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OBJECTIVE To establish the characteristic chromatogram of Chaenomeles sinensis， determine the contents of rutin， hyperin and quercitrin， and to identify C. sinensis and C. speciosa. METHODS HPLC method was performed on Agilent 5 TC-C18 column， with acetonitrile-0.2% formic acid solution as the mobile phase for gradient elution， at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ . The detection wavelength was 330 nm in characteristic chromatogram and 350 nm in content determination. The characteristic chromatogram of C. sinensis was established and similarity was evaluated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. Hierarchical cluster analysis of 15 batches of C. sinensis （S1-S15） was performed by using SPSS 23.0 software. The contents of 3 flavones in 15 batches of C. sinensis and 7 batches of C. speciosa （S16-S22） were determined， while their characteristic chromatograms were compared. RESULTS The similarities of the characteristic chromatogram for 15 batches of C. sinensis ranged from 0.783 to 0.969， and 11 characteristic peaks were confirmed. Four constituents were identified as chlorogenic acid， rutin， hyperin and quercitrin. The medicinal materials in 15 batches of C. sinensis could be divided into 2 categories： S5-S8 were one category， and the others belonged to one category. The characteristic chromatogram of C. sinensis was obviously different from C. speciosa. The contents of rutin， hyperin and quercitrin in 15 batches of C. sinensis were 48.99-294.45， 3.49-102.55， 31.98-149.49 μg/g， respectively. The content of rutin in C. speciosa was lower than that in C. sinensis. None of hyperin （except for S20） and quercitrin were detected in C. speciosa. CONCLUSIONS The characteristic chromatogram and the method for content determination of 3 flavones in C. sinensis are established successfully and can be used for the quality control of C. sinensis and its identification from C. speciosa.
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OBJECTIVE To establish thin-layer chromatography （TLC） identification method and high-performance liquid chromatography （HPLC） fingerprint of Inonotus obliquus， and to evaluate the quality of I. obliquus by chemical pattern recognition. METHODS TLC method was used to identify trametenolic acid and inotodiol in I. obliquus qualitatively. HPLC fingerprint of I. obliquus was established； Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition） was used to determine the common peaks and evaluate the similarity； chemical pattern recognition analysis [cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis （OPLS-DA）] of 22 batches of I. obliquus was performed with SPSS 23.0 software and SIMCA14.1 software. RESULTS In the TLC， the same color spots were found at the same position in the chromatograms of test sample and substance control. A total of 10 common peaks were marked in the HPLC fingerprints of 22 batches of I. obliquus， with similarities of 0.942-0.995. No. 3 peak was identified as trametenolic acid， No.4 peak as inotodiol， No. 9 peak as ergosterol and No. 10 peak as lanosterol. Results of cluster analysis showed that S1-S15， S19， S21 and S22 could be clustered into the first category， and S16-S18 and S20 were clustered into the second category. Results of principal component analysis showed that top 4 samples in the list of comprehensive score were S17， S18， S16 and S20. Results of OPLS-DA showed that three marking components that may affect the quality of I. obliquus were screened according to the standard of VIP＞1， i.e. No. 4 peak （inotodiol， VIP value of 1.86）， No. 3 peak （trametenolic acid， VIP value of 1.62） and No. 7 peak （VIP value of 1.27）. CONCLUSIONS This study establishes TLC method and HPLC fingerprint of I. obliquus successfully， which can provide reference for the quality control of I.obliquus by combining with chemical pattern recognition.
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Iron-chelating peptides have been widely considered as one of the best iron supplements to alleviate the iron deficiency. In this study, a novel oat peptides-ferrous (OP-Fe2+) chelate was prepared from antioxidant oat peptides obtained in the laboratory of the authors. The optimal preparation condition was obtained through the single-factor and response surface methodology, and the chelating rate could reach up to 62.6%. After chelation, the OP-Fe2+ chelate exhibited a significantly higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity than oat peptides. It was discovered that the hemoglobin concentration and the number of red blood cell levels in OP-Fe2+-treated iron-deficient anemic (IDA) rats were significantly higher than untreated IDA rats. The OP-Fe2+ chelate could also improve the hypertrophy of the spleen, serum iron (SI), total iron and binding capacity, and serum ferritin levels in the IDA rats. In addition, the OP-Fe2+ treatment significantly increased the antioxidant activities of super oxidase and glutathione in the liver homogenate of the IDA rats. Therefore, the OP-Fe2+ chelate is an effective type of iron supplement for IDA rats, which could be a promising source with anti-anemia and antioxidant activity.
