Abstract:
The present invention discloses an alga essence nucleic acid fluid concentration method, to apply the leaching function principle with the operation of freezes defreeze to achieve the goal of extract nucleic acid from the concentrated  chlorella.  Because of above operation steps with this invention, it will obtain better density of alga nucleic acid fluid. This invention is also to prepare above mentioned particular method of concentrate the alga nucleic acid and produce the alga nucleic acid fluid. The alga nucleic acid fluid may maintain it sweet flavor, and the alga nucleic acid fluid assumes the amber leaning to green color, it is sweet flavor and purely natural taste.

Description:
BACKGROUND OF THE INVENTION 
       [0001]    1. Field of the Invention 
         [0002]    The present invention is generally related to the nucleic acid fluid from the kind of  chlorella  (the  chlorella  hot water to extract “ Chlorella  Extracts; C.E.; C.G. F.”) Making use of the quality of the solute change in which is produced from leaching action, extraction of the essence in the alga contain the nucleic acid is designated as achievement purpose from operation of freezing and thawing. 
         [0003]    2. Description of the Prior Art 
         [0004]      Chlorella  ( Chlorella  sp.) It is a kind of unicellular alga plant in fresh water, it was discovered by the Dutch microbiologist “M W Bei Yonick” in 1890, the size is similar to the red corpuscle of the mankind, it can be observed through the microscope, generally being called “green blood corpuscle”, it is the unicellular plant which does not have maneuverability. According to the classification in botany, the  chlorella  is spherical monad plankton from the fresh water; mainly it is globose or is elliptical shape. Photosynthesis efficiency of the alga is several dozen faster or more than the other plant, also the chlorophyll which contained more abundant than the other plant. Simultaneously, the natural  chlorella  having the special separation structure which the other photosynthesis living things does not have, the new born natural alga absorbs nutrition and light energy from underwater and to grow, which it matures to blast cell from 20 to 24 hours and furthermore it will separates to four new cells, such a quick speed propagation, is because where the abundant special growth stimulus hormone is included. 
         [0005]    The alga includes 55% or more quantity of the vegetable protein, it is something rejoices by the vegetarians. In the alga it included nutrition component such as the chlorophyll and the vegetable fiber, vitamin A, B, C, D and E, the nicotinic acid, folic acid, calcium, iron, magnesium, and various mineral substance and amino acid etc. Furthermore as for the  chlorella  it will offer the vegetarian whom easily to lack the vitamin B group and, especially vitamin B12; The vitamin B12 it is the important substance which maintains the health of the red corpuscle and nervous system, it is difficult to find in the general vegetables and fruits, but in 5 grams of green algae it contains 4 milligrams of vitamin B12, in which, green algae nucleic acid included the higher quantity than the other food, an attached table as follows. 
         [0000]    
       
         
               
             
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 The quantity of nucleic acid is included of various foods 
               
             
          
           
               
                   
                 Food name 
               
             
          
           
               
                   
                 Green 
                 Sea 
                   
                 Bonito 
                   
                   
                 Green 
                   
                 Salmon 
                 Blue 
               
               
                   
                 algae 
                 cucumber 
                 Sardine 
                 paragraph 
                 Salmon 
                 Yeast 
                 onion 
                 Egg 
                 extract 
                 alga 
               
               
                   
                   
               
             
          
           
               
                 Content 
                 13000 
                 3605 
                 539 
                 907 
                 289 
                 1399 
                 78 
                 86 
                 10600 
                 4600 
               
               
                 nucleic 
                 DNA&amp;RNA 
                   
                   
                   
                   
                 RNA 
                   
                   
                 DNA 
               
               
                 acid 
               
               
                 mg/100 g 
               
               
                   
               
             
          
         
       
     
