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15 August 2009, Volume 30 Issue 16
Preparation and Characterization of Salidroside Proliposomes
XIA Shu-qin,FAN Ming-hui,XU Shi-ying*,ZHANG Xiao-ming
2009, 30(16):  35-40.  doi:10.7506/spkx1002-6630-200916001
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To enhance the stability of salidroside liposomes, freeze-drying method was used to prepare salidroside proliposomes. The particle morphology, size distribution and encapsulation efficiency of salidroside liposomes before freezedrying and after rehydration were studied in order to evaluate the protective effects of cryoprotectants. Mannitol was the optimal cryprotectant to prepare salidroside proliposomes. Salidroside proliposomes prepared with the mass ratio of mannitol to phospholipids of 20:1 looked plumpy and compact, the encapsulation efficiency reached above 40% after rehydration, its retention rate was higher than 90%, and the average particle size of salidroside liposomes after rehydration was 223.9 nm, which was similar to that of samples before freeze-drying. Fourier transform infrared (FTIR) spectral analysis indicated that mannitol and polar group of lipid formed hydrogen bonds which could protect the integrality of liposome membrane from being destroyed during freeze-drying.

Ultra High Pressure Extraction of Polysaccharides from Cordyceps militaris (L.ex Fr.) Link
GAO Feng1,ZHANG Shou-qin2,LIU Jing-bo1,YU Ya-li1,2,*
2009, 30(16 ):  41-43.  doi:10.7506/spkx1002-6630-200916002
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Ultra high pressure (UHP) technology was adopted to extract polysaccharides from Cordyceps militaris (L.ex Fr.) Link at normal temperature. The extraction of polysaccarides by UHP technology was optimized to obtain the maximum yield of polysaccarides by single-factor method and orthogonal array design. A polysaccharide yield of 10.13% was found under the optimized conditions for the extraction of polysaccarides from Cordyceps militaris (L.ex Fr.) Link as follows: extraction for 5 min using 45% as the extraction solvent at 400 MPa with the material/liquid ratio of 1:35 (g/ml). Polysaccharides can be extracted rapidly and effectively by this method without losing their physiological activity, thereby providing a novel and robust approach for extracting active components from Cordyceps militaris (L.ex Fr.) Link.

Extraction and Separation of Total Polyphenols from Herba gei
OUYANG Yu-zhu1,CHEN Xiao-dong1,2,TANG Hong-yu1,TANG Sai-yan1,XU Qiu-yan1
2009, 30(16 ):  44-47.  doi:10.7506/spkx1002-6630-200916003
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Ultrasonic-assisted extraction followed by macroporous resin separation was adopted to prepare total polyphenols from Herba gei. Four crucial extraction parameters such as alcohol concentration, material/solvent ratio, extraction temperature were optimized to obtain the highest yield of total polyphenols by orthogonal array design. Under the optimized conditions of extraction at 60 ℃ for 60 min with 60% ethanol as the solvent and the material/solvent ratio of 1:10 (g/ml), the yield of total polyphenols reached 1.77%.

Optimization of Microencapsulation Formulation of Jade Perch Oil and Its Effects on Fatty Acid Compositions
TAO Ning-ping,WANG Xi-chang*, KANG Yin
2009, 30(16 ):  48-51.  doi:10.7506/spkx1002-6630-200916004
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The microencapsulation formulation of refined oil from jade perch viscera was optimized to attain the maximum microencapsulation efficiency (MEE) by orthogonal array design, and the microstructure of microcapsules were observed under SEM and their fatty acid composition was analyzed by GC-MS. The optimum formula for microencapsulating jade perch oil was as follows: modified starch 50%, jade perch oil 35% and lecithin used as an emulsifier 0.75% and an MEE of 94.1% was achieved by the microencapsulation with this formula. The sensory and physical-chemical indexes of microcapsule products both met the fish oil microcapsule standard stipulated by the Chinese aquatic industry. The ultrastructure observation revealed hat the microcapsules had particle sizes ranging from 6 to 8 μm, and the surface and inside structures of the microcapsules both were good. Little differences of fatty acid composition were found among crude oil, refined oil and microencapsulated oil from jade perch viscera, suggesting that the processes of fish oil extraction, refinement and spray-drying microencapsulation all are designed soundly. As a result, jade perch oil with high concentration of unsaturated fatty acids vulnerable to the influence of environmental exposure is well protected by microencapsulation.

Ultrasonic-assisted Extraction of Alkaloids from Lotus Leaves
LIU Shu-xing1,GUO Rui-xia2,ZHAO Fang1
2009, 30(16 ):  52-55.  doi:10.7506/spkx1002-6630-200916005
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Ultrasonic-assisted extraction was applied to prepare alkaloids from lotus leaves. The extraction conditions of alkaloids were optimized to maximize the yield of alkaloids by single-factor and orthogonal array methods. Under the optimized extraction conditions as follows: extraction for 0.5 h with 65% ethanol as the extraction solvent and the solid to liquid ratio of 1:50, at 45 kHz, 80 ℃ and pH 5, the yield of alkaloids reached 0.17%.

Immobilized Malate Dehydrogenase-catalyzed Synthesis and Biological Activities of L-Ascorbyl Malate
MIAO Jing-zhi,LU Zhao-qi*,CAO Ze-hong,DONG Yu-wei
2009, 30(16 ):  56-59.  doi:10.7506/spkx1002-6630-200916006
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Using immobilized malate dehydrogenase as the catalyst, L-ascorbyl malate was synthesized in an acetone system, and the product content was determined by HPLC. The results showed that when the catalyst amount was 4 g/L, the molar ratio of L-ascorbic acid to malic acid was 1:3, and the reaction temperature was 55 ℃, the highest L-ascorbyl malate yield of 69.30% could be obtained. In addition, when the addition of L-ascorbyl malate to soybean oil was 0.05%, the POV value on the 7th day was only 8.41 meq/kg, indicating that L-ascorbyl malate has significant antioxidant activity. When the addition of L-ascorbic malate was 0.18%, its inhibitory rates against Escherichia coli, Staphyloccus aureus, and Bacillius subitills were 92.13%, 90.84% and 84.63%, respectively. It was concluded that L-ascorbyl malate is a potential antioxidant and bacteriostat.

Development of A Bifidobacterium Milk Beer
YAN Cheng
2009, 30(16 ):  60-64.  doi:10.7506/spkx1002-6630-200916007
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A new-type drink, which is a low-alcohol milk beer containing carbon dioxide, was made through the fermentation of fresh milk with bifidobacterium, Lactobacillus bulgaricus, Streptococcus thermophilus and yeast. Single-factor method and orthogonal array design were adopted to optimize the fermentation parameters to attain the highest sensory and proper alcohol degree. Compared with other three Angel brand dry yeast products such as wine active dry yeast, high-temperature resistant and highly-active dry yeast, and highly-active dry yeast, beer active dry yeast was more suitable for the production of milk beer because product yielded using it has a pleasant sour-sweet taste and elegant mouthfeel and is a fabulous combination of milk and beer. The best flavor of product was achieved by the simultaneous inoculation of yeast and Lactobacillus starter for fermentation. The optimal fermentation conditions were confirmed as follows: inoculation of 5% yeast and 3% Lactobacillus starter, and fermentation at 28 ℃ for 16 h.

Extraction and Properties of Pigment from Fruits of Basella rubra L.
ZHAO Jian-fen,DONG Ji,WEI Shou-lian
2009, 30(16):  65-68.  doi:10.7506/spkx1002-6630-200916008
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The optimal conditions for extracting pigment from fruits of Basella rubra L. were determined by orthogonal array design. The physico-chemical properties of the pigment such as solubility, spectral characteristics, thermal and light stability, recovering trait and color value were studied, and the effects of pH, metal ions and common food additives on the pigment were investigated. The highest pigment yield could be obtained under the optimized conditions as follows: using 0.01 mol/L hydrochloride as the extractant, extraction at 45 ℃ for 40 min. The pigment was easy to disperse in water and had good recovering trait and the average color value was 11.39. Its stability was nice below 60 ℃ and in pH 2 - 10, but light could accelerate its decomposition. Except Fe2+, Fe3+ and Cu2+, other metal ions such as K+, Na+, Ca2+, Mg2+, Zn2+ and food additives such as NaCl, VC, glucose, sodium benzoate, citric acid and Na2SO3 had no influence on the stability. Altogether, this pigment has good stability under certain conditions.

