turmeric (Curcuma longa), java turmeric (Curcuma xanthorrhiza) and cassumunar ginger (Zingiber cassumunar) are widely used in traditional Indonesian medicines (jamu). They have similar color for their rhizome and possess some similar uses, so it is possible to substitute one for the other. The identification and discrimination of these closely-related plants is a crucial task to ensure the quality of the raw materials. Therefore, an analytical method which is rapid, simple and accurate for discriminating these species using Fourier transform infrared spectroscopy (FTIR) combined with some chemometrics methods was developed. FTIR spectra were acquired in the mid-IR region (4000-400 cm(-1)). Standard normal variate, first and second order derivative spectra were compared for the spectral data. Principal component analysis (PCA) and canonical variate analysis (CVA) were used for the classification of the three species. Samples could be discriminated by visual analysis of the FTIR spectra by using their marker bands. Discrimination of the three species was also possible through the combination of the pre-processed FTIR spectra with PCA and CVA, in which CVA gave clearer discrimination. Subsequently, the developed method could be used for the identification and discrimination of the three closely-related plant species.

Capillary liquid chromatographic fingerprint used for discrimination of Zingiber montanum from related species.

Fingerprint analysis using capillary liquid chromatography (CLC) has been developed for discrimination of Zingiber montanum (ZM) from related species, for example Z. americans (ZA) and Z. zerumbet (ZZ). By comparing the fingerprint chromatograms of ZM, ZA, and ZZ we could identify ZM samples and discriminate them from ZA and ZZ by using their marker peaks. We also combined CLC fingerprint with multivariate analysis, including principal-component analysis (PCA) and canonical variate analysis (CVA); all three species were discriminated successfully. This result indicates that CLC fingerprint analysis in combination with PCA and CVA can be used for discrimination of ZM samples from samples of related species.