         [0006]    The alga is a kind of alkaline food, long term in take it, can adjust one&#39;s physique, many nutrition component which are included in it can helps adjustment of physiological function, maintains digestion performance, promotion of health and beauty care, strength physical ability, health maintenance and longevity prolongation of life, furthermore may take as nourishment for pre-natal, post-natal or after illness, at present time many countries are doing research of the  chlorella,  to mass produce it and develop into the products to sale. 
         [0007]    In the  chlorella  includes alga essence ( Chlorella  Extracts C.E.; C.G. F.), is also known as the alga growth stimulation factor, it is the essence of the alga. In every 100 grams of alga usually contains only 4˜5 grams of alga concentrated essence fluid. And in the alga concentrated essence fluid includes the Nucleic Acid, Nucleotide, Folic Acid, Niacin, Lysine, Alanine, Glycine, Praline, Glutamic Acid, Aspartic Acid, Punting Ten, Small Molecular Protein, the water soluble Vitamin and Mineral Substance, is a similar material and ingredient with the animal placenta element. Therefore, it is also named as plant placenta element; and it may stimulate the alga to grow faster. 
         [0008]    Generally during the process of concentration and extraction of alga essence, this concentration procedural is a key method to increase the application scope and the commercial value of the alga. 
         [0009]    The present technology of concentration procedural is to heat up the alga directly, and evaporate part of moisture to achieve the goal of concentration, and it may be used with the related auxiliary depressurize equipment to increase the efficiency of this part of process. 
         [0010]    Due to the long heating process of alga essence fluid, the destruction of the complicated nutrition ingredient is unavoidable. There is unpredictable reaction between various ingredients during the process. Furthermore the long heating process reduces the luster and flavor of the alga concentration fluid. The fact that to solve the fault of the above-mentioned traditional alga essence fluid concentration method and to offer the concentrated method to extracting the nucleic acid include within alga is immediate urgent matter. 
       SUMMARY OF THE INVENTION 
       [0011]    According to the background of this application, an alga essence nucleic acid fluid concentration method is developed and disclosed. 
         [0012]    The main purpose of this invention is to solve the fault from above mentioned traditional alga essence fluid concentration method. It is to offer the concentration method to extract its content of nucleic acid from alga, by applying the leaching function will create the characteristic of solute transformation phenomena, the refrigeration method will achieve it purpose of extract the nucleic acid ingredient from the concentrated alga essence fluid. This invention produces the alga nucleic acid fluid with it color and it luster leaning green with the pure natural flavor, and sweet taste. 
         [0013]    In order to achieve above goal, by applying the method with this invention, from  chlorella  to extract nucleic acid, include following; First, the formation of the  chlorella  mixture fluid, which made possible by at least intermix with the raw materials of  chlorella  and to infiltration with the mixed liquid to form the  chlorella  mixture fluid. Next, process of extraction is proceed, to heats the  chlorella  mixed liquid evenly at approximately from between 90 degree centigrade up to 120 degrees, and to maintain it for a period of time, then cooling the heated  chlorella  mixed fluid, it forms the  chlorella  extracted fluid. Continuously, the dregs and the suspended matter in the  chlorella  extract are removed and only the chlorophyll is left to form the prototype product of alga essence nucleic acid. Lastly, at least once the leaching process is done, depending on this leaching action to freezes and at the same time nature melting the prototype product of the alga essence nucleic acid fluid, and to complete the concentrated alga essence nucleic acid fluid. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0014]      FIG. 1  is a flow chart of the alga essence nucleic acid fluid concentrated production method according to the first embodiment of the present invention; 
           [0015]      FIG. 2  is a flow chart of the alga essence nucleic acid fluid concentrated production method according to the second embodiment of the present invention; 
           [0016]      FIG. 3  is a flow chart of the alga essence nucleic acid fluid concentrated production method according to the third embodiment of the present invention; 
           [0017]      FIG. 4  is a operating procedure of the alga essence nucleic acid fluid concentrated production method according to the present invention; and 
           [0018]      FIG. 5  is during the concentration stages where collected the leaching solution at the time of each stage the related figure of the solution include it quantity and the time according to the present invention. 
       
