Displaying publications 1 - 20 of 35 in total

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  1. A Protein Decomplexation Strategy in Snake Venom Proteomics
    Tan CH, Tan KY, Tan NH
    Methods Mol Biol, 2019;1871:83-92.
    PMID: 30276733 DOI: 10.1007/978-1-4939-8814-3_5
    Snake venoms are complex mixtures of proteins and peptides that play vital roles in the survival of venomous snakes. As with their diverse pharmacological activities, snake venoms can be highly variable, hence the importance of understanding the compositional details of different snake venoms. However, profiling venom protein mixtures is challenging, in particular when dealing with the diversity of protein subtypes and their abundances. Here we described an optimized strategy combining a protein decomplexation method with in-solution trypsin digestion and mass spectrometry of snake venom proteins. The approach involves the integrated use of C18 reverse-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and nano-electrospray ionization tandem mass spectrometry (nano-ESI-LC-MS/MS).
    Matched MeSH terms: Chromatography, Reverse-Phase
  2. A reversed phase high performance liquid chromatography method for the determination of fumonisins B1 and B2 in food and feed using monolithic column and positive confirmation by liquid chromatography/tandem mass spectrometry
    Khayoon WS, Saad B, Salleh B, Ismail NA, Abdul Manaf NH, Abdul Latiff A
    Anal Chim Acta, 2010 Oct 29;679(1-2):91-7.
    PMID: 20951862 DOI: 10.1016/j.aca.2010.09.008
    The development of a reversed phase high performance liquid chromatography fluorescence method for the determination of the mycotoxins fumonisin B(1) and fumonisin B(2) by using silica-based monolithic column is described. The samples were first extracted using acetonitrile:water (50:50, v/v) and purified by using a C(18) solid phase extraction-based clean-up column. Then, pre-column derivatization for the analyte using ortho-phthaldialdehyde in the presence of 2-mercaptoethanol was carried out. The developed method involved optimization of mobile phase composition using methanol and phosphate buffer, injection volume, temperature and flow rate. The liquid chromatographic separation was performed using a reversed phase Chromolith(®) RP-18e column (100 mm × 4.6 mm) at 30 °C and eluted with a mobile phase of a mixture of methanol and phosphate buffer pH 3.35 (78:22, v/v) at a flow rate of 1.0 mL min(-1). The fumonisins separation was achieved in about 4 min, compared to approximately 20 min by using a C(18) particle-packed column. The fluorescence excitation and emission were at 335 nm and 440 nm, respectively. The limits of detections were 0.01-0.04 μg g(-1) fumonisin B(1) and fumonisin B(2), respectively. Good recoveries were found for spiked samples (0.1, 0.5, 1.5 μg g(-1) fumonisins B(1) and B(2)), ranging from 84.0 to 106.0% for fumonisin B(1) and from 81.0 to 103.0% for fumonisin B(2). Fifty-three samples were analyzed including 39 food and feeds and 14 inoculated corn and rice. Results show that 12.8% of the food and feed samples were contaminated with fumonisin B(1) (range, 0.01-0.51 μg g(-1)) and fumonisin B(2) (0.05 μg g(-1)). The total fumonisins in these samples however, do not exceed the legal limits established by the European Union of 0.8 μg g(-1). Of the 14 inoculated samples, 57.1% contained fumonisin B(1) (0.16-41.0 μg g(-1)) and fumonisin B(2) (range, 0.22-50.0 μg g(-1)). Positive confirmation of selected samples was carried out using liquid chromatography-tandem mass spectrometry, using triple quadrupole analyzer and operated in the multiple reaction monitoring mode.
    Matched MeSH terms: Chromatography, Reverse-Phase/methods*
  3. Fulltext Anti-angiotensin converting enzyme (ACE) proteins from mycelia of Ganoderma lucidum (Curtis) P. Karst
    Mohamad Ansor N, Abdullah N, Aminudin N
    BMC Complement Altern Med, 2013;13:256.
