Displaying publications 501 - 520 of 1728 in total

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  1. UHPLC-MS phytochemical profiling, biological propensities and in-silico studies of Alhagi maurorum roots: a medicinal herb with multifunctional properties
    Saleem H, Sarfraz M, Khan KM, Anwar MA, Zengin G, Ahmad I, et al.
    Drug Dev Ind Pharm, 2020 May;46(5):861-868.
    PMID: 32352878 DOI: 10.1080/03639045.2020.1762199
    The biological, chemical, and in silico properties of methanol and dichloromethane (DCM) extracts of Alhagi maurorum roots with respect to the antioxidant, enzyme inhibition, and phytochemical composition were evaluated. Total bioactive contents were determined spectrophotometrically, and the individual secondary metabolites composition was assessed via ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) analysis. Antioxidant capacities were evaluated using a panoply of assays (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical scavenging, ferric reducing antioxidant power (FRAP), cupric reducing antioxidant power (CUPRAC), phosphomolybdenum total antioxidant capacity (TAC), and metal chelating activity (MCA)). The enzyme inhibition potential was studied against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase, tyrosinase, urease and lipoxygenase (LOX) enzymes. The methanol extract was found to contain higher total phenolic (105.91 mg GAE/g extract) and flavonoid (2.27 mg RE/g extract) contents which can be correlated to its more substantial antioxidant potential as well as AChE, BChE, tyrosinase and α-glucosidase inhibition. However, the DCM extract was the most effective against α-amylase (1.86 mmol ACAE/g extract) enzyme inhibition. The UHPLC-MS analysis of methanol extract identified the tentative presence of a total of 18 secondary metabolites, including flavonoids, saponins, phenolic and terpenoid derivatives. Three compounds named emmotin A, luteolin 5,3'-dimethyl ether, and preferrugone were further investigated for their in silico molecular docking studies against the tested enzymes. The selected compounds were found to have higher binding interaction with AChE followed by BChE, α-glucosidase, α-amylase, and tyrosinase. The results of the present study have demonstrated A. mauroram to be considered as a lead source of natural antioxidant and enzyme inhibitor compounds.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods
  2. A high-performance liquid chromatography analysis of plasma artemisinin using a glassy carbon electrode for reductive electrochemical detection
    Chan KL, Yuen KH, Jinadasa S, Peh KK, Toh WT
    Planta Med, 1997 Feb;63(1):66-9.
    PMID: 9063097
    A high-performance liquid chromatography assay equipped with a glassy carbon electrode for electrochemical detection (HPLC-ECD) was developed at reductive mode for the analysis of artemisinin, the antimalarial drug from Artemisia annua (Asteraceae) in human plasma. This method was selective, sensitive, and produced satisfactory recovery, precision, and accuracy. Analysis of plasma samples from 8 male volunteers given 10 mg kg-1 of artemisinin orally as an aqueous suspension showed a mean peak plasma concentration (Cmax) of 580.89 ng ml-1 +/- 88.64 SD at 2.5 h +/- 0.5 SD after dosing, and the mean area under the plasma concentration-time curve (AUC0-infinity) was 2227.57 ng h ml-1 +/- 677.22 SD. In addition, the elimination rate constant (Ke), elimination half-life (t1/2), and apparent volume of distribution (Vd) were calculated to be 0.2971 h-1 +/- 0.0644 SD, 2.42 h +/- 0.46 SD, and 16.26 l kg-1 +/- 3.44 SD, respectively.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods*
  3. Liquid Chromatographic Analysis of α-Dicarbonyls Using Girard-T Reagent Derivatives
    Lawrence GD, Rahmat R, Makahleh A, Saad B
    J Chromatogr Sci, 2017 Nov 01;55(10):1043-1050.