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
       [0019]    What is probed into the invention is an alga essence nucleic acid fluid concentration method. Detail descriptions of the structure and elements will be provided as followed in order to make the invention thoroughly understood. The application of the invention is not confined to specific details familiar to those who are skilled in the art. On the other hand, the common structures and elements that are known to everyone are not described in details to avoid unnecessary limits of the invention. Some preferred embodiments of the present invention will now be described in greater detail as followed. However, it should be recognized that the present invention can be practiced in a wide range of other embodiments besides those explicitly described, that is, this invention can also be applied extensively to other embodiments, and the scope of the present invention is expressly not limited except as specified in the accompanying claims. 
         [0020]    The invention is to provide an alga essence nucleic acid fluid concentration method, to eliminate traditional direct heating method, but to extract the concentration by carry out the freezes, and defrost operation, this method may further achieve the simplification and maintain establishing of the production equipment, and then reduces the production cost, the concentrates extract of the  chlorella  essence nucleic acid fluid by this invention, may maintain its sweet flavor, and to improve the luster by join the tradition facture to obtained the concentrated  chlorella  essence. 
         [0021]    The alga nucleic acid fluid referred in this invention is a kind of  chlorella  extracted from hot water concentration, the ingredient of this extracts are the protein, the polysaccharide, the Vitamin and the mineral substance etc the aqueous solution, long-term edible has the function of adjust one&#39;s physique, and to improve health. Moreover, in the  chlorella  nucleic acid fluid&#39;s special nutrition ingredient the alga essence accelerate growth factor (CGF) has many biological activity function, therefore the alga hot water extraction receives special attention, therefore, to judge the  chlorella  nucleic acid fluid quality is fit or unfit, usually rests on its nature quality (luster) and takes it CGF quantity (density). 
         [0022]    To determinate its content of  chlorella  CGF and its legal expression system act according to Taiwan Commodity Inspection Bureau the CNS4202 and N5134 stipulation method. Naturally raised alga its CGF content majorities are situated between 1.3˜2.5; it will be different along with the different kind of plants and growthraise condition. The alga nucleic acid fluid&#39;s CGF content by the nature most density indicated the absorbance quality under light wave length 260 nm, for example OD200 or OD400. (OD: Optical Density). 
         [0023]    As shown in  FIG. 1 , a first embodiment of the present application discloses an alga essence nucleic acid fluid concentrated production method  100 , comprising: forming a  Chlorella  mixed fluid  110 , performing a extraction process  120 , removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  130 , and performing a leaching process  140 . Depending on above description, forming a  Chlorella  mixed fluid  110 , wherein the  Chlorella  mixed fluid is mixed and dampened by at least one of  Chlorella  raw materials and a mixed fluid. As for mixed liquid it can be water, secondary water, deionizer water or all nonpoisonous solvents etc. The above-mentioned the OD (Optical Density) value of the  Chlorella  raw materials is between 1.5 and 2.5. The Best, the OD (Optical Density) value of the  Chlorella  raw materials is approximately 2.3. 
         [0024]    Continuously, an extraction process  120  is performed to heat the  Chlorella  mixed fluid evenly with first temperature and maintaining fixed time, and the heated  Chlorella  mixed fluid is to cooling down with the second temperature to form a  Chlorella  extract fluid. The above-mentioned first temperature is between 80 degree centigrade and 120 degree centigrade. As for being better, the above-mentioned first temperature is between 80 degrees centigrade to 95 degree centigrade. The above-mentioned  chlorella  mixed fluid is heated evenly at the first temperature, particular time is maintained, the particular time is between 30˜40 minutes. The above-mentioned second temperature is between 20 degree centigrade and 30 degree centigrade. 
         [0025]    After the completing the extraction process  120 , removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  130  to form an alga essence nucleic acid prototype product. Finally, a leaching process  140  is performed, the alga essence nucleic acid prototype product is frozen and then thawed naturally by leaching action to form a concentrated alga essence nucleic acid fluid. The above-mentioned concentrated alga essence nucleic acid fluid is more than or equal to 400. To repeat it at least 3 times as for relatively better ones with the above-mentioned leaching process  140 . 
         [0026]    As shown in  FIG. 2 , a second embodiment of the present application discloses an alga essence nucleic acid fluid concentrated production method  200 , comprising: forming a  Chlorella  mixed fluid  210 , performing a extraction process  220 , removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  230 , performing a frozen process  240 , and performing a thawing process  250 . Depending on above description, forming a  Chlorella  mixed fluid  210 , wherein the  Chlorella  mixed fluid is mixed and dampened by at least one of  Chlorella  raw materials and a mixed fluid. Then, an extraction process  220  is performed to heat the  Chlorella  mixed fluid evenly with first temperature and maintaining fixed time, and the heated  Chlorella  mixed fluid is to cooling down with the second temperature to form a  Chlorella  extract fluid. Next, removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  230  to form an alga essence nucleic acid prototype product. Next, a frozen process  240  is performed, the alga essence nucleic acid prototype product is frozen below freezing point of zero centigrade. Finally, a thawing process  250  is performed, the frozen alga essence nucleic acid fluid prototype product is naturally thawed approximately between 24 centigrade degree to 28 degrees to form said concentrated alga essence nucleic acid fluid. The above-mentioned  chlorella  raw materials OD value, first temperature, maintain a particular time at first temperature, second temperature, essence nucleic acid prototype product OD value and concentrated alga essence nucleic acid fluid OD value are same condition with the first embodiment of the present application. 
