Genistein 98%,Sophora japonica extract

Product Name: Genistein

Latin name:Genista tinctoria Linn.

Quality standard: National Standard

Specification: 98%

Test method:HPLC

CAS NO: 446-72-0

Appearance: Off- White powder

Storage method:  Store in cool and dry place. Keep away from strong light and heat.

Shelf life: 24 month

Source of extraction: Soybeans/Sophora japonica 

Package specification: 1kg / aluminum foil bag, 25kg/Drum or as required by customers

Product introduction: Genistein is slightly soluble in water

Sophora japonica Extract Genistein

1. Range

   This technical requirement specifies the technical requirements, test method, test rules and label of Sophora japonica extract Genistein.

   The requirements of this standard apply to the raw material of Sophora japonica made by the processing of extracts.

2. Standardization of reference documentThe terms in the following documents become terms of this technical requirement by reference to them. All subsequent amendments (excluding errata) or revisions to dated references shall not apply to this Technical Requirement; however, the parties to an agreement under this Technical Requirement are encouraged to study the availability of the latest version of these documents. For undated references, the latest version is applicable to this technical requirement.GB/T 5009.11 Determination of total arsenic and inorganic arsenic in food componentGB/T 5009.12 Determination of lead in foodsGB/T 5009.15 Determination of cadmium in foods

GB/T 5009.17 Determination of total and organic mercury in foodstuffs

GB/T 5009.19 Determination of multi – residues of organochlorine pesticides in food

GB 4789.2 Microbiological testing of food hygiene – determination of total number of colonies

GB 4789.3 Microbiological testing of food hygiene – determination of coliform bacteria

GB 4789.15 Food hygiene microbiology test for mold and yeast counts

Pharmacopoeia of the People’s Republic of China (2015 edition)

1. Structural formula

2. Genistein Molecular formula C₁₅H₁₀O₅  Molecular weight  270.23.

3. Technical requirements

1. Technical requirement
   1. Raw material
   2. Plant raw materials are harvested in winter to remove impurities and dry.

4.2 Sensory requirement

Conform to the requirement of chart 1

Chart 1  Sensory index

4.3 Symbolic ingredient

Conform to chart 2 requirement

Chart 2   Symbolic ingredient

4.4 Physicochemical requirement

Conform to chart 3 requirement

Chart 3 Physicochemical requirement

4.5 Microbiological indicator

Conform to chart 4 requirement

Chart 4 Microbiological indicators

5 Test method

5.1 After opening the sample, smell its smell and taste its taste immediately ;In addition, appropriate amount of the sample was placed in a white porcelain plate to observe its color and appearance, and to check for foreign bodies.

5.2 Physicochemical requirement

5.2.1 The purity of genistein The method was determined according to Appendix A

5.2.2 Mesh

The test was carried out according to the screening method stipulated in the General Rule 0982(Second Method) of the Fourth Part of the Pharmacopoeia of the People’s Republic of China (2015).

5.2.3 Moisture

The assay was conducted according to the method specified in General Rule 0832(second method) of the fourth part of Pharmacopoeia of the People’s Republic of China (2015 edition).

5.2.4 Ash content

The assay was carried out according to the method specified in General Rule 1106 of the fourth part of Pharmacopoeia of the People’s Republic of China (2015 edition)

5.2.5 Arsenic

The total arsenic and inorganic arsenic in food were determined according to GB/T 5009.11 method.

5.2.6 Lead

According to GB/T 5009.12 for the determination of lead in food.

5.2.7 Cadmium

Cadmium in food was determined according to GB/T 5009.15.

5.2.8 Hg

The total mercury and inorganic mercury in food were determined according to GB/T 5009.17.

5.2.9 Hexachloro-cyclohexane soprocide  DDT  Huintozene 

Execute according to GB/T 5009.19

5.4 Bacteriological examination

5.4.1 Total Plate Count

In accordance with the “national food hygiene standard” GB4789.2

5.4.2 Yeast & Mold

In accordance with the “national food hygiene standard” GB4789.15

5.4.3 E.Coli

In accordance with the “national food hygiene standard” GB4789.3

5.4.4 Salmonella

In accordance with the “national food hygiene standard” GB4789.4

5.4.5 Staphylococcus

In accordance with the “national food hygiene standard” GB4789.10

6 Test rule

6.1 Test

Samples were taken according to the method specified in the General Rule 0211 of the Four Parts of the Pharmacopoeia of the People’s Republic of China (2015 edition), and the products that were evenly mixed and had the same nature and quality within a certain limit were tested as the same batch.

6.2 Inspection Classification

6.2.1 Delivery inspection

Before leaving the factory, the quality inspection department of the factory shall inspect the products batch by batch according to the requirements of 4.2 ~ 4.5, and the products that pass the inspection and issue the quality certificate can be sold before leaving the factory.

6.2.2 Test type

Carry out type inspection according to chapter 4.Type inspection shall be carried out every six months in case of any of the following situations:

A) when there is a big change in raw materials;

B) Adjustment of key processes;

C) Resuming production after replacing equipment or stopping production;

D) There is a big difference between the results of ex-factory inspection and the last type inspection.

6.3 Decision rules

6.3.1 If one of the inspection results does not meet the technical requirements, samples of twice the quantity shall be randomly selected from the same batch of products for re-inspection, and the re-inspection result shall prevail. If there is still one index unqualified in the reinspection, the batch of products will be judged as unqualified.

