Optimization of gelatine extraction from grass carp (Catenopharyngodon idella) fish skin by response surface methodology

To establish the optimum gelatine extraction conditions from grass carp fish skin, response surface methodology (RSM) was adopted in this study. The effects of concentration of HCl (%, A), pre-treatment time (h, B), extraction temperature ( degrees C, C) and extraction time (h, D) were studied. The responses were yield (%) and gel strength (g). A=1.19%, B=24 h, C=52.61 degrees C and D=5.12h were determined as the optimum conditions while the predicted responses were 19.83% yield and 267 g gel strength. Gelling and melting points were 19.5 degrees C and 26.8 degrees C, respectively. Moreover, grass carp gelatine showed high contents of imino acids (proline and hydroxyproline) 19.47%. RSM provided a powerful tool to optimize the extraction parameters and the results may be adapted for industrial extraction of gelatine from grass carp fish skins.     

银条水苏糖提取条件研究

唇形科水苏属植物银条,也称银苗,药食两用,主产于偃师县(市),产量约有95%。本文以偃师产银条为原料,通过单因素、响应面分析法,研究了乙醇提取银条中水苏糖的最佳提取条件。以银条中水苏糖提取率为指标,通过单因素实验确定了银条水苏糖提取条件的参数范围,通过响应面分析法确定了其最佳提取条件。结果表明,最佳提取条件为乙醇浓度60%,液固比10:1,提取温度60℃,提取时间40 min,该条件下银条水苏糖提取率为46.8924%。     

Evaluation of enantiomeric purity of selected amino acids in honey

Highly sensitive and accurate HPLC methods were used for the determination of total amounts of proline, leucine and phenylalanine and their enantiomeric ratios in a variety of different honey samples. Significant amounts of D -leucine and D -phenylalanine and relatively low concentrations of D -proline have been found in honeys of different botanical and geographical origins. It is suggested that the enantiomeric ratios of amino acids could be used to test for storage effects, age, and the quality of the processing of the honey. © 1994 Wiley-Liss, Inc.     

Unifloral Honeys: Chemical Conversion and Pollen Reduction

Physico-chemical and pollen analyses were made of unifloral samples of nectar, crop content and honey. The samples were obtained by carrying out cage experiments exclusively with one plant species. The investigations of four plant species are presented: Centaurea montana, Coriandrum sativum. Euphorbia lathyris and Rubus idaeus. During the conversion of nectar to honey its chemical composition changes. The changes in carbohydrate composition cannot be explained by those enzymes described as specific for honey. During the period of time when the bees are working on gathered nectar, the chemical components of the honey, whose origin is the secretion from the bees, is increasing, while the amount of pollen is decreasing. There are negative correlations between the quantity of pollen grains and both enzymatic activity and the amount of proline. With respect to the botanical origin it is suggested that the absolute amount of pollen should be determined and compared with the chemical analysis of the honey.     

响应面设计法优化仙鹤草总多酚的超声提取工艺

在单因素实验的基础上,采用响应面分析法对影响超声辅助提取仙鹤草总多酚得率的主要因素(超声温度、料液比和乙醇浓度)进行优化,建立了影响因素与总多酚之间的函数关系。获得最佳工艺条件为:71%乙醇,料液比1∶24,超声提取温度60℃,提取时间20 min。在此最佳条件下,总多酚得率为3.56%,试验结果与模型预测值相符。     

A variety of volatile compounds as markers in unifloral honey from dalmatian sage (Salvia officinalis L.)

