Application of Chromatographic and Spectroscopic-Based Methods for Analysis of Omega-3 (ω-3 FAs) and Omega-6 (ω-6 FAs) Fatty Acids in Marine Natural Products.

Omega-3 fatty acids v(ω-3 FAs) such as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) and omega-6 fatty acids (ω-6 FAs) such as linoleic acid and arachidonic acid are important fatty acids responsible for positive effects on human health. The main sources of ω-3 FAs and ω-6 FAs are marine-based products, especially fish oils. Some food, supplements, and pharmaceutical products would include fish oils as a source of ω-3 FAs and ω-6 FAs; therefore, the quality assurance of these products is highly required. Some analytical methods mainly based on spectroscopic and chromatographic techniques have been reported. Molecular spectroscopy such as Infrared and Raman parallel to chemometrics has been successfully applied for quantitative analysis of individual and total ω-3 FAs and ω-6 FAs. This spectroscopic technique is typically applied as the alternative method to official methods applying chromatographic methods. Due to the capability to provide the separation of ω-3 FAs and ω-6 FAs from other components in the products, gas and liquid chromatography along with sophisticated detectors such as mass spectrometers are ideal analytical methods offering sensitive and specific results that are suitable for routine quality control.

Hydroxypropyl Methylcellulose as Hydrogel Matrix and Citric Acid-Locust Bean Gum as Negative Matrix for Controlled Release Tablet.

Purpose: This study aimed at determining the optimum concentration of hydroxypropyl methylcellulose (HPMC) as hydrogel matrix and citric acid-locust bean gum (CA-LBG) as negative matrix for controlled release tablet formulation. In addition, the study was to determine the effect of CA-LBG and HPMC. CA-LBG accelerates the disintegration of tablets into granules so that the HPMC granule matrix swells immediately and controls drug release. The advantage of this method is that the tablets do not produce large HPMC gel lumps without drug (ghost matrix) but form HPMC gel granules, which can be rapidly degraded after all of the drug is released. Methods : The experiment followed the simplex lattice design to obtain the optimum tablet formula with CA-LBG and HPMC concentrations as optimization factors. Tablet production by the wet granulation method and ketoprofen is the model of the active ingredient. The kinetics of ketoprofen release was studied using several models. Results : Based on the coefficients of each polynomial equation that HPMC and CA-LBG increased the value of angle of repose (29.91:27.87), tap index (18.99:18.77), hardness (13.60:13.32), friability (0.41:0.73), and release of ketoprofen (52.48:99.44). Interaction of HPMC and CA-LBG increased the value of angle of repose (3.25), tap index (5.64), and hardness (2.42). Interaction of HPMC and CA-LBG too decreased the friability value (-1.10) and release of ketoprofen (-26.36). The Higuchi, Korsmeyer-Peppas, and Hixson-Crowell model is the kinetics of eight experimental tablet formulas. Conclusions : The optimum concentrations of HPMC and CA-LBG for controlled release tablets are 32.97 and 17.03%, respectively. HPMC, CA-LBG, and a combination of both affect the physical quality of tablet and tablet mass. CA-LBG is a new excipient candidate that can control drug release from tablets by the matrix disintegration mechanism on the tablet.

The Combination of Vibrational Spectroscopy and Chemometrics for Analysis of Milk Products Adulteration.

Milk products obtained from cow, goat, buffalo, sheep, and camel as well as fermented forms such as cheese, yogurt, kefir, and butter are in a category of the most nutritious foods due to their high contents of high protein contributing to total daily energy intake. For certain reasons, high price milk products may be adulterated with low-quality ones or with foreign substances such as melamine and formalin which are added into them; therefore, a comprehensive review on analytical methods capable of detecting milk adulteration is needed. The objective of this narrative review is to highlight the use of vibrational spectroscopies (near infrared, mid infrared, and Raman) combined with multivariate analysis for authentication of milk products. Articles, conference reports, and abstracts from several databases including Scopus, PubMed, Web of Science, and Google Scholar were used in this review. By selecting the correct conditions (spectral treatment, normal versus derivative spectra at wavenumbers region, and chemometrics techniques), vibrational spectroscopy is a rapid and powerful analytical technique for detection of milk adulteration. This review can give comprehensive information for selecting vibrational spectroscopic methods combined with chemometrics techniques for screening the adulteration practice of milk products.