6.3.2 If any item in the microbial project does not meet the technical requirements, it will be judged as unqualified and will not be re-inspected.

6.3.3 Determination of type inspection is the same as that of factory inspection.

Labeling, packaging, transportation and storage

7.1 The label

7.1.1 The packaging label shall mark: sample name, date of production, batch number of production, specification, gross weight, net weight and execution standard.

7.1.2 The label content is clearly visible and the label is firmly pasted.

7.2 The packing

Packaging materials should meet the requirements of food hygiene.The packaging materials shall be inspected strictly before use.After the barrel is packed, it should be sealed with a seal label.

7.3 Transportation

7.3.1 The means of transport shall be clean and hygienic, and shall not be mixed with poisonous, harmful, corrosive or odor articles.

7.3.2 Light load and light discharge shall be carried out, and extrusion, exposure and rain shall be prevented during transportation.

7.4 Storage

7.4.1 The product shall not be stored mixed with toxic, harmful, corrosive or peculiar smell.

7.4.2 The product should be sealed and stored in a cool and dry place.

7.5 Shelf life

Shelf life is 24 months. 

Testing method of genistein purity by HPLC

1. PurposeTo establish the standard operating instruction for testing method of genistein purity by HPLC.
2. RangeApply to the test method of genistein purity.
3. Comparison productsGenistein (standard  product)
4. Reductant4.1Acetonitrile ( chromatographically pure)4.2 Methyl  alcohol ( chromatographically pure)4.3 Methyl  alcohol(analytically  pure)4.4 Analytically pure(analytically  pure)4.5 Water (ultrapure water)5. Instrument5.1HPLC  : Waters 2695  Infusion pump: Waters 2487 UV-detector :Empower

Chromatographic data working system.

5.2 Analytical balance: Sensitive quality.

5.3 USC instrument: Power: 250W  Frequency 40KHz. Glass moving phase filter（0.45μm）

5.4 Needle micro hole filter membrance.

5.5 Volumetric glass:mearing cylinder (100ml), Volumetric flask(25、50ml).

6. Solution preparation.

6.1 Solution preparation of comparison products.

Get the genistein comparison products precisely about 10mg into the volumetric  flask of 50ml, add the methyl alcohol  about 40ml, after ultrasonic dissolving, place it under the indoor temperature, use methyl alcohol metered volume to the scale, shake up.

6.2 The preparation of sample solution.

Get the sample about 10mg into the volume falsk of 50mg precisely, add the methyl about 40ml,  after ultrasonic vibrating extraction about 30min, place under the indoor temperature, use methyl metered volume to the scale, use the 0.45µm needle filter instrument to filter, then get the sample soution.

7.Chromatographic condition.

Chromatographic condition:Intersil ODS-C18 (150mm×4.6 mm ,5µm)  

Wave length:262nm

Moving phase: methyl alcohol: acetonitrile: 0.1% phosphoric acid solution

Column temperature: indoor input sample 10µl

Sensitivity:2.000AUFS  

The adaptable of system:According to the genistein number of theoretical plates can not less than 3000

8.Sample test:

Under the above chromatographic conditions, the comparison product solution and sample solution are absorbed precisely after the instrument is stabilized and the baseline is stabled

The retention time of genistein is about 6min. The purity of the sample is calculated by area external standard method.

            A1×W0× V1× K        ×100

^{Purity（ %）=}    A₀×V₀×W₁

A1:  Peak area of genistein in sample solution.

A0：Peak area of genistein in comparison product solution

W1:  Weighing sample weight mg W0: Weighing sample weight mg of comparison product

V1:    Sample dimension ml

V0： Comparison products dimensionml

K：Comparison products purity

Sample purity results are take the average of two parallel sample purity.

A brief introduction to the process of genistine

1、Extract

After crushing, the raw material of horse bean reached about 20 mesh, and was extracted 3 times with 75% ethanol by hot reflux, then concentrated into an extract with a specific gravity of 1.2

2、Extraction From

The extracted extractumis dissolved by adding an appropriate concentration of hydrochloric acid, the insoluble matter was filtered by standing, the PH value was adjusted by sodium hydroxide, and then trichloromethane was extracted for three times, and the extraction solution was concentrated.

3、Chromatography

Dissolve the extract and add appropriate amount of silica gel and mix it thoroughly. After drying, use 200-300 silica gel ethyl acetate ring/hexane system gradient chromatography, TLC detection and stage concentration.

4、Crystal

More than 95% of genitine was collected by chromatography, which was crystallized by ethanol and acetone, and the content of genitine reached more than 98%

5、Crushing dry

The product was put into phosphorus pentoxide and dried in a vacuum drying oven at a constant temperature for 48 hours

6、Detection of the packing

The finished product is sampled and tested according to the standard of the enterprise.

After passing the test, the product is packed and stored in the warehouse. 

Process description:

This process is our company’s original technology, in the process does not use benzene, carbon tetrachloride and other organic solvents which are very harmful to human body, only the main organic solvents used are ethanol, cyclohexane, ethyl acetate, acetone and so on, the rest are pure water.

The above process is for reference only. The specific parameters of our company have reservations, please understand!