Volatile compounds of unifloral Salvia officinalis L. honey has been investigated for the first time. The botanical origin of ten unifloral Salvia honey samples has been ascertained by pollen analysis (the honey samples displayed 23-60% of Salvia pollen). Fifty-four volatile compounds were identified by GC and GC/MS in ten Salvia honey extracts obtained by ultrasound-assisted extraction (USE) with pentane/Et(2)O 1 : 2. The yield of isolated volatiles varied from 25.7 to 30.5 mg kg(-1). Salvia honey could be distinguished on the basis of the high percentage of benzoic acid (6.4-14.8%), and especially phenylacetic acid (5.7-18.4%). Minor, but floral-origin important volatiles were identified such as shikimate pathway derivatives, 'degraded-carotenoid-like' structures (3,5,5-trimethylcyclohex-2-ene derivatives) and 2,6,6-trimethylcyclohex-2-ene derivatives. Compounds from other metabolic pathways such as aliphatic acids and higher linear hydrocarbons, as well as heterocycles (pyrans, furans, and pyrroles), were also present. Most of the identified compounds do not constitute specific Salvia honey markers, due to their presence in honeys of other botanical origins; however, their ratio in different honeys could be useful to distinguish floral origin. Salvia-honey volatile markers were: benzoic acid, phenylacetic acid, p-anisaldehyde, alpha-isophorone, 4-ketoisophorone, dehydrovomifoliol, 2,6,6-trimethyl-4-oxocyclohex-2-ene-1-carbaldehyde, 2,2,6-trimethylcyclohexane-1,4-dione, and coumaran.     

Optimization of enzymatic hydrolysis of visceral waste proteins of Catla (Catla catla) for preparing protein hydrolysate using a commercial protease

Protein hydrolysate was prepared from visceral waste proteins of Catla (Catla catla), an Indian freshwater major carp. Hydrolysis conditions (viz., time, temperature, pH and enzyme to substrate level) for preparing protein hydrolysates from the fish visceral waste proteins were optimized by response surface methodology (RSM) using a factorial design. Model equation was proposed with regard to the effect of time, temperature, pH and enzyme to substrate level. An enzyme to substrate level of 1.5% (v/w), pH 8.5, temperature of 50 degrees C and a hydrolysis time of 135 min were found to be the optimum conditions to obtain a higher degree of hydrolysis close to 50% using alcalase. The amino acid composition of the protein hydrolysate prepared using the optimized conditions revealed that the protein hydrolysate was similar to FAO/WHO reference protein. The chemical scores computed indicated methionine to be the most limiting amino acid. The protein hydrolysate can well be used to meet the amino acid requirements of juvenile common carp and hence has the potential for application as an ingredient in balanced fish diets.     

Response surface approach to optimize temperature,pH and time on antioxidant properties of wild bush (Plectranthus rugosus) honey from high altitude region (Kashmir Valley) of India

In this study, the combined effect of temperature (60 to 80 °C) time (10 to 15 min.) and pH (3 to 6) was employed on the anti-oxidant potential (1,1-diphenyl-2-picrylhydrazyl-radical scavenging activity-DPPH-RSA, total phenolic content-TPC, and total flavonoid content-TFC) of wild bush Indian honey from high altitudes of Kashmir Valley by using response surface methodology (RSM). The statistical analysis showed that all the process variables had a substantial effect on the responses related to DPPH-RSA, TFC, and TPC, all of which increased as temperature and time increased. With an increase in pH, the antioxidant activity of wild bush honey was significantly decreased. The heat treatment of honey at high temperature (80 °C) was found to be more efficacious than at 70 and 60 °C, respectively. The findings showed that at higher temperature, browning pigments were formed which enhanced considerably the antioxidant activity of honey.     

Ribonuclease of Bacillus intermedius 7 P. Purification by chromatography on phosphocellulose and several characteristics of the homogeneous enzyme]

A new procedure for isolation of homogenous ribonuclease of Bac. intermedius from a commercial source is described. The yields of 140 mg of RNAse from 200 g of the enzymic powder were attained. The amino acid composition of the enzyme was determined. The RNAse contains neither the sulfhydryl groups nor the disulfide bonds and has only one histidine residue. At the same time the amount of aromatic amino acid residues is relatively high. The enzyme is highly resistant to heat and acid treatment but is less stable in an alkaline solution. The pH optimum of the RNAse for the RNA digestion is 8,5; the temperature optimum for this reaction is 37 degrees. A spectrophotometric method for the RNAse activity assay using polyA as a specific substrate was developed. The purified product provides a suitable starting material for structural studies.     