    PMID: 24093919 DOI: 10.1186/1472-6882-13-256
    Ganoderma lucidum has been purported as a potent remedy in the treatment and prevention of several ailments, including hypertension. This study aimed to explore the anti-ACE potential of protein fractions from the mycelia of G. lucidum.
    Matched MeSH terms: Chromatography, Reverse-Phase
  4. Fulltext Antimycobacterial susceptibility evaluation of rifampicin and isoniazid benz-hydrazone in biodegradable polymeric nanoparticles against Mycobacterium tuberculosis H37Rv strain
    Hakkimane SS, Shenoy VP, Gaonkar SL, Bairy I, Guru BR
    Int J Nanomedicine, 2018;13:4303-4318.
    PMID: 30087562 DOI: 10.2147/IJN.S163925
    INTRODUCTION: Tuberculosis (TB) is the single largest infectious disease which requires a prolonged treatment regime with multiple drugs. The present treatment for TB includes frequent administration of a combination of four drugs for a duration of 6 months. This leads to patient's noncompliance, in addition to developing drug-resistant strains which makes treatment more difficult. The formulation of drugs with biodegradable polymeric nanoparticles (NPs) promises to overcome this problem.

    MATERIALS AND METHODS: In this study, we focus on two important drugs used for TB treatment - rifampicin (RIF) and isoniazid (INH) - and report a detailed study of RIF-loaded poly lactic-co-glycolic acid (PLGA) NPs and INH modified as INH benz-hydrazone (IH2) which gives the same therapeutic effect as INH but is more stable and enhances the drug loading in PLGA NPs by 15-fold compared to INH. The optimized formulation was characterized using particle size analyzer, scanning electron microscopy and transmission electron microscopy. The drug release from NPs and stability of drug were tested in different pH conditions.

    RESULTS: It was found that RIF and IH2 loaded in NPs release in a slow and sustained manner over a period of 1 month and they are more stable in NPs formulation compared to the free form. RIF- and IH2-loaded NPs were tested for antimicrobial susceptibility against Mycobacterium tuberculosis H37Rv strain. RIF loaded in PLGA NPs consistently inhibited the growth at 70% of the minimum inhibitory concentration (MIC) of pure RIF (MIC level 1 µg/mL), and pure IH2 and IH2-loaded NPs showed inhibition at MIC equivalent to the MIC of INH (0.1 µg/mL).

    CONCLUSION: These results show that NP formulations will improve the efficacy of drug delivery for TB treatment.

    Matched MeSH terms: Chromatography, Reverse-Phase
  5. Fulltext Characterisation of potential antidiabetic-related proteins from Pleurotus pulmonarius (Fr.) Quél. (grey oyster mushroom) by MALDI-TOF/TOF mass spectrometry
    Wahab NA, Abdullah N, Aminudin N
    Biomed Res Int, 2014;2014:131607.
    PMID: 25243114 DOI: 10.1155/2014/131607
    Pleurotus pulmonarius has been reported to have a potent remedial effect on diabetic property and considered to be an alternative for type 2 diabetes mellitus treatment. This study aimed to investigate the antidiabetic properties of ammonium sulphate precipitated protein fractions from P. pulmonarius basidiocarps. Preliminary results demonstrated that 30% (NH4)2SO4 precipitated fraction (F30) inhibited Saccharomyces cerevisiae α-glucosidase activity (24.18%), and 100% (NH4)2SO4 precipitated fraction (F100) inhibited porcine pancreatic α-amylase activity (41.80%). Following RP-HPLC purification, peak 3 from F30 fraction demonstrated inhibition towards α-glucosidase at the same time with meagre inhibition towards α-amylase activity. Characterisation of proteins using MALDI-TOF/TOF MS demonstrated the presence of four different proteins, which could be implicated in the regulation of blood glucose level via various mechanisms. Therefore, this study revealed the presence of four antidiabetic-related proteins which are profilin-like protein, glyceraldehyde-3-phosphate dehydrogenase-like protein, trehalose phosphorylase-like (TP-like) protein, and catalase-like protein. Hence, P. pulmonarius basidiocarps have high potential in lowering blood glucose level, reducing insulin resistance and vascular complications.