         [0027]    As shown in  FIG. 3 , a third embodiment of the present application discloses an alga essence nucleic acid fluid concentrated production method  300 , comprising: forming a  Chlorella  mixed fluid  310 , performing a extraction process  320 , removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  330 , performing a first frozen process  340 A, performing a first thawing process  350 A, performing a second frozen process  340 B, and performing a second thawing process  350 B. Depending on above description, forming a  Chlorella  mixed fluid  310 , wherein the  Chlorella  mixed fluid is mixed and dampened by at least one of  Chlorella  raw materials and a mixed fluid. Then, an extraction process  320  is performed to heat the  Chlorella  mixed fluid evenly with first temperature and maintaining fixed time, and the heated  Chlorella  mixed fluid is to cooling down with the second temperature to form a  Chlorella  extract fluid. Next, removing the dregs and the suspended matter and keeping the Chlorophyll in the  Chlorella  extract  330  to form an alga essence nucleic acid prototype product. Next, a first frozen process  340 A is performed, the alga essence nucleic acid prototype product is frozen below freezing point of zero centigrade. Next, a first thawing process  350 A is performed, the frozen alga essence nucleic acid fluid prototype product is naturally thawed approximately between 24 centigrade degree to 28 degrees. Next, a second thawing process  340 B is performed, the defrost alga essence nucleic acid fluid prototype product to freeze it again below the freezing point of the centigrade. Finally, a second thawing process  350 B is performed, the frozen alga essence nucleic acid fluid prototype product is naturally thawed approximately between 24 centigrade degree to 28 degrees to form the concentrated alga essence nucleic acid fluid. The above-mentioned  chlorella  raw materials OD value, first temperature, maintain a particular time at first temperature, second temperature, essence nucleic acid prototype product OD value and concentrated alga essence nucleic acid fluid OD value are same condition with the first embodiment of the present application. The above-mentioned frozen process ( 340 A;  340 B) and with thawing process ( 350 A;  350 B) in between it can proceed to repeat several time. 
       EXAMPLE 1 
       [0028]    With regular  chlorella  powder raw material ( Chlorella  sp. ) and adds on 1:8˜10 time (w/w) water to infiltration mix with the alga raw material powder, to evenly add heating process until reach to 90˜100° C., and maintains 50˜60 minutes then carries on to cool downit to the normal temperature (room temperature), will obtain the mixture and after the separation of the solid material (wreckage of algae body), and after the steps of extracting suspension to keep the chlorophyll etc, it clarifies the amber leaning green liquid, namely the  chlorella  extracted liquid include the alga essence accelerate growth factor (CGF), I.e. the alga essence of nucleic acid fluid has not concentrated is the initial prototypeend product (the usual OD value situated between 130˜170), placeput this has not yet concentrated alga essence nucleic acid fluid of the prototype initial end product into a regular vessel, store it in the refrigerator to freeze it, this frozen prototypeinitial end product of un-concentrated alga essence nucleic acid fluid, take it out from the refrigerator and causes it to dissolve under the room temperature, because of the dissolved state continues to carry on (coexistent and mix with a solid condition) has the leaching function, After gathering this dissolution, the liquid state part (to be called leaching fluid or drop filtrate liquates), namely the concentrated alga essence of nucleic acid fluid; the un-dissolved portion content extremely few solute; may get rid of it or to redo it, to repeat the above step. 
         [0029]    So-called “the leaching function” is refers to the ice in the initial dissolve liquid state portion, the solute content is much higher than average content in the ice solute; the ice dissolves the water is the function to cause the chemical composition to have the migration phenomenon in the ice. 
         [0030]    As show in  FIG. 4 , to operate the above steps repeatedly altogether by 3 times, at the prototype product of alga essence nucleic acid fluid it OD value is 170, it may be concentrates the OD value over above 400 of the green alga nucleic acid fluid. 
         [0031]    In which, takes the raw material of  chlorella  to target for OD value 1.5, after depends on above step to manufacture, obtains the OD130 alga essence nucleic acid fluid at the prototype initial end product, after carry out the first freeze, defrosting, and the collection procedure, to obtains the density is above OD170; if this raw material  chlorella  OD value target is 2.3, after depend on above manufacture step, obtains the initial prototypeend product of the alga essence nucleic acid fluid at OD170, after carries on again to freeze it, defrosting, withand the collection procedure, the final product density of the OD value is above 230, repeatedly operates the above freezing, the dissolvinged and collection steps reaches to 3 times, the alga essence nucleic acid fluid may result in the OD value is above 400. 
         [0032]    The concentration method apply to the production line operation has the extremely good flexibility, and ease to operate, the prototype initial end product of this alga essence nucleic acid fluid and place itat put in a regular 25˜35 liter barrel, place it in the refrigerator to freezecause it freeze, and take out this frozen prototype initial end product of alga essence nucleic acid fluid from the refrigerator and inverts it, causes it to dissolve naturally under the room temperature, continues to carry on with the dissolved state to forms the liquid-solid coexistent condition, to produce the leaching function, and then to collecting this leaching fluid (or calls drop filtrate), namely alga essence nucleic acid fluid from this invention. 
         [0033]    As show in  FIG. 5 , when carries on the concentration step again with this invention, is also works as the leaching function that to produce, continually dissolves in the process, during various stages the collection of leaching fluid (or called drop filtrate) is the solute content and the time relational graph. 
       EXAMPLE 2 
       [0034]    The Food Analysis Center of Japan in Aug. 6, 2007 (Heisei 19) analyzed assay with result report based on the concentrated alga essence nucleic acid fluid report send in for exam arranged by the applicant, as show in below: 
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Analytical Investigation Result 
               