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Objective To investigate the expression of LncRNA DLEU1 in ESCC tissues, and its effect on the proliferation and migration of ESCC cells. Methods We collected 58 cases of ESCC tissues and corresponding para-cancerous tissues. RT-qPCR was used to detect the relative expression levels of DLEU1 in ESCC tissues and cells. Log-rank test was used to analyze the relation between the expression of DLEU1 and clinicopathological features. Kaplan-Meier analysis was used to investigate the correlation between the expression of DLEU1 and the survival of ESCC patients. Multivariate Cox regression model was used to evaluate the effect of DLEU1 on the prognosis of ESCC patients. Effects of DLEU1 on the proliferation and migration of Eca9706 cells were evaluated by CCK-8 and wound healing assays, respectively. Results DLEU1 was highly expressed in ESCC tissues (P < 0.01) and significantly correlated with tumor size, TNM stage and lymph node metastasis (all P < 0.05). High expression of DLEU1 was negatively correlated with poor prognosis of ESCC patients (P < 0.01), and DLEU1 was also an independent prognostic risk factor (P < 0.05). Moreover, knockdown of DLEU1 significantly inhibited the proliferation and migration of Eca9706 cells, compared with the control group (P < 0.01). Conclusion DLEU1 is highly expressed in ESCC tissues. The expression of DLEU1 is an independent risk factor for the prognosis of ESCC patients and promotes ESCC cell proliferation and migration.
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This study evaluated the effects of ultrasound treatment on walnut meal protein (WMP) extraction and techno-functional properties. The Box-Behnken Design (BBD) was adopted for the optimization of the traditional and ultrasound-assisted extraction (UAE) processes. Standard protocols were used to assay the techno-functional characteristics. The extraction models' statistical results exhibited adequacy with the least desirability index of 95.8%. The UAE enhanced the WMP extraction yield, purity, and chemical score by 30.15%, 16.27%, and 9.74%, respectively, while reducing the extraction time by 25% over the control. The emulsion and foam stabilities and bulk density increased by 34.5%, 39.8%, and 6.1%, respectively, over the control. The α-helix decreased while ß-sheet, ß-turns and random coil secondary structure components increased significantly (p < .05) by 95.76%, 101.3%, 105.1%, and 85.7% correspondingly. The dual-frequency combination (20/40 kHz/kHz) was the best frequency mode. WMP could serve as a functional additive in manufactured foods as texture and flavor enhancer. PRACTICAL APPLICATIONS: Walnut meal protein (WMP) has a well-balanced amino acid profile and its economic use could be practically increased as a food ingredient by ultrasound-assisted extraction. By this technique, WMP could be employed for the development of enhanced food ingredients rather than being discarded as animal feed. This study showed a positive effect of ultrasonic-assisted alkaline pretreatment on WMP extraction, functionality and structure characteristics. In addition to process improvement, ultrasound is energy efficient and environmentally friendly. Therefore, the applicability of this technique to improve the functionality of plant proteins from industrial by-products to be included in food products is promising.
Subject(s)
Juglans , Amino Acids , Animals , Nuts , Plant Proteins , Protein Structure, SecondaryABSTRACT
To improve the utility of walnut meal protein, influences of ultrasonication on the kinetic and thermodynamic constraints of enzyme (trypsin) hydrolysis were assessed using a multi-frequency counter current S-type ultrasound machine. Results revealed that both the ultrasonication treatment and control hydrolysis obeyed the first-order kinetics within the study constraints. After 90 min hydrolysis time, the uppermost percentage conversion rate (57.5%) of substrate to product and the percentage increase (25.42%) in hydrolyzed protein concentration were obtained at 323 K and 10% substrate concentration, respectively. The Michaelis constant (KM ) in ultrasonic enzymolysis declined by 58.66% over the control. The reaction rate coefficient (k) for the proteolysis improved by 84.75%, 52.43%, 48.25%, and 37.79% at 219, 303, 313, and 323 K, correspondingly. Generally, the bond energy (Ea ), enthalpy change (ΔH), entropy change (ΔS), and Gibbs free energy (ΔG) were reduced by the ultrasound pretreatment with 36.61%, 28.05%, 18.22%, and 5.24%, respectively. PRACTICAL APPLICATIONS: Walnut meal protein has a well-balanced amino acid profile and its economic utilization could be increased practicably as a food ingredient via production of hydrolysates/peptides for the production of improved food ingredients instead of being cast off as animal feed. This study demonstrated a positive bearing of ultrasonication-assisted alkaline pretreatment on proteolytic reaction characteristics and its energy efficiency for walnut meal protein, which makes this technique applicable to the enhancement of plant proteins for inclusion in food products, especially tree nut and oil seed waste products from the oil industry.
Subject(s)
Food Handling/methods , Juglans/chemistry , Plant Proteins/chemistry , Trypsin/chemistry , Alkalies/chemistry , Amino Acids/chemistry , Biocatalysis , Entropy , Food Handling/instrumentation , Kinetics , Nuts/chemistry , Proteolysis , Thermodynamics , UltrasonicsABSTRACT
Objective: To probe into the antioxidation mechanism of lotus seedpod in vivo.Methods:Nineteen 4-month old rats were randomly divided into control group and procyanidins group. After feeding 100 mg/kg LSPC for 35 d, MDA and SOD in skin and serum, GSH-Px and Hyp in blood and skin were measured.Results: MDA levels in skin and serum in the procyanidins group were obviously lower than that of the control(P
