Determination of tryptophan in feedstuffs: Comparison of two methods of hydrolysis prior to HPLC analysis

The tryptophan content of nine samples of commercial feed was determined using two procedures (I and II) differing mainly in the hydrolysis conditions used prior to analysis. With I, hydrolysis was performed with 4 M LiOH in the presence of maltodextrin with 5-methyltryptophan used as internal standard at 110°C for 24 h; with II. oxygen-free 1.3 M Ba(OH)₂ was used at 125°C for 16 h. The two procedures gave identical results for a given sample indicating that the corrections for the incomplete recovery of tryptophan 5-methyltryptophan quantitation were satisfactory.     

固体石蜡作粘合剂的纯碳糊电极在流动注射分析中的应用

本文以固体石蜡作粘合剂的纯碳糊电极在流动注射分析中用电流法直接测定色氨酸，以HAc-NaAc(pH=3.5)缓冲液作载流液，选择电位1．4V,测定电流，峰高与被测物在一定浓度范围内成线形关系，实验测定了色氨酸的工作曲线，并讨论了各条件对结果的影响。     

How important is tryptophan in human health?

Tryptophan (Trp) is an amino acid and an essential component of the human diet. It plays a crucial role in many metabolic functions. Clinicians can use Trp levels in the course of diagnosing various metabolic disorders and the symptoms associated with those diseases. Furthermore, supplementation with this amino acid is considered in the treatment of depression and sleep disorders, mainly due to the Trp relationship with the synthesis of serotonin (5-HT) and melatonin. It is also used in helping to resolve cognitive disorders, anxiety, or neurodegenerative diseases. Reduced secretion of serotonin is associated with autism spectrum disorder, obesity, anorexia and bulimia nervosa, and other diseases presenting peripherals symptoms. The literature strongly suggests that Trp has a significant role in the correct functionality of the brain-gut axis and immunology. This information leads to the consideration of Trp as an essential dietary component due to its role in the serotonin pathway. A reduced availability of Trp in diet and nutraceutical supplementation should be considered with greater concern than one might expect. This paper constitutes a review of the more salient aspects gleaned from the current knowledge base about the role of Trp in diseases, associated nutritional disorders, and food science, in general.     

Direct determination of free tryptophan contents in soy sauces and its application as an index of soy sauce adulteration

A simple and rapid HPLC method using UV detector for determination of tryptophan (Trp) contents in pure fermented soy sauces, acid-HVP, and commercial soy sauces was developed. The method requires only simple sample pretreatment. The limit of detection (LOD) is estimated at 1 mg/L (signal-to-noise ratio of 3), and the recovery yields ranged from 92% to 108%. The analysis showed that Trp in all of the analysed acid-HVP samples was not detected and there was obvious detection of Trp (ranged from 136.4 to 261.8 mg/L) in all of the analysed fermented soy sauce samples. The observations suggest that Trp is a practical index of pure fermented soy sauce. The absence of Trp contents or lower level of Trp contents than that in pure fermented soy sauces suggests the soy sauces are not pure fermented soy sauces or presence of soy sauce adulteration. Simultaneous determination of levulinic acid (LV) in samples may be a good help in making assessment of soy sauce adulteration with acid-HVP.     

大肠杆菌色氨酸操纵子基因突变株的构建与表达

目的构建大肠杆菌色氨酸操纵子基因突变株,提高邻氨基苯甲酸合成酶和色氨酸合成酶的产量。方法利用依赖于DpnⅠ酶的PCR方法突变表达载体pET-22b(+)-Trp Operon上的关键位点,PCR扩增Trp OperonM基因,构建pET-22b(+)-Trp OperonM重组表达质粒,经酶切及测序鉴定正确后,转化大肠杆菌BL21(DE3),IPTG诱导表达。制备粗酶液,经比色法测定邻氨基苯甲酸合成酶和色氨酸合成酶的活性。结果 PCR扩增产物可见约7000bp的Trp OperonM条带;所构建的重组表达质粒经酶切及测序鉴定正确;邻氨基苯甲酸合成酶和色氨酸合成酶的活性比大肠杆菌BL21(DE3)空菌分别提高了4.5倍和5.2倍。结论已成功构建了大肠杆菌色氨酸操纵子基因突变株BL21(DE3)/pET-22b(+)-Trp OperonM,邻氨基苯甲酸合成酶和色氨酸合成酶的活性均有提高,为高产色氨酸基因工程菌的构建奠定了基础。     

Prediction of the rheology parameters of ripened semi-hard cheeses using fluorescence spectra in the UV and visible ranges recorded at a young stage

