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GB 1886.172-2016 English PDF

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GB 1886.172-2016: National Food Safety Standard -- Food Additives -- Rosemary Extract
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Basic data

Standard ID GB 1886.172-2016 (GB1886.172-2016)
Description (Translated English) National Food Safety Standard -- Food Additives -- Rosemary Extract
Sector / Industry National Standard
Classification of Chinese Standard X40
Word Count Estimation 9,998
Date of Issue 2016-08-31
Date of Implementation 2017-01-01
Regulation (derived from) State Health and Family Planning Commission Notice No.11 of 2016
Issuing agency(ies) National Health and Family Planning Commission of the People's Republic of China, State Food and Drug Administration

GB 1886.172-2016: National Food Safety Standard -- Food Additives -- Rosemary Extract

---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
(Food Safety National Standard Food Additive Rosemary Extract) National Standards of People's Republic of China National Food Safety Standard Food additive Rosemary Extract Published 2016-08-31 2017-01-01 implementation People's Republic of China National Health and Family Planning Commission issued National Food Safety Standard Food additive Rosemary Extract

1 Scope

This standard applies to the stem rosemary (RosmarinusofficinalisL.), Leaves as raw material by solvent extraction or supercritical carbon dioxide Extraction, refining process to produce food additives rosemary extract. The extraction solvent is water, methanol, ethanol, acetone, and (or) hexane.

2 Technical Requirements

2.1 Sensory requirements Sensory requirements shall comply with the requirements in Table 1. Table 1 Sensory requirements Project requires test methods Powder or a liquid state Proper amount of sample is placed in a clean, dry beaker or porcelain dish, from the Then observe the state of light 2.2 Physical and Chemical Indicators Physical and chemical indicators should be consistent with the provisions of Table 2. Table 2. Physical and chemical indicators project index Fat-soluble water-soluble Testing method The total antioxidant component (in terms carnosic acid and carnosol), w /% ≥ 10.0 - Appendix A A.2 Rosmarinic acid, w /% ≥ - 5.0 Appendix A A.3 Moisture a, w /% ≤ - 5.0 GB 5009.3 distillation or Karl Fischer method Lead (Pb)/(mg/kg) ≤ 2.0 GB 5009.75 or GB 5009.12 Arsenic (As)/(mg/kg) ≤ 3.0 GB 5009.76 Residual solvent b Hexane/(mg/kg) ≤ Methanol/(mg/kg) ≤ Appendix A A.4 For only a water-soluble powder products. b only for the extraction solvent is n-hexane or methanol product. NOTE. commercial rosemary extract product should comply with the standard rosemary extract as a raw material may be added for the processing, storage, standardization, dissolution Process purpose edible vegetable oils, maltodextrin, sodium chloride and other food materials and (or) meet the requirements of Food Additive Specifications emulsifiers, anti-caking Agent.