             
          
           
               
                   
                   
                 Detection 
                   
                   
               
               
                 Assay Items 
                 Result 
                 limit 
                 Note 
                 Method 
               
               
                   
               
               
                 Moisture 
                 97.4 g/100 g 
                   
                   
                 Normal pressure 
               
               
                   
                   
                   
                   
                 heating drying method 
               
               
                 Protein 
                 1.6 g/100 g 
                   
                 1 
                 Kjeldahl method 
               
               
                 Lipid 
                 0.1 g/100 g 
                   
                   
                 Acid resolution 
               
               
                   
                 Under 
               
               
                 Ash 
                 0.4 g/100 g 
                   
                   
                 Directly ashing 
               
               
                   
                   
                   
                   
                 method 
               
               
                 Carbohydrate 
                 0.6 g/100 g 
                   
                 2 
               
               
                 Energy 
                 9 kcal/100 g 
                   
                 3 
               
               
                   
               
               
                 Note 1. Nitrogen, protein conversion factor: 6.25 
               
               
                 Note 2. Formula: 100 − (Moisture + Protein + Lipid + Ash) 
               
               
                 Note 3. Nutrition indicatory standard [Heisei 15(2003) Public Welfare Ministry of Labor Public Notice No. 176] About energy conversion factor: Protein, 4; Lipid, 9; Carbohydrate, 4 
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Analytical Investigation Result 
               
             
          
           
               
                   
                   
                 Detection 
                   
                   
               
               
                 Assay Items 
                 Result 
                 limit 
                 Note 
                 Method 
               
               
                   
               
               
                 Moisture 
                 97.4 g/100 g 
                   
                   
                 Normal pressure 
               
               
                   
                   
                   
                   
                 heating drying method 
               
               
                 Protein 
                 1.6 g/100 g 
                   
                 1 
                 Kjeldahl method 
               
               
                 Lipid 
                 0.1 g/100 g 
                   
                   
                 Acid resolution 
               
               
                   
