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Formulation of maize- and peanut-based semi-synthetic growth media for the ecophysiological studies of aflatoxigenic Aspergillus flavus in maize and peanut agro-ecosystems

Yazid SNE, Thanggavelu H, Mahror N, Selamat J, Samsudin NIP

Int J Food Microbiol, 2018 Oct 03;282:57-65.
PMID: 29913332 DOI: 10.1016/j.ijfoodmicro.2018.06.007

In studying the ecophysiology of fungal phytopathogens, several stages are involved (in vitro, greenhouse, in planta). Most in vitro studies extensively utilise the general growth media such as Potato Dextrose Agar and Malt Extract Agar. Although the crop components in these media serve as excellent carbon sources and yield luxuriant growth, they are not naturally contaminated with Aspergillus flavus and thus might result in under- or overestimation of its actual toxigenic potentials. Empirical data on the formulation of semi-synthetic growth medium mimicking the natural crop commonly contaminated by A. flavus for the ecophysiological studies in vitro are scarce. The present work was aimed at investigating the ecophysiology of A. flavus on commercial growth media (PDA, MEA); formulating maize- and peanut-based semi-synthetic growth media using two methods of raw material preparation (milling, hot water extraction) at different concentrations (1, 3, 5, 7, 9% w/v), and comparing the ecophysiological parameters between commercial and formulated growth media. Growth rates were obtained by computing the hyphal expansion data into y = mx + c equation. AFB1 was quantified using high performance liquid chromatography with fluorescence detector. Formulated media were found to yield significantly higher growth rates when compared to commercial media. However, commercial media yielded significantly higher AFB1 when compared to all formulated media. Between the two formulations, milling yielded significantly higher growth rates and AFB1 when compared to hot water extraction. Although in vitro data cannot directly extrapolate in planta performance, results obtained in the present work can be used to gauge the actual toxigenic potential of A. flavus in maize and peanut agro-ecosystems.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Enzymatic synthesis of polyphenol glycosides catalyzed by transglycosylation reaction of cyclodextrin glucanotransferase derived from Trichoderma viride

Nazir S, Sulistyo J, Hashmi MI, Ho AL, Khan MS

J Food Sci Technol, 2018 Aug;55(8):3026-3034.
PMID: 30065412 DOI: 10.1007/s13197-018-3223-x

Present study was conducted to evaluate the ability of Trichoderma viride as a source of cyclodextrin glucanotransferase that has shown transglycosylation activity in the presence of polyphenolic constituents extracted from Moringa oleifera leaves as its acceptor and wheat flour as its substrate to catalyze synthesis of polyphenolic glycosides as transglycosylation (transfer) reaction products. The enzymatic synthesized polyphenolic glycosides were then purified using octa-dodecyl-functionalized silica gel column chromatography prior to analysis using thin layer chromatography and high performance liquid chromatography and identified using nuclear magnetic resonance (NMR) spectroscopy. The high performance liquid chromatogram performed that the isolated transglycosylation products had retention times and concentration at 1.446 min (0.0017 mg/ml), 1.431 min (0.14 mg/ml), and 1.474 min (0.012 mg/ml), respectively, compared to the retention time of arbutin (1.474 min) that was applied as authentic standard for polyphenol glycoside. Moreover, observation using 1H NMR as well as 13C NMR showed that structures of the transglycosylation products were identified as gallic acid-4-O-β-glucopyranoside, ellagicacid-4-O-β-glucopyranoside, and catechin-4'-O-glucopyranoside, respectively.