    Matched MeSH terms: Chromatography, Reverse-Phase
  6. Characterization of apigenin and luteolin derivatives from oil palm (Elaeis guineensis Jacq.) leaf using LC-ESI-MS/MS
    Tahir NI, Shaari K, Abas F, Parveez GK, Ishak Z, Ramli US
    J Agric Food Chem, 2012 Nov 14;60(45):11201-10.
    PMID: 23116142 DOI: 10.1021/jf303267e
    The palm oil industry generates several byproducts, and more than half of the dry weight of the waste is of oil palm leaf whereby the tissue is underutilized. Recently, several research studies found promising potential of oil palm fronds as a source of nutraceutical due to its bioactive properties. However, the chemical composition of the tissue is still not deciphered. Using reversed-phase liquid chromatography (LC) electrospray mass spectrometry (ESI-MS), glycosylated apigenin and luteolin were separated and identified from oil palm (Elaeis guineensis Jacq.) leaf and structures of the constituents were elucidated by collision-induced dissociation (CID) tandem MS. From 28 derivatives of the flavones, 9 compounds were conjugated with hydroxymethylglutaric (HMG) acid. Improved knowledge on oil palm especially on bioactive component of the leaf tissue will allow correlation of its beneficial effects and further promotes efficient utilization of this agriculture byproduct.
    Matched MeSH terms: Chromatography, Reverse-Phase/methods
  7. Fulltext Cloud point extraction of parabens using non-ionic surfactant with cylodextrin functionalized ionic liquid as a modifier
    Noorashikin MS, Raoov M, Mohamad S, Abas MR
    Int J Mol Sci, 2013;14(12):24531-48.
    PMID: 24351832 DOI: 10.3390/ijms141224531
    A cloud point extraction (CPE) process using non-ionic surfactant (DC193C) to extract selected paraben compounds from water samples was investigated using reversed phase high performance liquid chromatography (RP-HPLC). The CPE process with the presence of β-cyclodextrin (βCD) functionalized ionic liquid as a modifier (CPE-DC193C-βCD-IL) is a new extraction technique that has been applied on the optimization of parameters, i.e., pH, βCD-IL concentration and phase volume ratio. This CPE-DC193C-βCD-IL method is facilitated at 30 °C, showing great losses of water content in the surfactant-rich phase, resulting in a high pre-concentration factor and high distribution coefficient. The developed method CPE-DC193C-βCD-IL did show enhanced properties compared to the CPE method without the modifier (CPE-DC193C). The developed method of CPE-DC193C-βCD-IL gives an excellent performance on the detection of parabens from water samples with the limit of detection falling in the range of 0.013-0.038 µg mL-1. Finally, the inclusion complex formation, hydrogen bonding, and π-π interaction between the βCD-IL, benzyl paraben (ArP), and DC 193C were proven using 1H NMR and 2D NOESY spectroscopy.
    Matched MeSH terms: Chromatography, Reverse-Phase
  8. Combination of saponification and dispersive liquid-liquid microextraction for the determination of tocopherols and tocotrienols in cereals by reversed-phase high-performance liquid chromatography
    Shammugasamy B, Ramakrishnan Y, Ghazali HM, Muhammad K
    J Chromatogr A, 2013 Jul 26;1300:31-7.
    PMID: 23587317 DOI: 10.1016/j.chroma.2013.03.036
    A simple sample preparation technique coupled with reversed-phase high-performance liquid chromatography was developed for the determination of tocopherols and tocotrienols in cereals. The sample preparation procedure involved a small-scale hydrolysis of 0.5g cereal sample by saponification, followed by the extraction and concentration of tocopherols and tocotrienols from saponified extract using dispersive liquid-liquid microextraction (DLLME). Parameters affecting the DLLME performance were optimized to achieve the highest extraction efficiency and the performance of the developed DLLME method was evaluated. Good linearity was observed over the range assayed (0.031-4.0μg/mL) with regression coefficients greater than 0.9989 for all tocopherols and tocotrienols. Limits of detection and enrichment factors ranged from 0.01 to 0.11μg/mL and 50 to 73, respectively. Intra- and inter-day precision were lower than 8.9% and the recoveries were around 85.5-116.6% for all tocopherols and tocotrienols. The developed DLLME method was successfully applied to cereals: rice, barley, oat, wheat, corn and millet. This new sample preparation approach represents an inexpensive, rapid, simple and precise sample cleanup and concentration method for the determination of tocopherols and tocotrienols in cereals.