Volatile compounds of Asphodelus microcarpus Salzm. et Viv. Honey obtained by HS-SPME and USE analyzed by GC/MS

Chemical analysis of Asphodelus microcarpus Salzm. et Viv. honey is of great importance, since melissopalynology does not allow the unambiguous determination of its botanical origin. Therefore, the volatile compounds of eight unifloral asphodel honeys have been investigated for the first time. The honey extracts were obtained by headspace solid-phase microextraction (HS-SPME) and ultrasonicsolvent extraction (USE) and analyzed by GC and GC/MS. In the honey headspace, 31 volatile compounds were identified with high percentages of 2-phenylacetaldehyde (2; 14.8–34.7%), followed by somewhat lower percentages of methyl syringate (1; 10.5–11.5%). Compound 2 is not a specific marker of the botanical origin of the honey, but its high percentage can be emphasized as headspace characteristic of asphodel honey. The extraction solvent for all the samples was selected after extracting a representative sample with pentane, Et(2)O, pentane/Et(2)O 1:2 (v/v), and CH(2)Cl(2) . Compound 1 was the major constituent of all the USE extracts (46.8–87.0%). According to these preliminary results, all the honey samples were extracted by USE with the solvent pentane/Et(2)O 1:2. A total of 60 volatile compounds were identified with 1 as predominant compound (69.4–87.0%), pointing out 1 as Asphodelus honey volatile marker.     

响应面分析法优化超声波辅助酶法提取南五味子多糖工艺的研究

为了建立超声波辅助酶法提取南五味子多糖的方法,并研究其提取工艺。按照Box-Benhnken中心组合试验设计原理,以多糖得率为响应值,以单因素实验数据为基础,进行响应面分析实验,考察提取pH、提取温度和提取时间对多糖得率的影响。结果显示：最佳提取工艺条件为：提取pH为4.8、提取温度57℃、提取时间66 min、复合酶用量为3.0%,最佳工艺条件下南五味子多糖得率为8.73%±0.25%。研究结果表明,该提取方法可减少多糖提取时间和提取溶剂用量,降低温度,有效提高了多糖的提取得率,可应用于南五味子多糖的制备。研究结果为南五味子多糖的开发应用提供了较好的依据。     

Optimization of microwave-assisted transesterification of dry algal biomass using response surface methodology

The effect of microwave irradiation on the simultaneous extraction and transesterification (in situ transesterification) of dry algal biomass to biodiesel was investigated. A high degree of oil/lipid extraction from dry algal biomass and an efficient conversion of the oils/lipids to biodiesel were demonstrated in a set of well-designed experimental runs. A response surface methodology (RSM) was used to analyze the influence of the process variables (dry algae to methanol (wt/vol) ratio, catalyst concentration, and reaction time) on the fatty acid methyl ester conversion. Based on the experimental results and RSM analysis, the optimal conditions for this process were determined as: dry algae to methanol (wt/vol) ratio of around 1:12, catalyst concentration about 2 wt.%, and reaction time of 4 min. The algal biodiesel samples were analyzed with GC-MS and thin layer chromatography (TLC) methods. Transmission electron microscopy (TEM) images of the algal biomass samples before and after the extraction/transesterification reaction are also presented.     

