CN102783657A - Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave - Google Patents
Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave Download PDFInfo
- Publication number
- CN102783657A CN102783657A CN2012103095451A CN201210309545A CN102783657A CN 102783657 A CN102783657 A CN 102783657A CN 2012103095451 A CN2012103095451 A CN 2012103095451A CN 201210309545 A CN201210309545 A CN 201210309545A CN 102783657 A CN102783657 A CN 102783657A
- Authority
- CN
- China
- Prior art keywords
- water
- microwave
- residue
- dietary fiber
- soybean dietary
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave, and belongs to the field of food processing. The method comprises the following steps: (1) the soybean is crushed, and pretreated by extrusion to obtain an extruded material; the extruded material is mixed with water to obtain mixed liquor; alkali protease is added in the mixed liquor to perform enzymolysis and centrifuged to obtain free oil, emulsion, hydrolysate and residue; (2) the residue obtained in the step (1) is dried, crushed and dispersed in water; pH value is adjusted to acidity, and microwave extraction is performed; then the extracted liquid is centrifuged to obtain supernatant; the pH value of the supernatant is adjusted to alkalinity; and the supernatant is placed still, and centrifuged to obtain precipitate; and the precipitate is washed with water, dried and crushed to obtain the water-insoluble soybean dietary fiber. The method has the advantages of simplicity in processing device, convenience in operation and control, great reduction in acid-alkali dosage, reduction in production cost and pollution, short extracting time, high extracting efficiency, and high purity of the water-insoluble soybean dietary fiber.
Description
Technical field
The invention belongs to foods processing technique, relate generally to microwave and combine acid to dissolve the method that the heavy method of alkali is extracted water-insoluble soybean dietary fiber in the aqueous enzymatic method residue.
Background technology
The residue major part that produces in the aqueous enzymatic extraction soybean oil process is handled or is let alone as just feed, fertilizer and rots, inreal its edibility of performance.Containing a large amount of water-insoluble soybean dietary fibers in the residue, is high a kind of of inexpensive matter in numerous natural dietary fiber sources.Therefore the soybean dietary fiber of Development and Production high-quality transforms the appreciation that realizes residue, alleviates residue and environment is polluted has far-reaching practical significance.
The method for distilling of water-insoluble soybean dietary fiber mainly is that the heavy method of alkali is dissolved in acid at present; Because dissolving heavy extraction of alkali, acid needs with a large amount of strong acid and highly basic; Not only require equipment acid, alkali tolerance high; And the discharging of soda acids will increase second environmental pollution in a large number, so reduce the extraction that the different assisted extraction methods of soda acid consumption progressively are used for the bean dregs water insoluble dietary fiber.Slow waves etc. (2002) dissolve the heavy method of alkali through adding protease with the lipase auxiliary acid, under nearly neutral pH value condition, from bean dregs, are extracting 70%～75% water-insoluble soybean dietary fiber, and recovery rate and traditional acid are dissolved alkali to sink method suitable.Shang Yongbiao (2007) and Li Wenjia (2010) all adopt acid-base method to extract water-insoluble soybean dietary fiber in the bean dregs, and dietary fiber content reaches about 75% in the product.Pan Lihua etc. (2011) adopt ultrasonic auxiliary acid to dissolve the heavy method of alkali and extract the water-insoluble soybean dietary fiber, and effect is superior to acid-base method, but has the shortcoming that extraction time is long, extraction efficiency is low.Existing water-insoluble soybean dietary fiber extractive technique exist extraction time long, extraction efficiency is low, product purity is lower, to problems such as environment pollute.
Summary of the invention
Technical problem to be solved by this invention is the deficiency that overcomes above-mentioned prior art; Provide microwave to combine acid to dissolve the method that the heavy method of alkali is extracted water-insoluble soybean dietary fiber in the aqueous enzymatic method residue, reach shorten extraction time, reduce cost, reduce pollutions, the purpose of raising product recovery rate and purity.