Matched MeSH terms: Chromatography, High Pressure Liquid
Design and development of dry powder sulfobutylether-β-cyclodextrin complex for pulmonary delivery of fisetin

Mohtar N, Taylor KM, Sheikh K, Somavarapu S

Eur J Pharm Biopharm, 2017 Apr;113:1-10.
PMID: 27916704 DOI: 10.1016/j.ejpb.2016.11.036

This study has investigated complexation of fisetin, a natural flavonoid, with three types of cyclodextrins to improve its solubility. Sulfobutylether-β-cyclodextrin (SBE-β-CD) showed the highest complexation efficiency while maintaining the in vitro antioxidant activity of fisetin. Addition of 20%v/v ethanol in water improved the amount of solubilized fisetin in the complex 5.9-fold compared to the system containing water alone. Spray drying of fisetin-SBE-β-CD complex solution in the presence of ethanol produced a dry powder with improved aerosolization properties when delivered from a dry powder inhaler, indicated by a 2-fold increase in the fine particle fraction (FPF) compared to the powder produced from the complex solution containing water alone. The pitted morphological surface of these particles suggested a more hollow internal structure, indicating a lighter and less dense powder. Incorporation of 20%w/w leucine improved the particle size distribution of the powder and further increased the FPF by 2.3-fold. This formulation also showed an EC50 value equivalent to fisetin alone in the A549 cell line. In conclusion, an inhalable dry powder containing fisetin-SBE-β-CD complex was successfully engineered with an improved aqueous solubility of fisetin. The dry powder may be useful to deliver high amounts of fisetin to the deep lung region for therapeutic purposes.

Matched MeSH terms: Chromatography, High Pressure Liquid
Novel metabolites from Trichoderma atroviride against human prostate cancer cells and their inhibitory effect on Helicobacter pylori and Shigella toxin producing Escherichia coli

Saravanakumar K, Mandava S, Chellia R, Jeevithan E, Babu Yelamanchi RS, Mandava D, et al.

Microb Pathog, 2018 Oct 10;126:19-26.
PMID: 30316006 DOI: 10.1016/j.micpath.2018.10.011

The present study aimed to purify and identify the metabolites from T. atroviride using high-performance liquid chromatography (HPLC) and 1H and 13C nuclear magnetic resonance spectrometer (NMR) followed by analyzing their toxicological, antibacterial and anticancer properties. This work identified two metabolites - TM1 and TM2. TM1 was in two forms: (i) 1, 3-dione-5, 5-dimethylcyclohexane; and, (ii) 2-enone-3hydroxy -5,5-dimethylcylohex, while TM2 was 4H-1,3-dioxin-4-one-2,3,6-trimethyl. These metabolites did not exhibit any irritant or allergic reaction as revealed by HET- CAM test. TM2 significantly inhibited the growth of H. pylori and Shigella toxin producing Escherichia coli (STEC) as evident by in vitro and microscopic observations of bacterial cell death. TM2 also induced the cell death and cytotoxicity, as revealed by cell viability test and western blot analysis. According to microscopic, flow cytometer and western blot analysis, TM2 treated cells displayed higher ROS, cell death, and apoptosis-related protein expression than TM1 and control. This study concluded that TM2 derived from T. atroviride was a potential therapeutic agent for anti-prostate cancer and antibiotic agent against MDR- H. pylori and STEC and it is also recommended to carry out further in vivo animal model experiments with improved stability of the metabolites for future pharmaceutical trails.

Matched MeSH terms: Chromatography, High Pressure Liquid
Selective magnetic nanographene oxide solid-phase extraction with high-performance liquid chromatography and fluorescence detection for the determination of zearalenone in corn samples

Thongprapai P, Cheewasedtham W, Chong KF, Rujiralai T

J Sep Sci, 2018 Dec;41(23):4348-4354.
PMID: 30267469 DOI: 10.1002/jssc.201800441

A magnetic nanographene oxide sorbent as a selective sorbent for the magnetic solid-phase extraction combined with high-performance liquid chromatography and fluorescence detection was developed and proved to be a robust method for zearalenone determination in corn samples. Optimum extraction of zearalenone (20 mg magnetic nanographene oxide sorbent, extraction for 15 min, desorption time of 15 min using 1 mL of 0.5% formic acid in methanol) resulted in low limits of detection (05 mg/L) and quantitation (0.13 mg/L) and good linearity range of 0.13-1.25 mg/L with the correlation coefficient of 0.9957. Acceptable recoveries (79.3-80.6%) with relative standard deviations below 4% and satisfactory intra- and interday precisions (2-7.4%) were achieved. Additionally, the proposed method has been proved to be good in several aspects: easily prepared sorbent with high affinity to zearalenone, convenient and fast procedure, and high extraction efficiency.

