• CELLMED
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• v.11 no.3
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• pp.13.1-13.9
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• 2021

Background and Objective: Nux-vomica based traditional Unani formulation, Habb-e-Azaraqi (HAZ) is an important drug used by Unani physicians since several decades. It possesses Muqawwi-i-A'sab (nervine tonic), Muharrik-i-A'sab (nervine stimulant) properties and is an effective treatment option for diseases like Laqwa (facial palsy), Falij (paralysis), Niqris (gout) and Waja'al-Mafasil (arthritis) etc. The aim of the study is to access and provide information of HAZ for its TLC, HPTLC Fingerprinting defining its clear qualitative perspective and acute oral toxicity evaluation for its safety assessment which was not done earlier, thus contributing in the field of research. Materials and Methods: The chief ingredient, nux-vomica was detoxified as per method mentioned in Unani Pharmacopeia before its use in formulation. TLC and HPTLC was developed under four detection system i.e., UV 366nm, UV 254nm, exposure to iodine vapours and after derivatization with anisaldehyde sulphuric acid. Acute toxicity studies were performed as per OECD Guidelines 425 at a limit dose of 2000 mg/kg. Observations were done for signs of toxicity, body weight, and feed consumption at regular intervals followed by haematological and biochemistry evaluation. Results: The generated data proved the authenticity and established the TLC and HPTLC profile of the formulation. Acute toxicity revealed no significant differences in HAZ-treated animals with respect to body weight gain, feed consumption, haematology, clinical biochemistry evaluation. No significant gross pathological observation was noticed in necropsy. Conclusion: Data of the present study is substantial and scientific proof of HAZ in terms of standardization and toxicity study that can be utilize in future research activities.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.559-565
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• 2021

This study aimed to optimize gamma-aminobutyric acid (GABA) production by employing five strains of lactic acid bacteria (LAB) that were capable of high cell growth and GABA production using a modified synthetic medium. GABA production in the strains was qualitatively confirmed via detection of colored spots using thin layer chromatography. Lactobacillus plantarum SGL058 and Lactococcus lactis SGL027 were selected as the suitable strains for GABA production. The conditions of the carbon and nitrogen sources were determined as 5 g/l glucose (L. plantarum SGL058), 5 g/l lactose (L. lactis SGL027), 10 g/l yeast extract (L. plantarum SGL058), and 20 g/l yeast extract (L. lactis SGL027) for GABA production. The cell growth, monitored by optical density at 600 nm, was 5.93 for L. plantarum SGL058. This value was higher than the 3.04 produced by L. lactis SGL027 at 36 h using a 5-L fermenter. The highest concentration of GABA produced was 546.7 ㎍/ml by L. plantarum SGL058 and 404.6 ㎍/ml by L. lactis SGL027, representing a GABA conversion efficiency of (%, w/w) of 4.0% and 3.4%, respectively. The fermentation profiles of L. plantarum SGL058 and L. lactis SGL027 provide a basis for the utilization of LAB in GABA production using a basal synthetic medium.

• Analytical Science and Technology
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• v.19 no.1
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• pp.107-114
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• 2006

Early diagnosis and medical intervention are critical for the treatment of patients with metabolic disorders. A rapid analytical method was developed for simultaneous quantification of organic acids and amino acids in urine without labor-intensive pre-extraction procedure showing high sensitivity and specificity. A new method consisted of simple two-step trimethylsilyl (TMS)-trifluoroacetyl (TFA) derivatization using GC/MS-selective ion monitoring (SIM). Filter paper urine specimens were dried under nitrogen after being fortified with internal standard (tropate) in a mixture of distilled water and methanol. Methyl orange was added to the residue as indicator reagent. Silyl derivative of carboxylic functional group was followed by trifluoroacetyl derivative for amino functional group. N-methyl-N-(trimethylsilyl-trifluoroacetamide) and N-methyl-bistrifluoroacetamide were consecutively added and heated for 15-20 min at $65^{\circ}C-70^{\circ}C$, for TMS-TFA derivative, respectively. This reactant was analyzed by GC/MS-SIM. Linear dynamic range showed 0.001-50 mg with the detection limit of (S/N=3) 10-200 ng, and the quantification limit of 80-900 ng in urine. Correlation coefficient of regression line was 0.994-0.998. When the method was applied to the patients 'urine, it clearly differentiated the normal from the patient with metabolic disorder. The study showed that the developed method could be the method of choices in rapid and sensitive screening for organic aciduria and amino acidopathy.