    Matched MeSH terms: Chromatography, Reverse-Phase/methods*
  9. Fulltext Cytotoxicity of Snake Venoms and Cytotoxins From Two Southeast Asian Cobras (Naja sumatrana, Naja kaouthia): Exploration of Anticancer Potential, Selectivity, and Cell Death Mechanism
    Chong HP, Tan KY, Tan CH
    Front Mol Biosci, 2020;7:583587.
    PMID: 33263003 DOI: 10.3389/fmolb.2020.583587
    Venoms of cobras (Naja spp.) contain high abundances of cytotoxins, which contribute to tissue necrosis in cobra envenomation. The tissue-necrotizing activity of cobra cytotoxins, nevertheless, indicates anticancer potentials. This study set to explore the anticancer properties of the venoms and cytotoxins from Naja sumatrana (equatorial spitting cobra) and Naja kaouthia (monocled cobra), two highly venomous species in Southeast Asia. The cytotoxicity, selectivity, and cell death mechanisms of their venoms and cytotoxins (NS-CTX from N. sumatrana: NS-CTX; N. kaouthia: NK-CTX) were elucidated in human lung (A549), prostate (PC-3), and breast (MCF-7) cancer cell lines. Cytotoxins were purified through a sequential fractionation approach using cation-exchange chromatography, followed by C18 reverse-phase high-performance liquid chromatography (HPLC) to homogeneity validated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identified by liquid chromatography-tandem mass spectrometry (LCMS/MS). The cobra venoms and their respective cytotoxins exhibited concentration-dependent growth inhibitory effects in all cell lines tested, with the cytotoxins being more potent compared to the corresponding whole venoms. NS-CTX and NK-CTX are, respectively, P-type and S-type isoforms of cytotoxin, based on the amino acid sequences as per LCMS/MS analysis. Both cytotoxins exhibited differential cytotoxic effects in the cell lines tested, with NS-CTX (P-type cytotoxin) being significantly more potent in inhibiting the growth of the cancer cells. Both cytotoxins demonstrated promising selectivity only for the A549 lung cancer cell line (selectivity index = 2.17 and 2.26, respectively) but not in prostate (PC-3) and breast (MCF-7) cancer cell lines (selectivity index < 1). Flow cytometry revealed that the A549 lung cancer cells treated with NS-CTX and NK-CTX underwent necrosis predominantly. Meanwhile, the cytotoxins induced mainly caspase-independent late apoptosis in the prostate (PC-3) and breast (MCF-7) cancer cells lines but lacked selectivity. The findings revealed the limitations and challenges that could be faced during the development of new cancer therapy from cobra cytotoxins, notwithstanding their potent anticancer effects. Further studies should aim to overcome these impediments to unleash the anticancer potentials of the cytotoxins.
    Matched MeSH terms: Chromatography, Reverse-Phase
  10. Fulltext Determination of Vitamin C, β-carotene and Riboflavin Contents in Five Green Vegetables Organically and Conventionally Grown
    Malays J Nutr, 2003;9(1):-.
    MyJurnal
    As consumer interest in organically grown vegetables is increasing in Malaysia, there is a need to answer whether the vegetables are more nutritious than those conventionally grown. This study investigates commercially available vegetables grown organically and conventionally, purchased from retailers to analyse â-carotene, vitamin C and riboflavin contents. Five types of green vegetables were selected, namely Chinese mustard (sawi) (Brassica juncea), Chinese kale (kailan) (Brassica alboglabra), lettuce (daun salad) (Lactuca sativa), spinach (bayam putih) (Amaranthus viridis) and swamp cabbage (kangkung) (Ipomoea aquatica). For vitamin analysis, a reverse-phase high performance liquid chromatography was used to identify and quantify β-carotene, vitamin C and riboflavin. The findings showed that not all of the organically grown vegetables were higher in vitamins than that conventionally grown. This study found that only swamp cabbage grown organically was highest in β-carotene , vitamin C and riboflavin contents among the entire samples studied. The various nutrients in organically grown vegetables need to be analysed for the generation of a database on nutritional value which is important for future research.