    PMID: 28977384 DOI: 10.1093/chromsci/bmx073
    The measurement of α-dicarbonyls and other degradation products of sugars has become important in view of their toxicity. Although there are several methods used for their analysis, most require long reaction times to form UV absorbing or fluorescent derivatives and the nonpolar nature of commonly used derivatives necessitates relatively high concentrations of organic solvents for elution in reverse phase liquid chromatography. The present method describes the use of Girard-T reagent in a simple, one step derivatization of α-dicarbonyls and conjugated aldehydes and analysis using ion-pair reverse phase liquid chromatography. The limit of detection was in the range of 0.06-0.09 μM (4-12 ng/mL) for glyoxal, methylglyoxal, 3-deoxyglucosone and 5-hydroxymethylfurfural with good linear response and reproducibility using UV detection. The hydrazone derivatives were stable for several days in solution. The method was used to study degradation of several sugars and quantification of the target α-dicarbonyls and 5-hydroxymethylfurfural in several soft drinks.
    Matched MeSH terms: Chromatography, Liquid/methods*
  4. Discovery of Antimalarial Drugs from Streptomycetes Metabolites Using a Metabolomic Approach
    Ahmad SJ, Abdul Rahim MBH, Baharum SN, Baba MS, Zin NM
    J Trop Med, 2017;2017:2189814.
    PMID: 29123551 DOI: 10.1155/2017/2189814
    Natural products continue to play an important role as a source of biologically active substances for the development of new drug. Streptomyces, Gram-positive bacteria which are widely distributed in nature, are one of the most popular sources of natural antibiotics. Recently, by using a bioassay-guided fractionation, an antimalarial compound, Gancidin-W, has been discovered from these bacteria. However, this classical method in identifying potentially novel bioactive compounds from the natural products requires considerable effort and is a time-consuming process. Metabolomics is an emerging "omics" technology in systems biology study which integrated in process of discovering drug from natural products. Metabolomics approach in finding novel therapeutics agent for malaria offers dereplication step in screening phase to shorten the process. The highly sensitive instruments, such as Liquid Chromatography-Mass Spectrophotometry (LC-MS), Gas Chromatography-Mass Spectrophotometry (GC-MS), and Nuclear Magnetic Resonance ((1)H-NMR) spectroscopy, provide a wide range of information in the identification of potentially bioactive compounds. The current paper reviews concepts of metabolomics and its application in drug discovery of malaria treatment as well as assessing the antimalarial activity from natural products. Metabolomics approach in malaria drug discovery is still new and needs to be initiated, especially for drug research in Malaysia.
    Matched MeSH terms: Chromatography, Liquid; Gas Chromatography-Mass Spectrometry
  5. Cyclodextrin based polymer sorbents for micro-solid phase extraction followed by liquid chromatography tandem mass spectrometry in determination of endogenous steroids
    Manaf NA, Saad B, Mohamed MH, Wilson LD, Latiff AA
    J Chromatogr A, 2018 Mar 30;1543:23-33.
    PMID: 29478831 DOI: 10.1016/j.chroma.2018.02.032
    Sorbents were prepared by cross-linking β-cyclodextrin (β-CD) using two different types of cross-linker units at variable reactant mole ratios. The resulting polymers containing β-CD were evaluated as sorbents in micro-solid phase extraction (μ-SPE) format for the extraction of the endogenous steroids testosterone (T), epitestosterone (E), androsterone (A), etiocholanolone (Etio), 5α-androstane-3α,17β-diol (5αAdiol) and 5β-androstane-3α,17β-diol (5βAdiol). The best sorbent (C1; cyclodextrin polymer) showed superior extraction characteristics compared with commercial sorbents (C18 and Bond Elut Plexa). Parameters influencing the extraction efficiency of the C1 sorbent such as extraction and desorption times, desorption solvent and volume of sample were investigated. The extracts were separated using a Hypersil Gold column (50 × 2.1 mm, 1.9 μm) under gradient elution coupled to a LC-MS/MS. The compounds were successfully separated within 8 min. The method offers good repeatability (RSD  0.995) were within the range of 1-200 ng mL-1 for T and E, 250-4000 ng mL-1 for A and Etio and 25-500 ng mL-1 for 5αAdiol and 5βAdiol, respectively. The method was applied for the determination of steroid profile of urine from volunteers.
