The increasing prevalence of penicillin-resistant Streptococuus pneumoniae urges for fast and accurate susceptibility testing methods. This study evaluated the comparability of three commonly used techniques; disk diffusion, E-test and agar dilution, to detect penicillin susceptibility in clinical isolates of S. pneumoniae. Fifty pneumococcal isolates, obtained from patients at the University of Malaya Medical Centre, were selected to include both penicillin-susceptible strains and those that had decreased susceptibility (resistant and intermediate) to penicillin. The minimum inhibitory concentration (MIC) values of penicillin to serve as the reference was determined by the agar dilution method in which, based on the MIC breakpoints recommended by the National Committee for Clinical Laboratory Standards (NCCLS), 27 strains had decreased susceptibility to penicillin with 17 strains resistant and 10 intermediate. Comparing to the agar dilution method, oxacillin disk diffusion test detected all strains with decreased penicillin susceptibility as such while E-test showed a close agreement of susceptibility (92%) of the isolates to penicillin. This confirmed that oxacillin is a good screening test for S. pneumoniae isolates with decreased susceptibility to penicillin while E-test is very reliable for rapid and accurate detection of penicillin susceptibility.
The polymerase chain reaction (PCR) is useful for amplifying specific mRNAs, particularly those present in low copy numbers. However, due to the exponential nature of the amplification process, PCR cannot readily be used to quantify gene expression. A competitive PCR technique was developed to address this shortcoming. An internal standard that is 100% homologous to, but shorter than, the target gene was constructed. The practicality of the method was demonstrated by determining the expression levels of a human transcription factor, peroxisome proliferator-activated receptor gamma 1 (hPPARgamma1) which is normally present in low copy numbers in selected cells. A mock system was used to test the accuracy and sensitivity of the method, which was subsequently used to determine the expression of this receptor in lipopolysaccharide (LPS)-activated monocytes, which are known to express hPPARgamma1 differentially during cellular activation. Densitometric analysis showed that the competitive PCR method reliably estimated the expression levels of hPPARgamma1 at the attomole (10(-18)) level in monocytes.
One of the non-destructive methods used for the identification and verification of metals is by the energy-dispersive X-ray fluorescence (EDXRF) technique. EDXRF analysis provides several important advantages such as simultaneous determination of the elements present, enable to analyse a very wide concentration range, fast analysis with no tedious sample preparation. The paper shows how this technique is developed and applied in the identification and verification of different grades of stainless steels. Comparison of the results obtained from this analysis with certified reference standards show very small differences between them.
The DCA (Drug Control Authority), Malaysia, has implemented the phase 3 registration of traditional medicines on 1 January 1992, with special emphasis on the quality, efficacy, and safety (including the presence of heavy metals) in all pharmaceutical dosage forms of traditional medicine preparations. As such, a total of 100 products in various pharmaceutical dosage forms of a herbal preparation, containing Tongkat Ali, were analyzed for mercury content using cold vapor atomic absorption spectrophotometer. Results showed that 36% of the above products possessed 0.52 to 5.30 ppm of mercury and, therefore, do not comply with the quality requirement for traditional medicines in Malaysia. Out of these 36 products, 5 products that possessed 1.05 to 4.41 ppm of mercury were in fact have already registered with the DCA, Malaysia. However, the rest of the products that contain 0.52 to 5.30 ppm of mercury still have not registered with the DCA, Malaysia. Although this study showed that only 64% of the products complied with the quality requirement for traditional medicines in Malaysia pertaining to mercury, they cannot be assumed safe from mercury contamination because of batch-to-batch inconsistency.
Crude palm oil contains 600 to 1000 ppm of tocols in the form of tocopherols and tocotrienols. These palm tocols have been isolated and analyzed in the past by various chromatographic techniques such as open column chromatography, high-performance liquid chromatography, as well as thin-layer chromatography. Supercritical fluid chromatography (SFC) has emerged as a more advanced chromatographic technique in recent years. The tocols present in palm oil are successfully isolated using SFC. Identification of these tocols is supported by various spectroscopic techniques such as 1H NMR, 13C NMR, and mass spectrometry.