Technical problem to be solved by this invention realizes through following technical scheme:
The method of soybean dietary fiber in a kind of Microwave Extraction aqueous enzymatic method residue; This method may further comprise the steps: (1) soybean is pulverized the back and adopts expelling-expansion pretreatment to obtain expanded material; Expanded material mixed with water obtain mixed liquor; In mixed liquor, add alkali protease and carry out enzymolysis, centrifugation obtains free oil, emulsion, hydrolyzate and residue behind the enzymolysis; (2) residue that step (1) is obtained is dry, be scattered in the water after pulverizing, and using the 0.1mol/L lactic acid solution to regulate pH value is 4.5 to carry out Microwave Extraction, and the Microwave Extraction condition is a water and the residue ratio 10-30mL/g of drying; Microwave power is 400-800W; Crossing grit number is the 20-100 order, and the microwave time is 10-30min, with Microwave Extraction liquid centrifugalize supernatant; After using 0.1mol/LNaOH adjusting supernatant pH value to be 9; Leave standstill, centrifugalize sediment, with sediment washing, dry, after pulverizing the water-insoluble soybean dietary fiber.
Described Microwave Extraction optimum condition is: water is 18.1mL/g with dry residue ratio, and microwave power 666W crosses grit number 60 orders, microwave time 19.9min.
This method adopts microwave to combine acid to dissolve the heavy method of alkali and extracts the water-insoluble soybean dietary fiber, and microwave has the very strong penetrability and the very high thermal efficiency, in the microwave electromagnetic field of fast vibration; Produce a large amount of heats; Temperature is risen rapidly, and the pressure that the aqueous water vaporization produces makes dietary fiber be discharged in the acid flux material cell membrane and cell wall rupture; Utilize the insoluble character of its alkalescence that dietary fiber is precipitated out again, accomplish from the soybean residue, to extract and separate the water-insoluble soybean dietary fiber; This method has the advantages that required process equipment is simple, cost is low, extraction time is short, and product extraction efficiency height and purity are high.
Description of drawings
Fig. 1 is an overall process route map of the present invention;
Fig. 2 be the liquid material than with microwave power alternately to the response surface of recovery rate;
Fig. 3 be the liquid material than with cross grit number alternately to the response surface of recovery rate;
Fig. 4 be the liquid material than with the microwave time alternately to the response surface of recovery rate;
Fig. 5 is a microwave power and cross grit number alternately to the response surface of recovery rate;
Fig. 6 is that microwave power and microwave time are alternately to the response surface of recovery rate;
Fig. 7 was that grit number and microwave time are alternately to the response surface of recovery rate.
The specific embodiment
Below in conjunction with accompanying drawing the specific embodiment of the invention is described in detail.
The method of soybean dietary fiber in a kind of Microwave Extraction aqueous enzymatic method residue; This method may further comprise the steps: (1) soybean is pulverized the back and adopts expelling-expansion pretreatment to obtain expanded material; Expanded material mixed with water obtain mixed liquor; In mixed liquor, add alkali protease and carry out enzymolysis, centrifugation obtains free oil, emulsion, hydrolyzate and residue behind the enzymolysis; (2) residue that step (1) is obtained is dry, be scattered in the water after pulverizing, and using the 0.1mol/L lactic acid solution to regulate pH value is 4.5 to carry out Microwave Extraction, and the Microwave Extraction condition is a water and the residue ratio 10-30mL/g of drying; Microwave power is 400-800W; Crossing grit number is the 20-100 order, and the microwave time is 10-30min, with Microwave Extraction liquid centrifugalize supernatant; After using 0.1mol/LNaOH adjusting supernatant pH value to be 9; Leave standstill, centrifugalize sediment, with sediment washing, dry, after pulverizing the water-insoluble soybean dietary fiber.
Described Microwave Extraction optimum condition is: water is 18.1mL/g with dry residue ratio, and microwave power 666W crosses grit number 60 orders, microwave time 19.9min.
