Mycolic acids (MAs), i.e. 2-alkyl, 3-hydroxy long-chain fatty acids, are the hallmark of the cell envelope of Mycobacterium tuberculosis and are related with antibiotic resistance and host immune escape. Nowadays, they've become hot target of new anti-tuberculosis drugs. There are two main methods to detect MAs, ¹⁴C metabolic labeling thin-layer chromatography (TLC) and liquid chromatograph mass spectrometer (LC-MS). However, the user qualification of ¹⁴C or the lack of standards for LC-MS hampered the easy use of this method. TLC is a common way to analyze chemical substance and can be used to analyze MAs. In this study, we used tetrabutylammonium hydroxide and methyl iodide to hydrolyze and formylate MAs from mycobacterium cell wall. Subsequently, we used diethyl ether to extract methyl mycolate. By this method, we can easily extract and analyze MA in regular biological labs. The results demonstrated that this method could be used to compare MAs of different mycobacterium in different growth phases, MAs of mycobacteria treated by anti-tuberculosis drugs or MAs of mycobacterium mutants. Therefore, we can use this method as an initial validation for the changes of MAs in researches such as new drug screening without using radioisotope or when the standards are not available.
Mycolic Acids/metabolism* ; Chromatography, Thin Layer ; Mycobacterium tuberculosis ; Fatty Acids ; Antitubercular Agents/pharmacology*

Eleven compounds were isolated from the 95% ethanol extract of the stems of Dendrobium officinale after water extraction by various modern chromatographic techniques, such as silica gel column chromatography(CC), octadecyl-silica(ODS) CC, Sephadex LH-20 CC, preparative thin layer chromatography(PTLC) and preparative high performance liquid chromatography(PHPLC). According to spectroscopic analyses(MS, 1D-NMR, 2D-NMR) combined with optical rotation data and calculated electronic circular dichroism(ECD), their structures were identified as dendrocandin Y(1), 4,4'-dihydroxybibenzyl(2), 3-hydroxy-4',5-dimethoxybibenzyl(3), 3,3'-dihydroxy-5-methoxybibenzyl(4), 3-hydroxy-3',4',5-trimethoxybibenzyl(5), crepidatin(6), alternariol(7), 4-hydroxy-3-methoxypropiophenone(8), 3-hydroxy-4,5-dimethoxypropiophenone(9), auriculatum A(10) and hyperalcohol(11). Among them, compound 1 was a new bibenzyl derivative; compounds 2 and 7-11 have not been previously reported from Dendrobium plants; compound 6 was reported from D.officinale for the first time. Compounds 3-6 exhibited potent antioxidant activity with IC_(50) values of 3.11-9.05 μmol·L~(-1) in ABTS radical scavenging assay. Compound 4 showed significant inhibitory effect on α-glucosidase, with IC_(50) value of 17.42 μmol·L~(-1), indicating that it boasted hypoglycemic activity.
Dendrobium ; Biological Assay ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Bibenzyls

Bile acids facilitate the absorption of lipids, and affect the development of various diseases by regulating intestinal flora structure and modulating immunity and metabolism. It is therefore important to quantitatively detect bile acids. Current analytical methods are still immature due to constituent complexity, structural heterogeneity and bioactive variability of bile acids. Detection of individual bile acids is of significance for pharmacological research, clinical diagnosis and disease prevention. Advances have been made in bile acid analysis from multiple sources including serum, bile, urine and feces, although several limitations still exist for bile acid quantification. Here we review research progress in conventional bile acid assays, including spectrophotometry, thin-layer chromatography, liquid/gas chromatography and liquid/gas chromatography-mass spectrometry. Moreover, we emphasize the development of bile acid biosensors that may have promising prospects.
Bile ; Bile Acids and Salts ; Biosensing Techniques ; Chromatography, Thin Layer ; Gas Chromatography-Mass Spectrometry

The fingerprint technology could reflect the internal chemical characteristics of Chinese herbal medicine or preparation, which has the characteristics of "wholeness" and "fuzziness". It is suitable for evaluating the quality of intermediate and finished products in the production process of traditional Chinese medicine formula granules. In this paper, the applications of high performance liquid chromatography (HPLC), thin layer chromatography (TLC), gas chromatography (GC) and infrared spectrum (IR) fingerprint technology in the quality control of traditional Chinese medicine formula granules were reviewed, and their advantages and disadvantages were analyzed. The aim of this article is to enhance the combined application of various fingerprint technologies in traditional Chinese medicine formula granules. It could provide technical reference for realizing the stability of production process and improving the overall quality of formula granules.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; standards ; Medicine, Chinese Traditional ; Quality Control

OBJECTIVETo test the in vitro antimicrobial efficacy of water and methanol extracts of 23 plant species that are commonly used in Libyan folk medicine.