    Matched MeSH terms: Chromatography, Liquid*
  6. Estimation of uncertainty from method validation data: Application to a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatin using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent
    Azilawati MI, Dzulkifly MH, Jamilah B, Shuhaimi M, Amin I
    J Pharm Biomed Anal, 2016 Sep 10;129:389-397.
    PMID: 27454091 DOI: 10.1016/j.jpba.2016.07.012
    A detailed procedure for estimating uncertainty according to the Laboratory of Government Chemists/Valid Analytical Measurement (LGC/VAM) protocol for determination of 18 amino acids in gelatin is proposed. The expanded uncertainty was estimated using mainly the method validation data (precision and trueness). Other sources of uncertainties were contributed by components in standard preparation measurements. The method scope covered a single matrix (gelatin) under a wide range of analyte concentrations. The uncertainty of method precision, μ(P) was 0.0237-0.1128pmolμl(-1) in which hydroxyproline and histidine represented the lowest and highest values of uncertainties, respectively. Proline and phenylalanine represented the lowest and highest uncertainties value for method recovery, μ(R) that was estimated within 0.0064-0.0995pmolμl(-1). The uncertainties from other sources, μ(Std) were 0.0325, 0.0428 and 0.0413pmolμl(-1) that were contributed by hydroxyproline, other amino acids and cystine, respectively. Hydroxyproline and phenylalanine represented the lowest and highest values of expanded uncertainty, U(y) that were determined at 0.0949 and 0.2473pmolμl(-1), respectively. The data were accurately defined and fulfill the technical requirements of ISO 17025:2005.
    Matched MeSH terms: Chromatography, Reverse-Phase/methods
  7. Simultaneous determination of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in human serum by ultra performance liquid chromatography: An economical and validated method with bovine serum albumin
    Chin SF, Osman J, Jamal R
    Clin Chim Acta, 2018 Oct;485:60-66.
    PMID: 29935177 DOI: 10.1016/j.cca.2018.06.024
    A simple and economical method has been developed for simultaneous determination of human serum 25-hydroxyvitamin D2 (25OHD2) and 25-hydroxyvitamin D3 (25OHD3) using Ultra Performance Liquid Chromatography (UPLC). Non-human matrix of 4% BSA was used to construct the calibration curve and in quality control samples' preparation to avoid interference of the endogenous 25-hydroxyvitamin D (25OHD) present in the human serum. 25OHD2, 25OHD3 and dodecanophenone (internal standard, IS) were separated on a CORTECS solid-core particle column and monitored by photodiode array detector at wavelength of 265 nm within five min run time. The relationship between 25OHD concentration and peak area ratio (25OHD:IS) was linear over the range of 12.5 - 200 nM with mean correlation coefficients (r2) >0.998. The limit of detection (LOD) for 25OHD2 and 25OHD3 was 3.00 nM and 3.79 nM, while the lower limit of quantification (LLOQ) was 9.11 nM and 11.48 nM, respectively. High repeatability was obtained for both isomers with intra-day CV% <5.6% and <5.3% for inter-day assay. This method was further tested with a commercial lyophilized serum control with an accuracy of 92.87-108.31% and applied on 214 human serum samples. In summary, this validated method with BSA can be reliably applied for routine quantification of 25OHD in adults.
    Matched MeSH terms: Chromatography, High Pressure Liquid/economics
  8. Fulltext MECP2 mutations in Malaysian Rett syndrome patients
    Fong CB, Thong MK, Sam CK, Mohamed Noor MN, Ariffin R
    Singapore Med J, 2009 May;50(5):529-33.
    PMID: 19495527
    Rett syndrome (RS) is a severe neurodevelopmental disorder characterised by normal neurological development followed by progressive developmental regression. The X-linked dominant inheritance of RS has been mapped to the gene that encodes the methyl-CpG-binding protein-2 (MECP2) at Xq28. In the present study, denaturing high-performance liquid chromatography (DHPLC) was used to detect mutations in the MECP2 gene in 20 Malaysian RS patients.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods
  9. Production of gasoline range hydrocarbons from catalytic cracking of linoleic acid over various acidic zeolite catalysts
    Gurdeep Singh HK, Yusup S, Quitain AT, Kida T, Sasaki M, Cheah KW, et al.