Storage modulus (G′), loss modulus (G″), strain, tan (δ) and complex viscosity (η*) of 20 semi-hard cheeses were measured by dynamic oscillatory analysis after 2, 30 and 60 days of ripening. On the same cheeses and at the same ages, tryptophan and riboflavin fluorescence spectra were recorded. The aim was to predict the rheology parameters of ripened cheeses from spectra recorded on these cheeses at a young stage. Using partial least square, tryptophan fluorescence spectra recorded at 20 °C on 2-days-old cheeses predicted G′, G″, strain, tan (δ) and η* measured at 80 °C on the 60-days-old cheeses with correlation coefficients (R) of 0.98, 0.97, 0.98, 0.98 and 0.97, respectively. Riboflavin fluorescence spectra gave slightly lower correlation coefficients of 0.88, 0.88, 0.92, 0.87 and 0.88, respectively. Dependent only on visible light, the riboflavin fluorescence spectra potentially provide viable and economic prediction of the rheology of ripe cheese.     

pH tuning of Nafion for selective detection of tryptophan

Selective and sensitive detection of the amino acid tryptophan is of importance in food processing, pharmaceutical formulations and in biological fluids. Electrochemical methods of detection of tryptophan are hampered by sluggish electron transfer kinetics and in complex matrices through overlapping peaks from interferents. This study examines the potential of the cation exchange membrane Nafion^® to enhance selectivity and sensitivity of this analyte through a seldom explored feature of this membrane: pH manipulation. A detailed examination of the effect of pH on the selectivity afforded by Nafion^® as a function of the analyte charge is presented. Selective detection of tryptophan and significant increases in sensitivity of its detection was observed in the presence of melatonin, dopamine and other interferents present in a pharmaceutical formulation through manipulation of the pH of the solution. At pH 3.0 at a Nafion^®-modified electrode, changes in the protonation of melatonin and tryptophan lowered the anodic potential of the analytes in a non-uniform manner increasing the peak resolution and permitting analyses with detection limits of 1.6 ± 0.1 nM and 1.6 ± 0.2 nM, respectively.     

Second-order data obtained from differential pulse voltammetry: Determination of tryptophan at a gold nanoparticles decorated multiwalled carbon nanotube modified glassy carbon electrode

Three-way data obtained from different pulse heights of differential pulse voltammetry (DPV) was analyzed using multivariate curve resolution by alternating least squares (MCR-ALS) algorithm. Differential pulse voltammograms of tryptophan were recorded at a gold nanoparticles decorated multiwalled carbon nanotube modified glassy carbon electrode (GCE/MWCNTs-nanoAu). The determination of tryptophan was performed even in the presence of unexpected electroactive interference(s). Both the simulated and experimental data were non-bilinear. Therefore a potential shift algorithm was used to correct the observed shift in the data. After correction, the data was augmented and MCR-ALS was applied to the augmented data. A relative error of prediction of less than 8% for the determination of the simulated analyte of interest and tryptophan in synthetic samples indicated that the methodology employing voltammetry and second-order calibration could be applied to complex analytical systems.     

Identification and occurrence of the bioactive β-carbolines norharman and harman in coffee brews

Norharman and harman, two heterocyclic β-carboline alkaloids with biological activity, were found in brewed coffee. Identification and analysis were carried out by HPLC-MS and RP-HPLC-fluorescence, respectively. All tested samples of brewed coffee including ground coffee, decaffeinated coffee, instant coffee and espresso contained both norharman and harman in variable amounts. Norharman was the major β-carboline alkaloid in brewed coffee at levels up to 9.34 μg g^-1 in instant ground coffee compared with harman, which had levels up to 1.67 μg g^-1. The two β-carbolines appeared to be formed during roasting of the coffee beans. It is concluded that drinking coffee is a major exogenous dietary source of these bioactive β-carboline alkaloids previously reported as mild psychoactive compounds in animal studies and in vitro co-mutagens. These results support our previous conclusion that foods containing β-carbolines are an important exogenous source of these alkaloids in humans.     

色氨酸、组氨酸羰基锝配合物的制备及其在荷S180瘤小鼠体内的分布

采用两步法合成了色氨酸及组氨酸的羰基锝配合物。纸上电泳结果表明，在pH为4．7、7．4和9．2三种缓冲体系中配合物不带电荷。从实验结果推测了其可能的结果。在荷肉瘤S180的小鼠体内分布实验表明，组氨酸羰基锝配合物在体内稳定，清除速率也较快，在肿瘤中有较高的摄取，肿瘤／内达到3－4。而色氨酸的羰基锝配合物在体内不稳定，各组织和器官内滞留时间较长、摄取较高，肿瘤中也有一定的摄取，肿瘤／肌肉为2－3，但其效果远不如组氨酸羰基锝配合物。讨论了利用羰基锝标记多肽和蛋白质的途径。