Comprehensive Review on Application of FTIR Spectroscopy Coupled with Chemometrics for Authentication Analysis of Fats and Oils in the Food Products.

Currently, the authentication analysis of edible fats and oils is an emerging issue not only by producers but also by food industries, regulators, and consumers. The adulteration of high quality and expensive edible fats and oils as well as food products containing fats and oils with lower ones are typically motivated by economic reasons. Some analytical methods have been used for authentication analysis of food products, but some of them are complex in sampling preparation and involving sophisticated instruments. Therefore, simple and reliable methods are proposed and developed for these authentication purposes. This review highlighted the comprehensive reports on the application of infrared spectroscopy combined with chemometrics for authentication of fats and oils. New findings of this review included (1) FTIR spectroscopy combined with chemometrics, which has been used to authenticate fats and oils; (2) due to as fingerprint analytical tools, FTIR spectra have emerged as the most reported analytical techniques applied for authentication analysis of fats and oils; (3) the use of chemometrics as analytical data treatment is a must to extract the information from FTIR spectra to be understandable data. Next, the combination of FTIR spectroscopy with chemometrics must be proposed, developed, and standardized for authentication and assuring the quality of fats and oils.

The Authentication of Java Turmeric (Curcuma xanthorrhiza) Using Thin Layer Chromatography and 1H-NMR Based-Metabolite Fingerprinting Coupled with Multivariate Analysis.

The identification of adulteration practices of medicinal plants used as herbal medicine is very important to ensure the quality, safety, and efficacy. In this study, thin layer chromatography (TLC) and proton nuclear magnetic resonance (1H-NMR)-based metabolite fingerprinting coupled with multivariate analysis were used for authentication of Curcuma xanthorrhiza extract from Curcuma aeruginosa. Curcumin contents obtained from C. xanthorrhiza extract from various regions were in the range of 0.74%-1.23%. Meanwhile, curcumin contents obtained from C. xanthorrhiza extract adulterated with 0%, 10%, 25%, 40%, 50%, and 75% of C. aeruginosa were 1.02%, 0.96%, 0.86%, 0.69%, 0.43%, and 0.27%, respectively. The decreasing of curcumin contents in adulterant concentrations of 40% and more in C. xanthorrhiza rhizome could indicate the adulteration with other rhizomes. Multivariate analysis of PCA (principal component analysis) using data set obtained from 1H-NMR spectra clearly discriminated pure and adulterated C. xanthorrhiza with C. aeruginosa. OPLS-DA (orthogonal projections to latent structures-discriminant analysis) successfully classified pure and adulterated C. xanthorrhiza with higher R2X (0.965), R2Y (0.958), and Q2(cum) (0.93). It can be concluded that 1H-NMR-based metabolite fingerprinting coupled with PCA and OPLS-DA offers an adequate method to assess adulteration practice and to evaluate the authentication of C. xanthorrhiza extracts.

Terpenoid-lupeol of red dragon fruit (Hylocereus polyrhizus) and its immunomodulatory activity.