METHODSThe antimicrobial activity was determined using the well-diffusion method. Four test microorganisms were used namely, Escherichia coli, Salmonella species, Staphylococcus aureus and Bacillus subtilis. The minimum inhibitory concentration (MIC) was determined for the high biologically active crude plant extracts.

RESULTSAmong 23 medicinal plants used in the study, only 5 methanolic extracts [Rosmarinus offcinalis L., Carduus marianium L., Lantana camara L., Rhus tripartite (ueria) Grande, and Thymus capitatus (L.) Hoffm (link)] showed the highest antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Salmonella species, while 22 methanolic and aqueous extracts showed moderate to weak antimicrobial activity on all tested organisms. However 19 of the extracts showed no activity at all against Gram-ve and Gram +ve microorganisms. MIC was found to be 1.25 mg/mL (Thymus capitatus), 3 mg/mL (Rhus tripartite), 4 mg/mL (Carduus marianium), 5 mg/mL (Rosamarinus officinalis) and 5 mg/mL (Lantana camara), respectively.

CONCLUSIONSThe present results revealed that, crude methanolic extracts of the investigated Libyan folk medicinal plants exhibited mild to high in vitro antibacterial activities against Gram-positive and Gram-negative microorganisms.

Anti-Infective Agents ; pharmacology ; Bacteria ; drug effects ; Chromatography, Thin Layer ; Libya ; Microbial Sensitivity Tests ; Plant Extracts ; pharmacology ; Plants, Medicinal ; chemistry

OBJECTIVES: This study aims to identify and partially purify antibacterial compounds against Streptococcus mutans from Galla Rhois extract. METHODS: Galla Rhois was extracted with n-hexane or ethanol and concentrated in a rotary evaporator. The antibacterial effect of the Galla Rhois extract against S. mutans was determined by the paper discdiffusion method with n-hexane, ethanol, methanol, ethyl acetate, dimethyl sulfoxide (DMSO), acetone, and distilled water as the solvents. The active compounds were purified by partition chromatography, thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC). RESULTS: The antibacterial effect of the n-hexane extract was more effective against S. mutans than the ethanol extract (P<0.05). The antibacterial component of Galla Rhois was partially purified using partition chromatography and HPLC, and the antibacterial activity was confirmed. CONCLUSIONS: The partially purified component of Galla Rhois showed strong antibacterial effect against S. mutans. These results confirm that the antibacterial compounds of Galla Rhois can be used for the prevention of dental caries.
Acetone ; Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Chromatography, Thin Layer ; Dental Caries ; Dimethyl Sulfoxide ; Ethanol ; Methanol ; Solvents ; Streptococcus mutans* ; Streptococcus* ; Water

The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through α-amylase and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in α-amylase and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that α-amylase activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.
Aflatoxins ; Alcoholic Beverages ; Amylases ; Asia ; Aspergillus* ; Calmodulin ; Chromatography, Thin Layer ; DNA, Intergenic ; Fermentation* ; Glucan 1,4-alpha-Glucosidase ; Mass Screening ; Polymerase Chain Reaction ; Triticum

OBJECTIVETo study and compare the conventional extraction procedure with microwave assisted extraction (MAE) for some Ayurvedic Rasayana drugs and to evaluate their antioxidant potential and carry out the characterization of extracts by thin layer chromatography.

METHODSThree Ayurvedic rasayana plants Allium sativum Linn., Bombax ceiba Linn. and Inula racemosa Hook. were evaluated for an improved MAE methodology by determining the effects of grinding degree, extraction solvent, effect of dielectric constant and duration of time on the extractive value. Antioxidant potential of all three drugs was evaluated with 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power was determined by using Gallic acid as standard. Further thin layer chromatographic (TLC) analysis was performed on pre-activated Silica Gel G plates and Rf value were compared with those reported for the important biomarkers.