                 Under 
               
               
                 Ash 
                 0.4 g/100 g 
                   
                   
                 Directly ashing method 
               
               
                 Sugar 
                 0.4 g/100 g 
                   
                 2 
               
               
                 Dietary Fiber 
                 0.2 g/100 g 
                   
                   
                 Enzyme - Gravimetric 
               
               
                   
                   
                   
                   
                 method 
               
               
                 Energy 
                 8 kcal/100 g 
                   
                 3 
               
               
                   
               
               
                 Note 1. Nitrogen, protein conversion factor: 6.25 
               
               
                 Note 2. Nutrition indicatory standard [Heisei 15(2003) Public Welfare Ministry of Labor Public Notice No. 176] About Formula: 100 − (Moisture + Protein + Lipid + Dietary Fiber) 
               
               
                 Note 3. Nutrition indicatory standard [Heisei 15(2003) Public Welfare Ministry of Labor Public Notice No. 176] About energy conversion factor: Protein, 4; Lipid, 9; Suger, 4; Dietary fiber, 2 
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Analytical Investigation Result 
               
             
          
           
               
                 Assay Items 
                 Result 
                 Detection limit 
                 Note 
                 Method 
               
               
                   
               
               
                 Sodium 
                 2.7 mg/100 g 
                   
                   
                 Atom extinction brightness method ICP 
               
               
                 Iron 
                 0.44 mg/100 g 
                   
                   
                 luminous analysis 
               
               
                 Calcium 
                 9.9 mg/100 g 
                   
                   
                 ICP luminous analysis 
               
               
                 Kalium 
                 117 mg/100 g 
                   
                   
                 Atom extinction brightness method ICP 
               
               
                 Magnesium 
                 20.5 mg/100 g 
                   
                   
                 luminous analysis 
               
               
                 Zinc 
                 0.14 mg/100 g 
                   
                   
                 ICP luminous analysis 
               
               
                 Selenium 
                 Not detect 
                   5 μg/100 g 
                   
                 Fluorescent brightness method 
               
               
                 Entire chrome 
                 Not detect 
                 0.05 mg/100 g  
                   
                 ICP luminous analysis 
               
               
                 Vitamin A(Retinol equivalence) 
                 3 μg/100 g 
                   
                 1 
               
               
                 α-Carotin 
                 8 μg/100 g 
                   
                   
                 High speed liquid chromatograph method 
               
               
                 β-Carotin 
                 27 μg/100 g 
                   
                   
                 High speed liquid chromatograph method 
               
               
                 Thiamine (Vitamin B 1 ) 
                 0.03 mg/100 g 
                   
                 2 
                 High speed liquid chromatograph method 
               
               
                 Ribo Flavin (Vitamin B 2 ) 
                 0.42 mg/100 g 
                   
                   
                 High speed liquid chromatograph method 
               
               
                 VitaminB6 
                 0.13 mg/100 g 
                   
                 3 
                 Bio assay method 
               
               
                 VitaminB 12   
                 1.1 μg/100 g 
                   
                 4 
                 Bio assay method 
               
               
                 Ascorbic acid(Total Vitamin C) 
                 Not detect 
                   1 mg/100 g 
                 5 
                 High speed liquid chromatograph method 
               
               
                 Vitamin D 
                 Not detect 
                 0.1 μg/100 g 
                   
                 High speed liquid chromatograph method 
               
               
                 Vitamin D (International unit) 
                 — 
               
               
                 Vitamin E           
                 Not detect 
                   
                   
                 High speed liquid chromatograph method 
               
               
                 Folic acid 
                 82 μg/100 g 
                 0.1 mg/100 g 
                 6 
                 Bio assay method 
               
               
                   
               
               
                 —: Because it does not detect, it does not calculate. Induction embody it measured 
               
               
                 Note 1. α-Carotin 24 μg and β-Carotin 12 μg, respectively take the retinol equivalence of 1 μg 
               
               
                 Note 2. As a thiamine salt 
               
               
                 Note 3. Fungus strain used:  Saccharomyces cerevisiae  ( S. uvarum ) ATCC 9080 
               
               
                 Note 4. Fungus strain used:  Lactobacillus delbrueckii  subsp.  Lactis  ( L. leichmannii ) ATCC 7830 
               
               
                 Note 5. Apply hydrazine and measured after changes into induction embody. 
               