Extraction and Biological Activities of Polysaccharides from Fermented Mycelia of Lachnum calyculiforme
YE Ming,PENG Wei,FANG Cong,LI Shi-yan,YANG Liu
2009, 30(16):  69-72.  doi:10.7506/spkx1002-6630-200916009
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The fermented mycelia of Lachnum calyculiforme YM-262 were extracted with hot water to produce polysaccharides. The effects of temperature of hot water, extraction duration and liquid/material ratio (V/m) on yield of polysaccharides were investigated by one-factor-at-a-time method. Subsequently, the factors were optimized to attain the maximum yield of polysaccharides using a three-factor and three-level orthogonal array design leading to a set of 9 experiments. Furthermore, polysaccharides obtained by optimized extraction procedure were analyzed for their antioxidant activity and antimicrobial activity. The optimum conditions for extraction of polysaccharides were determined as follows: with a ratio of liquid to material of 70:1, extraction at 80 ℃ for 4 h, and a yield of polysaccharides of 9.16% was achieved by the optimized extraction procedure. The polysaccharides were found to possess considerable antioxidant activity—their scavenging rate to DPPH radicals was 18.88% at 30 mg/L, to hydroxyl radicals 50.43% at 40 mg/L, and to superoxide anion radicals 85.53% at 100 mg/L. The polysaccharides also displayed certain antimicrobial activities to Escherichia.coli, Bacillus subtilis, Staphylococcus aureus and Saccharomyces cerevisiae, with a minimum inhibitory concentration (MIC) in the range of 15.61-62.5 μg/ml.

Preparation and Sulfation of Polysaccharides from Pumpkin
KONG Qian,ZHOU Ting,WU Gai-lan,HUANG Jing-rong,LI Quan-hong*
2009, 30(16):  73-77.  doi:10.7506/spkx1002-6630-200916010
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Crude water-soluble polysaccharides were separated from pumpkin by hot water extraction and gradient ethanol precipitation, and purified/fractionated by DEAE-Sepharose Fast Flow anion-exchange chromatography and a acidic polysaccharide PL-2 was obtained. Sepharose CL-6B gel filtration chromatography and HPLC analysis demonstrated that the component of PL-2 was homogenous and its molecular weight was more than 2000 kD. The polysaccharide was chemically modified by chlorosulfuric acid-pyridine method, sulfur trioxide-pyridine method and sulfamic acid, respectively. The identification of BaCl2-gelatin turbidimetry and infrared spectra revealed that sulfur trioxide-pyridine method displayed the best sulfation effect.

Use of Membrane Separation Combined with Alcohol Precipitation for the Separation and Purification of Isoflavones from Low-denatured Soybean Meal
XU Fu-ping1,LIANG Zhi-jia2,TIAN Juan-juan2
2009, 30(16):  78-82.  doi:10.7506/spkx1002-6630-200916011
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A new technology of combined membrane separation and alcohol precipitation was applied for the purification of isoflavones from the ethanol extract of low-denatured soybean meal. The method for the determination of isoflavone content was developed, and the membrane separation conditions of isoflavones including membrane material type and membrane hole number, membrane regeneration, and the relationship of membrane flux with operation pressure and temperature were investigated. Compared with resin adsorption method, alcohol precipitation was easier to operate and had less process steps and led to higher purity of isoflavones. Alcohol precipitation parameters for the purification of isoflavones were optimized by single-factor and orthogonal array methods. Under the optimized conditions as follows: mass fraction of isoflavone extract 30%, 70% ethanol/material ratio 7:1, stirring time 10 min, adjusting soluble solid content in the supernant after recover of ethanol to 5%, and standing time 5 h, the content of isoflavones in final product was above 60%. An isoflavone yield of 0.7‰ was obtained under the optimized conditions as follows: mass fraction of isoflavone extract 30%, 70% ethanol/material ratio 5:1, soluble solid content in the supernant 8%, and standing time 4 h.

Production Process of Calcium-rich Germinated Brown Rice
JIANG Hu1,FU Jin-heng1,2,*,SU Hu3,FANG Lan-kun1,CAI Yan1,HAN Xiao-yun1
2009, 30(16 ):  83-85.  doi:10.7506/spkx1002-6630-200916012
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With the brown rice produced in Jiangxi as the raw material, the best soaking and cultural conditions of brown rice germination were determined, and the effects of different concentrations of CaCl2 solution on the germination rate and the content of γ-aminobutyric acid (GABA), one of important nutrients in brown rice were investigated. The results indicated that when the brown rice was soaked in 0.2% CaCl2 solution at 28 ℃ for 9 h and then cultured in constant temperature and humidity incubator at 30 ℃ for 21 h, the germination rate was over 80%, and the calcium and GABA content in the germinated brown rice were 0.62 and 0.36 g/kg, respectively.

Microencapsulation of Hawthorn Flavonoids
GAO Han,SUN Jun-liang,LIU Ben-guo,KONG Jin,ZHOU Quan-xia
2009, 30(16 ):  86-88.  doi:10.7506/spkx1002-6630-200916013
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The technology for producing microcapsules of hawthorn flavanoids was studied. The key factors affecting microcapsulation were confirmed by orthogonal test and the optimal conditions obtained were as follows: with the ratio of material core to wall of 1:5 (g/g) and wall material concentration of 3%, grinding at 25 ℃ for 15 min. Under these conditions, the microencapsulation of product could reach 76.3%.

Preparation of A Food-grade Preservative, Calcium Propionate
GAO Xin1,YANG De-yu2,WANG Xiao-gang1,LI Feng1,MEI Xiao-bing1,FAN Zheng1
2009, 30(16):  89-93.  doi:10.7506/spkx1002-6630-200916014
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A new technology for producing calcium propionate from oyster shell under normal pressure and temperature was studied. Based on the single factor tests of water/shell ratio, shell-acid ratio, reaction time and neutralization time, the optimum production conditions were determined by response surface methodology (RSM) as follows: particle diameter of oyster shell 0.125 -0.160 mm, water/shell ratio 16.7, shell/acid ratio 1.08, reaction time under normal temperature 172 min, pH of solution after reaction adjusted to 7-8, and neutralization time 38 min. The verification test showed that the content of calcium propionate reached 99.61%.

Response Surface Methodology as An Approach to Optimizing Ultrasonic-assisted Extraction of Flavonoids from Navel Orange Peel
YANG Jia,ZHANG Guo-wen*,WANG Jia-rong,FENG Li-hui
2009, 30(16 ):  94-97.  doi:10.7506/spkx1002-6630-200916015
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The ultrasonic-assisted extraction process of flavonoids from navel orange peel was studied. On the basis of single factor tests, response surface analysis was designed by Design-Expert Software 7.1.3.1, and the effects of extraction time, ethanol concentration, ratio of liquid to solid, and temperature on the yield of flavonoids were investigated. The results showed that the optimum extraction conditions were as follows: with 58.91% ethanol as the extraction solvent and a ratio of liquid to solid of 30.68 ml/g, twice extraction at 75 ℃ for 20.79 min. The predicted yield of flavonoids was 3.298%, close to the practical yield of flavonoids of 3.351%.

Optimization of Ultrasonic-assisted Extraction Technology of Ganoderma lucidum Polysaccharides by Response Surface Methodology
DONG Yan-hong1,LI Shu-jing1,ZHENG Hui-hua2,LI Xue1,CHEN Gui-tang3,ZHAO Li-yan1,*, XIN Zhi-hong1, HU Qiu-hui1
2009, 30(16):  98-101.  doi:10.7506/spkx1002-6630-200916016
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Three key factors affecting the extraction yield of Ganoderma lucidum polysaccharides such as ultrasonic power, ultrasonic extraction time and ratio of water to solid were optimized by response surface methodology. Design Expert analysis indicated that the best ultrasonic power, ultrasonic extraction time and the ratio of water to solid were 680 W, 17 min and 28:1, respectively. Under these conditions, the yield of polysaccharide was (20.79 ± 0.11) mg/g, which was consistent with the predicted value.

Ultrasonic-assisted Extraction of Flavonoids from Navel Orange Peel
HUANG Yun-hong,GAO Xing-qiang,LI Wei-wei,LONG Zhong-er
2009, 30(16 ):  102-105.  doi:10.7506/spkx1002-6630-200916017
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Ultrasonic treatment was applied to the ethanol extraction of flavonoids from navel orange peel. The effects of four crucial variables such as ultrasonic power, ethanol concentration, material/liquid ratio and ultrasonic treatment time on the yield of flavonoids were investigated by single-factor method. An orthogonal array design leading to a set of 9 combinations of these variables was performed to obtain the highest yield of flavonoids. Under the optimized conditions as follows: ultrasonic power 125 W, 60% alcohol as the extraction solvent, material/liquid ratio 1: 35, and extraction time 60 min, a yield of flavonoids of 49.3 mg/g was achieved.

