Content of multiple components (neochlorogenic acid,L-tryptophan,vicenin-2,isoquercitrin,and astragalin) in Moringa oleifera leaves was determined by high-performance liquid chromatography (HPLC),and the absolute content-time curves were plotted.Based on Fick's law of diffusion and Higbie's penetration theory,the parameters of the equations were calculated,and the measured results were substituted into the mathematical model to fit the equations.The n and a obtained from the equations on the decocting time factor and the solvent volume were close to each other.The dynamic models of the five components are as follows:■.The variation of the content of multiple components in M.oleifera leaves with time and solvent volume was explored.It was found that the content of the components was the highest when the leaves were decocted for 30 min with solvent volume 12 folds of the medicinal material.The dissolution and destruction of components and the diffusion movement of components are the main causes of the content change of M.oleifera leaves at different time and with different solvent volumes.The R~2of the linear equations on the content and the equations on the decocting process (5-30min and solvent volume 12-20 folds of the medicinal materials) was≥0.999 8 and≥0.9,respectively.Thus,the content determination and the decocting kinetic model had high accuracy,which can reflect the change law of the content of key components in M.oleifera leaves during the decoction.This study is expected to serve as a reference for optimizing the decocting technology.
Kinetics ; Moringa oleifera/chemistry* ; Plant Leaves/chemistry* ; Solvents ; Tryptophan/analysis*

OBJECTIVETo establish a method for comparing the differences between fresh and dried ginger juice.

METHODThe RP-HPLC fingerprint method was performed on an Alltech C18 column (4.6 mm x 250 mm, 5 microm) with mobile phase in gradient elution composed of A-acetonitrie and B-water at a flow rate: 0.8 mL x min(-1). The detecting wavelength was 280 nm, and the column temperature 25 degrees C.

RESULTThere was no significant difference among the same breed ginger juice of different batches. But there was significant difference between crushed ginger juice and the boiled juice. Trytophan, 6-gingerol were common constituents of the three kinds of ginger juice, the fresh ginger and the dry ginger. Besides, 6-shogaol emerged in the boiled juice.

CONCLUSIONThe RP-HPLC fingerprints spectrum can be used to distinguish different ginger juices. And the crushed juice of fresh ginger have the same chemical consititents with the fresh ginger.

Catechols ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Fatty Alcohols ; chemistry ; Ginger ; chemistry ; Plant Extracts ; chemistry ; Tryptophan ; chemistry

Ecdysis is a common phenomenon that happens throughout the life phase of the giant freshwater prawn Macrobrachium rosenbergii. It is vital to better understand the correlation between cannibalism and biochemical compound that exists during the moulting process. The objective of the present study was to determine the amino acid profile released by M. rosenbergii during the ecdysis process that promotes cannibalism. To accomplish this, changes in amino acid levels (total amino acid (TAA) and free amino acid (FAA)) of tissue muscle, exoskeleton, and sample water of culture medium from the moulting (E-stage) and non-moulting (C-stage) prawns were analysed using high-performance liquid chromatography (HPLC). Comparison study revealed that among the TAA compounds, proline and sarcosine of tissues from moulting prawn were found at the highest levels. The level of FAA from water that contains moulting prawns (E-stage) was dominated by tryptophan and proline. Significant values obtained in the present study suggested that these amino acid compounds act as a chemical cue to promote cannibalism in M. rosenbergii during ecdysis. The knowledge of compositions and compounds that were released during the moulting process should be helpful for better understanding of the mechanism and chemical cues that play roles on triggering cannibalism, and also for future dietary manipulation to improve feeding efficiencies and feeding management, which indirectly impacts productivity and profitability.
Amino Acids/chemistry* ; Animals ; Aquaculture ; Cannibalism ; Chromatography, High Pressure Liquid ; Feeding Behavior ; Fresh Water ; Malaysia ; Molting ; Palaemonidae/physiology* ; Proline/chemistry* ; Tryptophan/chemistry*

OBJECTIVETo screen the active component of Wuzhuyutang (WZYT, Evodiae prescription) and investigate the regulatory effects of the components in WZYT on the TPH2 promoter, and to explore the possible molecular mechanism of WZYT on migraine.

METHODBy transfecting a TPH2 promoter regulating Red Fluorescent Protein expressing plasmid into PC12 cell, the global fluorescence intensities and calculations of fluorescent cells after components treatment were statistically evaluated.