A selective and sensitive reversed-phase (RP) high-performance liquid chromatographic method is developed for the quantitative analysis of five naturally occurring flavonoids of Blumea balsamifera DC, namely dihydroquercetin-7,4'-dimethyl ether (DQDE), blumeatin (BL), quercetin (QN), 5,7,3',5'-tetrahydroxyflavanone (THFE), and dihydroquercetin-4'-methyl ether (DQME). These compounds have been isolated using various chromatographic methods. The five compounds are completely separated within 35 min using an RP C18, Nucleosil column and with an isocratic methanol-0.5% phosphoric acid (50:50, v/v) mobile phase at the flow rate of 0.9 mL/min. The separation of the compounds is monitored at 285 nm using UV detection. Identifications of specific flavonoids are made by comparing their retention times with those of the standards. Reproducibility of the method is good, with coefficients of variation of 1.48% for DQME, 2.25% for THFE, 2.31% for QN, 2.23% for DQDE, and 1.51% for BL. The average recoveries of pure flavonoids upon addition to lyophilized powder and subsequent extraction are 99.8% for DQME, 99.9% for THFE, 100.0% for BL, 100.6% for DQDE, and 97.4% for QN.
The paper relates a study on the development of an analysis procedure for measuring the gold coating thickness using EDXRF technique. Gold coating thickness was measured by relating the counts under the Au Lα peak its thickness value. In order to get a reasonably accurate result, a calibration graph was plotted using five gold-coated reference standards of different thicknesses. The calibration graph shows a straight line for thin coating measurement until 0.9μm. Beyond this the relationship was not linear and this may be resulted from the selfabsorption effect. Quantitative analysis was also performed on two different samples of goldcoated jewelry and a phone connector. Result from the phone connector analysis seems to agree with the manufacturer’s gold coating value. From the analysis of gold-coated jewelry it had been able to differentiate the two articles as gold wash and gold electroplated.
Single and double dichotic digit tests in Malay language were developed and standardized as an initial attempt to incorporate tests of auditory processing within the scope of audiology practice in Malaysia. Normative data under free recall, directed right-ear first, and directed left-ear first listening conditions were determined using 120 Malay children between the ages of 6 and 11 years old with normal hearing and normal academic performance. Test-retest reliability was assessed in 15 of the study subjects. In general, the double dichotic digit test produced greater differences in scores between age groups, and a greater right-ear advantage than the single dichotic digit test. In addition, the double dichotic digit test also had higher test-retest reliability. These findings suggest the double dichotic digit test is more clinically applicable.
In this study, the development and validation of a high-performance liquid chromatography (HPLC) assay for determination of repaglinide concentration in human plasma for pharmacokinetic studies is described. Plasma samples containing repaglinide and an internal standard, indomethacin were extracted with ethylacetate at pH 7.4. The recovery of repaglinide was 92%+/-55.31. Chromatographic separations were performed on Purospher STAR C-18 analytical column (4.8 mm x 150 mm; 5 microm particle size). The mobile phase composed of acetonitrile-ammonium formate (pH 2.7; 0.01 M) (60:40, v/v). The flow rate was 1 ml/min. The retention time for repaglinide and indomethacin were approximately 6.2 and 5.3 min, respectively. Calibration curves of repaglinide were linear in the concentration range of 20-200 ng/ml in plasma. The limits of detection and quantification were 10 ng/ml and 20 ng/ml, respectively. The inter-day precision was from 5.21 to 11.84% and the intra-day precision ranged from 3.90 to 6.67%. The inter-day accuracy ranged 89.95 to 105.75% and intra-day accuracy ranged from 92.37 to 104.66%. This method was applied to determine repaglinide concentration in human plasma samples for a pharmacokinetic study.
This study is a result of an analysis of free and conjugated phytoestrogens daidzein, genistein, daidzin, genistin and coumesterol in human cord blood plasma using LCMS. Cord blood was collected from urban and rural populations of Malaysia (n=300) to establish a simple preliminary database on the levels of the analyzed compounds in the collected samples. The study also aimed to look at the levels of phytoestrogens in babies during birth as this may have a profound effect on the developmental process. The sample clean up was carried out by solid-phase extraction using C18 column and passed through DEAE sephadex gel before analysis by LCMS. The mean concentrations of total phytoestrogens were daidzein (1.4+/-2.9 ng/ml), genistein (3.7+/-2.8 ng/ml), daidzin (3.5+/-3.1 ng/ml), genistin (19.5+/-4.2 ng/ml) and coumesterol (3.3+/-3.3 ng/ml). Distribution of phytoestrogen was found to be higher in samples collected from rural areas compared to that of urban areas.