Optimization of Extraction and Physico-chemical Properties of Tilapia Skin Collagen
YANG Xian-qing1,ZHANG Shuai1,2,HAO Shu-xian1,LI Lai-hao1,WU Yan-yan1,SHI Hong1,HUANG Hui1
2009, 30(16 ):  106-110.  doi:10.7506/spkx1002-6630-200916018
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The hot water extraction of collagen from tilapia skin was optimized using Box-Behnken design and response surface methodology and the collagen was analyzed for its physicochemical properties. The optimum conditions for the improved extraction of collagen from tilapia skin were determined as follows: soaking tilapia skin in 0.213 mol/L HCl solution with a ratio of material to liquid of 1:20 (m/V) at 4 ℃ for 21 min prior to the hot water extraction with a ratio of material to liquid of 1:7 (m/V) at 42 ℃ for 12.6 h, and the optimized extraction led to a collagen content of 293.018 mg/g. The collagen was rich in glycine, proline and hydroxyproline. SDS-PAGE analysis showed that this collagen contained α1, α2 and β chains indicating that it is a typical type I collagen. Its gel strength and characteristic viscosity were 682 g and 1.461dl/g, respectively.

Ultrasonic-assisted Extraction of Alkaloids from Semen Strychni by Response Surface Analysis
ZOU Jian-guo,XU Xiao-long,LIU Yan-yan,PENG Hai-long,XU Biao,LUO Ping,XIAO Jian-mou
2009, 30(16 ):  111-114.  doi:10.7506/spkx1002-6630-200916019
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Response surface methodology (RSM) was applied to optimize the ultrasonic-assisted extraction conditions of alkaloids from Semen Strychni. Based on the signal factor test, ethanol concentration, ultrasonic power and extraction time were chosen as casual factors, and a three-factor and three-level Box-Behnken central composite design leading to a set of 15 combinations of these variables was performed to attain the highest weighed value of yields of strychnine and brucine (their weighed coefficients both were 0.5 ). The optimal ultrasonic-assisted extraction conditions were determined as follows: particle size of Semen Strychni 20-40 mesh, ethanol concentration 32.3%, pH 5, ultrasonic power 300 W, ultrasonic-assisted extraction time 37 min, and ratio of liquid to solid 10:1. Under these conditions, the yields of strychnine and brucine were 18.75 mg/g and 9.24 mg/g respectively, which were consistent with the predicted values. These results demonstrate the validity of this extraction process.

Optimization of Water Extraction of Cochineal Dye
ZHANG Hong1,LU Yan-min1,2,ZHENG Hua1,ZHOU Mei-cun2,FENG Ying1, CHEN Xiao-ming1,*,ZHANG Zhong-he1
2009, 30(16 ):  115-118.  doi:10.7506/spkx1002-6630-200916020
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The effects of extraction temperature, extraction time and ratio of raw material to liquid on the extraction of cochineal dye were investigated by single-factor method. Subsequently, these factors were optimized by orthogonal array design. The optimal conditions for the maximum extraction of cochineal dye were determined as follows: with the ratio of raw material to liquid of 1:4 (m/V), extraction temperature 80 ℃ for an extraction duration of 5 h, and extraction number 6. Under these conditions, the yield of red dye was 41.89%, and its purity was 32.15%.

Use of Freezing Wall-breaking Method for the Extraction of Barley Green from Buckwheat Seedlings
WANG Wei-jian1,GUO Li-quan2,LENG Jin-song1
2009, 30(16 ):  119-122.  doi:10.7506/spkx1002-6630-200916021
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A liquid product of barley green was prepared from buckwheat seedlings by the steps of freezing wall breaking, extraction with 1/150 mmol/L phosphate buffer containing 0.01% sodium erythorbate by grinding, vacuum concentration and instantaneous ultra-high temperature sterilization. The freezing wall breaking extraction of buckwheat seedlings was optimized by orthogonal array design. Under optimized conditions, the contents of protein, chlorophyll and rutin and the SOD activity of barley green extract were 0.26%, 9.37 mg/100 ml, 1.03 mg/100 ml, and 242 U/100 ml, respectively. Other indexes of the product obtained were in accordance with the national standards for plant health products.

Microencapsulation of Arachidonic Acid
YANG Jing,ZHENG Wei-wan*,ZHOU Xiao-juan,SHI Hao,LONG Ji-yun,LIAO He-jing
2009, 30(16 ):  123-126.  doi:10.7506/spkx1002-6630-200916022
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Spray-drying method was used to prepare a type of stable arachidonic acid (AA) product, AA microcapsules, which could slow down the process of auto-oxidation effectively, and the properties of AA microcapsules were tested and evaluated. The AA microcapsules prepared using maltodextrin as the wall material and complex emulsifier L had the best stability. No layer phenomenon and particles being linked to the wall were found in reconstituted emulsions. The embedding rate reached 93.48%.

Quality Control of Apple-Roxburgh Rose Cloudy Juice during the Processing
ZHAO Guang-yuan,GAO Zhi-song,ZHENG Jian-qiang
2009, 30(16 ):  127-130.  doi:10.7506/spkx1002-6630-200916023
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During the processing of apple-roxburgh rose cloudy juice, the L* value was increased at first and then decreased, while a* value was increased, however these changes were not significant. The turbidity was increased in the processes of illing enzyme inactivation, homogenization and sterilization. The amount of polyphenols in juice was decreased because of oxidization, and the amounts of total phenols, tanninin in apple-roxburgh rose cloudy juice were retained at 59.39% and 65.12%, respectively. The reducing VC and total VC were decreased by 57% and 26.34%, respectively. GC-MS analysis indicated that the volatile components were mainly esters and alcohols. Alcohols decreased and esters increased after enzyme inactivation, while alcohols and esters both changed conversely after sterilization.

Ultrasonic-assisted Extraction of Total Flavonoids from Leaves of Wild Xanthium L. Grown in Xinjiang
ABULIKEMU Abulizi,ABULIMITI Abudoukadeer,DILINUER Talipu*
2009, 30(16 ):  131-134.  doi:10.7506/spkx1002-6630-200916024
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An ultrasonic-assisted extraction of total flavonoids from leaves of wild Xanthium L. grown in Xinjiang was developed. Single-factor method was adopted to investigate the effects of ethanol concentration, extraction temperature, ultrasonic treament time and material/liquid ratio on the extraction yield of total flavonoids. Subsequently, a four-variable/three-level orthogonal array design was carried out to maximize the extraction of total flavonoids. An extraction yield of total flavonoids of 3.10 mg/g was attained under the optimum extraction conditions as follows: ethanol concentration 50%, extraction temperature 30 ℃, ultrasonic treatment time 30 min, and material/liquid ratio 1:50. The ultrasonic-assisted extraction can be regarded as a purely physical process without any pollution, thus providing an ideal approach to extracting flavonoids from leaves of Xanthium L.

Microwave Extraction and Content Determination of Polysaccharides from Rubia cordifolia L.
TIAN Chun-lian1,XIANG Bin2
2009, 30(16 ):  135-136.  doi:10.7506/spkx1002-6630-200916025
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The microwave extraction of polysaccharides from Rubia cordifolia L. was optimized to attain the maximum extraction yield by orthogonal array design, and the contents of polysaccharides in different parts of Rubia cordifolia L. were measured by anthrone-sulfuric acid method. The medium-fire microwave extraction for 60 s with the ratio of material to water of 1:40 led to the highest extraction yield of 10.20%. Old leaves of Rubia cordifolia L. had the highest content of polysaccharides of 13.34%, followed by old stems, 10.28%.

Optimization of Supercritical CO2 Extraction of Allicin from Garlic Cloves
YANG Liu,TAO Ning-ping*
2009, 30(16 ):  137-141.  doi:10.7506/spkx1002-6630-200916026
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We optimized the supercritical CO2 extraction of allcin from garlic cloves to obtain the highest extraction yield by single-factor and orthogonal array methods. The optimal conditions for the improved supercritical CO2 extraction of allicin were determined as follows: extraction press 15 MPa, extraction temperature 40 ℃, static extraction time 30 min and dynamic extraction time 105 min. Under these conditions, the extraction yield could reach 76.70%, which was higher than 63.16% by traditional solvent extraction and 68.12% by ultrasonic-assisted extraction.