• Analytical Science and Technology
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• v.22 no.5
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• pp.407-414
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• 2009

A simultaneous determination method for cholesterol, lanosterol and desmosterol was developed using gas chromatography/mass spectrometry. Urine was enzymatically hydrolyzed with ${\beta}$-glucuronidase/arylsulfatase. Samples were prepared using extractions with a mixture of ethyl acetate-hexane (2:3, v/v), followed by derivatization with a mixture of MSTFA/TMSI/TMCS (100:2:5 v/v/v). All analyses were performed using GC/MS in selective ion monitoring mode. Good linearities ($r^2=0.998{\sim}0.999$) in calibration curve and a satisfactory recovery (80.0%~113%) were achieved. Accuracy and precision values within ${\pm}15%$ in the concentration range of 5 to 200 ng/mL were also observed for all compounds. The developed method was applied to pravastatin-treated (70 and 250 mg/kg/day for 7 days, oral) hyperlipidemia rats. Those sterols were significantly lower in drug-treated rats compared to the controls, which justifies the drug efficacy. Therefore, these results indicate that the developed method was successfully applied to examine statin drug efficacy with urine sample.

• Analytical Science and Technology
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• v.28 no.5
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• pp.323-330
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• 2015

Alkaptonuria, a rare inherited metabolic disease, is characterized by a lack of homogentisate dioxygenase and accumulation of homogentisic acid (HGA), leading to homogentisic aciduria, arthritis, and ochronosis. In this study, a rapid analytical method, without an expensive and tedious solid phase extraction step, was developed to quantify HGA in plasma using GC-MS. HGA-spiked pooled plasma samples were subjected to liquid-liquid extraction (LLE) with ethyl acetate, followed by trimethylsilyl derivatization (TMS) and GC-MS quantification using selected ion monitoring. The formation of TMS derivative of the 1 carboxylic and 2 hydroxyl functional groups was performed by reacting BSTFA (with 10% TMCS) for 5 min at 80 ℃. For selected ion monitoring, quantification and confirmation ions were determined based on specific ions (m/z 384, m/z 341 and m/z 252) of the TMS derivative of HGA. Calibration curves of pooled normal plasma specimens showed a linear relationship in the range of 1-100 ng/µL. The precision and accuracy were within a relative standard deviation (RSD) of 1 to 15% and a bias of -5 to 25%. Recoveries were obtained in the range of 99-125% and 95-115% for intra-day and inter-day assay, respectively, at 2, 20 and 80 ng/µL. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/µL and 4 ng/µL, respectively. No homogentisic acid was excreted from normal Korean plasma samples. Collectively, the results from the present study suggest that this method could be useful for routine diagnosis and therapeutic monitoring of alkaptonuria patients with excellent sensitivity and rapidity.

• Analytical Science and Technology
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• v.24 no.2
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• pp.69-77
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• 2011

Ethylene glycol (EG) is produced commercially in large amounts and is widely used as antifreeze or deicing solution for cars, boats, and aircraft. EG poisoning occurs in suicide attempts and infrequently, either intentionally through misuse or accidental as EG has a sweet taste. EG has in itself a low toxicity, but is in vivo broken down to higher toxic organic acids which are responsible for extensive cellular damage in various tissues caused principally by the metabolites glycolic acid and oxalic acid. The most conclusive analytical method of diagnosing EG poisoning is determination of EG concentration. However, victims are sometimes admitted at a late stage to hospitals or died during emergency treatment like a gastric lavage or found rotten dead, when blood EG concentrations are low or not detected. Therefore, in this study, the identification of EG was not only performed by gas chromatograpyc-mass spectrometry (GC-MS) following derivatization but also further toxicological analyses of metabolites, glycolic acid (GA) and oxalic acid (OA), were performed by ion chromatography in various biological specimens. A ranges of blood concentrations (3 cases) was $10\sim2,400\;{\mu}g/mL$ for EG, $224\sim1,164\;{\mu}g/mL$ for GA and ND $\sim40\;{\mu}g/mL$ for OA, respectively, In other biological specimens (liver, kidney, bile and pleural fluid), a range of concentrations (3 cases) was ND $\sim55,000\;{\mu}g/mL$ for EG, ND $\sim1,124\;{\mu}g/mL$ for GA and ND $\sim60\;{\mu}g/mL$ for OA, respectively. Liver and kidney tissues were recommended specimens including blood because OA, a final metabolite of EG, was identified large amounts in these despite no detectable EG caused by some therapy.

• The Korean Journal of Pharmacology
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• v.24 no.2
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• pp.233-240
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• 1988

A simple and selective isocratic HPLC method for the analysis of tissue polyamine contents is described and applied on the study of the changes of the hepatic polyamine contents after partial hepatectomy in male rats. The hepatic polyamines are extracted with 0.4 M perchloric acid containing 2 mM disodium EDTA, and then the extract is redissolved in 100 ul of 1 M sodium carbonate and incubated with 300 ul of FNBT-dimethylsulfoxide (1: 100) mixture. The N-2'-nitro-4'-trifluoromethylphenyl drivatives of polyamines are separated through a ERC-ODS column in an isocratic mode with an acetonitrile-water (80:20) mobile phase within 20 min. per a sample, while monitoring the effluent at 242 nm. This improved method which could detect subnanogram of each polyamines is highly specific and reproducible as evidenced by the application of it on the study of the changes of polyamine contents in the regenerating rat liver after partial hepatectomy.