Matched MeSH terms: Chromatography, High Pressure Liquid
γ-Aminobutyric acid production by selected lactic acid bacteria isolate of an Indonesian indigenous fermented buffalo milk (dadih) origin

Harnentis H, Nurmiati N, Marlida Y, Adzitey F, Huda N

Vet World, 2019 Aug;12(8):1352-1357.
PMID: 31641319 DOI: 10.14202/vetworld.2019.1352-1357

Aim: This study aimed at optimizing γ-aminobutyric acid (GABA) production using lactic acid bacteria (LAB) of an Indonesian indigenous fermented buffalo milk (dadih) origin. This study utilized LAB previously cultured from dadih that has the ability to produce GABA.
Materials and Methods: The study started with the identification of selected LAB by 16S rRNA, followed by optimization of GABA production by culture conditions using different initial pH, temperature, glutamate concentration, incubation time, carbon, and nitrogen sources. 16S rRNA polymerase chain reaction and analysis by phylogenetic were used to identify Lactobacillus plantarum (coded as N5) responsible for the production of GABA.
Results: GABA production by high-performance liquid chromatography was highest at pH of 5.5, temperature of 36°C, glutamate concentration of 500 mM, and incubation time of 84 h. Peptone and glucose served as the nitrogen and carbon sources, respectively, whereas GABA was produced at optimum fermentation condition of 211.169 mM.
Conclusion: Production of GABA by L. plantarum N5 was influenced by initial pH of 5.5, glutamic acid concentration, nitrogen source, glucose as carbon source, and incubation temperature and time.

Matched MeSH terms: Chromatography, High Pressure Liquid
Electrochemically Modulated Liquid-Liquid Extraction for Sample Enrichment

Jajuli MN, Hussin MH, Saad B, Rahim AA, Hébrant M, Herzog G

Anal Chem, 2019 06 04;91(11):7466-7473.
PMID: 31050400 DOI: 10.1021/acs.analchem.9b01674

A new sample preparation method is proposed for the extraction of pharmaceutical compounds (Metformin, Phenyl biguanide, and Phenformin) of varied hydrophilicity, dissolved in an aqueous sample. When in contact with an organic phase, an interfacial potential is imposed by the presence of an ion, tetramethylammonium (TMA+), common to each phase. The interfacial potential difference drives the transfer of ionic analytes across the interface and allows it to reach up to nearly 100% extraction efficiency and a 60-fold enrichment factor in optimized extraction conditions as determined by HPLC analysis.

Matched MeSH terms: Chromatography, High Pressure Liquid
Tissue-specific and maturity-dependent distribution of pyridine alkaloids in Areca triandra

Wu J, Zhang H, Wang S, Yuan L, Grünhofer P, Schreiber L, et al.

J Plant Res, 2019 Jul;132(4):531-540.
PMID: 31127431 DOI: 10.1007/s10265-019-01115-9

Areca nuts (seeds of Areca catechu L.) are a traditional and popular masticatory in India, Bangladesh, Malaysia, certain parts of China, and some other countries. Four related pyridine alkaloids (arecoline, arecaidine, guvacoline, and guvacine) are considered being the main functional ingredients in areca nut. Until now, A. catechu is the only known species producing these alkaloids in the Arecaceae family. In the present study, we investigated alkaloid contents in 12 Arecaceae species and found that only Areca triandra Roxb. contained these pyridine alkaloids. We further analyzed in more detail tissue-specific and development-related distribution of these alkaloids in leaves, male and female flowers and fruits in different stages of maturity in A. triandra by ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry. Results revealed that the alkaloids were most abundant in young leaves, the pericarp of ripe fruits and the endosperm of unripe fruits in developmental stage 2. Abundance of the 4 different alkaloids in A. triandra fruits varied during maturation. Pericarps of ripe fruits had the highest arecaidine concentration (4.45 mg g-1) and the lowest guvacoline concentration (0.0175 mg g-1), whereas the endosperm of unripe fruits of developmental stage 2 contained the highest guvacoline concentration (3.39 mg g-1) and the lowest guvacine concentration (0.245 mg g-1). We conclude that A. triandra is useful in future as a further valuable source of Areca alkaloids.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Preparation and Evaluation Pumpkin Seed Oil-based Vitamin E Cream Formulations for Topical Application