Red dragon fruit (Hylocereus polyrhizus, (F.A.C. Weber) Britton and Rose) has been reported to have various biological activities such as antimicrobial, anti-hypercholesterolemia, anti-diabetes mellitus, cardiovascular risk reduction, health supplement, and melanoma cell inhibitory. The red thick peel of this fruit is just practically a waste that is possibly utilized to maintain health, therefore this research aimed to isolate and identify active compounds of H. Polyrhizus peels which can improve the immune system of body. In order to simplify methanol extract was partition and fractionation. The active compounds of petroleum ether fraction were separated and purified using preparative thin layer chromatography. The identification of the compounds structure was conducted through spectroscopic techniques, including UV, FT-IR, 13CNMR and 1HNMR spectroscopy. The data of spectra revealed that the isolate is lupeol. The statistical analysis of macrophage activity showed that the isolate with concentrations of 100, 50, 25, 12.5 and 6.25µg/mL could activate the macrophages higher than control negative. Terpenoid generated from the isolation of Hylocereus polyrhizus was identified as lupeol (1-isopropenyl-3a,5a,5b,8,8,11a-hexamethyl-eicosahydrocyclopenya [α] chrysen-9ol. In vitro test shows that the isolated compound had an immunomodulatory activity by increases macrophage phagocytosis of latex beads.

Esterification of citric acid with locust bean gum.

Esterification of citric acid (CA) with locust bean gum (LBG) was prepared by hydrochloric acid (HCl) as a catalyst and UV irradiation (254 nm) as esterification energy. This study aims to determine the best conditions of esterification. Other than that, it is to know the effect of amount HCl and UV irradiation time for the esterification process of CA with LBG. The amounts of HCl are 0.18 and 0.30 M, while the variations of UV irradiation time are 75 and 100 minutes. Polyester (CA-LBG) were characterized by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), X-ray diffractometer (XRD), esterification degree, and viscosity. Parameters for determining the best conditions for esterification are esterification degree and viscosity. The best conditions of esterification were obtained by using 0.30 M mL HCl and 100 minutes of UV irradiation time resulted in CA-LBG having a value of esterification degree 9.69 % and viscosity 7.46 cPs. HCl accelerates protonation on the O atoms and the formation of positive C atoms of carbonyl groups of citric acid. The time of UV irradiation gives the longer energy for the bond formation between the positive C atoms of the carbonyl group and the O atoms of the hydroxyl group at C-6 atoms of mannose and galactose.

Origins and spread of novel genetic variants of sulfadoxine-pyrimethamine resistance in Plasmodium falciparum isolates in Indonesia.

BACKGROUND: While malaria incidence in Indonesia has decreased threefold in the last decade, more than 200,000 cases were reported in 2016. Different endemicity of Plasmodium falciparum malaria among several islands in Indonesia has been recognized and two unique mutations of P. falciparum dihydropteroate synthase (pfdhps) affecting sulfadoxine-pyrimethamine (SP) resistance were detected from the research of SP efficiency and genotype analysis in South Kalimantan. In this study, geographical distribution and origin of these pfdhps K540T and I588F mutations were analysed. METHODS: Malaria parasites DNA from several endemic areas in Indonesia; Sumatera, Java, Kalimantan, Lombok, Sumbawa, Timor, Sulawesi, and Papua islands; in two periods, 2004-2006 and 2009-2012 were subjected for pfdhfr and pfdhps sequence analysis. RESULTS: Different genotype polymorphisms of pfdhfr and pfdhps were observed in the parasites from various regions in Indonesia and relatively more divergent genotypes were determined from Kalimantan isolates in both 2004-2006 and 2009-2012. The parasites containing K540T mutation were identified in 2004-2006 isolates from East Kalimantan, East Java and Sumbawa as an SGTGA haplotype. The other I588F mutation was also determined in 2004-2006 parasites, isolated from Lombok and Sumbawa islands as an SGEAA(588F) haplotype. The parasites with pfdhfr/pfdhps quintuple or sextuple mutation, a genotype marker of SP resistance, were determined mostly in Kalimantan in both 2004-2006 and 2009-2012. CONCLUSION: Analysis of the prevalence and pfdhfr/pfdhps combined genotypes of K540T or I588F mutations suggested that K540T might be origin in Kalimantan Island and I588F in Sumbawa Island and then these were spread to other areas along with people movement. This research indicates regular monitoring of drug efficacy and parasite genotype analysis is important to keep efficiency and prevent the spread of resistance. It is also essential for the latest anti-malarial drug artemisinin-based combination therapy.