Embodiment: water-insoluble soybean dietary fiber optimized parameter screening test in the Microwave Extraction aqueous enzymatic method residue
1 materials and methods
1.1 material, reagent
| Soybean | Heilungkiang Academy of Agricultural Sciences cultivates cultivates farming 42, and wherein protein content is 41.6%, and fat content is 21.3%, and moisture content is 10.6%, ash content 4.3%. |
| Alcalase alkaline endo protease | Novo company (1.2 * 10 5U/mL) |
1.2 key instrument equipment
| PHS-25 type acidometer | Shanghai great achievement instrument plant |
| Electronic analytical balance | Mei Lete-Tuo benefit instrument (Shanghai) Co., Ltd. |
| Centrifuge | Beijing Medical Centrifugal Machine Factory |
| Accurate electric blender | Jintan City, Jiangsu Province high honour instrument Manufacturing Co., Ltd |
| Electric-heated thermostatic water bath | Yuyao City east electric instrument factory |
| Nitrated appearance | Shanghai Qianjian Instrument Co., Ltd. |
| Planetary ball mill | Nanda Tianzun Electronics Co., Ltd., Nanjing |
| Soxhlet extractor | Tianjin Glass Instrument Factory |
| Micro-wave oven | China Shuande Glanz micro-wave oven Electrical Appliances Co., Ltd |
1.3 experimental technique
1.3.1 the composition measurement of soybean
The mensuration of crude protein: the GB6432-94 standard method is carried out
The mensuration of crude fat: the Suo Shi extraction process is measured among the GB5512-85
The mensuration of moisture: GB304-87 measures
The mensuration of ash content: GB5009.4-85
The mensuration of dietary fiber: GB/T2222.4-2008
1.3.2 technological process
1.3.3 computing formula
2 results and discussion
2.1 empirical factor level code table
On the basis of single factor research; Choosing liquid material ratio, microwave power, mistake grit number and microwave time 4 factors is independent variable, is response with the recovery rate, according to the center combination design principle; The experiment of design response surface analysis, its factor level coding schedule is seen table table 2-1.
Table 2-1 factor level coding schedule
2.2 response surface EE and experimental result
This experimental applications response surface optimization carries out process optimization.With A, B, C, D is independent variable, is response R with the recovery rate, and response surface experimental program and result see table 2-2.Experiment 1-24 is factorial experiment, and 25-36 is 12 center tests, in order to the estimating experiment error.
Table 2-2 test arrangement and result
| The experiment number | The liquid material is than A (mL/g) | Microwave power B (W) | Cross grit number C (order) | Microwave time D (min) | Recovery rate R (%) |
| 1 | -1 | -1 | -1 | -1 | 48.21 |
| 2 | 1 | -1 | -1 | -1 | 41.79 |
| 3 | -1 | 1 | -1 | -1 | 69.24 |
| 4 | 1 | 1 | -1 | -1 | 48.74 |
| 5 | -1 | -1 | 1 | -1 | 47.41 |
| 6 | 1 | -1 | 1 | -1 | 44.97 |
| 7 | -1 | 1 | 1 | -1 | 74.93 |
| 8 | 1 | 1 | 1 | -1 | 57.32 |