    Environ Sci Pollut Res Int, 2019 Nov;26(33):34039-34046.
    PMID: 30232774 DOI: 10.1007/s11356-018-3223-4
    Employment of edible oils as alternative green fuel for vehicles had raised debates on the sustainability of food supply especially in the third-world countries. The non-edible oil obtained from the abundantly available rubber seeds could mitigate this issue and at the same time reduce the environmental impact. Therefore, this paper investigates the catalytic cracking reaction of a model compound named linoleic acid that is enormously present in the rubber seed oil. Batch-scale experiments were conducted using 8.8 mL Inconel batch reactor having a cyclic horizontal swing span of 2 cm with a frequency of 60 cycles per minute at 450 °C under atmospheric condition for 90 min. The performance of HZSM-5, HBeta, HFerrierite, HMordenite and HY catalysts was tested for their efficiency in favouring gasoline range hydrocarbons. The compounds present in the organic liquid product were then analysed using GC-MS and classified based on PIONA which stands for paraffin, isoparaffin, olefin, naphthenes and aromatics respectively. The results obtained show that HZSM-5 catalyst favoured gasoline range hydrocarbons that were rich in aromatics compounds and promoted the production of desired isoparaffin. It also gave a higher cracking activity; however, large gaseous as by-products were produced at the same time.
    Matched MeSH terms: Gas Chromatography-Mass Spectrometry/methods
  10. Fulltext Characterization of high-value bioactives in some selected varieties of Pakistani Rice (Oryza sativa L.)
    Zubair M, Anwar F, Ashraf M, Uddin MK
    Int J Mol Sci, 2012;13(4):4608-4622.
    PMID: 22605998 DOI: 10.3390/ijms13044608
    The present study reports the composition and variation of fatty acids, sterols, tocopherols and γ-oryzanol among selected varieties namely Basmati Super, Basmati 515, Basmati 198, Basmati 385, Basmati 2000, Basmati 370, Basmati Pak, KSK-139, KS-282 and Irri-6 of Pakistani rice (Oryza sativa L). Oil content extracted with n-hexane from different varieties of brown rice seed (unpolished rice) ranged from 1.92% to 2.72%. Total fatty acid contents among rice varieties tested varied between 18240 and 25840 mg/kg brown rice seed. The rice tested mainly contained oleic (6841-10952 mg/kg) linoleic (5453-7874 mg/kg) and palmitic acid (3613-5489 mg/kg). The amounts of total phytosterols (GC and GC-MS analysis), with main contribution from β-sitosterol (445-656 mg/kg), campesterol (116-242 mg/kg), Δ(5)-avenasterol (89-178 mg/kg) and stigmasterol (75-180 mg/kg) were established to be 739.4 to 1330.4 mg/kg rice seed. The content of α-, γ- and δ-tocopherols as analyzed by HPLC varied from 39.0-76.1, 21.6-28.1 and 6.5-16.5 mg/kg rice seed, respectively. The amounts of different γ-oryzanol components (HPLC data), identified as cycloartenyl ferulate, 24-methylene cycloartanyl ferulate, campesteryl ferulate and β-sitosteryl ferulate, were in the range of 65.5-103.6, 140.2-183.1, 29.8-45.5 and 8.6-10.4 mg/kg rice seed, respectively. Overall, the concentration of these bioactives was higher in the Basmati rice cultivars showing their functional food superiority. In conclusion, the tested varieties of Pakistani rice, especially the Basmati cultivars, can provide best ingredients for functional foods.
    Matched MeSH terms: Chromatography, High Pressure Liquid; Gas Chromatography-Mass Spectrometry
  11. Fulltext Assessing product adulteration of Eurycoma longifolia (Tongkat Ali) herbal medicinal product using DNA barcoding and HPLC analysis
    Abubakar BM, Salleh FM, Shamsir Omar MS, Wagiran A
    Pharm Biol, 2018 Dec;56(1):368-377.