Ultrasonic Extraction of Cochineal Dye
LU Yan-min1,2,ZHENG Hua1,ZHANG Hong1,*,ZHOU Mei-cun2,GAN Jin1,MA Li-yi1,ZHANG Zhong-he1
2009, 30(16 ):  142-145.  doi:10.7506/spkx1002-6630-200916027
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Single-factor and orthogonal array methods were adopted to optimize the ultrasonic extraction of cochineal dye. The optimal conditions for the improved extraction of cochineal dye were determined as follows: ratio of material to solvent 1:6, ultrasonic power 1400 W, ultrasonic treatment time 12 min, pulse on time 10 min, pulse off time 4 min, and extraction number 5. Under these conditions, the yield of cochineal dye was 42.08%.

Preparation and Identification of Octenylsuccinate Sucrose Ester and Its Application in Microencapsulation of Soybean Oil
ZHOU Xiao-juan,ZHENG Wei-wan*,YANG Jing,SHI Hao,LIAO He-jing
2009, 30(16 ):  146-148.  doi:10.7506/spkx1002-6630-200916028
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Octenylsuccinate sucrose ester was synthesized via esterification with sucrose and octenylsuccinate as raw materials. IR spectral analysis revealed that the C = O and C = C vibration absorption peaks appeared at 1724.05 and 1569.57 cm-1, respectively. A new microencapsulation system was established and an embedding rate of 96.2% was obtained by replacing sodium caseinate with octenylsuccinate sucrose ester in the microencapsulation. It is illustrated that octenylsuccinate sucrose este has good surface activity.

Application of Microwave Technology in Protein Extraction from Swim Bladder of Silver Carp
DUAN Zhen-hua1,2,SUN Xiao-ling1,WANG Ju-lan2
2009, 30(16 ):  149-152.  doi:10.7506/spkx1002-6630-200916029
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The microwave extraction of protein from swim bladders of silver carps was investigated. Four crucial extraction parameters were optimized to attain the highest extraction yield of protein by single-factor and orthogonal array designs. The optimal conditions for attain the maximum extraction yield of 22.79% were determined as follows: with the ratio of material to liquid (water) of 1:30, extraction at 300 W for 2.5 min. This study will provide a practical reference for the application of  microwave extraction in the preparation of protein from swim bladders.

Response Surface Optimization of Neutrase-catalyzed Hydrolysis of Cottonseed Protein
GAO Dan-dan,CHANG Tong,CAO Yu-sheng*,WANG La-mei
2009, 30(16 ):  153-157.  doi:10.7506/spkx1002-6630-200916030
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Cottonseed protein was hydrolyzed by neutrase to produce active peptides. Single-factor method was adopted to investigate the effects of temperature, pH, enzyme amount and hydrolysis time on degree of hydrolysis (DH) of cottonseed protein. Subsequently, a central composite design (CCD) involving 17 experiments of three variables (i.e., temperature, pH and hydrolysis time) at three levels combined with response surface methodology was employed to attain the highest DH. When the amount of enzyme was 6000 U, the optimal temperature, pH value and hydrolysis time were 39.5 ℃, 7.5 and 5.5 h, respectively. Under the above conditions, the DH was 38.09%.

Purification of Saponins from Aralia continentalis Leaves Using Macroporous Adsorption Resin and Its Bioactivity
FENG Ying1,2,LI Tian-lai2,*,MENG Xian-jun1,FAN Wen-li2
2009, 30(16 ):  158-161.  doi:10.7506/spkx1002-6630-200916031
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Static absorption and desorption experiments were carried out to screen the optimum resin from 4 types of macroporous adsorption resins and macroporous resin HPD-100 was found to have the best performance for the purification of crude extract of saponins from Aralia continentalis leaves. In dynamic adsorption and desorption experiments, the optimum purification conditions of saponins by HPD-100 were determined as follows: saponin concentration 0.3 mg/ml, sampling volume 18 times of bed volume (BV), pH value of the sample 5, and gradient elution water → 10% ethanol → 30% ethanol → 50% ethanol. The eluates of 30% and 50% ethanol both had certain scavenging activity to superoxide radical and antibacterial activity.

Extraction and Preliminary Purification of Cochineal Wax
ZHENG Hua,ZHANG Hong*,GAN Jin,TANG Li-ying,ZHAO Hong,LU Yan-min,MA Li-yi,ZHANG Zhong-he
2009, 30(16 ):  162-165.  doi:10.7506/spkx1002-6630-200916032
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For the comprehensive utilization of cochineal and the development of new insect wax resource, several common organic solvents were taken to extract cochineal wax. Xylene and carbon tetrachloride were two efficient solvents for the extraction of cochineal wax with a yield of 2.95% - 2.97%. A cochineal wax product with 85.0 ℃ melting point and 80.4% whiteness could be obtained from the xylene extract by the single-stage treatment with absolute ethanol, while the three-stage treatment with xylene, petroleum ether and ethanol yielded a product with 84.3 ℃ melting point and 82.2% whiteness. The melting point and the whiteness of the two products both were similar to those of other common insect wax products.

Optimization of Quick-frozen Production Technology of Chinese Northern Steamed Bread
ZHANG Jian,LI Meng-qin,REN Hong-tao,ZHENG Xiang-yang
2009, 30(16 ):  166-168.  doi:10.7506/spkx1002-6630-200916033
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The quick-frozen production technology of Chinese northern steamed bread was investigated by orthogonal array design and fuzzy comprehensive evaluation. The high-quality quick-frozen steamed bread could be produced when the addition amount of water was 46%–48%, the quick-frozen wind velocity was 3–4 m/s, and the formula of modifier was as follows: monoglyceride 0.3%, lipase 30 mg/kg, complex phosphate 0.2%, and vitamin C 10 mg/kg.

Use of Wet Superfine Comminution Technology for the Extraction of Total Flavonoids from Flos Chrysanthemi
ZONG Wei,ZHANG Huan-huan
2009, 30(16 ):  169-171.  doi:10.7506/spkx1002-6630-200916034
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Total flavonoids in Flos Chrysanthemi were extracted by wet superfine comminution technology. Single-factor method was adopted to investigate the effects of gap between colloid mill teeth, ethanol concentration, extraction temperature and extraction time on the extraction yield of total flavonoids was investigated. Subsequently, an orthogonal array design involving 9 experiments of the four factors at three levels was employed to attain the highest extraction yield of total flavonoids. The optimum extraction conditions were determined as follows: gap between colloid mill teeth 20 μm, ethanol concentration 75%, temperature 70 ℃ for an extraction duration of 20 min. Under these conditions, the extraction yield of total flavonoids was 3.45%. A high extraction yield can be achieved in a short period of time using wet superfine comminution. In general, this technology is a promising approach for the extraction of total flavonoids from Flos Chrysanthemi.

Response Surface Methodology as An Approach to Optimize Preparation Process of Antihypertensive Peptides from Egg White Proteins
LIU Bo-qun,LIN Song-yi,YU Zhi-peng,YANG Yu-jun,ZHAO Wen-zhu,LIU Jing-bo*
2009, 30(16 ):  172-176.  doi:10.7506/spkx1002-6630-200916035
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Egg white proteins were hydrolyzed by Alcalase to prepare antihypertensive peptides. The effects of four crucial factors on the degree of hydrolysis (DH) of egg white proteins were analyzed by single-factor method. Subsequently, a three factor/three-level Box-Behnken experimental design combining with response surface methodology (RSM) and quadratic programming (QP) was employed for maximizing the Alcalase-catalyzed hydrolysis of egg white proteins and a quadratic regression model was obtained using the Design-Expert software. The optimal hydrolysis conditions were determined as follows: substrate concentration 1.14 %, temperature 58 ℃, pH 10.10, and ratio of enzyme to substrate 6%. Under these conditions, the angiotensin-converting enzyme (ACE) inhibitory activities of 1 mg/ml peptide hydrolysates after hydrolysis for 1, 1.5, 2, 2.5 and 3 h were determined by HPLC to be 25.72% , 66.85%, 36.00% , 43.31% , 50.84%, respectively. It is very clear that the peptide hydrolysate after 1.5 h of hydrolysis has the highest ACE inhibitory activity among the 5 peptide hydrolysates.

Extraction and Separation of Ursolic Acid from Hawthorn Fruits
LI Hai-bin
2009, 30(16 ):  177-180.  doi:10.7506/spkx1002-6630-200916036
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This study investigated the ethanol extraction and the purification using macroporous adsorption resin of ursolic acid from hawthorn fruits. An orthogonal array design involving 16 experiments of four crucial factors at four levels was employed to maximize the ethanol extraction. The optimal macroporous adsorption resin and solvent for purifying ursolic acid were determined and the dynamic adsorption and desorption of ursolic acid on macroporous adsorption resin AB-8 were investigated. The optimum extraction conditions of ursolic acid were as follows: using 90% ethanol as the extractant and a ratio of solid to liquid of 1:10, extraction at 80 ℃ for 2.5 h, and the extraction yield of ursolic acid was 1.07% under these conditions. The optimum purification conditions with macroporous adsorption resin AB-8 were as follows: ursolic acid concentration 3.01 –4.01 mg/ml and eluent pH 10.0 for a stepwise elution with water and different concentrations of ethanol solution at a flow rate of 1.5 ml/min. A purity of 97.0% and a recovery of 98.1% were achieved under the optimized purification conditions. An ursolic acid product with high purity and yield can be obtained by this extraction and purification process and therefore it is a promising approach for the industrialized production of ursolic acid.