To explore how many millimeters of tooth size discrepancy (TSD) are clinically significant, to determine what percentage of a representative orthodontic population has such a tooth size discrepancy, and to determine the ability of simple visual inspection to detect such a discrepancy.
Chitosan (beta-1, 4-D-glucosamine) is a deacetylated form of chitin with excellent biological properties in wound management. The natural properties of chitosan have the physical and chemical limitations to be widely used in biomedical fields. The improvement of the physical and chemical properties of chitosan with some additional chemicals will alter its biocompatibility. Therefore, the biological attribute of the modified chitosan must be evaluated. In this study, the cytotoxicity of oligo-chitosan (OC) and N, O- carboxymethyl-chitosan (NO-CMC) derivatives (O-C 1%, O-C 5%, NO-CMC 1% and NO-CMC 5%) was evaluated using primary normal human epidermal keratinocyte (pNHEK) cultures as an in vitro toxicology model at standardized cell passages (fourth passages). 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl tetrazolium bromide (MTT) was used as a cell viability assay. The O-C 1% is one of the most compatible chitosan derivatives because it steadily sustained >70% of viable cells until 72 hr post-treatment. This was followed by O-C 5%, NO-CMC 5% and NO-CMC 1%. Therefore, oligo-chitosan had the ideal properties of a biocompatible material compared to N, O- carboxymethyl-chitosan in this study.
The Drug Control Authority (DCA) of Malaysia implemented the phase three registration of traditional medicines on 1 January, 1992. A total of 100 products in various pharmaceutical dosage forms of a herbal preparation, containing Eugenia dyeriana, either single or combined preparations (more than one medicinal plant), were analyzed for the presence of lead contamination, using atomic absorption spectrophotometry. These samples were bought from different commercial sources in the Malaysian market, after performing a simple random sampling. Results showed that 22% of the above products failed to comply with the quality requirement for traditional medicines in Malaysia. Although this study showed that 78% of the products fully complied with the quality requirement for traditional medicines in Malaysia pertaining to lead, however, they cannot be assumed safe from lead contamination because of batch-to-batch inconsistency.
A simple high-performance liquid chromatography (HPLC) method to determine the content of betulinic acid (BA) in rat plasma collected at different times (0-8 h) after oral administration of Orthosiphon stamineus leaf extract was developed. The features of the assay include protein precipitation using acetonitrile and isocratic elution using reverse phase C-18 column with ultraviolet (UV) detection. The recovery of BA from plasma varied from 98.4 to 102.5%. The R.S.D of intra- and inter-day precision from rat plasma ranged from 4.2 to 9.8%. The maximum concentration of BA in the plasma was 1.2 +/- 0.3 microg/ml at 1 h after oral administration of the extract.
A simple high-performance liquid chromatography (HPLC) method was developed to determine the content of andrographolide (AP) and 14-deoxy-11,12-dideoxyandrographolide (DIAP) in a pooled urine of rat obtained within 24h after an oral dose of Andrographis paniculata leaf extract at 1g/kg body weight. Cumulative urinary excretion of AP and DIAP in 24h after oral administration of the extract was 0.88% and 1.61% of oral dose administered, respectively. The extract showed significant reduction (p<0.05) of MDA levels and elevation of total antioxidant status in rat urine samples collected in 24 after oral administration.
Characterisation of fatty acids composition of three palm-based reference materials was carried out through inter-laboratory proficiency tests. Twelve laboratories collaborated in these tests and the fatty acids compositions of palm oil, palm olein and palm stearin were determined by applying the MPOB Test Methods p3.4:2004 and p3.5:2004. Determination of consensus values and their uncertainties were based on the acceptable statistical agreement of results obtained from the collaborating laboratories. The consensus values and uncertainties (%) for each palm oil reference material produced are listed as follows : 0.20% (C12:0), 1.66+/-0.05% (C14:0), 43.39+/-0.39% (C16:0), 0.14+/-0.06% (C16:1), 3.90+/-0.11% (C18:0), 40.95+/-0.23% (C18:1), 9.68+/-0.21% (C18:2), 0.16+/-0.07% (C18:3) and 0.31+/-0.08% (C20:0) for fatty acids composition of palm oil; 0.23+/-0.04% (C12:0), 1.02+/-0.04% (C14:0), 39.66+/-0.19% (C16:0), 0.18+/-0.07% (C16:1), 3.81+/-0.04% (C18:0), 44.01+/-0.08% (C18:1), 10.73+/-0.08% (C18:2), 0.20+/-0.06% (C18:3) and 0.34+/-0.04% (C20:0) for fatty acids composition of palm olein; and 0.20% (C12:0), 1.14+/-0.05% (C14:0), 49.42+/-0.25% (C16:0), 0.16+/-0.08% (C16:1), 4.15+/-0.10% (C18:0), 36.14+/-0.77% (C18:1), 7.95+/-0.29% (C18:2), 0.11+/-0.07% (C18:3) and 0.30+/-0.08% (C20:0) for fatty acids composition of palm stearin.