    PMID: 30058427 DOI: 10.1080/13880209.2018.1479869
    CONTEXT: Eurycoma longifolia Jack (Simaroubaceae) commonly known as Tongkat Ali is one of the most important plants in Malaysia. The plant extracts (particularly roots) are widely used for the treatment of cough and fever besides having antimalarial, antidiabetic, anticancer and aphrodisiac activities.

    OBJECTIVES: This study assesses the extent of adulteration of E. longifolia herbal medicinal products (HMPs) using DNA barcoding validated by HPLC analysis.

    MATERIALS AND METHODS: Chloroplastic rbcL and nuclear ITS2 barcode regions were used in the present study. The sequences generated from E. longifolia HMPs were compared to sequences in the GenBank using MEGABLAST to verify their taxonomic identity. These results were verified by neighbor-joining tree analysis in which branches of unknown specimen are compared to the reference sequences established from this study and other retrieved from the GenBank. The HMPs were also analysed using HPLC analysis for the presence of eurycomanone bioactive marker.

    RESULTS: Identification using DNA barcoding revealed that 37% of the tested HMPs were authentic while 27% were adulterated with the ITS2 barcode region proven to be the ideal marker. The validation of the authenticity using HPLC analysis showed a situation in which a species which was identified as authentic was found not to contain the expected chemical compound.

    DISCUSSION AND CONCLUSIONS: DNA barcoding should be used as the first screening step for testing of HMPs raw materials. However, integration of DNA barcoding with HPLC analysis will help to provide detailed knowledge about the safety and efficacy of the HMPs.

    Matched MeSH terms: Chromatography, High Pressure Liquid/methods
  12. Fulltext Rapid resolution liquid chromatography method development and validation for simultaneous determination of homocysteine, vitamins B(6), B(9), and B(12) in human serum
    Shaik MM, Gan SH
    Indian J Pharmacol, 2013 Mar-Apr;45(2):159-67.
    PMID: 23716893 DOI: 10.4103/0253-7613.108303
    Hyperhomocysteinemia and vitamins B(6), B(9), and B(12) deficiencies usually result in various neurological, vascular, ocular, renal, and pulmonary abnormalities. However, to date, there are no simultaneous detection methods available for determining homocysteine, vitamins B(6), B(9), and B(12) levels in various biological fluids. In this study, we aim to develop a new validated simultaneous detection method for all four compounds to save both cost and time of analysis.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods*
  13. Phytochemical quantification and HPLC analysis of Parkia speciosa pod extract
    Cheong NDH, Mohamed E, Haron N, Camalxaman SN, Abdullah A, Mohamad Yusof MI, et al.
    Med J Malaysia, 2024 Mar;79(Suppl 1):34-39.
    PMID: 38555883
    INTRODUCTION: Parkia speciosa Hassk., commonly known as bitter bean or twisted cluster bean, is a tropical leguminous plant species native to Southeast Asia. The plant's edible pods have been traditionally used in various cuisines, particularly in Malaysian, Thai, and Indonesian cooking. Apart from being used as a food ingredient, the pods of P. speciosa also have a range of potential applications in other fields, including medicine, agriculture, and industry. The pods are said to have several phytochemicals that hold great therapeutic values such as reducing inflammation, improving digestion, and lowering blood sugar levels. However, there is limited information on the specific phytochemical contents of the pods in the literature. Thus, the aim of this study is to quantify the total phenolic and flavonoid compounds and to determine the concentrations of four selected phytochemical compounds in the P. speciosa pod extract (PSPE).

    MATERIALS AND METHODS: Quantification of the total phenolic (TPC) and flavonoid contents (TFC) in PSPE were done via colourimetric methods; and the determination of the concentrations of four specific phytochemicals (gallic acid, caffeic acid, rutin, and quercetin) were done via High- Performance Liquid Chromatography (HPLC).

    RESULTS: Colourimetric determination of PSPE showed TPC and TFC values of 84.53±9.40 mg GAE/g and 11.96±4.51 mg QE/g, respectively. Additional analysis of the phytochemicals using HPLC revealed that there were 6.45±3.36 g/kg, 5.91±1.07 g/kg, 0.39±0.84 g/kg, and 0.19±0.47 g/kg of caffeic acid, gallic acid, rutin, and quercetin, respectively.