Extrusion Technology and Raw Material System of High-moisture Textured Soy Protein
LIU En-qi1,HE Ju-ping1,LIU Quan-de1,CHEN Zhen-jia2,WANG Wei2,LI Yu-e2
2009, 30(16 ):  181-184.  doi:10.7506/spkx1002-6630-200916037
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Soy protein isolate and potato starch were partially added to cold pressed soybean meal to develop high-moisture textured soy protein products by optimized extrusion technology and raw material system. The results showed that extrusion temperature, material moisture, content of soy protein isolate and potato starch had significant effects on product quality. The texturization degree, brilliance index, effective lysine content and water keeping ability were increased gradually with the increase of material moisture content and high forming temperature was needed at the same time, and the water binding capacity and tissue integrity were increased gradually with the addition of soy protein isolate as well. Starch was helpful to bond and extrusion moulding among textured soy protein, but the addition quantity should be strictly controlled, because high starch content would decrease the texturization degree of textured soybean protein. The optimum technology of raw material system and extrusion temperature was determined in this study as follows: soy protein isolate 20%, starch 5%, material moisture content 60%, and extrusion temperature 185 ℃.

Optimization of Ethanol Extraction of Rutin from of Chinese Jujube Residue by Response Surface Methodology
WANG Na1,PAN Zhi-li1,XIE Xin-hua1,XING Wei-wei2,AI Zhi-lu1,*
2009, 30(16 ):  185-188.  doi:10.7506/spkx1002-6630-200916038
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The effects of ethanol concentration, temperature, extraction time and ratio of material to liquid on the extraction of rutin from Chinese jujube residue were investigated. Subsequently, a central composite design leading to 17 experiments of the other three variables expect ratio of material to liquid at three levels combined with response surface methodology was employed to maximize rutin extraction. The optimum extraction conditions were determined as follows: ratio of liquid to solid 1:40 (m/V), ethanol concentration 69%, and temperature 70 ℃ for an extraction duration of 8.5 h, and under these conditions, the maximum extraction rate of 155.46 mg/100 g was obtained.

Response Surface Optimization of Extraction Technology of Procyanidins from Buckwheat Shells
ZHA Yang-chun1,YANG Yi-ting1,HU Xiao-han1,LI Tao1,2,LIU Rui1,*
2009, 30(16 ):  189-192.  doi:10.7506/spkx1002-6630-200916039
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Procyanidins in buckwheat shells was extracted with a mixture of 70% ethanol and water. In order to maximize the extraction of procyanidins, a rotatable orthogonal central composite design combined with response surface and ridge analyses was employed. Extraction temperature and time affected the yield of procyanidins significantly (P < 0.05), while the effect of ratio of material to liquid was not significant. The optimum extraction conditions were as follows: ratio of material to liquid 1: 16.9, temperature 73 ℃, and extraction time 118.9 min. Under these conditions, the actual yield of procyanidins was 0.56%, which accounts for 94.9% of the theoretically predicted value of 0.59%.

Preparation and Quality Evaluation of Coix Seed Oil Liposomes
LIU Yu-zhen,XIONG Hua*,JIANG Yan,HAN Dan,XU Jing-shui
2009, 30(16 ):  193-197.  doi:10.7506/spkx1002-6630-200916040
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Coix seed oil liposomes were prepared by a technology of combined ethanol injection and ultrasonic. An orthogonal array design involving 9 experiments of four crucial factors at 3 levels was employed to obtain the highest embedding rate. The morphology of coix seed oil liposomes was observed under SEM, their particle size and Zeta potential were measured, and their storage stability was evaluated in terms of appearance, T500nm, pH value and retention rate of coix seed oil. The optimum preparation conditions were as following: concentration of lecithin 20 mg/ml, ratio of lecithin to coix seed oil to cholesterol 3:2:1 (m/m), and using phosphate buffer solution (pH 6.7) as the aqueous phase. The coix seed oil liposomes prepared under the above conditions were observed to be uniform and round with a mean particle size of 207.4 nm and their Zeta potential and embdeding efficiency were - 26.4 mV and 72.48%, respectively. The stability of bilayer structure of coix seed oil liposomes is related to storage temperature, and they have the longest storage life in a light-avoiding environment at 4 ℃.

Microwave Extraction and Stability of Brown Pigment from Camellia oleifera Shells
QIU He-yuan,CHEN Jie-man,HU Jing-ru
2009, 30(16 ):  198-202.  doi:10.7506/spkx1002-6630-200916041
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The microwave extraction technology of brown pigment from Camellia oleifera shells was optimized by L9(34) orthogonal array design and the stability of the pigment was studied. The optimal technological conditions were determined as follows: microwave irradiation time 240 s, microwave power 320 W, extraction solvent 70% ethanol and ratio of material to solution 1:10 (g/ml). A yield of brown pigment of 5.92% was achieved under these conditions. This pigment was found to have a brown-red color and stable to natural light, ultraviolet radiation, temperature, oxidizing agent, reducing agent and some metal ions such as K+, Ca2+, Cu2+, Zn2+, Ba2+ and Mg2+. In pH 2-7, it was also stable. Alkali had hyperchromic effects on the pigment, while Fe3+ had hypochromic effect.

Synthesis of Glyceryl Butyrate and Its Application in Microencapsulation of Soybean Salad Oil
LIAO He-jing1,ZHENG Wei-wan1,*,YI Lu-yao1,2,ZHOU Xiao-juan1,YANG Jing1,SHI Hao1
2009, 30(16 ):  203-205.  doi:10.7506/spkx1002-6630-200916042
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Glyceryl butyrate was synthesized via esterification from glycerol (0.5 mol) and butanoic acid (0.4 mol) with carbon tetrachloride (25 ml) as the water carrying agent and phosphoric acid as the catalyzer. The structural analysis by FT-IR spectroscopy showed that the target product was obtained, which contained 60.2% glyceryl monobutyrate. The use of this synthesized glyceryl butyrate as an emulsifier led to a high microencapsulation efficiency of soybean salad oil.

Removal of Undesirable Flavor from Chinese Traditional Cheese
XUE Lu,HU Zhi-he
2009, 30(16 ):  206-209.  doi:10.7506/spkx1002-6630-200916043
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The undesirable flavor such as bitterness, acerbity, and rancidity is usually formed during storage of Chinese traditional cheese, which restricts the industrialization of Chinese traditional cheese. In an attempt to explore convenient and economical methods for the removal of undesirable flavor from Chinese traditional cheese, the effects of coagulating procedures and addition of masking agents on the sensory quality of Chinese traditional cheese were investigated. The undesirable flavor of Chinese traditional cheese could be remarkably reduced by the coagulation at 55-65 ℃ for 80-100 min with the addition of glutinous rice wine of 6%-8%. The single addition of 0.15% β-cyclodextrin, 0.03% malic acid, 0.1% gelatin and 0.03% citric acid all could debitterize Chinese traditional cheese, and did not impact its tissue. No improvement was found in the flavor of cheese by the single addition of Ca2+, CMC and taurine.

GC-MS Determination of Monosaccharide Composition of Oligosaccharides from Fruits of Chinese Jujube (Ziziphus jujuba Mill var. muzao)
LIN Qin-bao1,JIANG Mei-feng1,YANG Chun2
2009, 30(16 ):  210-212.  doi:10.7506/spkx1002-6630-200916044
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Oligosaccharides were produced from fruits of Chinese jujube by extraction using water and membrane separation followed by purification by Sephadex-G15 column chromatography and were analyzed for their monosaccharide composition by GC-MS and monosaccharides obtained in the Sephadex-G15 column purification were also analyzed by GC-MS. The oligosaccharides were composed of arabinose, rhamnose, ribose, mannose, galactose and glucose. The monosaccharides from fruits of Chinese jujube were found to be mainly glucose and fructose.