    Matched MeSH terms: Chromatography, Reverse-Phase
  11. Determination of radical scavenging activity and Vitamin A, C and E in organically grown Red Pitaya (Hylocereus sp.)
    Mohd Adzim Khalili, R.,, Norhayati, A.H., Rokiah,M. Y., Asmah, R., Siti Muskinah, M., Abdul Manaf, A.
    International Food Research Journal, 2010;17(2):405-409.
    MyJurnal
    This study was conducted to determine radical scavenging activity and vitamin antioxidant composition in red pitaya from organic plantation. For antioxidant vitamins analysis, a reverse-phase high performance liquid chromatography was used and radical scavenging activity of methanolic and water extract were determined using 1,1-diphenyl-2-pircrylhydrazyl assay. Results for radical scavenging activity, red pitaya methanolic extract achieved the highest percentage 70.13% compared with water extract (47.13%). Antioxidant vitamins composition in red pitaya showed that the concentration of vitamin A is 120.13 ± 0.69 μg/100 g freeze-dried sample, vitamin C is 540.27 ± 0.59 μg/100 g fresh samples and vitamin E is 105.67 ± 0.56 μg/100 g freeze-dried samples. This shows that red pitaya may become an alternative and potential source of natural antioxidant.
    Matched MeSH terms: Chromatography, Reverse-Phase
  12. Fulltext Determination of types of fat ingredient in some commercial biscuit formulations
    Yanty, N.A.M., Marikkar, J.M.N., Abdulkarim, S.M.
    International Food Research Journal, 2014;21(1):277-282.
    MyJurnal
    A study was carried out to compare the composition and thermal profiles of the fat component of six brands of commercial biscuits (BA, BB, BC, BD, BE & BF) with those of lard and palm oil. Extraction of fat from biscuit samples was done using petroleum ether according to the soxhlet extraction procedure. The isolated fat samples along with lard and palm oil were analyzed using gas liquid chromatography (GLC), reversed-phase high performance liquid chromatography (RP-HPLC), and differential scanning calorimetry (DSC). According to GLC analysis, palm oil, lard and all six biscuit brands had either palmitic or oleic acid as major fatty acids. Sn-2 positional analysis of fatty acids showed that oleic (> 60%) as the most dominant fatty acid of palm oil and biscuit brands BA, BB, BC, and BD while palmitic (> 60%) as the most dominant fatty acid of lard and biscuit brands BE and BF. RP-HPLC analysis showed that the triacylglycerol (TAG) profiles of lard and biscuit brands BE and BF were closely similar while those of brands BA, BB, BC, and BD and palm oil were similar. DSC analysis showed that the cooling and heating profiles of lard and brands BE and BF were similar, while those of palm oil and brands BA, BB, BC, and BD were similar. Hence, this study concluded that biscuit brands BE and BF are not suitable for consumers whose religious restriction prohibit the use of lard as food ingredient.
    Matched MeSH terms: Chromatography, Reverse-Phase
  13. Determination of underivatized long chain fatty acids using RP-HPLC with capacitively coupled contactless conductivity detection
    Makahleh A, Saad B, Siang GH, Saleh MI, Osman H, Salleh B
    Talanta, 2010 Apr 15;81(1-2):20-4.