    CONCLUSION: The findings show that PSPE contains substantial amounts of caffeic acid, gallic acid, rutin, and quercetin, which may indicate its potential as antibacterial, anti-inflammatory, anti-lipid, and antiviral medicines.

    Matched MeSH terms: Chromatography, High Pressure Liquid/methods
  14. Fulltext From the Front or Back Door? Quantitative analysis of direct and indirect extractions of α-mangostin from mangosteen (Garcinia mangostana)
    Eukun Sage E, Jailani N, Md Taib AZ, Mohd Noor N, Mohd Said MI, Abu Bakar M, et al.
    PLoS One, 2018;13(10):e0205753.
    PMID: 30321238 DOI: 10.1371/journal.pone.0205753
    The pulp and pericarp of mangosteen (Garcinia mangostana) fruit are popular food, beverage and health products whereby 60% of the fruit consist of the pericarp. The major metabolite in the previously neglected or less economically significant part of the fruit, the pericarp, is the prenylated xanthone α-mangostin. This highly bioactive secondary metabolite is typically isolated using solvent extraction methods that involve large volumes of halogenated solvents either via direct or indirect extraction. In this study, we compared the quantities of α-mangostin extracted using three different extraction methods based on the environmentally friendly solvents methanol and ethyl acetate. The three solvent extractions methods used were direct extractions from methanol (DM) and ethyl acetate (DEA) as well as indirect extraction of ethyl acetate obtained via solvent partitioning from an initial methanol extract (IEA). Our results showed that direct extraction afforded similar and higher quantities of α-mangostin than indirect extraction (DM: 318 mg; DEA: 305 mg; IEA: 209 mg per 5 g total dried pericarp). Therefore, we suggest that the commonly used method of indirect solvent extraction using halogenated solvents for the isolation of α-mangostin is replaced by single solvent direct extraction using the environmentally friendly solvents methanol or ethyl acetate.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods
  15. Effect of Gibberellin and Heat Shock on the Lipid Composition of Endoplasmic Reticulum in Barley Aleurone Layers
    Grindstaff KK, Fielding LA, Brodl MR
    Plant Physiol, 1996 Feb;110(2):571-581.
    PMID: 12226205
    The heat-shock responses of barley (Hordeum vulgare L. cv Hi- malaya) aleurone layers incubated with or without gibberellic acid (GA3) were compared. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that heat shock blocked the synthesis and secretion of secretory proteins from GA3-treated layers but not untreated layers. This suppression of secretory protein synthesis has been correlated with changes in endoplasmic reticulum (ER) membranes (F.C. Belanger, M. R. Brodl, T.-h.D. Ho [1986] Proc Natl Acad Sci USA 83: 1354-1358; L. Sticher, A.K. Biswas, D.S. Bush, R.L. Jones [1990] Plant Physiol 92: 506-513). Our secretion data suggested that the ER membranes of aleurone layers incubated without GA3 may be more heat shock tolerant. To investigate this, the lipid profiles of membrane extracts in aleurone layers labeled with [14C]glycerol were examined. Heat shock markedly increased [14C]glycerol incorporation into phosphatidylcholine (PC), and gas chromatography revealed an increase in the amount of saturated fatty acids associated with thin layer chromatography-purified PC in GA3-treated layers. In contrast, aleurone layers incubated without GA3 at normal temperature contained PC-associated fatty acids with a greater degree of saturation than GA3-treated layers. Heat shock modestly increased the degree of fatty acid saturation in untreated aleurone layers. This same trend was noted in fatty acids isolated from ER membranes purified by continuous sucrose density centrifugation. We propose that increased fatty acid saturation may help sustain ER membrane function in heat-shocked aleurone layers incubated in the absence of GA3.