Accelerated Oxidation of Methyl Blue for Determination of Trace Nitrite in Foods
WU Ding,LU Gui-hong,LIU Chang-peng,GAO Yu-long,YAO Ming-lan
2009, 30(16 ):  213-215.  doi:10.7506/spkx1002-6630-200916045
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Nitrite can accelerate the oxidation reaction of methyl blue with potassium bromate in acidic medium, which causes the color fading of methyl blue. Based on it, a detection method for the trace nitrite in food was developed in the present study. The maximum absorption peak of the reaction system was presented at 550 nm, and in the nitrite concentration of 0.2-1.2 μg/ml there was a good linear correlation (r = 0.994). The average spike recovery in meat products was 93.16%. The nitrite contents in Chinese sausage, red sausage, nitrite pork, nitrite beef and nitrite pork chop were (15.27 ± 0.08), (4.15±0.13), (90.57 ± 1.69), (99.83 ± 2.75) and (52.13 ± 1.10) mg/kg, respectively. The nitrite content in Chinese cabbage increased with the prolongation of storage time, and reached 6.97 mg/kg when the decay of Chinese cabbage happened.

Extraction and Determination of Different Forms of Oxalate in Vegetables
ZENG Fang,LIU Zhong-zhen,XU Gui-zhi,WANG Rong-hui,OU Jun
2009, 30(16):  216-219.  doi:10.7506/spkx1002-6630-200916046
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The effects of temperature and amount of added HCl on the extraction of water-soluble oxalate and total oxalate from four vegetables (spinach, amaranth, coriander and Chinese chive) were investigated by ion chromatographic method. The watersoluble oxalate was extracted using water at different temperatures, and total oxalate was extracted using different amounts of 0.5 mol/L HCl solution. The centrifugation supernatant of extracts were supernatant diluted and filtrated well enough prior to ion chromatographic detection of oxalate. The water-soluble oxalate contents of spinach and amaranth increased with the increase of temperature, and their total oxalate contents increased with the increase of addition amount of 0.5 mol/L HCl in the range of 0.5-2 ml. While no significant changes in both the two forms of oxalate in Chinese chive and coriander were found when temperature and HCl addition amount were changed. The best extraction efficiency was obtained when total and water-soluble oxalates were extracted using 2 ml of 0.5 mol/L HCl and boiled water for 20 min respectively. The recoveries of water-soluble and total oxalate in the four vegetables were between 95.5% and 103.5%, and the contents of total and water-soluble oxalates both decreased in the following order: spinach > amaranth > Chinese chive > coriander. This method is easy to operate, with little interference as well as high sensitivity and recovery and therefore is applicable to the determination of different forms of oxalate in various vegetables.

GC-MS Analysis of Fatty Acid Composition of Litchi Pulp
ZHONG Hui-zhen1,XU Yu-juan2,*,LI Chun-mei1,WEN Jing2,WU Ji-jun2,LIU Liang2
2009, 30(16):  220-222.  doi:10.7506/spkx1002-6630-200916047
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A GC-MS method was developed for the fatty acid composition analysis of litchi pulp. Fatty acids were extracted from litchi pulp using a mixture of chloroform and methanol (2:1, V/V) and esterfied with methanol prior to GC-MS. A total of 16 fatty acids were isolated and identified, and most of them were unsaturated fatty acids, especially the content of 9-octadecenoic acid was the highest (44.405% of total fatty acids) followed by hexadecanoic acid (25.144%) and 9,12-octadecadienoic acid (18.905%). On the whole, straight-chain saturation fatty acids (SFA) accounted for 28.44%, monounsaturated fatty acids (MUFA) 52.46% and polyunsaturated fatty acids (PUFA) 19.11%.

High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry for the Determination of Arsenic Content and Speciation in Enteromorpha
YU Jing-jing1,CAO Xuan1,2,CUI Wei-gang1,3,YU Zhen-hua1,4,LEE Frank Sen-Chun1,WANG Xiao-ru1,5,*
2009, 30(16):  223-227.  doi:10.7506/spkx1002-6630-200916048
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This study analyzed arsenic content and speciation in Enteromorpha sampled from various coastal zones of Qindao using high performance liquid chromatography coupled to inductively coupled plasma-mass spectrometry (HPLC-ICP-MS). The results showed that the total arsenic content in Enteromorpha was ranged from 3.0 to 9.7 μg/g. The speciation analysis of hot water extract of Enteromorpha indicated that the major arsenic forms in Enteromorpha were inorganic quinquevalence arsenate and a substance suspected as arsenic sugar. Besides, there were small amounts of inorganic trivalence arsenite and dimethylarsenate (DMA) in the extract. Due to the toxicity or potential toxicity of these arsenic forms, it should be paid great caution in the application of Enteromorpha as a material for food and medicine.

HPLC Determination of Nucleosides in Different Tissues of Xylaria nigripes
LIAO Qiong,ZHOU Rong,CHEN Zuo-hong*
2009, 30(16):  228-230.  doi:10.7506/spkx1002-6630-200916049
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Objective: To determine the kinds and contents of nucleosides in different tissues (natural sclerotium, natural stroma, cultivated stroma and fermented mycelium) of Xylaria nigripes. Methods: Samples were extracted using water in an ultrasonic field, and the extracts were filtrated through 0.45 μm membrane prior to HPLC analysis. The chromatographic separation was achieved on a YWG C18 column (4.0 mm × 300 mm,10 μm) at 40 ℃ using a mobile phase consisting of a mixture of methanol and water (0 min: 0% → 8 min: 10% → 10 min:15%) at a flow rate of 1 ml/min, and the analytes were detected at 259 nm. Results: The natural sclerotium, natural stroma, cultivated stroma and fermented mycelium of Xylaria nigripes mainly contained uridine and adenosine, and the content of nucleosides in sclerotium was higher than that in stroma and fermented mycelium. Besides, the content of nucleosides in natural stroma was similar to that in cultivated stroma. Conclusion: nucleosides were rich in Xylaria nigripes, and the cultivated stroma can replace the natural stroma and sclerotium.

Adsorption of Cadmium onto Cross-linked Chitosan and Its Application in Atomic Absorption Spectrometric Determination of Trace Cadmium in Food Industrial Water
WANG He-cai
2009, 30(16):  231-234.  doi:10.7506/spkx1002-6630-200916050
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The adsorption behavior of cross-linked chitosan prepared with epichlorohydrin to trace cadmium (Ⅱ) in water was investigated. One-factor-at-a-time and orthogonal array methods were applied to maximize adsorption of cross-linked chitosan to cadmium. Range analysis proved that the most significant factor affecting adsorption was pH of solution, followed by amount of added cross-linked chitosan. The average removal rate of Cd2+ was 97.4% with a RSD of 8.4% (n = 6) and the average saturated absorption capacity was 45.6 mg/g, suggesting good precision. The enrichment of cadmium with cross-linked chitosan was applied to the atomic absorption spectrometric determination of trace cadmium in brewery industrial water. This method is fast, effective and simple for the determination of trace cadmium in food industrial water with a high recovery of 97.2%-103.3%.

Solid Phase Extraction Followed by Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Determination of Eight Anabolic Steroids in Milk
LU Hui-qing1,CHEN Hui-hua2,WEI Min-jue2,YING Yong-fei2
2009, 30(16):  235-239.  doi:10.7506/spkx1002-6630-200916051
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This study developed a quick and specific method for the identification and trace quantification of 8 anabolic steroids (TS, PG, NT, MTS, BOL, TB, DIA, and STA) in milk. Samples were extracted using methyl tert-butyl ether, and the extracts were cleaned up by solid-phase extraction on a C18 (SPE) cartridge, blowed to dryness by nitrogen, and dissolved in a mixture of 10% methanol and water (10:90, V/V) prior to LC-MS/MS analysis in a multiple reaction monitoring (MRM) mode. The limits of detection (LODs) ranged in 0.2 - 0.5 μg/L, and the limits of quantification (LOQs) were between 0.5 and 1.0 μg/L. The regression equations were well linear between 2.0 and 200.0 μg/L for these steroids (r > 0.9990). The recoveries ranged from 59.1% to 97.7% with a RSD of 3.2%-11.9% (n = 3) at three spiked levels of 1.0-5.0 μg/L. The method is simple, sensitive and rapid and therefore can be used for the simultaneous determination of 8 anabolic steroids in milk.