Protein hydrolysate from visceral waste proteins of Catla (Catla catla): optimization of hydrolysis conditions for a commercial neutral protease

Protein hydrolysate was prepared from visceral waste proteins of an Indian freshwater major carp, Catla catla. Hydrolysis conditions (viz., time, temperature and enzyme to substrate level) for preparing protein hydrolysates from the fish visceral waste proteins using in situ pH of the visceral mass were optimized by response surface methodology (RSM) by employing a factorial design. The regression coefficient close to 1.0, observed during both experimental and validation runs, indicated the validity of prediction model. An enzyme to substrate level of 1.25 % (v/w), temperature of 55 degrees C and a hydrolysis time of 165 min were found to be the optimum conditions to obtain a higher degree of hydrolysis of >48% using multifect-neutral. The amino acid composition of the protein hydrolysate prepared using the optimized conditions revealed that the protein hydrolysate was similar to FAO/WHO reference protein. The chemical scores computed indicated methionine to be the most limiting amino acid. The protein hydrolysate has the potential for application as an ingredient in balanced fish diets.     

响应面法优化低温豆粕大豆分离蛋白提取工艺

采用响应面分析法(RSM)对低温豆粕大豆分离蛋白(soybean protein isolated,SPI)的碱提工艺进行优化。在单因素实验的基础上,选取了影响SPI提取率的4个关键因素(pH、温度、时间和液料比)进行四因素五水平的中心组合旋转实验设计(CCRD)。通过RSM建立了响应值(SPI提取率)与各影响因素之间的回归方程,并获得了SPI的最优提取条件:pH 8.5,提取温度55℃,提取时间42.8 min,料水比1∶9.7(g/mL),此条件下SPI提取率预测值为37.12%,与实验值(36.69%)的误差为1.16%。将一次碱提后残渣进行二次碱提,二次提取率为10.16%。2次碱提上清液等电点沉淀的蛋白沉淀率分别为84.03%和85.84%。     

响应面法优化红景天总黄酮提取工艺的研究

研究了红景天中总黄酮提取率的影响因素,先经单因素实验初步确定影响因素,再用Box-Behnken响应面法建立影响因素的二次回归模型,通过对二次回归模型求解编码转换得知,液料比40:1(ml/g),酶添加量1.2%,酶解时间4.5 h,酶解温度45℃,pH5,超声时间为12 min,超声功率为250w中的高档,超声次数为3次,在这最优条件下总黄酮提取率为3.99%。     

水酶法提取紫苏籽油脂工艺

采用中心组合设计(CCD)-响应面(RSM)优化紫苏籽油脂的水酶法提取工艺。在单因素试验的基础上采用中心组合设计方法,研究了酶的种类、酶解温度、pH、液(mL)固(g)比、加酶量、以及时间相互作用对紫苏油脂提取率的影响。结果显示,拟合得到方程显著,确定的紫苏油脂提取最优条件为:碱性蛋白酶在pH9.5条件下液(mL)固(g)比9.97∶1、加酶量2.75%、温度56.1℃、时间5.25h,该条件下紫苏油脂的提取率可达到37.65%,与理论值38.3%十分接近,建立的模型真实可靠,确定了紫苏油脂的最佳提取工艺。经气相色谱检测紫苏籽油中含有棕榈酸、硬脂酸、油酸、亚油酸、α-亚麻酸等脂肪酸,水酶法提取紫苏油脂的α-亚麻酸相对含量最高67.9%,且相对溶剂法及冷榨法理化指标最好。     

Extraction,development and validation of HPLC-UV method for rapid and sensitive determination of colchicine from Colchicum autumnale L. Bulbs

Colchicum autumnale L. also known as the autumn crocus, contains colchicine alkaloid having antifungal properties. The tuber of this plant is rich in terms of colchicine. In this research an ultrasound-assisted extraction (UAE) method was optimized for the extraction of colchicine from Colchicum autumnale L. bulbs before high-performance liquid chromatography with UV detection (HPLC-UV). Optimization of various extraction parameters was performed using response surface methodology (RSM) to evaluate the maximum colchicine yield from Colchicum autumnale L. bulbs. The Box-Behnken design (BBD) and RSM were used to investigate the effect of three key parameters (extraction time (20–60?min), extraction temperature (40–80?°C) and ultrasound power (500–700?W) on extraction efficiency. The variance analysis suggested that the dependent response variable of yield of colchicine may be expressed by a quadratic polynomial model. The optimal theoretical extraction conditions were found to be an ultrasonication power of 602.4?W, an extraction time of 42?min and a temperature of 64?°C. Under these conditions, the optimum foreseen yield was 0.237%. The experimental colchicine yield obtained by following the optimized conditions was found to be 0.238%. These values are very well compatible with each other.     