RESULTDifferent regulatory effects of different components in WZYT with different concentrations on TPH2 promoter were observed.

CONCLUSIONTPH2 promoter drove Red Fluorescent Protein expressing cell line can be used as system screening components targeting TPH2 promoter activity. The possible mechanism of WZYT on migraine may due to its stimulating effects on TPH2 promoter, and promote the synthesis and release of 5-HT in cerebral.

Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Evodia ; chemistry ; Humans ; Migraine Disorders ; drug therapy ; enzymology ; genetics ; PC12 Cells ; Promoter Regions, Genetic ; drug effects ; Rats ; Tryptophan Hydroxylase ; genetics ; metabolism

Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.
Amino Acids ; chemistry ; Caseins ; chemistry ; metabolism ; Dietary Proteins ; analysis ; Evaluation Studies as Topic ; Hydrolysis ; Milk Proteins ; chemistry ; metabolism ; Models, Statistical ; Nutritive Value ; Protein Hydrolysates ; chemistry ; Solubility ; Temperature ; Time Factors ; Tryptophan ; chemistry ; Whey Proteins

OBJECTIVETo explore the differences of metabolic footprint in the conditioned culture medium of placental explants between early-onset and late-onset severe preeclampsia.

METHODSIn 13 cases of early-onset severe preeclampsia and 14 cases of late-onset severe preeclampsia, the placentas were sampled at the surface of the maternal placenta. High performance liquid chromatography-mass spectrometry (HPLC-MS) was used to determine the differences in the metabolites in the conditioned culture medium of the placental villous explants cultured in 6% atmospheric O(2) for 96 h. Standard samples were used to establish the tryptophan and kynurenine chromatography library by HPLC-MS to analyze the concentration of tryptophan and kynurenine in the conditioned culture medium.

RESULTSThirty-six metabolites showed statistically significant differences between early-onset and late-onset severe preeclampsia (P<0.05). The concentration of kynurenine was significantly higher in early-onset severe preeclampsia than in late-onset severe preeclampsia (P<0.05).

CONCLUSIONEarly-onset and late-onset severe preeclampsia may have different pathogeneses. By detecting the concentration of metabolites, metabolomic strategies provide a new means for predicting the onset time of severe preeclampsia.

Chorionic Villi ; metabolism ; Culture Media, Conditioned ; chemistry ; Female ; Humans ; In Vitro Techniques ; Kynurenine ; metabolism ; Ornithine ; metabolism ; Placenta ; metabolism ; Pre-Eclampsia ; metabolism ; Pregnancy ; Tryptophan ; metabolism

A FAST (fluorescence of advanced Maillard products and Soluble Tryptophan) method for identification of reconstituted milk made from skim milk powder in the fresh milk was developed. Considering milk and skim milk powders variations from different seasons and countries, milk was collected from different dairy farms in different seasons and skim milk powders were collected from different countries to measure the Tryptophan (Trp), advanced Maillard products (AMP) fluorescence values. The results showed that there were differences (P<0.01) between raw and reconstituted milk. The plot of values in each mixed level of raw and reconstituted milk had a correlation coefficient >0.97. The FAST method is a simple, rapid, low-cost and sensitive method enabling the detection of 5% reconstituted milk in fresh milk. The measurement of the Trp, AMP fluorescence values and calculation of the FAST index is a suitable method for large-scale monitoring of fresh milk samples.
Animals ; Cattle ; Food Analysis ; methods ; Food Contamination ; prevention & control ; Glycation End Products, Advanced ; analysis ; Maillard Reaction ; Milk ; chemistry ; classification ; Powders ; Spectrometry, Fluorescence ; methods ; Tryptophan ; analysis

OBJECTIVETo investigate the effects of nitrogen forms on the camptothecin (CPT) content, tryptophan synthase (TSB) and tryptophan decarboxylase (TDC) activities in Camptotheca acuminata seedlings.

METHODThe seedlings of C. acuminata with 6 pairs of leaves were subjected to 5 different NH4(+) -N/NO3(-) -N ratio (0 : 100, 75 : 25, 50 : 50, 25 : 75, 100 : 0) treatments by sand culture in a greenhouse. The CPT content, TSB activity in the young leaves and TDC in the stem barks of the seedlings were determined by HPLC on the 15th, 30th, 45th, 60th and 75th day, respectively.