HPLC Determination of Resveratrol in Roots, Stems and Leaves of Peanut
WU Xiang-yang1,WANG Cai-xia2,DA Zu-lin1,YANG Liu-qing3,*,ZHANG Rong-xian1, YANG Ling-ling2,ZHOU Ye3
2009, 30(16):  240-242.  doi:10.7506/spkx1002-6630-200916052
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Objective: To determine the contents of resveratrol in roots, stems and leaves of peanut. Methods: A HPLC determination combined with one-step pretreatment by silica gel column chromatography was established. The analysis was achieved by separation on an Hypersil ODS-2 column (4.6 mm × 250 mm, 5 μm) at 35 ℃ using a methanol-water (45:55, V/ V) mobile phase at a flow rate of 0.8 ml/min followed by UV detection at 306 nm. Results: The contents of resveratrol in the mixtures of roots, stems and leaves of peanuts from Shandong, Jiangsu and Hebei provinces were 903.69, 116.21 and 88.69 μg/g, respectively. The contents of resveratrol in roots, stems and leaves of peanut from Shandong province were 223.08, 1211.19 and 429.79 μg/g, respectively. The calibration curve was well linear over the concentration range from 0.228 to 1.14 μg (R = 0.9999), and the limit of detection was 4.3 ng (3σ). The average recovery was 99.6%with a RSD of 3.09% (n = 5). Conclusions: This method is simple, fast and accurate.

Solid-phase Extraction and HPLC Determination of Six Sudan Reds and Para Red in Pepper Oil
WU Yin-liang1,YANG Ting1,ZHAO Jian1,HUANGFU Wei-guo1,SHEN Jian-zhong2
2009, 30(16):  243-246.  doi:10.7506/spkx1002-6630-200916053
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A method was presented for determining six Sudan reds and Para red in pepper oil using solid-phase extraction (SPE) and HPLC. Samples were extracted with acetonitrile and cleaned up on a C18 SPE column using n-hexane as the eluant, and the eluate was evaporated to dryness and dissolved in methanol prior to HPLC analysis. The analytes were detected at 510 nm, and quantified using an external calibration method. The detection limits were 7.0-7.2 μg/kg for six Sudan reds and Para red in pepper oil. At three spiked levels of 25-250 μg/kg, the average recoveries for six Sudan reds and Para red in pepper oil were 78.3% -93.5%, and the intra and inter CVs were 3.3%-9.2% and 4.6%-10.6%, respectively. There was a good linear correlation (r > 0. 9997) between the peak area and the concentrations of Sudan reds and Para red in the range of 15 to 2000 μg/L.

GC-MS Analysis of Chemical Constituents of Volatile Oil from Lingdera thomsonii Allen Fruits
ZHANG Xian-jun,DU Ping
2009, 30(16 ):  247-250.  doi:10.7506/spkx1002-6630-200916054
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The chemical constituents of volatile oil from Lingdera thomsonii Allen was analyzed by GC-MS. The volatile oil was extracted by steam distillation with a yield of 0.55%. A total of 29 constituents were isolated and among them 24 compounds were identified, content of which accounted for 98.89% of total constituents. Most of these constituents were sesquiterpenes or oxygenated compounds, such as 1,7,7-trimethyl-bicyclo[2.2.1]-hept-2-ylester. The peaks appearing at retention time of 45.16 min correspond to acetate retinal and oxidation ledene, suggesting they may exist in the volatile oil.

GC-MS Analysis of Fatty Acid Composition of Wild Chicory Chicory Grown in Plastic Shed
DING Xu-guang,HOU Dong-yan,GUAN Chong-xin,LIU Xiao-yuan
2009, 30(16 ):  251-253.  doi:10.7506/spkx1002-6630-200916055
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A method using GC-MS was presented for the analysis of fatty acid composition of wild chicory and chicory grown in plastic shed. Fatty acids were extracted from samples by Soxhlet extraction method and esterfied with methanol prior to GCMS. A total of 10 fatty acids containing 30.87% unsaturated fatty acids were found in chicory grown in plastic shed and 7 fatty acids from wild chicory were isolated and identified, in which unsaturated fatty acids accounted for 57.3%.

FTA Filter Paper-based Template Preparation for the PCR Detection of Salmonella in Meat
LI Wei-hao,ZHANG Hui-yan,LIU Wei-hua,LI Yan,LI Ying-jun,ZHANG Wei*
2009, 30(16):  254-257.  doi:10.7506/spkx1002-6630-200916056
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An assay of PCR was developed for the detection of Salmonella in meat. Based on flotation and solvent extraction technology, FTA filter was used to extract Salmonella DNA from fresh meat to eliminate the inhibitory factors of PCR reaction. An invA gene in surface protein of epithelial cells absorbed and invaded by Salmonella was used to amplify a 376 bp DNA fragment, which could be confirmed by DNA sequencing. The detection limits of Salmonella in pork, beef, mutton and chicken homogenates all were 7×101 CFU/ml, and the detection could be finished in 6 h, which was shorter than that of the conventional method of combined enrichment and PCR by 12-24 h. In this study, 72 samples were detected and the results were compared with the GB/T 4789.4—2003 method. The detection rates of Salmonella by this method and the GB/T 4789.4—2003 method were 27.8% and 22.2%, and the detection by the latter method cost 5 days. In general, this method is a rapid and sensitive approach to the Salmonella detection in meat. Meanwhile, the DNA template preparation using FTA filter paper can constructs a technological platform for the fast detection of pathogens in food.

RP-HPLC Detection of 1-Deoxynojirimycin in Different Varieties and Sites of Mulberry Leaves
WEI Zhao-jun,JIANG Lei,ZHOU Le-chun,WANG Ai-min,ZHANG Song
2009, 30(16 ):  258-261.  doi:10.7506/spkx1002-6630-200916057
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A method of pre-column derivatization with FMOC-Cl followed by RP-HPLC with UV detection was established for the determination of 1-deoxynojirimycin (DNJ) content in different varieties and sites of mulberry leaves. DNJ was extracted from mulberry leaves using 0.05 mol/L HCl and derivatized using FMOC-Cl in borate buffer solution (pH 8.50) prior to RP-HPLC detection. The chromatographic separation was achieved on a Waters X-Terra column using a mobile phase consisting of a mixture of acetonitrile and 0.1% aqueous acetic acid (55:45, V/V). There was a high positive correlation between the peak area and concentration of DNJ-FMOC with a correlation coefficient of 0.9852. The DNJ content was varied in different varieties of mulberry leaves. Among 15 varieties of mulberry, Qingyang Qing mulberry leaves contained the highest DNJ (0.1486%), while Pi mulberry leaves the lowest (0.04291%). Also, mulberry leaves in different sites contained different DNJ contents. It decreased from the top to bottom gradually, indicating DNJ content is reduced with the maturation of mulberry leaves.

Matrix Solid-phase Dispersion Extraction and Gas Chromatography-Mass Spectrometry Determination of Multi-residue Pesticides in Fruits
ZHANG Li-jin,WANG Xiao*
2009, 30(16 ):  262-265.  doi:10.7506/spkx1002-6630-200916058
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An analysis procedure was developed for multi-residue pesticides in fruits using matrix solid-phase dispersion extraction combined with gas chromatography-mass spectrometry (GC-MS) detection. Fruit samples were shattered and grinded together with florisil and Na2SO4, and the extracts were cleaned up on a graphitized carbon black column using ethyl acetate as the eluant and concentrated prior to GC-MS detection in a selected ion monitoring mode. The recoveries of 61 pesticides were ranged from 73.6%-113.7%, and the relative standard deviations all were below 13.7%. The detection limits were ranged from 0.002 to 0.01 mg/kg. Generally, the proposed method is proved to be quick, economic,time-saving and labour-saving.

Determination of Cadmium Content in Meat Foodstuffs by EDTA Decrement Titration
WANG Rui-bin,MA Ya-jun
2009, 30(16 ):  266-268.  doi:10.7506/spkx1002-6630-200916059
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A new method for the rapid determination of cadmium content in meat foodstuffs by EDTA decrement titration was developed. Samples were digested with a mixture of HNO3 and H2SO4 (2:1, V/V), and one portion of the digestion solution was added with thiourea to mask Hg2+ at pH 5-6 prior to EDTA titration of total metal ions with xylenol orange as the indicator. Cd2+ in another portion was masked by the addition of KI, and other ions except Cd2+ in the solution were titrated by standard EDTA solution. Finally, the amount of cadmium was calculated though the volume difference of EDTA solution consumed in the titration of two portions. The relative standard deviation of this method was less than 2.0% and the recovery was between 97.8% and 100.2%. It is indicated that the improved method is an easy-operating way with high accuracy and little disturbance, and its results are consistent with those of national standard method

SPME-GC-MS Analysis of Flavor Components in Dried Capsicum before and after Baking
LI Da1,WANG Zhi-song1,DING Zhu-hong1,*,DENG Sheng-yi2
2009, 30(16 ):  269-271.  doi:10.7506/spkx1002-6630-200916060
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The flavor components in dried capsicum before and after baking were analyzed by SPME-GC-MS. A total of 29 volatile components were found in dried capsicum and 31 in baked capsicum. Five volatile components were lost after baking, most of which were ester compounds; meanwhile 5 new components were produced, most of which were alkane compounds. After baking, an obvious increase was found in the relative contents of himachalene, nerolidol, 5-epi-aristolochene and α-longip, while those of hexyl-2-methylbutyrate and 2-butyl-1-decene changed conversely. These results demonstrated that baking can significantly improve the flavor quality of capsicum.