    Matched MeSH terms: Chromatography, Gas; Chromatography, Thin Layer
  16. Fulltext The Role of Semipurified Fractions Isolated from Quercus infectoria on Bone Metabolism by Using hFOB 1.19 Human Fetal Osteoblast Cell Model
    Abdullah AR, Hapidin H, Abdullah H
    Evid Based Complement Alternat Med, 2018;2018:5319528.
    PMID: 29861772 DOI: 10.1155/2018/5319528
    Background. Quercus infectoria (QI) is a plant used in traditional medicines in Asia. The plant was reported to contain various active phytochemical compounds that have potential to stimulate bone formation. However, the precise mechanism of the stimulation effect of QI on osteoblast has not been elucidated. The present study was carried out to isolate QI semipurified fractions from aqueous QI extract and to delineate the molecular mechanism of QI semipurified fraction that enhanced bone formation by using hFOB1.19 human fetal osteoblast cell model. Methods. Isolation of QI semipurified fractions was established by means of column chromatography and thin layer chromatography. Established QI semipurified fractions were identified using Liquid Chromatography-Mass Spectrometry (LC-MS). Cells were treated with derived QI semipurified fractions and investigated for mineralization deposition and protein expression level of BMP-2, Runx2, and OPN by ELISA followed gene expression analysis of BMP-2 and Runx2 by RT-PCR. Results. Column chromatography isolation and purification yield Fractions A, B, and C. LC-MS analysis reveals the presence of polyphenols in each fraction. Results show that QI semipurified fractions increased the activity and upregulated the gene expression of BMP-2 and Runx2 at day 1, day 3, and day 7. OPN activity increased in cells treated with QI semipurified fractions at day 1 and day 3. Meanwhile, at day 7, expression of OPN decreased in activity. Furthermore, the study showed that combination of Fractions A, B, and C with osteoporotic drug (pamidronate) further increased the activity and upregulated the gene expression of BMP-2 and Runx2. Conclusions. These findings demonstrated that polyphenols from semipurified fractions of QI enhanced bone formation through expression of the investigated bone-related marker that is its potential role when combined with readily available osteoporotic drug.
    Matched MeSH terms: Chromatography, Liquid; Chromatography, Thin Layer
  17. Fulltext Isolation, Characterization, Crystal Structure Elucidation, and Anticancer Study of Dimethyl Cardamonin, Isolated from Syzygium campanulatum Korth
    Memon AH, Ismail Z, Aisha AF, Al-Suede FS, Hamil MS, Hashim S, et al.
    Evid Based Complement Alternat Med, 2014;2014:470179.
    PMID: 25530783 DOI: 10.1155/2014/470179
    Syzygium campanulatum Korth is an equatorial, evergreen, aboriginal shrub of Malaysia. Conventionally it has been used as a stomachic. However, in the currently conducted study dimethyl cardamonin or 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC) was isolated from S. campanulatum Korth, leaf extract. The structural characterization of DMC was carried out by making use of various techniques including UV, IR, NMR spectral followed by LC-MS, and X-ray crystallographic techniques. For determining the purity of compound, highly effective techniques including TLC, HPLC, and melting point were used. The cytotoxicity of DMC and three different extracts of S. campanulatum was evaluated against human colon cancer cell line (HT-29) by three different assays. DMC and ethanolic extract revealed potent and dose-dependent cytotoxic activity on the cancer cell line with IC50 12.6 and 90.1 µg/mL, respectively. Quite astonishingly to our knowledge, this is the very first report on S. campanulatum as being a rich source (3.5%) of DMC, X-ray crystallography, and anticancer activity on human colon cancer cells.
    Matched MeSH terms: Chromatography, High Pressure Liquid; Chromatography, Liquid
  18. A highly selective two-way purification method using liquid chromatography for isolating αS2-casein from goat milk of five different breeds
    Mohsin AZ, Sukor R, Selamat J, Meor Hussin AS, Ismail IH, Jambari NN, et al.
    J Chromatogr B Analyt Technol Biomed Life Sci, 2020 Dec 01;1160:122380.