    PMID: 20188881 DOI: 10.1016/j.talanta.2009.11.030
    A reversed-phase high-performance liquid chromatographic method with capacitively coupled contactless conductivity detector (C(4)D) has been developed for the separation and the simultaneous determination of five underivatized long chain fatty acids (FAs), namely myristic, palmitic, stearic, oleic, and linoleic acids. An isocratic elution mode using methanol/1mM sodium acetate (78:22, v/v) as mobile phase with a flow rate of 0.6 mL min(-1) was used. The separation was effected by using a Hypersil ODS C(18) analytical column (250 mm x 4.6 mm x 5 microm) and was operated at 45 degrees C. Calibration curves of the five FAs were well correlated (r(2)>0.999) within the range of 5- 200 microg mL(-1) for stearic acid, and 2-200 microg mL(-1) for the other FAs. The proposed method was tested on four vegetable oils, i.e., pumpkin, soybean, rice bran and palm olein oils; good agreement was found with the standard gas chromatographic (GC) method. The proposed method offers distinct advantages over the official GC method, especially in terms of simplicity, faster separation times and sensitivity.
    Matched MeSH terms: Chromatography, Reverse-Phase*
  14. Fulltext Development of HPLC Method for Quantification of Sinigrin from Raphanus sativus Roots and Evaluation of Its Anticancer Potential
    Nair AB, Gandhi D, Patel SS, Morsy MA, Gorain B, Attimarad M, et al.
    Molecules, 2020 Oct 26;25(21).
    PMID: 33114598 DOI: 10.3390/molecules25214947
    Sinigrin, a precursor of allyl isothiocyanate, present in the Raphanus sativus exhibits diverse biological activities, and has an immense role against cancer proliferation. Therefore, the objective of this study was to quantify the sinigrin in the R. sativus roots using developed and validated RP-HPLC method and further evaluated its' anticancer activity. To achieve the objective, the roots of R. sativus were lyophilized to obtain a stable powder, which were extracted and passed through an ion-exchange column to obtain sinigrin-rich fraction. The RP-HPLC method using C18 analytical column was used for chromatographic separation and quantification of sinigrin in the prepared fraction, which was attained using the mobile phase consisting of 20 mM tetrabutylammonium: acetonitrile (80:20%, v/v at pH 7.0) at a flow rate of 0.5 mL/min. The chromatographic peak for sinigrin was showed at 3.592 min for pure sinigrin, where a good linearity was achieved within the concentration range of 50 to 800 µg/mL (R2 > 0.99), with an excellent accuracy (-1.37% and -1.29%) and precision (1.43% and 0.94%), for intra and inter-day, respectively. Finally, the MTT assay was performed for the sinigrin-rich fraction using three different human cancer cell lines, viz. prostate cancer (DU-145), colon adenocarcinoma (HCT-15), and melanoma (A-375). The cell-based assays were extended to conduct apoptotic and caspase-3 activities, to determine the mechanism of action of sinigrin in the treatment of cancer. MTT assay showed IC50 values of 15.88, 21.42, and 24.58 µg/mL for DU-145, HCT-15, and A-375 cell lines, respectively. Increased cellular apoptosis and caspase-3 expression were observed with sinigrin-rich fraction, indicating significant increase in overexpression of caspase-3 in DU-145 cells. In conclusion, a simple, sensitive, fast, and accurate RP-HPLC method was developed for the estimation of sinigrin in the prepared fraction. The data observed here indicate that sinigrin can be beneficial in treating prostate cancer possibly by inducing apoptosis.
    Matched MeSH terms: Chromatography, Reverse-Phase
  15. Fulltext Effect of different drying methods on the degradation of selected flavonoids in Centella asiatica
    Mohd Zainol, M.K., Abdul-Hamid A., Abu Bakar, F., Pak Dek, S.
    International Food Research Journal, 2009;16(4):531-537.