Flow Injection Analysis of Nitrate and Nitrite in Vegetables
REN Nai-lin,LI Hong
2009, 30(16 ):  272-274.  doi:10.7506/spkx1002-6630-200916061
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Based on the reduction from nitrate to nitrite through a Zn-C powder mixture column, a new method for the determination of nitrate and nitrite in vegetables using flow injection analysis were proposed. Samples were homogenized and extracted, and the extract was decolorized and divided into two portions. The nitrite content in one portion was directly determined by hydrochloride naphthalene ethylenediamine colorimetry, and another was determined by the same method after nitrate was reduced to nitrite. The content of nitrate was calculated by subtraction method. The linear range of nitrite was 0.1- 2.4μg/ml, the detection limit 0.01 μg/ml, the range of relative standard deviation from 0.03% to1.8%, and the sample determination frequency 10 samples/h.

Analysis of Volatile Compounds of Essential Oil from Origanum vulgare L. by GC-MS and GC-O
ZHANG Yu-yu,SUN Bao-guo,ZHU Jun*
2009, 30(16 ):  275-277.  doi:10.7506/spkx1002-6630-200916062
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A method based on distillation extraction followed by GC-MS coupled to GC-olfactometry was presented for the analysis of volatile compounds of essential oil from Origanum vulgare L. Thirty-five compounds were identified, accounting for 97.50% of total peak area, and consisted of 22 hydrocarbons (60.66%), 6 ketones (26.56%), 2 esters (0.64%), 2 phenols (9.18%), 2 alcohols (0.44%) and 1 aldehyde (0.02%). All these results can provide valuable and scientific information for the development and utilization of Origanum vulgare L. essential oil.

Effects of Different Lethal Methods and Partial Freezing Treatment on Fresh-keeping Properties of Tilapia
YANG Guang1,GUO Juan2,LIN Xiang-dong2,*
2009, 30(16):  278-281.  doi:10.7506/spkx1002-6630-200916063
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This study investigated the effects of different lethal methods on the freshness of tilapia, and discussed the freshness changes and part of physical properties of tilapia in a state of partial freezing. The results showed that the initial freezing point of tilapia was -1.5 ℃. The lethal time of tilapia kept in 0 ℃ ice water was 30 min, and this method could prolong the time required to enter rigidity period, which was beneficial to the fresh-keeping of tilapia. This study demonstrates that conductivity method is an accurate, rapid and convenient method for measuring freezing point.

Effects of 6-Benzyladenine Treatment on Postharvest Respiration and Quality of Toona sinensis Roem Sprout
LI Zhi-xun,SHEN Lin,XU Lu-feng,WAN Li,ZHAO Dan-ying,SHENG Ji-ping*
2009, 30(16 ):  282-285.  doi:10.7506/spkx1002-6630-200916064
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Fresh sprouts of Toona sinensis Roem treated with 100 mg/L 6-benzyladenine (6-BA) were stored at (25 ± 2) ℃ with RH (50 ± 5)% for 4 d, and then the postharvest respiratory rate, browning rate, nutritional and functional ingredients were analyzed. The results indicated that the respiratory rate and browning level were inhibited and the appearance of respiratory peak was postponed in Toona sinensis Roem sprouts treated with 6-BA. The nutritional and functional ingredients such as soluble protein, soluble sugar, vitamin C, flavonoids, saponins and total polyphenols declined slower in treated sample than those in control. Altogether, this study demonstrat that the postharvest quality decline of Toona sinensis Roem sprouts is associated closely with their respiration and 6-BA treatment could suppress their respiration during postharvest storage to keep their quality.

Effects of Cassava Modified Starch and Sucrose Ester on Water-holding Capacity of Chilled Meat
ZHAO Gai-ming,SUN Ling-xia,LIU Yan-xia,HAO Hong-tao,ZHU Chao-zhi,ZHANG De-chuan
2009, 30(16 ):  286-289.  doi:10.7506/spkx1002-6630-200916065
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The effects of concentration of cassava modified starch and immersion time on water-holding capacity of chilled meat were investigated by single-factor method. Subsequently, a three-variable and three-level quadratic regression orthogonal rotatable composite design combined with response surface methodology (RSM) was performed to maximize moisture content and minimize loss rates of drip and storage and two stepwise regression models for moisture content and storage loss rate were built up. The optimized parameters for achieving the best water-holding capacity of chilled meat were as follows: concentration of cassava modified starch 0.30%, concentration of sucrose esters 0.44%, and immersion time 270 s.

Development of A Chinese Gooseberry Root Beverage
LI Jia-xing1,SUN Jin-yu2,LIU Fei3,CHEN Shuang-ping2,QIN Yi2,YAN You-bing3
2009, 30(16 ):  290-292.  doi:10.7506/spkx1002-6630-200916066
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A novel beverage was developed from Chinese gooseberry root. This study mainly focused on the optimization of ratio of material to water, boiling time of material, complex ratio of stabilizers and proportion of main and auxiliary materials. The decoction obtained had good quality when Chinese gooseberry root was boiled for 3 h with the ratio of material to water of 1:20 (g/ml). A mixture of carrageenan gum and CMC-Na at the ratio of 1:1 presented the best stabilizing effect. The beverage composed of 60% decoction, 1‰ complex stabilizer, 5.0% honey and 6.0% fructose syrup had nice flavor, taste and stability.

Development of A Lactic Acid Bacteria-fermented Beverage from Chinese Wolfberry Fruits and Milk
HUANG Yu,ZHOU Qing-feng,HAI Yang,WU Chun-yan,ZHAO Li,LIU Jian-tao,SU Wei*
2009, 30(16 ):  293-295.  doi:10.7506/spkx1002-6630-200916067
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A new-type lactic acid bacteria-fermented beverage was developed from Chinese wolfberry fruits and milk by the following procedures of protein removal from Chinese wolfberry fruit slurry by enzymolysis, fermentation of mixed hydrolysate of Chinese wolfberry fruit slurry and fresh milk using a Lactobacillus starter composed a mixture of Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus and Lactobacillus acidophilus and formulation of final product. Our results showed that the largest protein removal from Chinese wolfberry fruit slurry could be attained when the enzymolysis conditions were as follows: material/water ratio 3:1, papain dose 0.04%, pH 7.0-7.5, and temperature 50 ℃ for 90 min. The optimal formula of final beverage product to obtain the highest sensory score was composed of 25% hydrolysate of Chinese wolfberry fruit slurry, 65% fresh milk, 7% sucrose, 0.1% Lactobacillus starter, and 0.2% compound emulsion stabilizer.

Development of A Fermented Vinegar Beverage from Oleaster Fruits
PAN Lan1,JIA Xiao-guang2,*,WANG Ji-guo1,JIA Sheng-jie2,SHEN Tong1
2009, 30(16):  296-298.  doi:10.7506/spkx1002-6630-200916068
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Oleaster fruits grown in Xinjiang area were processed into a full-bodied fermented vinegar beverage by alcohol fermentation by yeast preserved in our lab, acetic acid fermentation by a strain of acetic acid bacteria natural vinegar cake and beverage blending. The best brewing conditions of vinegar were determined as follows: alcohol fermentation was carried out at 30 ℃ with the sugar content in oleaster juice of 10-12°Bx and yeast inoculation of 10%; when alcohol volume fraction reached 5.5%-6%, acetic acid fermentation was done with acetic acid bacteria amount of 10% at 32 ℃ for 5 days.

Development of A Wheat Germ Yogurt
YAO Ming-lan,WU Ding,WAN Xing,XING Guan-nan,ZHANG Li-li,LU Gui-hong
2009, 30(16 ):  299-301.  doi:10.7506/spkx1002-6630-200916069
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In order to achieve the scientific and rational nutrition matching, a wheat germ yogurt was developed using Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivaricus subsp. thermophilus isolated and purified from a commercial yoghurt branded “Weigang”. A product with nice flavor and taste could be obtained by adding 2 ml of Lactobacillus delbrueckii subsp.bulgaricus and 2 ml of Streptococcus salivaricus subsp. thermophilus to a mixture of 6 g of skimmed milk powder, 4-5 g of wheat germ powder, 6 g of sugar and 100 ml of distilled water for fermentation at 37 ℃ for 7 h.