RESULTSThe total extractive value for Allium sativum Linn. was 36.95% (w/w) and 49.95% (w/w) for ethanol extraction respectively. In case of Bombax ceiba Linn. the yield of aqueous extract by MAE was 50% (w/w) compared to 42% (w/w) in ethanol (50% v/v). Percent yield of Inula racemosa Hook. in aqueous extract was found to be 27.55% (w/w) which was better than ethanol extract (50%) where the yield was 25.95% (w/w). Upon antioxidant activity evaluation. sativum extract showed an absorbance of 0.980±0.92 at concentration of 500 μg with maximum reducing capacity. This was followed by. ceiba Linn. 0.825±0.98 and. racemosa Hook. with 0.799±2.01 at a concentration of 500 μg. TLC based standardization of. sativum Linn. extract shows single spot with Rf value of 0.38, B. ceiba Linn. extract shows Rf values were 0.23, 0.58, 0.77, 0.92 and I. racemosa Hook. extract spot had a Rf value of 0.72.

CONCLUSIONSA significant improvement in extractive values was observed as a factor of time and other advantages by using MAE technology. All three drugs have high antioxidant potential and a TLC profiling similar to reported ones. The presence of fructan type polysaccharide can be further utilized for bioactivity directed fractionation and evaluation of immunomodulatory activity.

Antioxidants ; pharmacology ; Biphenyl Compounds ; chemistry ; Chromatography, Thin Layer ; methods ; Ethanol ; chemistry ; Free Radical Scavengers ; pharmacology ; Inhibitory Concentration 50 ; Medicine, Ayurvedic ; Microwaves ; Oxidation-Reduction ; drug effects ; Pharmaceutical Preparations ; isolation & purification ; Picrates ; chemistry ; Plants, Medicinal ; chemistry ; Solvents ; Time Factors

Tibetan medicine Asteris Flos is the flowers of Aster souliei, A. flaccidus, and A. asteroides, with the function of clearing away heat and toxic matter, relieving cough, and removing phlegm. In order to control the quality of Asteris Flos, the morphological and chemical methods were established for identification of three origins. The morphological features of three species were described and photographed, and the microscopic characteristics of three drug powders were also described in detail and pictured. The results showed that three origins of Asteris Flos could be easily distinguished by their macro- and micro-morphologic features, and a key for distinguishing the three origins was given. Moreover, a TLC method, with apigenin-7-O-β-D-glucurono pyranoside and chlorogenic acid as chemical references, was also established for the identification of three origins. The results showed that the TLC chromatograms of the flowers of A. souliei and A. flaccidus were very similar, but different from that of A. asteroides. The established macroscopic, powder microscopic and TLC methods for identification of three origins of AF were simple, accurate, and reproducible, and also effective and easy to operate.
Aster Plant ; anatomy & histology ; chemistry ; cytology ; Chlorogenic Acid ; analysis ; Chromatography, Thin Layer ; methods ; Flowers ; anatomy & histology ; chemistry ; cytology ; Medicine, Tibetan Traditional

To improve bond selectivity of recombinant β-glucuronidase in Escherichia coli (PGUS-E), based on the PGUS-E structure guidance, three key points R329, T369 and N467 were identified to be responsible for the bond selectivity of PGUS-E, and further saturation mutagenesis was conducted. Two positive mutants R329K and T369V were obtained by a combined selection technique of thin-layer chromatography and high performance liquid chromatography. Compared to PGUS-E, the bond selectivity of mutants R329K and T369V increased by 26.9% and 34.3%, respectively; whereas the biochemical properties such as pH and temperature profile were unchanged. Nevertheless, the activity was decreased compared to PGUS-E. These results further confirmed that sites R329 and T369 played important roles for the bond selectivity and activity. In summary, this study significantly increased the bond selectivity of PGUS-E by structure guided saturation mutagenesis, providing experimental support for elucidating the relationship between the structure and function of PGUS-E.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Escherichia coli ; metabolism ; Glucuronidase ; chemistry ; Industrial Microbiology ; Mutagenesis ; Recombinant Proteins ; chemistry ; Temperature