Simultaneous Determination of Metamizole, Thiamin and Pyridoxin Using UV-Spectroscopy in Combination with Multivariate Calibration.

PURPOSE: Analysis of drugs in multicomponent system officially is carried out using chromatographic technique, however, this technique is too laborious and involving sophisticated instrument. Therefore, UV-VIS spectrophotometry coupled with multivariate calibration of partial least square (PLS) for quantitative analysis of metamizole, thiamin and pyridoxin is developed in the presence of cyanocobalamine without any separation step. METHODS: The calibration and validation samples are prepared. The calibration model is prepared by developing a series of sample mixture consisting these drugs in certain proportion. Cross validation of calibration sample using leave one out technique is used to identify the smaller set of components that provide the greatest predictive ability. The evaluation of calibration model was based on the coefficient of determination (R(2)) and root mean square error of calibration (RMSEC). RESULTS: The results showed that the coefficient of determination (R(2)) for the relationship between actual values and predicted values for all studied drugs was higher than 0.99 indicating good accuracy. The RMSEC values obtained were relatively low, indicating good precision. The accuracy and presision results of developed method showed no significant difference compared to those obtained by official method of HPLC. CONCLUSION: The developed method (UV-VIS spectrophotometry in combination with PLS) was succesfully used for analysis of metamizole, thiamin and pyridoxin in tablet dosage form.

Two novel mutations of pfdhps K540T and I588F, affecting sulphadoxine-pyrimethamine-resistant response in uncomplicated falciparum malaria at Banjar district, South Kalimantan Province, Indonesia.

BACKGROUND: Mutations in pfdhfr and pfdhps genes have been shown to associate with sulphadoxine-pyrimethamine (SP) resistance of Plasmodium falciparum parasites. However, pfdhfr, pfdhps genotypes and the correlations to SP treatment outcome in Indonesia has not yet been well analysed. METHODS: After obtaining informed consent, 61 uncomplicated falciparum malaria patients were recruited in Banjar district, South Kalimantan Province, Indonesia, from October 2009 to August 2010. They were treated by a single oral dose of SP and its effects on clinical and parasitological status were followed until day 28 after treatment. Occasionally, a thick smear blood film for microscopy observation and blood spot on a filter paper for pfdhfr and pfdhps genotype analysis were collected. RESULTS: Pfdhfr and pfdhps genotypes from 24 P. falciparum-infected patients consisting of adequate clinical parasitological response (ACPR) (n = 6; 25.0%) and early treatment failure (ETF) (n = 10; 41.7%) or late parasitological failure (LPF) (n = 8; 33.3%) were obtained by sequencing. Two novel mutations of pfdhps gene, K540T and I588F, were determined in ten and five isolates, respectively. These mutations were present in the pfdhfr/pfdhps combined haplotypes of ANRNI/SGTGA (n = 6), ANRNL/SGTGA (n = 4), and ANRNI/SGEAA(588F) (n = 5), (mutation codons are bold typed); these haplotypes were mostly belonging to parasitological failure (ETF or LPF). The parasites acquiring five mutations in pfdhfr/pfdhps haplotypes and four mutations with additional I588F did not respond adequately to SP treatment. CONCLUSION: Many of Plasmodium falciparum infected patients in Banjar district, South Kalimantan, Indonesia did not respond adequately to SP treatment and these low ineffectiveness of SP in this area was associated with two novel mutations of pfdhps, K540T and I588F.

Effects of marmin, a compound isolated from Aegle marmelos Correa, on contraction of the guinea pig-isolated trachea.