Appendix A

Testing method A.1 General Provisions At time of the reagents in water and does not indicate other requirements, refer to three analytical reagent water and GB/T 6682 provisions. Test When the standard solution, the standard solution, the determination of impurities, formulations and articles not indicate other requirements, are by GB/T 601, GB/T 602, GB/T 603 Preparation predetermined. The test does not indicate when the solution which is formulated with a solvent, refer to the aqueous solution. A.2 total antioxidants (to carnosic acid and carnosol meter) measured A.2.1 Method summary The main antioxidants (carnosol and carnosic acid) of the rosemary extract liposoluble maximum absorption at a wavelength of 280nm Detection Peak received, in accordance with standard concentration of carnosic acid and the corresponding peak area of the standard curve equation can be derived carnosic acid, and a standard curve Sample peak area corresponding to the sample solution to give a concentration. Meanwhile, since at 280 nm, the peak area of the same concentration of carnosol is sage Acid 1.36 times. Thus the concentration can be calculated based on the standard curve carnosol carnosic acid. A.2.2 Reagents and materials A.2.2.1 Water. in line with a water GB/T 6682 provisions. A.2.2.2 acetone. chromatography. A.2.2.3 Acetonitrile. chromatographically pure. A.2.2.4 carnosic acid standard. purity ≥98%. Phosphoric acid solution A.2.2.5. 1 → 1000. A.2.3 Instruments and Equipment HPLC. equipped with a UV detector. A.2.4 Reference chromatographic conditions A.2.4.1 Column. C18 reversed-phase column (φ4.6mm × 250mm, 5μm); or other equivalent column. A.2.4.2 Mobile phase A. aqueous phosphoric acid solution. A.2.4.3 mobile phase B. acetonitrile phosphoric acid solution. A.2.4.4 Column temperature. 30 ℃. A.2.4.5 Detection wavelength. 280nm. A.2.4.6 mobile phase flow rate. 1.0mL/min. A.2.4.7 Injection volume. 10μL. A.2.4.8 gradient conditions. see Table A.1. Table A.1 gradient elution Time/min Mobile phase A /% Mobile Phase B /% flow rate/(mL/min) 0.0 7723 1.0 1.0 7723 1.0 0 100 25.0 1.0 0 100 30.0 1.0 7723 1.0 30.5 7723 1.0 35.0 A.2.5 Analysis step A.2.5.1 standard curve Carnosic acid was dissolved in acetone standard, prepared a mixed standard solution gradient, a concentration gradient that the carnosic acid 0.010mg/mL ~ 1.000mg/mL. In reference A.2.4 chromatographic conditions, the standard solution is measured, a repetitive injections. The concentration of the standard solution and Peak area of the standard curve. Linear relationship should reach R2≥0.99. Recording standard curve linear equation Y = a × cb. Where, c Is the concentration of carnosic acid, Y is the peak area corresponding to the concentration, a and b are the slope and intercept of the standard curve. Preparation of sample solution A.2.5.2 Sample Weigh 140mg ~ 180mg (accurate to 0.0001g), was dissolved in 20mL acetone and 25mL volumetric flask to volume, mixed well After filtered through a 0.22μm uniform microporous membrane. A.2.5.3 Determination In reference A.2.4 chromatographic conditions, the sample was subjected to measurement, a repeated injection. Referring to FIG B.1 rosemary extract identify spectrum Carnosic acid and carnosol relative positions of the two peaks determined in response to, and record the peak area of both Y1 and Y2. A.2.6 calculation results A.2.6.1 carnosic acid concentration c1 Carnosic acid concentration c1, in milligrams per milliliter (mg/mL), calculated according to formula (A.1). c1 = Y1-b (A.1) Where. Peak area of sample solution Yl --- carnosic acid; B --- sage standard curve formula intercept acid; A --- carnosic acid in the slope of the standard curve formula. A.2.6.2 carnosol concentration c2 Carnosol concentration c2, in milligrams per milliliter (mg/mL), is calculated according to equation (A.2). c2 = Y2-b 1.36a (A.2) Where. Y2 --- sample solution carnosol peak area; B --- sage standard curve formula intercept acid; 1.36 --- at 280nm, the peak area of the same concentration was 1.36 times carnosol carnosic acid; A --- carnosic acid in the slope of the standard curve formula. Fraction of the total mass of the antioxidant A.2.6.3 w1 The total antioxidant component (in terms carnosol and carnosic acid) of mass fraction w1, according to equation (A.3) is calculated. w1 = c1 c2 () × V m × 100% (A.3) Where. C1 --- sample solution carnosic acid concentration, in milligrams per milliliter (mg/mL); c2 sage --- phenol concentration in the sample solution, in milligrams per milliliter (mg/mL); V --- sample constant volume in milliliters (mL); --- m sample mass, in milligrams (mg). The test result to the arithmetic mean of replicates results. Obtained in two independent determination results under repeatability conditions of absolute difference Value is not more than 5% of the arithmetic mean. Determination of rosmarinic acid A.3 A.3.1 Method summary Water-soluble rosemary extract was dissolved in methanol, methanol - phosphoric acid solution as the mobile phase, octadecylsilane filler for liquid chromatography of Column and UV spectrum detector or diode array detector, sample rosmarinic acid RP-HPLC separation and determination, and standard Retention time qualitative comparison of the quasi-peak area external standard. A.3.2 Reagents and materials A.3.2.1 Methanol. HPLC grade. A.3.2.2 rosmarinic acid standard. purity ≥98%. Phosphoric acid solution A.3.2.3. 0.5mL phosphate was added 100mL of water. A.3.2.4 standard stock solution. Accurately weigh 10 mg of rosmarinic acid standard (accurate to 0.0001g), dissolved in mobile phase and dilute to 10mL, mixing, home refrigerator. 1mL of this solution containing rosmarinic acid 1.0mg. A.3.3 Instruments and Equipment HPLC. equipped with UV detector or diode array detector. A.3.4 Reference chromatographic conditions A.3.4.1 Column. octadecyl silane column filler (φ4.6mm × 250mm, 5μm); or other equivalent color Spectrum column. A.3.4.2 Mobile phase. methanol. phosphoric acid = 45. 