• Korean Journal of Weed Science
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• v.30 no.4
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• pp.361-370
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• 2010

This study was carried out to establish the analytical method of MCPA residue in brown rice and rice straw by HPLC/UVD. When MCPA was extracted from sample under the pH 3.6 by adding acetone 200 mL and 1N-HCl 100 mL, the extraction efficiency was high by 87%. And purification efficiency was high by 83% when 5 mL of 1% methanol/acetonitrile was eluated by the florisil Sep-pak cartridge column. From spiking of $0.1{\mu}g\;mL^{-1}$ and $0.25{\mu}g\;mL^{-1}$ of MCPA to control sample, respectively, average recovery rate of MCPA in brown rice was 96.0% and 94.9% and that in rice straw was 92.5% and 88.2%, respectively. Precision of experiment was very high by relative standard deviation of 1.5% to 5.7%. In brown rice and rice straw treated with bentazone+MCPA (11+1.2%) of 30 kg and 60 kg per ha at 30 days after rice transplanting, respectively, maximum residue limit was under $0.05{\mu}g\;mL^{-1}$ of the recommended rate of Korean Food and Drug Administration. From the above results, the analytical procedure of MCPA in plants such as hydrolysis, saponification and derivatization were ommited, and retention time was faster and recovery rate was higher than the existed results of HPLC/UVD. Therefore, these results were greatly improved and seemed to be usefully applied for residue analysis of MCPA in plants.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.424-434
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• 2011

The increase in the consumption of herb medicines have made their use a public health problem due to the potential fungal contamination and the risk of the presence of my cot ox ins. 360 samples of herb medicines were evaluated for the aflatoxin contamination. The natural occurrence of aflatoxins in these samples were determined using immunoaffinity column clean up and high performance liquid chromatography (HPLC) with post-column derivatization. For samples analyzed, mean levels (incidence) of AFB1, AFB2, AFG1 and AFG2 in positive samples were $1.4\;{\mu}g/kg$(46.4%), $0.4\;{\mu}g/kg$(25.4%), $1.1\;{\mu}g/kg$(37.8%) and $0.9\;{\mu}g/kg$(24.3%), respectively. Recoveries of the full analytical procedure were 71.7~99.7% for AFB1, 88.1~99.2% for AFB2, 82.8~95.5% for AFG1 and 77.9~90.0% for AFG2. The excess cancer risk estimated using the cancer potency of aflatoxin B1 (7 $(mg/kg/day)^{-1}$ for $HBsAg^-$ and 230 $(mg/kg/day)^{-1}$ for $HBsAg^+$) were $1.30{\times}10^{-5}{\sim}1.22{\times}10^{-7}$ for hepatits B surface antigen negative ($HBsAg^-$) and $3.31{\times}10^{-4}{\sim}3.12{\times}10^{-6}$ for hepatits B surface antigen positive ($HBsAg^+$) respectively. In conclusion, although the contamination levels of samples used in the study were low, further actions are also required to undertake a program of herbal surveys in order to access mycotoxin contamination overall so that the safety of public will be protected.

• Journal of Applied Biological Chemistry
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• v.60 no.3
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• pp.283-291
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• 2017

Rice that the half of population in the world eats as a staple food is mostly produced and consumed in Asia. However, its consumption is nowadays decreasing mainly due to diet diversity. Accordingly, some attempts are in demand to enhance the utilization of rice. In this study, profiling of volatile and non-volatile flavor components in rice pastes obtained by ${\alpha}$-amylase was performed and compared according to nine different rice cultivars domestically cultivated in Korea using gas chromatography-mass spectrometry combined by solid phase microextraction and gas chromatography-time of flight-mass spectrometry after a derivatization, respectively. In total, 46 volatile compounds identified included 6 alcohols, 6 aldehydes, 4 esters, 4 furan derivatives, 4 ketones, 1 acid, 1 sulfur-containing compound, 7 hydrocarbons, 5 aromatics and 8 terpenes. The non-volatile flavor components found were composed of 12 amino acids, 6 sugars and 4 sugar alcohols. In principal component analysis, rice paste samples could be discriminated according to cultivars on the score plots of volatile and non-volatile flavor compounds. In particular, some volatile compounds such as pentanal and 4,7-dimethylundecane could contribute to distinguish Senong 17 white and Senong 17 brown, whereas ethanol, 6-methylhep-5-en-2-one, and tridecane could be highly related to the discrimination of Iipum from other cultivars. Among non-volatile compounds, some amino acids such as glycine, serine and ${\gamma}$-aminobutyric acid and some sugars such as sucrose and fructose were mainly responsible for the discrimination of Danmi from the other cultivars. On the other hand, galactose, arabitol and mannose were more closely related to Senong 17 white than Senong 17 brown.