Ong TS, Chu CC, Tan CP, Nyam KL

J Oleo Sci, 2020;69(4):297-306.
PMID: 32249259 DOI: 10.5650/jos.ess19250

Plant seed oil is often incorporated into the cream emulsions to provide multifunctional effects on the skin. In the current study, pumpkin seed oil (PSO) was used to develop a stable oil-in-water emulsion. The study aimed to optimise PSO cream formulation and determine the synergistic effect of the PSO with vitamin E oil added. The physical properties, antioxidant activities and storage stability of the formulations were analysed. Besides, the synergistic effect of the best formulation was analysed based on α-tocopherol content using ultra-high performance liquid chromatography (UHPLC). The storage stability test was assessed upon storing at 25 ± 2°C and 40 ± 2°C for 12 weeks. The best formulation (20% PSO, vitamin E oil and beeswax) selected showed physically and microbiologically stable. The incorporation of vitamin E oil into the formulation produced with PSO was found to be compatible, as it showed a synergistic effect in the amount of α-tocopherol content (combination index (CI) = 0.98). Thus, PSO had shown its potency to be incorporated into the topical products with a promising potential in delivering additional properties that can nourish the skin.

Matched MeSH terms: Chromatography, High Pressure Liquid
Initial screening of mangrove endophytic fungi for antimicrobial compounds and heavy metal biosorption potential

Ling Onn M, Teen Lim P, Aazani Mujahid, Proksch P, Müller M

Sains Malaysiana, 2016;45:1063-1071.

Endophytic fungi provide protection to their host plant and the fungi often produce antimicrobial compounds to aid the host
fighting off pathogens. These bioactive compounds were secondary metabolites which were often produced as waste- or
by-products. In the present study, endophytic fungi isolated from mangrove plants and soils were characterized and their
antimicrobial production and bioremediation potential of heavy metals copper (Cu) and zinc (Zn) were assessed. Twelve
(12) isolated and identified endophytic fungi belonged to seven species; Penicillium, Curvularia, Diaporthe, Aspergillus,
Guignardia, Neusartorya and Eupenicillium. Antimicrobial activities of these 12 fungal endophytes were tested against
Gram negative bacteria; Bacillus subtilis, Staphylococcus aureus, Gram positive bacteria; Escherichia coli and fungi;
Candida albicans and Aspergillus niger among others. Two isolates (related to Guignardia sp. and Neusartoya sp.) showed
strong antimicrobial (and antifungal) activity whereas the rest showed no activity. Compounds were isolated from both
isolates and screened using HPLC. Both isolates displayed chemically very interesting chromatograms as they possessed a
high diversity of basic chemical structures and peaks over a wide range of polarities, with structures similar to Trimeric
catechin and Helenalin among others. For bioremediation assessment, the results showed maximum biosorption capacity
for two isolates related to Curvularia sp. and Neusartorya sp., with the former removing 25 mg Cu/g biomass and the
latter removing 24 mg Zn/g biomass. Our results indicated the potential of mangrove endophytic fungi in producing
bioactive compounds and also highlighted their potential for the treatment of heavy metal-contaminated wastewater.

Matched MeSH terms: Chromatography, High Pressure Liquid
A validated size exclusion chromatography method coupled with fluorescence detection for rapid quantification of bevacizumab in ophthalmic formulations

Jirjees F, Soliman K, Wang Y, Sonawane R, Sheshala R, Jones D, et al.