55. A.3.4.3 flow rate. 0.6mL/min. A.3.4.4 Column temperature. 40 ° C. A.3.4.5 Injection volume. 10μL. A.3.4.6 Detection wavelength. 283nm. A.3.4.7 retention time. 18min. A.3.5 Analysis step Preparation of sample solution A.3.5.1 200 mg of sample was accurately weighed (accurate to 0.0001g), dissolved in mobile phase and set the volume to 100mL, microporous membrane through 0.45μm Filtering, to obtain a sample solution. Draw standard curve A.3.5.2 Imbibe stock standard solution, formulated 0mg/mL, the standard solution series 0.25mg/mL, 0.5mg/mL, 1.0mg/mL, and In reference A.3.4 chromatographic conditions, chromatographic analysis, the content of rosmarinic acid in accordance with the standard solution and the peak area corresponding to peak Area of the vertical axis, rosmarinic acid content as abscissa, the standard curve. A.3.5.3 Determination Accurately draw sample solution 10 L, under predetermined conditions of chromatography, chromatographic analysis, qualitative retention time, peak area external standard Quantitative. A.3.6 calculation results Rosmarinic acid content w2, calculated according to formula (A.4). w2 = c × V 1000 × m × 100% (A.4) Where. C --- rosmarinic acid concentration in the sample solution results in a standard curve, in micrograms per milliliter (μg/mL); V --- constant volume of sample solution, in milliliters (mL); 1000 --- mass conversion factor; --- m sample mass, in milligrams (mg). The test result to the arithmetic mean of replicates results. Obtained in two independent determination results under repeatability conditions of absolute difference Value is not more than 2.5% of the arithmetic mean. A.4 Determination of residual solvents (hexane, methanol) of A.4.1 Reagents and materials A.4.1.1 water. a water GB/T 6682 provisions. Sodium chloride solution A.4.1.2. 10g of sodium chloride was dissolved in 100mL of water. A.4.1.3 standard component to be measured. n-hexane, methanol, HPLC grade. A.4.1.4 internal standard solution. 160μg/g n-propanol solution. A.4.2 Instruments and Equipment Gas chromatograph. with a flame ionization detector (FID) and a headspace sampler. A.4.3 Reference chromatographic conditions A.4.3.1 Column. Shi Ying capillary column (φ0.15μm × 15m), the coating is 6% and 94% cyanopropyl phenyl dimethylpolysiloxane, coated Layer thickness was 0.84μm. Column or the same performance. A.4.3.2 carrier gas. helium. A.4.3.3 carrier gas flow rate. 0.8mL/min. A.4.3.4 Column temperature. 40 ℃ maintained 5min, at 25 ℃/min was heated to 250 ℃, total run time of 13.4min. A.4.3.5 Inlet temperature. 250 ℃. A.4.3.6 Detector temperature. 300 ℃. A.4.3.7 injection volume. 1.0mL. A.4.4 Reference Headspace Conditions A.4.4.1 Sample heating temperature. 90 ℃. A.4.4.2 sample heating time. 10min. A.4.4.3 sample stirring rotational speed. 400r/min. A.4.4.4 Split ratio. 1.50 A.4.4.5 Injector temperature. 120 ℃. A.4.4.6 injection speed. 1mL/s A.4.5 Analysis step Preparation of standard stock solution A.4.5.1 Component to be measured with a standard (n-hexane, methanol), standard stock solution 260μg/g were prepared. A.4.5.2 Preparation of standard solution series Table A.2 sunflower oil weighed, added to standard stock solution (separately analyzed for each solvent), sodium chloride solution and internal standard solution, prepared standard Series of standard solutions were injected into 20mL sample vial. Table A.2 Preparation of the standard solution series Sunflower oil/mg standard stock solution/mg sodium chloride solution/mg internal standard solution/mg 2505027001000 25010026501000 25020025501000 25050022501000 250,100,017,501,000 Preparation of sample solution A.4.5.3 0.25g sample was weighed (accurate to 0.0001g), was added sodium chloride solution and internal standard solution 2750mg 1000mg, injection into 20mL Sample bottle. A.4.5.4 Determination A.4.3 and A.4.4 in the reference operation conditions, respectively, of the sample solution and the standard solution series chromatographed. A.4.6 calculation results A.4.6.1 average response factor f The average response factor of standard solution f, according to formula (A.5) Calculated. f = cs ci × Ri Rs (A.5) Where. --- CS concentration of each standard solution component to be measured in micrograms per gram (μg/g); --- CI concentration of each standard solution in n-propanol (internal standard), expressed in micrograms per gram (μg/g); --- RI peak area of each standard solution chromatogram propanol (internal standard); and RS --- peak area of each component to be measured in the standard solution chromatogram; 5 --- Number of standard solution series. A.4.6.2 component content measured wi Component content measured wi, in milligrams per kilogram (mg/kg), calculated according to formula (A.6). wi = Ru × c0 × f R0 × cu (A.6) Where. Ru --- peak area component measured chromatogram of the sample solution; cO propanol --- sample solution (internal standard) concentration, in micrograms per gram (μg/g); f --- average response factor; --- R0 peak area in the chromatogram of the sample solution propanol (internal standard); and --- Cu sample solution of rosemary extract concentration, in micrograms per gram (μg/g). By the formula (A.6) calculated component to be measured (n-hexane and methanol) content of w3 and w4, and is the sum of the two residual solvent content in the sample. The test result to the arithmetic mean of replicates results. Obtained in two independent determination results under repeatability conditions of absolute difference Value is not more than 5% of the arithmetic mean.

Appendix B

Rosemary extract thereof with reference to chromatograms Rosemary extract thereof with reference to the chromatogram shown in Figure B.1. Description. 1 --- solvents; 2 --- carnosol; 3 --- Sage acid. Figure B.1 rosemary extract reference schematic chromatography

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