Solid-phase microextraction (SPME) coupled to gas chromatography has been applied for the headspace analysis (HS) of 12 target flavour compounds in a model orange beverage emulsion. The main volatile flavour compounds studied were: acetaldehyde, ethyl acetate, alpha-pinene, ethyl butyrate, beta-pinene, myrcene, limonene, gamma-terpinene, octanal, decanal, linalool and citral (neral plus geranial). After screening the fibre type, the effect of other HS-SPME variables such as adsorption temperature (25-55 degrees C), extraction time (10-40 min), sample concentration (1-100% w/w), sample amount (5-10 g) and salt amount (0-30% w/w) were determined using a two-level fractional factorial design (2(5-2)) that was expanded further to a central composite design. It was found that an extraction process using a carboxen-polydimethylsiloxane fibre coating at 15 masculineC for 50 min with 5 g of diluted emulsion 1% (w/w) and 30% (w/w) of sodium chloride under stirring mode resulted in the highest HS extraction efficiency. For all volatile flavour compounds, the linearity values were accurate in the concentration ranges studied (r(2) > 0.97). Average recoveries that ranged from 90.3 to 124.8% showed a good accuracy for the optimised method. The relative standard deviation for six replicates of all volatile flavour compounds was found to be less than 15%. For all volatile flavour compounds, the limit of detection ranged from 0.20 to 1.69 mg/L.
Extending the frying-life of oils is of commercial and economic importance. Due to this fact, assessment on the thermal stability of frying oils could provide considerable savings to the food processors. In this study, the physico-chemical properties of five palm products mainly palm oil, single-fractionated palm olein, double-fractionated palm olein, red palm olein and palm-based shortening during 80 hours of heating at 180 degrees C were investigated. Heating properties of these products were then compared with that of high oleic sunflower oil, which was used as reference oil. The indices applied in evaluating the quality changes of oils were free fatty acid, smoke point, p-anisidine value, tocols, polar and polymer compounds. Three palm products i.e. palm oil, single-fractionated palm olein and double-fractionated palm olein were identified to be the most stable in terms of lower formation of free fatty acid, polar and polymer compounds as well as preserving higher smoke point and tocols content compared to the other three oils. The low intensity of hydrolytic and oxidative changes due to prolonged heating, suggests that these palm products are inherently suitable for frying purposes.
An instrumental neutron activation analytical (INAA) technique is used for the determination of thirty elements in five coal samples collected from Kapar power station, imported from Indonesia and Australia. Analyses of the samples are being associated with standards. All irradiations were performed in the nuclear reactor of Malaysia Nuclear Agency (MNA). Samples were counted by Hyper Pure Germanium (HPGe) detector for short period irradiations at MNA, while for the long period irradiations the samples were counted at Universiti Kebangsaan Malaysia (UKM). The concentrations of thirty elements have been determined: The major components are Cl, Ca, Mg, K, Fe, Ti and Na with the mean concentrations in the range between 70±69 ppm- 6100±1639 ppm; and the trace elements are Zr, V, Mn , Sc, Cr, Co, As ,Br ,Rb ,Sb ,Ba , La, Ce, Nd, Sm, Eu, Tb, Yb, Lu, Hf, Th, U and Ta with the mean concentrations in the range between 0.1381±0.0202 - 69.0±2.8 ppm. The results have been compared to the reported data of eight coal samples from the United States and the reported data of Australian bituminous coal.
During the years 1985-2008, the Secondary Standards Dosimetry Laboratory of Malaysia (SSDL Malaysia) has participated 37 times in the IAEA/WHO intercomparison programmes. This paper reports an analysis of the intercomparison data and demonstrates that the quality of the SSDL calibration service is well within the limits required by IAEA.