    MyJurnal
    The effect of different drying methods on the degradation of flavonoids in Centella asiatica was evaluated. C. asiatica leaf, root and petiole were dried using air-oven, vacuum oven and freeze drier. Flavonoid was determined utilizing reverse-phase high performance liquid chromatography (RP-HPLC). Results of the study revealed the presence of high concentration of flavonoids in C. asiatica leaf, root and petiole, which include, naringin (4688.8 ± 69 μg/100 g, 3561.3 ± 205 μg/ 100 g, and 978.3 ± 96 μg/ 100 g), rutin (905.6 ± 123 μg/ 100 g, 756.07 ± 95 μg/ 100 g, and 557.25 ± 58 μg/ 100 g), quercetin (3501.1 ± 107 μg/ 100 g, 1086.31 ± 90 μg/ 100 g, and 947.63 ± 83 μg/ 100 g) and catechin (915.87 ± 6.01 μg/ 100 g, 400.6 ± 67 μg/ 100 g, and 250.56 ± 18 μg/ 100g). Luteolin, kaempferol and apigenin on the other hand, were inconsistently present in some parts of C. asiatica. Air-oven treatment resulted in the highest total flavonoids degradation followed by vacuum oven and freeze dried with percent degradation of 97%, 87.6% and 73%, respectively. Catechin and rutin were found to be the most stable flavonoids with percent degradation up to 35%, 66% and 76% for freeze dried, vacuum oven and air oven, respectively.
    Matched MeSH terms: Chromatography, Reverse-Phase
  16. 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
  17. Evaluation of dynamics of derivatization and development of RP-HPLC method for the determination of amikacin sulphate
    Shehzadi N, Hussain K, Khan MT, Salman M, Islam M
    Pak J Pharm Sci, 2017 Sep;30(5):1767-1777.
    PMID: 29084700
    The absence of chromophore and/or conjugated system, prerequisite for UV and florescent light detection, or absorbance at very low wavelength necessitates the development of simple and reliable methods for the determination of amikacin sulphate. Therefore, the present study describes for the first time dynamics of the drug derivatization using ninhydrin reagent and development and validation of a simple RP-HPLC method, using diode array detector (DAD). The variables such as heating time, heating type, drug-reagent ratio, reagent composition and storage temperature of the derivative were optimized. The analyte and aqueous ninhydrin solution upon heating for 2.00-5.00 min produced the colored drug-derivative which was stable for one month at refrigeration. The derivatized drug (20.00μL) was eluted through a column - Eclipse DB-C18 (5.00 µm, 4.60×150.00 mm), maintained at 25°C- using isocratic mobile phase comprising water and acetonitrile (70:30, v/v) at a flow rate of 1.00 mL/min, and detected at 400 nm. The method was found to be reliable (98.08-100.72% recovery), repeatable (98.02-100.72% intraday accuracy) and reproducible (98.47-101.27% inter day accuracy) with relative standard deviation less than 5%. The results of the present study indicate that the method is easy to perform, specific and sensitive, and suitable to be used for the determination of amikacin sulphate in bulk and pharmaceutical preparations using less expensive/laborious derivatization.
    Matched MeSH terms: Chromatography, Reverse-Phase/methods*; Chromatography, Reverse-Phase/standards
  18. Fulltext Formulation of chelating agent with surfactant in cloud point extraction of methylphenol in water
    Hazrina HZ, Noorashikin MS, Beh SY, Loh SH, Zain NNM
    R Soc Open Sci, 2018 Jul;5(7):180070.
    PMID: 30109066 DOI: 10.1098/rsos.180070
    Cloud point extraction (CPE) is a separation and preconcentration of non-ionic surfactant from one liquid phase to another. In this study, Sylgard 309 and three different types of additives for CPE, namely CPE-Sylgard, CPE-Sylgard-BMIMBr and CPE-Sylgard-GLDA, are investigated to extract methylphenol from water samples. The methylphenols are well separated by reversed-phase high-performance liquid chromatography (HPLC) with isocratic elution of acetonitrile : water; 60 : 40 (v/v) and detection at 260 nm. The optimized parameters for the effect of salt, surfactant, temperature, time of extraction, pH, interference study and the performance of different additives on methylphenol extraction are investigated. CPE-Sylgard-GLDA is chosen because it gives us a high peak and good peak area compared with CPE-Sylgard and CPE-Sylgard-BMIMBr. The recovery extractions of CPE-Sylgard-GLDA are obtained in the range of 80-99% as the percentage of relative standard deviation (RSD) is less than 10. The LOD and LOQ are 0.05 ppm and 0.18 ppm, respectively. The method developed for CPE-Sylgard-GLDA coupled with HPLC is feasible for the determination of methylphenol because it is simple, effective, cheap, and produces a high percentage of recovery.