    PMID: 32971369 DOI: 10.1016/j.jchromb.2020.122380
    The main challenges in the purification of αS2-casein are due to the low quantity in milk and high homology with other casein subunits, i.e., αS1-casein, β-casein, and κ-casein. To overcome these challenges, the aim of this study was to develop a two-step purification to isolate native αS2-casein in goat milk from five different breeds; British Alpine, Jamnapari, Saanen, Shami, and Toggenburg. The first step of the purification was executed by anion-exchange chromatography under optimal elution conditions followed by size exclusion chromatography. Tryptic peptides from in-gel digestion of purified αS2-casein were sequenced and analyzed by LC-ESI-MS/MS. From 1.05 g of whole casein, the highest yield of αS2-casein (6.7 mg/mL) was obtained from Jamnapari and the lowest yield (2.2 mg/mL) was from Saanen. A single band of pure αS2-casein was observed on SDS-PAGE for all breeds. The αS2-casein showed coverage percentage of amino acid sequence from 76.68 to 92.83%. The two-step purification process developed herein was successfully applied for isolating native αS2-casein from goat milk with high purity, which will allow for future in vitro studies to be conducted on this protein.
    Matched MeSH terms: Chromatography, Liquid/methods*
  19. LC-MS/MS quantitation of antimalarial drug piperaquine and metabolites in human plasma
    Aziz MY, Hoffmann KJ, Ashton M
    J Chromatogr B Analyt Technol Biomed Life Sci, 2017 Sep 15;1063:253-258.
    PMID: 28863865 DOI: 10.1016/j.jchromb.2017.06.035
    PURPOSE: This study aimed to develop a sensitive, quantitative assay for the antimalarial piperaquine (PQ) and its metabolites M1 and M2 in human plasma.

    RESULTS: Analytes were gradiently separated on a C18 column and detected with a Sciex API 4000 MS/MS with an ESI source operated in the positive ion mode with deuterated PQ as internal standard. The response was linear in the range 3.9-2508nM with a runtime of 7.0min per sample. The method was applied to clinical samples from healthy volunteers.

    CONCLUSION: This LC-MS/MS method for the simultaneous quantitation of PQ and two of its metabolites in plasma may prove helpful for assessment of metabolite safety issues in vivo.

    Matched MeSH terms: Chromatography, Liquid/methods*
  20. HPLC-MS/MS method for bioavailability study of bruceines D & E in rat plasma
    Man F, Choo CY
    J Chromatogr B Analyt Technol Biomed Life Sci, 2017 Sep 15;1063:183-188.
    PMID: 28869873 DOI: 10.1016/j.jchromb.2017.08.037
    Bruceines D and E are quassinoids from seeds of Brucea javanica (L.) Merr. exhibiting hypoglycemia effect. The crude drug is used as a traditional medicine by diabetes patients. The aim of this study is to understand the bioavailability and pharmacokinetics of both the bruceines D & E. A rapid and sensitive HPLC-MS/MS method was developed and validated for the quantification of both quassinoids, bruceines D & E in rat plasma. Both the bruceines D & E were separated with the Zorbax SBC-18 column with gradient elution and mobile phase system of acetonitrile and deionized water with 0.1% formic acid at a flow rate of 0.5mL/min. Analytes were detected in multiple reaction monitoring (MRM) mode with electrospray positive ionization. The quassinoids, namely bruceines D & E were detected with transitions of m/z 411.2→393.2 and m/z 395.2→377.2, respectively. Another quassinoid, eurycomanone was used as the internal standard with transition of m/z 409.2→391.2. The method was validated and conformed to the regulatory requirements. The validated method was applied to pharmacokinetic and bioavailability studies in rats. The pharmacokinetic study indicated both bruceine D and E were rapidly absorbed into the circulation system and reached its peak concentration at 0.54±0.34h and 0.66±0.30h, respectively. Bruceine E was eliminated slower than Bruceine D with t1/2 value almost increased two-fold compared to Bruceine D. In conclusion, a rapid, selective and sensitive HPLC-MS/MS method was developed for the simultaneous determination of both the bruceines D and E in rat plasma. Both bruceines D and E displayed poor oral bioavailability.
    Matched MeSH terms: Chromatography, High Pressure Liquid/methods*
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