| 9 | -1 | -1 | -1 | 1 | 60.13 |
| 10 | 1 | -1 | -1 | 1 | 50.15 |
| 11 | -1 | 1 | -1 | 1 | 81.07 |
| 12 | 1 | 1 | -1 | 1 | 70.38 |
| 13 | -1 | -1 | 1 | 1 | 51.98 |
| 14 | 1 | -1 | 1 | 1 | 59.63 |
| 15 | -1 | 1 | 1 | 1 | 88.97 |
| 16 | 1 | 1 | 1 | 1 | 70.53 |
| 17 | -2 | 0 | 0 | 0 | 63.81 |
| 18 | 2 | 0 | 0 | 0 | 43.41 |
| 19 | 0 | -2 | 0 | 0 | 49.83 |
| 20 | 0 | 2 | 0 | 0 | 89.42 |
| 21 | 0 | 0 | -2 | 0 | 46.15 |
| 22 | 0 | 0 | 2 | 0 | 53.81 |
| 23 | 0 | 0 | 0 | -2 | 70.36 |
| 24 | 0 | 0 | 0 | 2 | 92.11 |
| 25 | 0 | 0 | 0 | 0 | 90.27 |
| 26 | 0 | 0 | 0 | 0 | 83.44 |
| 27 | 0 | 0 | 0 | 0 | 88.53 |
| 28 | 0 | 0 | 0 | 0 | 85.47 |
| 29 | 0 | 0 | 0 | 0 | 82.81 |
| 30 | 0 | 0 | 0 | 0 | 83.43 |
| 31 | 0 | 0 | 0 | 0 | 86.45 |
| 32 | 0 | 0 | 0 | 0 | 88.37 |
| 33 | 0 | 0 | 0 | 0 | 89.2 |
| 34 | 0 | 0 | 0 | 0 | 92.18 |
| 35 | 0 | 0 | 0 | 0 | 86.94 |
| 36 | 0 | 0 | 0 | 0 | 85.46 |
2.3 response surface interpretation
Carry out data analysis through statistical analysis software Design-Expert, it is following to set up quadratic response face regression model:
R=86.88-4.97A+9.84B+1.72C+5.99D-3.50AB+1.05AC+0.97AD+1.16BC+1.33BD-0.45CD-8.86A
2-4.86B
2-9.77C
2-1.96D
2
Regression analysis and The results of analysis of variance are seen table 2-3, and the response surface analysis of mutual phase is seen Fig. 1-Fig. 7.
Table 2-3 returns and The results of analysis of variance
| Variable | The free degree | Quadratic sum | All square | The F value | Pr>;F |
| A | 1 | 592.32 | 592.32 | 52.02 | ＜0.0001 |
| |
1 | 2322.44 | 2322.44 | 203.96 | ＜0.0001 |
| |
1 | 71.24 | 71.24 | 6.26 | 0.0207 |
| |
1 | 860.76 | 860.76 | 75.60 | ＜0.0001 |
| AB | 1 | 196.35 | 196.35 | 17.24 | ＜0.0001 |
| AC | 1 | 17.54 | 17.54 | 1.54 | 0.2283 |
| |
1 | 15.04 | 15.04 | 1.32 | 0.2634 |
| BC | 1 | 21.65 | 21.65 | 1.90 | 0.1825 |
| BD | 1 | 28.12 | 28.12 | 2.47 | 0.1310 |
| |
1 | 3.30 | 3.30 | 0.29 | 0.5958 |
| |
1 | 2513.46 | 2513.46 | 220.74 | ＜0.0001 |
| |
1 | 755.47 | 755.47 | 66.35 | ＜0.0001 |
| |
1 | 3054.56 | 3054.56 | 268.26 | ＜0.0001 |
| |
1 | 122.47 | 122.47 | 10.76 | 0.0036 |
| Return | 14 | 10574.72 | 755.34 | 66.34 | ＜0.0001 |
| Residue | 21 | 239.12 | 11.39 | ? | ? |
| Lose and intend | 10 | 144.72 | 14.47 | 1.69 | 0.2020 |
| Error | 11 | 94.40 | 8.58 | ? | ? |
| Summation | 35 | 10813.84 | ? | ? | ? |
2-3 can know that the linear relationship between equation dependent variable and the independent variable is obvious by table, and this regression models is (p significantly<0.0001), lose and intend not remarkable (the p > of item; 0.05), and this model R
2=97.79%, R
2 Adj=96.31%, explain that this model and experimental fit are good, linear relationship is remarkable between independent variable and the response, and the theory that can be used for this reaction is inferred.Can obtain the factor contributions rate by the F check is: B>D>A>C, i.e. microwave power>the microwave time>liquid material ratio>the mistake grit number.