Ultrasound Assisted Extraction of Phenolic Compounds from Peaches and Pumpkins

The ultrasound-assisted extraction (UAE) method was used to optimize the extraction of phenolic compounds from pumpkins and peaches. The response surface methodology (RSM) was used to study the effects of three independent variables each with three treatments. They included extraction temperatures (30, 40 and 50°C), ultrasonic power levels (30, 50 and 70%) and extraction times (10, 20 and 30 min). The optimal conditions for extractions of total phenolics from pumpkins were inferred to be a temperature of 41.45°C, a power of 44.60% and a time of 25.67 min. However, an extraction temperature of 40.99°C, power of 56.01% and time of 25.71 min was optimal for recovery of free radical scavenging activity (measured by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) reduction). The optimal conditions for peach extracts were an extraction temperature of 41.53°C, power of 43.99% and time of 27.86 min for total phenolics. However, an extraction temperature of 41.60°C, power of 44.88% and time of 27.49 min was optimal for free radical scavenging activity (judged by from DPPH reduction). Further, the UAE processes were significantly better than solvent extractions without ultrasound. By electron microscopy it was concluded that ultrasonic processing caused damage in cells for all treated samples (pumpkin, peach). However, the FTIR spectra did not show any significant changes in chemical structures caused by either ultrasonic processing or solvent extraction.     

Production of activated carbon from coconut shell: optimization using response surface methodology

The production of activated carbon from coconut shell treated with phosphoric acid (H3PO4) was optimized using the response surface methodology (RSM). Fifteen combinations of the three variables namely; impregnation ratio (1, 1.5, and 2); activation time (10, 20, and 30 min); and activation temperature (400, 450, and 500 degrees C) were optimized based on the responses evaluated (yield, bulk density, average pore diameter, small pore diameter, and number of pores in a unit area). Pore diameters were directly measured from scanning electron microscope (SEM) images. Individual second-order response surface models were developed and contour plots were generated for the optimization analysis. The optimum range identified for impregnation ratio was from 1.345 to 2, while for the activation time was from 14.9 to 23.9 min. For the activation temperature it was from 394 to 416 degrees C. The optimum points are 1.725, 19.5 min, and 416 degrees C, respectively. The models were able to predict well the values of the responses when the optimum variable parameters were validated as proven by the generally acceptable values of the residual percentages. Direct characterization of the pores using the SEM was found to be a good technique to actually see the pores and get actual measurements. Additionally, RSM has also proven to be a good tool in optimization analysis to get not only optimum production condition points but ranges, which are crucial for the flexibility of the production process, as well.     

Temporal changes of nutrient composition from pollen patty to bee bread with special emphasis on amino and fatty acids composition

Bee bread is prepared from pollen sources and salivary secretions by honey bee workers to serves the nutritional purpose of colony members. However, changes in nutrient composition occurring in bee bread from the pollen sources including pollen patty are poorly understood. We, therefore, examined the nutrient contents of pollen patty, used as source pollen feed of honey bee colony, and bee bread prepared by honey bee from this source at different time interval. Results revealed that amino acid content was increased from pollen patty to bee bread but fatty acid content was decreased. Essential amino acids like valine, lysine, and nonessential amino acids such as proline and alanine increased significantly over time. Among fatty acids, oleic acid, linoleic acid and linolenic acid decreased significantly from pollen patty to bee bread. Minerals contents in bee bread did not show any significant change during the study period. These changes could be presumably justified due to addition of nectar, honey, gland secretions and also microbial symbionts which remains as a task of further study.