J Pharm Biomed Anal, 2019 Sep 10;174:145-150.
PMID: 31167158 DOI: 10.1016/j.jpba.2019.05.038

Bevacizumab is a full-length human monoclonal antibody used to treat various neovascular diseases such as wet age-related macular degeneration (AMD), diabetic eye disease and other problems of the retina. Monthly intravitreal injections of bevacizumab (Avastin®) are effective in the treatment of wet AMD. However, there is a growing demand in the development of sustained release ophthalmic formulations. Therefore, this study aims, for the first time, to develop a rapid, simple, and sensitive method using size exclusion chromatography coupled with fluorescence detection for routine quantification of bevacizumab in ophthalmic formulations and during in vitro release studies. The selected chromatographic conditions included an aqueous mobile phase composed of 35 mM sodium phosphate buffer and 300 mM sodium chloride (pH 6.8), a flow rate of 0.5 mL/min, and the fluorescence detector was operated at excitation and emission wavelengths of 280 and 340 nm, respectively. The peak area-concentration relationship maintained its linearity over concentration range of 0.1-20 μg/mL (R2 = 0.9993), and the quantitation limit was 100 ng/mL. The method was validated for specificity, accuracy, precision, and robustness. The developed method had a run time of 6 min at temperature 25 °C, making it a unique validated method for rapid and cost-effective quantification of bevacizumab.

Matched MeSH terms: Chromatography, High Pressure Liquid
Chitosan-Coated Alginate Nanoparticles Enhanced Absorption Profile of Insulin Via Oral Administration

Jaafar MHM, Hamid KA

Curr Drug Deliv, 2019;16(7):672-686.
PMID: 31250754 DOI: 10.2174/1567201816666190620110748

BACKGROUND: In this study, four nanoparticle formulations (F1 to F4) comprising varying ratios of alginate, Pluronic F-68 and calcium chloride with a constant amount of insulin and chitosan as a coating material were prepared using polyelectrolyte complexation and ionotropic gelation methods to protect insulin against enzymatic degradation.
METHODS: This study describes the formulation design, optimisation, characterisation and evaluation of insulin concentration via oral delivery in rats. A reversed-phase high-performance liquid chromatography (HPLC) method was developed and validated to quantify insulin concentration in rat plasma. The proposed method produced a linear response over the concentration range of 0.39 to 50 µg/ml.
RESULTS: In vitro release study showed that dissolution of insulin in simulated gastric juice of pH 1.2 was prevented by alginate core and chitosan coating but rapidly released in simulated intestinal fluid (pH 6.8). Additionally, Formulation 3 (F3) has a particle size of 340.40 ± 2.39 nm with narrow uniformity exhibiting encapsulation efficiency (EE) of 72.78 ± 1.25 % produced highest absorption profile of insulin with a bioavailability of 40.23 ±1.29% and reduced blood glucose after its oral administration in rats.
CONCLUSION: In conclusion, insulin oral delivery system containing alginate and chitosan as a coating material has the ability to protect the insulin from enzymatic degradation thus enhance its absorption in the intestine. However, more work should be done for instance to involve human study to materialise this delivery system for human use.

Matched MeSH terms: Chromatography, High Pressure Liquid
Characterization of Polar and Non-Polar Compounds of House Edible Bird's Nest (EBN) from Johor, Malaysia

Hun Lee T, Hau Lee C, Alia Azmi N, Kavita S, Wong S, Znati M, et al.

Chem Biodivers, 2020 Jan;17(1):e1900419.
PMID: 31721431 DOI: 10.1002/cbdv.201900419

This work investigated the polar (PC: protein, amino acid and metabolite) and non-polar (NPC: fatty acid) compounds and bioactivity characteristics of the EBN harvested from the state of Johor in Malaysia. The electrophoretic gels exhibited 15 protein bands (16-173 kD) with unique protein profile. Amino acids analysis by AccQ⋅Tag method revealed 18 types of amino acids in EBN. Metabolite profiling was performed using High-Performance Liquid Chromatography coupled with Quadrupole Time-of-Flight Mass Spectrometer (HPLC-QTOF/MS) technique and a total of 54 compounds belonging to different groups were detected and identified. These findings help to uncover the relation of therapeutic activity of EBN. The EBN was further extracted with AcOEt and BuOH. The AcOEt extract was fractionated into three fractions (F1 -F3 ), and the high triglyceride content in F2 was verified by gC-FID. The three groups of fatty acids discovered in EBN are 48.43 % of poly-unsaturated (PUFA), 25.35 % of saturated fatty acids (SFA) and 24.74 % of mono-unsaturated fat (MUFA). This is the first time to report results ofEBN, BuOH, and AcOEt extracts and of fraction F2 (TEBN) on their analysis for their antioxidant activities by DPPH, ABTS and catalase assay and for their paraoxonase and anti-tyrosinase activities. The results showed that TEBN exhibited the significant bioactivity in all assays. These findings suggest that TEBN is a good source for natural bioactive compounds in promoting body vigor. Current work widened the content of EBN especially on the triglyceride and also marked the content of specific location (Johor, Malaysia) of EBN origin.