    Matched MeSH terms: Chromatography, Reverse-Phase
  19. Fulltext Geraniin extracted from the rind of Nephelium lappaceum binds to dengue virus type-2 envelope protein and inhibits early stage of virus replication
    Abdul Ahmad SA, Palanisamy UD, Tejo BA, Chew MF, Tham HW, Syed Hassan S
    Virol J, 2017 11 21;14(1):229.
    PMID: 29162124 DOI: 10.1186/s12985-017-0895-1
    BACKGROUND: The rapid rise and spread in dengue cases, together with the unavailability of safe vaccines and effective antiviral drugs, warrant the need to discover and develop novel anti-dengue treatments. In this study the antiviral activity of geraniin, extracted from the rind of Nephelium lappaceum, against dengue virus type-2 (DENV-2) was investigated.

    METHODS: Geraniin was prepared from Nephelium lappaceum rind by reverse phase C-18 column chromatography. Cytotoxicity of geraniin towards Vero cells was evaluated using MTT assay while IC50 value was determined by plaque reduction assay. The mode-of-action of geraniin was characterized using the virucidal, attachment, penetration and the time-of-addition assays'. Docking experiments with geraniin molecule and the DENV envelope (E) protein was also performed. Finally, recombinant E Domain III (rE-DIII) protein was produced to physiologically test the binding of geraniin to DENV-2 E-DIII protein, through ELISA competitive binding assay.

    RESULTS: Cytotoxicity assay confirmed that geraniin was not toxic to Vero cells, even at the highest concentration tested. The compound exhibited DENV-2 plaque formation inhibition, with an IC50 of 1.75 μM. We further revealed that geraniin reduced viral infectivity and inhibited DENV-2 from attaching to the cells but had little effect on its penetration. Geraniin was observed to be most effective when added at the early stage of DENV-2 infection. Docking experiments showed that geraniin binds to DENV E protein, specifically at the DIII region, while the ELISA competitive binding assay confirmed geraniin's interaction with rE-DIII with high affinity.

    CONCLUSIONS: Geraniin from the rind of Nephelium lappaceum has antiviral activity against DENV-2. It is postulated that the compound inhibits viral attachment by binding to the E-DIII protein and interferes with the initial cell-virus interaction. Our results demonstrate that geraniin has the potential to be developed into an effective antiviral treatment, particularly for early phase dengue viral infection.

    Matched MeSH terms: Chromatography, Reverse-Phase
  20. Fulltext HMG-CoA reductase inhibitory activity and phytocomponent investigation of Basella alba leaf extract as a treatment for hypercholesterolemia
    Baskaran G, Salvamani S, Ahmad SA, Shaharuddin NA, Pattiram PD, Shukor MY
    Drug Des Devel Ther, 2015;9:509-17.
    PMID: 25609924 DOI: 10.2147/DDDT.S75056
    The enzyme 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase is the key enzyme of the mevalonate pathway that produces cholesterol. Inhibition of HMG-CoA reductase reduces cholesterol biosynthesis in the liver. Synthetic drugs, statins, are commonly used for the treatment of hypercholesterolemia. Due to the side effects of statins, natural HMG-CoA reductase inhibitors of plant origin are needed. In this study, 25 medicinal plant methanol extracts were screened for anti-HMG-CoA reductase activity. Basella alba leaf extract showed the highest inhibitory effect at about 74%. Thus, B. alba was examined in order to investigate its phytochemical components. Gas chromatography with tandem mass spectrometry and reversed phase high-performance liquid chromatography analysis revealed the presence of phenol 2,6-bis(1,1-dimethylethyl), 1-heptatriacotanol, oleic acid, eicosyl ester, naringin, apigenin, luteolin, ascorbic acid, and α-tocopherol, which have been reported to possess antihypercholesterolemic effects. Further investigation of in vivo models should be performed in order to confirm its potential as an alternative treatment for hypercholesterolemia and related cardiovascular diseases.
    Matched MeSH terms: Chromatography, Reverse-Phase
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