RESULTThe obvious relationship between CPT content and nitrogen forms was observed. When NH4(+) - N /NO3(-) -N ratio was 25 : 75, CPT accumulation in young leaves displayed the best advantages (the highest value is 5.69 per thousand) and increased in the early 30 days of treatment and then declined. There was no obvious relationship between TSB activity in the young leaves and nitrogen forms. TDC activity in the stem bark was the highest when NH4(+) -N /NO3(-) -N ratio was 25 : 75, and the change of TDC activity paralleled to CPT content in the young leaves.

CONCLUSIONA short-term treatment that NH4(+) -N /NO3(-) -N ratio was 25:75 may gain high CPT content in the young leaves through enhancing the TDC activity in the stem bark of C. acuminata seedlings.

Aromatic-L-Amino-Acid Decarboxylases ; metabolism ; Camptotheca ; drug effects ; enzymology ; metabolism ; Camptothecin ; metabolism ; Drugs, Chinese Herbal ; metabolism ; Nitrogen ; chemistry ; pharmacology ; Plant Leaves ; drug effects ; enzymology ; metabolism ; Seedlings ; drug effects ; enzymology ; metabolism ; Tryptophan Synthase ; metabolism

YfiBNR is a recently identified bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) signaling system in opportunistic pathogens. It is a key regulator of biofilm formation, which is correlated with prolonged persistence of infection and antibiotic drug resistance. In response to cell stress, YfiB in the outer membrane can sequester the periplasmic protein YfiR, releasing its inhibition of YfiN on the inner membrane and thus provoking the diguanylate cyclase activity of YfiN to induce c-di-GMP production. However, the detailed regulatory mechanism remains elusive. Here, we report the crystal structures of YfiB alone and of an active mutant YfiB(L43P) complexed with YfiR with 2:2 stoichiometry. Structural analyses revealed that in contrast to the compact conformation of the dimeric YfiB alone, YfiB(L43P) adopts a stretched conformation allowing activated YfiB to penetrate the peptidoglycan (PG) layer and access YfiR. YfiB(L43P) shows a more compact PG-binding pocket and much higher PG binding affinity than wild-type YfiB, suggesting a tight correlation between PG binding and YfiB activation. In addition, our crystallographic analyses revealed that YfiR binds Vitamin B6 (VB6) or L-Trp at a YfiB-binding site and that both VB6 and L-Trp are able to reduce YfiB(L43P)-induced biofilm formation. Based on the structural and biochemical data, we propose an updated regulatory model of the YfiBNR system.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; genetics ; metabolism ; Binding Sites ; Biofilms ; Crystallography, X-Ray ; Cyclic GMP ; analogs & derivatives ; metabolism ; Dimerization ; Molecular Dynamics Simulation ; Molecular Sequence Data ; Mutagenesis ; Protein Structure, Quaternary ; Pseudomonas aeruginosa ; metabolism ; Sequence Alignment ; Tryptophan ; chemistry ; metabolism ; Vitamin B 6 ; chemistry ; metabolism

A UPLC-MS/MS method based on metabonomic skills was developed to study the serum metabolic changes of rats after acute liver injury induced by CCl4 and to evaluate the action mechanism of Si-Ni-San. The integrated data were exported for principal components analysis (PCA) by using SIMCA-P software, in order to find the potential biomarkers. It showed that clear separation of healthy control group, model group, silymarin group, Si-Ni-San group was achieved by using the PCA method. Nine significantly changed metabolites were identified as potential biomarkers of acute liver injury. Compared with the health control group, the model group rats showed higher levels of phenylalanine, tryptophan and GCDCA together with lower levels of LPC 16 : 0, LPC 18 : 0, LPC 18 : 1, LPC 16 : 1, LPC 20 : 4 and LPC 22 : 6. These changes of serum metabolites suggested that the disorders of amino acid metabolism, lipid metabolism, bile acid biosynthesis and anti-oxidative damage were related to acute liver injury induced by CCl4. Si-Ni-San might have the anti-liver injury effect on all these four metabolic pathways.
Animals ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; blood ; etiology ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Glycodeoxycholic Acid ; blood ; Lysophosphatidylcholines ; blood ; Male ; Metabolomics ; Phenylalanine ; blood ; Plants, Medicinal ; chemistry ; Principal Component Analysis ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Tryptophan ; blood