Matched MeSH terms: Chromatography, High Pressure Liquid
Bioavailability and pharmacokinetic studies of eurycomanone from Eurycoma longifolia

Low BS, Ng BH, Choy WP, Yuen KH, Chan KL

Planta Med, 2005 Sep;71(9):803-7.
PMID: 16206032

A validated HPLC analysis of eurycomanone (1), a bioactive quassinoid, in rat plasma following oral and intravenous administration of Eurycoma longifolia Jack extract was developed for pharmacokinetic and bioavailability studies. Relatively high plasma eurycomanone concentrations were detected after an intravenous injection of 10 mg/kg extract F2 containing 1.96 mg/kg of the quassinoid. However, it declined rapidly to zero after 8 h. Its mean elimination rate constant (k(e)), biological half-life (t(1/2)), volume of distribution (V(d)) and clearance (CL) were 0.88 +/- 0.19 h (-1), 1.00 +/- 0.26 h, 0.68 +/- 0.30 L/kg and 0.39 +/- 0.08 L/h/kg, respectively. Following oral administration of eurycomanone, its Cmax and Tmax values were detected as 0.33 +/- 0.03 microg/mL and 4.40 +/- 0.98 h, respectively. The plasma concentration of the quassinoid after oral administration was much lower than after intravenous application in spite of the oral dose being 5 times higher. The results indicate that eurycomanone is poorly bioavailable when given orally. A comparison of the AUC (0-->infinity) obtained orally to that obtained after an intravenous administration (normalized for dose differences) revealed that the absolute bioavailability of the compound was low with 10.5 %. Furthermore, the compound appeared to be well distributed in the extravascular fluids because of its relatively high V(d) value. The poor oral bioavailability was not attributed to instability problems because eurycomanone has been shown to be stable under different pH conditions. Thus, its poor oral bioavailability may be due to poor membrane permeability in view of its low P value and/or high first-pass metabolism.

Matched MeSH terms: Chromatography, High Pressure Liquid
Identification of angiotensin-converting enzyme inhibitory proteins from mycelium of Pleurotus pulmonarius (oyster mushroom)

Ibadallah BX, Abdullah N, Shuib AS

Planta Med, 2015 Jan;81(2):123-9.
PMID: 25590365 DOI: 10.1055/s-0034-1383409

Pleurotus pulmonarius (grey oyster mushroom) has been acknowledged as a recuperative agent for many diseases in addition to its recognition as a nutritious provision. We performed a study on P. pulmonarius mycelium for an antihypertensive effect via the angiotensin-converting enzyme inhibitory activity. The preliminary assay on the mycelial water extract demonstrated that the angiotensin-converting enzyme inhibitory activity had an IC50 value of 720 µg/mL. Further protein purifications via ammonium sulphate precipitation and RP-HPLC resulted in 60× stronger angiotensin-converting enzyme inhibitory activity than that of the mycelial water extract (IC50 = 12 µg/mL). Protein identification and characterisation by MALDI-TOF/TOF, later corroborated by LC-MS/MS, indicated three proteins that are responsible for the blood pressure lowering effects via different mechanisms: serine proteinase inhibitor-like protein, nitrite reductase-like protein, and DEAD/DEAH box RNA helicase-like protein.