Marmin or 7-(6',7'-dihydroxygeranyl-oxy)coumarin is a compound isolated from Aegle marmelos Correa. In the study, we examined the effects of marmin on the contraction of guinea pig-isolated trachea stimulated by several inducers, namely histamine, metacholine, compound 48/80. We also evaluated its action against contraction induced by extracellular or intracellular calcium ion. The possibility of marmin to potentiate the relaxation effect of isoprenaline was also studied. Marmin added in the organ bath at 10 min prior to the agonist inhibited the contraction elicited by histamine and metacholine in a concentration-dependent manner. Moreover, marmin antagonized the histamine-induced contraction in competitive manner. Marmin mildly potentiated the relaxation effect of isoprenaline. In the study, marmin abrogated the contraction of tracheal smooth muscle induced by compound 48/80, an inducer of histamine release. Besides, marmin successfully inhibited CaCl(2)-induced contraction in Ca(2+)-free Krebs solution. Marmin also inhibited two phases of contraction which were consecutively induced by metacholine and CaCl(2) in Ca(2+)-free Krebs solution. Based on the results we concluded that marmin could inhibit contraction of the guinea-pig tracheal smooth muscle, especially by interfering histamine receptor, inhibiting the histamine release from mast, inhibiting intracellular Ca(2+) release from the intracellular store and the Ca(2+) influx through voltage-dependent Ca(2+) channels.

Authentication analysis of red fruit (Pandanus Conoideus Lam) oil using FTIR spectroscopy in combination with chemometrics.

INTRODUCTION: Red fruit (Pandanus conoideus Lam) is endemic plant of Papua, Indonesia and Papua New Guinea. The price of its oil (red fruit oil, RFO) is 10-15 times higher than that of common vegetable oils; consequently, RFO is subjected to adulteration with lower price oils. Among common vegetable oils, canola oil (CaO) and rice bran oil (RBO) have similar fatty acid profiles to RFO as indicated by the score plot of principal component analysis; therefore, CaO and RBO are potential adulterants in RFO. OBJECTIVE: To develop FTIR spectroscopy in combination with chemometrics of partial least square regression (PLSR) and discriminant analysis (DA) for authentication of RFO from CaO and RBO. RESULTS: The presence of CaO in RFO was better determined at frequency regions of 1200-1050 cm⁻¹; meanwhile, the combined frequency ranges of 1207-1078 and 1747-1600 cm⁻¹ were exploited for quantitative analysis of RBO with acceptable values of coefficient of determination (R²) and errors in calibration, prediction and during cross validation. DA based on Mahalanobis distance was able to discriminate between RFO and RFO adulterated with CaO and RBO. CONCLUSION: FTIR spectroscopy combined with PLSR and DA can be successfully used for quantification and classification of oil adulterants in RFO. The developed method is rapid and environmentally friendly and sample preparation is easy.

Effects of aegeline, a main alkaloid of Aegle Marmelos Correa leaves, on the histamine release from mast cells.

Aegeline or N-[2-hydroxy-2(4-methoxyphenyl) ethyl]-3-phenyl-2-propenamide is a main alkaloid isolated from Aegle marmelos Correa collected in Yogyakarta Indonesia. In our study, we investigated the effects of aegeline on the histamine release from mast cell. The study was performed by using (1) rat basophilic leukemia (RBL-2H3) cell line, and (2) rat peritoneal mast cells (RPMCs). DNP(24)-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine release from mast cell. In our study, aegeline inhibited the histamine release from RBL-2H3 cells induced by DNP(24)-BSA. Indeed, aegeline showed strong inhibition when RBL-2H3 cells induced by Ca(2+) stimulants such as thapsigargin and ionomycin. Aegeline is suggested to influence the intracellular Ca(2+) pool only since could not inhibit the (45)Ca(2+) influx into RBL-2H3 cells. Aegeline showed weak inhibitory effects on the histamine release from RPMCs, even though still succeed to inhibit when the histamine release induced by thapsigargin. These findings indicate that aegeline altered the signaling pathway related to the intracellular Ca(2+) pool in which thapsigargin acts. Based on the results, the inhibitory effects of aegeline on the histamine release from mast cells depended on the type of mast cell and also involved some mechanisms related to intracellular Ca(2+) signaling events via the same target of the action of thapsigargin or downstream process of intracellular Ca(2+) signaling in mast cells.