               
                 Note 6. Fungus strain used:  Lactobacillus rhamnosus  ( L. casei ) ATCC 7469 
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Analytical Investigation Result 
               
             
          
           
               
                 Assay Items 
                 Result 
                 Detection limit 
                 Note 
                 Method 
               
               
                   
               
               
                 Punting ten 
                 0.19 mg/100 g 
                   
                 1 
                 Bio assay method 
               
               
                 Biotin 
                 5.6 μg/100 g 
                   
                 1 
                 Bio assay method 
               
               
                 Inositol 
                 5 mg/100 g 
                   
                 2 
                 Bio assay method 
               
               
                 Niacin 
                 3.02 mg/100 g 
                   
                 3 
               
               
                 Niacin(Nicotine suitable quantity) 
                 2.85 mg/100 g 
                   
                 1 
                 Bio assay method 
               
               
                 Toriputohuan 
                 10 mg/100 g 
                   
                   
                 High speed liquid chromatograph method 
               
               
                 Choline 
                 Not detect 
                 0.03 g/100 g 
                 4 
               
               
                 Dextro glucose 
                 0.22 g/100 g 
                   
                 5 
                 High speed liquid chromatograph method 
               
               
                 Arabinose 
                 Not detect 
                 0.02 g/100 g 
                 5 
                 High speed liquid chromatograph method 
               
               
                 Xylose 
                 Not detect 
                 0.02 g/100 g 
                 5 
                 High speed liquid chromatograph method 
               
               
                 Rhamnose 
                 0.04 g/100 g 
                   
                 5 
                 High speed liquid chromatograph method 
               
               
                 Mannose 
                 Not detect 
                 0.02 g/100 g 
                 5 
                 High speed liquid chromatograph method 
               
               
                 Galactose 
                 0.05 g/100 g 
                   
                 5 
                 High speed liquid chromatograph method 
               
               
                 Lysine 
                 66 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Alanine 
                 112 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Glycine 
                 69 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Proline 
                 57 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Glutamic acid 
                 169 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Ceric 
                 45 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                 Aspartic acid 
                 99 mg/100 g 
                   
                   
                 Amino acid automatic analysis 
               
               
                   
               
               
                 Note 1. Fungus strain used:  Lactobacillus plantarum  ATCC 8014 
               
               
                 Note 2. Fungus strain used:  Saccharomyces cerevisiae  ( S. uvarum ) ATCC 9080 
               
               
                 Note 3. Niacin(Nicotine suitable quantity) and 1/60 of           total quantity was made by niacin equivalent 
               
               
                 Note 4. Based on            salt settling method. The room temperature mixes the approximately 1 hour later 
               
               
                 Note 5. Acid adding water to disassembled it and then measured. Hydrolysis condition: 72% of Sulfuric acid, after agitating it for one hour at room temperature, 4% sulfuric acid, put in autoclave (121° C.), one hour 
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Analytical Investigation Result 
               
             
          
           
               
                   
                   
                 Detection 
                   
                   
               
               
                 Assay Items 
                 Result 
                 limit 
                 Note 
                 Method 
               
               
                   
               
               
                 Total 
                 3.0 mg/100 g 
                   
                   
                 Extinction brightness 
               
               
                 Chlorophyll 
                   
                   
                   
                 method (Visible) 
               
               
                 Chlorophyll a 
                 2.7 mg/100 g 
               
               
                 Chlorophyll b 
                 0.3 mg/100 g 
               
               
                 Cobalt 
                 Not detect 
                 0.05 ppm 
                   
                 Atom extinction 
               
               
                   
                   
                   
                   
                 brightness method 
               
               
                 Germanium 
                 Not detect 
                   1 ppm 
                   
                 ICP luminous analysis 
               
               
                   
               
             
          
         
       
     
         [0035]    Other modifications and variations are possibly developed in light of the above demonstrations. It is therefore to be understood that within the scope of the appended claims the present invention can be practiced otherwise than as specifically described herein. Although specific embodiments have been illustrated and described herein, it is obvious to those skilled in the art that many modifications of the present invention may be made without departing from what is intended to be limited solely by the appended claims.