Matched MeSH terms: Chromatography, High Pressure Liquid
Antioxidant potential assessment of phenolic and flavonoid rich fractions of Clematis orientalis and Clematis ispahanica (Ranunculaceae)

Karimi E, Ghorbani Nohooji M, Habibi M, Ebrahimi M, Mehrafarin A, Khalighi-Sigaroodi F

Nat Prod Res, 2018 Aug;32(16):1991-1995.
PMID: 28774179 DOI: 10.1080/14786419.2017.1359171

The antioxidant activities of crude extract fractions using Hexane, Chloroform, Ethyl Acetate, Butanol and Water of Clematis orientalis and Clematis ispahanica were investigated. 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay and the ferric reducing/antioxidant potential (FRAP) were used to evaluate the antioxidant capacity. The total phenolics were found to be 4.37-9.38 and 1.32-11.37 mg gallic acid equivalents (GAE)/g in different fractions for C. orientalis and C. ispahanica, respectively. The ethyl acetate fraction of C. orientalis and chloroform fraction of C. ispahanica showed the highest DPPH and FRAP activities at a concentration of 300 μg/mL. The predominant phenolic compounds identified by HPLC in C. orientalis were Resorcinol (603.5 μg/g DW) in chloroform fraction and Ellagic acid (811.7 μg/g DW) in chloroform fraction of C. ispahanica.

Matched MeSH terms: Chromatography, High Pressure Liquid
Level of natural hepatotoxin (Indospicine) contamination in Australian camel meat

Tan ET, Al Jassim R, D'Arcy BR, Fletcher MT

Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2016 Oct;33(10):1587-1595.
PMID: 27575484

Camel meat production for human consumption and pet food manufacture accounts for a relatively small part of overall red meat production in Australia. Reliable statistical data for the Australian production and consumption of camel meat are not available; however, it is estimated that 300,000 feral camels roam within the desert of central Australia, with an annual usage of more than 3000 camels for human consumption, 2000 for pet food manufacture and a smaller number for live export. Despite a small Australian camel meat production level, the usage of camel meat for pet food has been restricted in recent years due to reports of serious liver disease and death in dogs consuming camel meat. This camel meat was found to contain residues of indospicine, a non-proteinogenic amino acid found in certain Indigofera spp., and associated with mild to severe liver disease in diverse animals after dietary exposure to this hepatotoxin. The extent of indospicine-contaminated Australian camel meat was previously unknown, and this study ascertains the prevalence of such residue in Australian camel meat. In this study, indospicine levels in ex situ (95 samples collected from an abattoir in Queensland) and in situ (197 samples collected from camels after field culling in central Australia) camel meat samples were quantitated using a validated ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The quantitation results showed 46.7% of the in situ- and 20.0% of the ex situ-collected camel meat samples were contaminated by indospicine (more than the limit of detection (LOD) of 0.05 mg kg(-1) fresh weight). The overall indospicine concentration was higher (p < 0.05) in the in situ-collected samples. Indospicine levels detected in the present study are considered to be low; however, a degree of caution must still be exercised, since the tolerable daily intake for indospicine is currently not available for risk estimation.

Matched MeSH terms: Chromatography, High Pressure Liquid
Guinea pig genital tract lipidome reveals in vivo and in vitro regulation of phosphatidylcholine 16:0/18:1 and contribution to Chlamydia trachomatis serovar D infectivity

Wali S, Gupta R, Yu JJ, Mfuh A, Gao X, Guentzel MN, et al.