Application responds face optimizing analytical method is analyzed regression model, seeks the optimal response result and is 18.1mL/g for liquid material ratio, and microwave power is 666W, and crossing grit number is 60 orders, and the microwave time is 19.9min, and response surface has optimal value to be approximately about 92.64%.
2.4 confirmatory experiment and contrast test
Under the optimum condition that the response surface analysis method is tried to achieve, promptly the liquid material is than being 18.1mL/g, and microwave power is 666W; Crossing grit number is 60 orders; The microwave time is 19.9min, carries out 3 parallel laboratory tests, and the mean value of 3 parallel laboratory test water-insoluble soybean dietary fiber recovery rates is 91.12%.Experiment value and regression equation predicted value that response is described are identical good.
Claims (2)
1. the method for soybean dietary fiber in the Microwave Extraction aqueous enzymatic method residue; This method may further comprise the steps: (1) soybean is pulverized the back and adopts expelling-expansion pretreatment to obtain expanded material; Expanded material mixed with water obtain mixed liquor; In mixed liquor, add alkali protease and carry out enzymolysis, centrifugation obtains free oil, emulsion, hydrolyzate and residue behind the enzymolysis; It is characterized in that: the residue that (2) obtain step (1) is dry, be scattered in the water after pulverizing, and using the 0.1mol/L lactic acid solution to regulate pH value is 4.5 to carry out Microwave Extraction, and the Microwave Extraction condition is a water and the residue ratio 10-30mL/g of drying; Microwave power is 400-800W; Crossing grit number is the 20-100 order, and the microwave time is 10-30min, with Microwave Extraction liquid centrifugalize supernatant; After using 0.1mol/LNaOH adjusting supernatant pH value to be 9; Leave standstill, centrifugalize sediment, with sediment washing, dry, after pulverizing the water-insoluble soybean dietary fiber.
2. the method for soybean dietary fiber in a kind of Microwave Extraction aqueous enzymatic method residue according to claim 1; It is characterized in that described Microwave Extraction optimum condition is: water is 18.1mL/g with dry residue ratio; Microwave power 666W crosses grit number 60 orders, microwave time 19.9min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103095451A CN102783657A (en) | 2012-08-28 | 2012-08-28 | Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103095451A CN102783657A (en) | 2012-08-28 | 2012-08-28 | Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102783657A true CN102783657A (en) | 2012-11-21 |
Family
ID=47149419
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103095451A Pending CN102783657A (en) | 2012-08-28 | 2012-08-28 | Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102783657A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103497822A (en) * | 2013-10-14 | 2014-01-08 | 铜陵瑞璞牡丹产业发展有限公司 | Method of extracting protein and dietary fibers from peony seeds or peony seed meal |
| CN106136012A (en) * | 2016-07-09 | 2016-11-23 | 东北农业大学 | A kind of method that aqueous enzymatic method residue prepares soybean dietary fiber beverage |
| CN106750568A (en) * | 2016-11-28 | 2017-05-31 | 东北农业大学 | The method that edible packing membrane is prepared using aqueous enzymatic method residue |
| CN107125331A (en) * | 2017-07-03 | 2017-09-05 | 东北农业大学 | The method that a kind of utilization soybean aqueous enzymatic method hydrolyzate and residue prepare the bean curd of page thousand |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101602979A (en) * | 2009-06-09 | 2009-12-16 | 东北农业大学 | A kind of extracting method of soybean oil |
| CN101627813A (en) * | 2009-08-11 | 2010-01-20 | 黄山旅游集团佳龙绿色食品有限公司 | Method for extracting soybean dietary fibre and soybean protein from soybean residue |
-
2012
- 2012-08-28 CN CN2012103095451A patent/CN102783657A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101602979A (en) * | 2009-06-09 | 2009-12-16 | 东北农业大学 | A kind of extracting method of soybean oil |