Metabolomics, 2016 Apr;12(4).
PMID: 27642272

INTRODUCTION: Chlamydia trachomatis (Ct), is the leading cause of sexually transmitted infections worldwide. Host transcriptomic- or proteomic profiling studies have identified key molecules involved in establishment of Ct infection or the generation of anti Ct-immunity. However, the contribution of the host metabolome is not known.
OBJECTIVES: The objective of this study was to determine the contribution of host metabolites in genital Ct infection.
METHODS: We used high-performance liquid chromatography-mass spectrometry, and mapped lipid profiles in genital swabs obtained from female guinea pigs at days 3, 9, 15, 30 and 65 post Ct serovar D intravaginal infection.
RESULTS: Across all time points assessed, 13 distinct lipid species including choline, ethanolamine and glycerol were detected. Amongst these metabolites, phosphatidylcholine (PC) was the predominant phospholipid detected from animals actively shedding bacteria i.e., at 3, 9, and 15 days post infection. However, at days 30 and 65 when the animals had cleared the infection, PC was observed to be decreased compared to previous time points. Mass spectrometry analyses of PC produced in guinea pigs (in vivo) and 104C1 guinea pig cell line (in vitro) revealed distinct PC species following Ct D infection. Amongst these, PC 16:0/18:1 was significantly upregulated following Ct D infection (p < 0.05, >twofold change) in vivo and in vitro infection models investigated in this report. Exogenous addition of PC 16:0/18:1 resulted in significant increase in Ct D in Hela 229 cells.
CONCLUSION: This study demonstrates a role for host metabolite, PC 16:0/18:1 in regulating genital Ct infection in vivo and in vitro.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Potential use of cord blood for Hb E hemoglobinopathy screening programme using capillary electrophoresis

Wan Mohd Saman WA, Hassan R, Mohd Yusoff S, Che Yaakob CA, Abdullah NA, Ghazali S, et al.

Malays J Pathol, 2016 Dec;38(3):235-239.
PMID: 28028293 MyJurnal

BACKGROUND: Thalassemia and hemoglobinopathies are inherited red blood cell disorders found worldwide. Hemoglobin (Hb) E disorder is one of the hemoglobinopathies known to have the high prevalence in South East Asia. Most of transfusion-dependent thalassemias were genotypically compound heterozygous Hb E/ β-thalassemia. In Malaysia, the national screening program for thalassemia was implemented for early pregnancy or secondary school girls; however many participants do not turn-up and missed the screening test. Screening for thalassemia using samples from cord blood is an alternative choice as it is a readily available source of blood and hence early detection of the disease. The purpose of this study was to determine the potential use of cord blood for the screening of HbE hemoglobinopathy by using capillary electrophoresis (CE).
METHODS: Cord blood samples were collected from 300 newborns of healthy mothers. Hematological parameters were determined and hemoglobin quantitation for all cord blood samples were performed using capillary electrophoresis system (CES) and high performance liquid chromatography (HPLC).
RESULTS: Majority of cord blood samples (63%) revealed Hb AF followed by Hb AFA2 (20%). Hb AFE was detected in 10.7% with the mean value of Hb E ranging from 2.3%-11.1%.
CONCLUSION: Hemoglobin E was detected in cord blood using capillary electrophoresis system. It can be recommended in areas where Hb E/β is prevalent. Implementation of a screening strategy using CE on cord blood sampling will identify the disease early. With regular follow-up on these patients, the status of their disease can be determined earlier and appropriate management implemented.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Subcritical Water Technology for Extraction of Phenolic Compounds from Chlorella sp. Microalgae and Assessment on Its Antioxidant Activity

Zakaria SM, Kamal SMM, Harun MR, Omar R, Siajam SI

Molecules, 2017 Jul 03;22(7).
PMID: 28671617 DOI: 10.3390/molecules22071105

Chlorella sp. microalgae is a potential source of antioxidants and natural bioactive compounds used in the food and pharmaceutical industries. In this study, a subcritical water (SW) technology was applied to determine the phenolic content and antioxidant activity of Chlorella sp. This study focused on maximizing the recovery of Chlorella sp. phenolic content and antioxidant activity measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay as a function of extraction temperature (100-250 °C), time (5-20 min) and microalgae concentration (5-20 wt. %) using response surface methodology. The optimal operating conditions for the extraction process were found to be 5 min at 163 °C with 20 wt. % microalgae concentration, which resulted in products with 58.73 mg gallic acid equivalent (GAE)/g phenolic content and 68.5% inhibition of the DPPH radical. Under optimized conditions, the experimental values were in close agreement with values predicted by the model. The phenolic content was highly correlated (R² = 0.935) with the antioxidant capacity. Results indicated that extraction by SW technology was effective and that Chlorella sp. could be a useful source of natural antioxidants.

Matched MeSH terms: Chromatography, High Pressure Liquid

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