| CN101627813A (en) * | 2009-08-11 | 2010-01-20 | 黄山旅游集团佳龙绿色食品有限公司 | Method for extracting soybean dietary fibre and soybean protein from soybean residue |
Non-Patent Citations (2)
| Title |
|---|
| 尹艳等: "微波提取水溶性大豆多糖工艺研究", 《食品研究与开发》 * |
| 潘利华等: "超声辅助提取水不溶性大豆膳食纤维及其物理特性", 《农业工程学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103497822A (en) * | 2013-10-14 | 2014-01-08 | 铜陵瑞璞牡丹产业发展有限公司 | Method of extracting protein and dietary fibers from peony seeds or peony seed meal |
| CN103497822B (en) * | 2013-10-14 | 2015-11-18 | 铜陵瑞璞牡丹产业发展有限公司 | The method of protein, food fibre and peony seed oil is extracted from peony seeds or the peony seeds dregs of rice |
| CN106136012A (en) * | 2016-07-09 | 2016-11-23 | 东北农业大学 | A kind of method that aqueous enzymatic method residue prepares soybean dietary fiber beverage |
| CN106750568A (en) * | 2016-11-28 | 2017-05-31 | 东北农业大学 | The method that edible packing membrane is prepared using aqueous enzymatic method residue |
| CN107125331A (en) * | 2017-07-03 | 2017-09-05 | 东北农业大学 | The method that a kind of utilization soybean aqueous enzymatic method hydrolyzate and residue prepare the bean curd of page thousand |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102805352A (en) | Method for extracting soybean dietary fibers from residues of water enzyme method | |
| CN102796613B (en) | Demulsification method by soybean grease microbe extraction through aqueous enzymatic method | |
| CN102224907B (en) | Method for extracting dietary fibers from bean pods | |
| CN102783657A (en) | Method of extracting soybean dietary fiber from aqueous enzymatic method residue by microwave | |
| CN102766533B (en) | Biological enzyme demulsification method for extracting soybean oil and fat by aqueous enzymatic method | |
| CN102206542B (en) | Method for preparing maize germ oil through enzymatic degumming | |
| CN102559365A (en) | Method and device for extracting rapeseed oil by extrusion expansion aqueous enzymatic method | |
| CN103497822B (en) | The method of protein, food fibre and peony seed oil is extracted from peony seeds or the peony seeds dregs of rice | |
| Yin et al. | Optimization of production conditions for β-mannanase using apple pomace as raw material in solid-state fermentation | |
| CN101224022A (en) | Processing method for simultaneously preparing sesame oil and protein by water enzymatic extraction | |
| CN102827687A (en) | Extraction method of soybean oligosaccharides | |
| CN103589505A (en) | Evening primrose oil extraction method | |
| CN102229852A (en) | Method for extracting fatty acid balanced soybean blend oil | |
| CN102816633A (en) | Method for synchronously extracting corn germ oil and separation protein by using alcohol-assisted enzyme method | |
| CN102911787A (en) | Method for assisting in subcritical extraction of soybean oil by using ultrasonic waves | |
| CN102864011B (en) | Method for demulsifying soybean oil and fat extracted by aqueous enzymatic method with supercritical CO2 | |
| CN102796608A (en) | Method for recovering soybean oil and protein in hydrolysate with aqueous enzymatic method | |
| Qi et al. | Extract dietary fiber from the soy pods by chemistry-enzymatic methods | |
| CN102827683A (en) | Method for extracting colza oil and protein by using solid state fermentation aqueous enzymic method | |
| Qin et al. | Extraction and utilization of active substances from edible fungi substrate and residue: A review | |
| CN103881115B (en) | A kind of method strengthening enzymolysis process extraction tobacco leaf xylogen | |
| CN105037744A (en) | Lignin solvent and preparation method therefor and application thereof | |
| CN103570794A (en) | Extracting method of camellia seed saponin | |
| CN103621920B (en) | Method for extracting melanoidin from soybean paste residue cake | |
| CN101558817B (en) | Method for producing protein concentrate by high temperature peanut meal |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121121 |