GHT1280-2019 English PDF
Basic dataStandard ID: GH/T 1280-2019 (GH/T1280-2019)Description (Translated English): Determination of cafferic acid,p-coumaric acid,ferulic acid,pinobanksin,pinocembrin,3-O-acetylpinobanksin,chrysin and galangin in propolis-Reversed-phase high performance liquid chromatography method Sector / Industry: Supply and Marketing Cooperatives Industry Standard (Recommended) Classification of Chinese Standard: B47 Date of Issue: 2019-11-28 Date of Implementation: 2020-03-01 Regulation (derived from): All China Supply and Marketing Cooperative Announcement 2019 No. 4 Issuing agency(ies): All-China Federation of Supply and Marketing Cooperatives GHT1280-2019: Determination of cafferic acid,p-coumaric acid,ferulic acid,pinobanksin,pinocembrin,3-O-acetylpinobanksin,chrysin and galangin in propolis-Reversed-phase high performance liquid chromatography method---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. (Determination of caffeic acid, p-coumaric acid, ferulic acid, luteolin, pinusin, luteolin 3-acetate, salicin and galangin in propolis.Reversed-phase high-performance liquid chromatography) ICS 65.140 B 47 People's Republic of China Supply and Marketing Cooperation Industry Standard Caffeic acid, p-coumaric acid, ferulic acid, pine leaf in propolis Leptin, Pinusin, Brassinolide 3-acetate, Aspen And galangin content determination Reversed-phase high performance liquid chromatography Determination of cafferic acid, p-coumaric acid, ferulic acid, pinobanksin, pinocembrin, 3-O-acetylpinobanksin, chrysin and galangin in propolis- Reversed-phase high performance liquid chromatography method 2019-11-28 Release 2020-03-01 Implementation Published by All China Supply and Marketing Cooperative ForewordThis standard complies with the drafting rules of GB/T 1.1-2009 "Guidelines for Standardization Work Part 1. Structure and Compilation of Standards". This standard was proposed by the All-China Supply and Marketing Cooperative. This standard is under the jurisdiction of the National Bee Product Standardization Working Group. This standard was drafted. Zhejiang University, Hangzhou Fengzhiyu Bee Industry Co., Ltd., Hangzhou Tianzhu Miyuan Health Products Co., Ltd., Jinyun Green Beekeeping Professional cooperatives. The main drafters of this standard. Hu Fuliang, Zhang Cuiping, Zhou Ping, Wang Lei, Hu Yuanqiang. Caffeic acid, p-coumaric acid, ferulic acid, pleurolidin, pinusin, Determination of phytoplanktin 3-acetate, chrysin and galangin Reversed-phase high performance liquid chromatography1 ScopeThis standard specifies caffeic acid, p-coumaric acid, ferulic acid, phloxanthin, pinusin, pterin 3-acetate, and astaxanthin in propolis. And Galangin by RP-HPLC Method. This standard applies to caffeic acid, p-coumaric acid, ferulic acid, phloxanthin, pinusin, pterodoxin in propolis and ethanol extracts of propolis. Determination of 3-O-acetate, chrysin and galangin. Detection limits of liquid chromatography in this standard. caffeic acid, p-coumaric acid, ferulic acid, phloxanthin, pinusin, phloxanthin 3-acetate, Chrysin and galangin are 0.41, 0.39, 1.20, 0.29, 1.11, 1.49, 0.86, 0.91 μg/mL. Limits of quantification for liquid chromatography in this standard. caffeic acid, p-coumaric acid, ferulic acid, phloxanthin, pinusin, phloxanthin 3-acetate, Chrysin and galangin are 1.35, 2.16, 4.00, 0.98, 3.69, 4.97, 2.88, 3.03 μg/mL, respectively.2 Normative referencesThe following documents are essential for the application of this document. For dated references, only the dated version applies to this document. For undated references, the latest version (including all amendments) applies to this document. GB/T 6682 Analytical laboratory water specifications and test methods3 terms and definitionsThe following terms and definitions apply to this standard. 3.1 Propolis propolis The worker bee collects adhesive substances formed by mixing plant secretions such as resin with secretions from the maxillary gland and wax gland.4 PrincipleThe sample was extracted with ethanol, concentrated, and then dissolved in methanol. The sample was detected at 280 nm using a reverse-phase high performance liquid chromatography UV detector. Standard quantification.5 Reagents and materialsLaboratory water shall comply with the primary water specified in GB/T 6682. Unless otherwise specified, all reagents are of analytical grade. 5.1 Methanol. chromatographically pure. 5.2 Ethanol. 5.3 Acetic acid, chromatographically pure. 5.4 Mobile phase. gradient elution with methanol (5.1) and 1% acetic acid (5.3). 5.5 Caffeic acid, p-coumaric acid, ferulic acid, luteolin, pinusin, luteolin 3-acetate, chrysin and galangin standards. Purity ≥98% 5.6 Standard reserves of caffeic acid, p-coumaric acid, ferulic acid, luteolin, pinusin, luteolin 3-acetate, chrysin and galangin Solution. Accurately weigh an appropriate amount of each standard substance (5.5), and prepare a 1.0 mg/mL standard stock solution with a methanol solution (5.1). Store at 4 ° C for 6 months. 5.7 Standard work for caffeic acid, p-coumaric acid, ferulic acid, luteolin, pinusin, luteolin 3-acetate, chrysin and galangin Solution. Pipette an appropriate amount of standard stock solution (5.6) at a concentration of 1.0 mg/mL, and dilute with methanol solution (5.1) to the appropriate concentration for standard work. Solution, ready to use. 5.8 Filter. 0.45 µm, organic.6 Instruments and equipment6.1 HPLC. equipped with UV detector. 6.2 Crusher. 6.3 Rotary evaporator. 6.4 Ultrasonic cleaner.7 Sample preparation and storage7.1 Preparation of test specimens Take an appropriate amount of a representative sample, freeze, pulverize, and mix as a sample for inspection. 7.2 Sample preservation The samples were stored at 0 ° C to 4 ° C.8 steps8.1 Propolis sample pretreatment Accurately weigh 10.0 g (accurate to 0.1 g) sample of propolis, and use 30 mL of 95% ethanol for ultrasonic extraction for 15 min each time. Three times, combine the filtrates and make up to 100 mL (V1). 10 mL (V2) was evaporated to dryness under reduced pressure, and 25 mL (V3) of methanol was used for ultrasonic dissolution and determination. Capacity, filtered by a 0.45 μm filter membrane for determination. 8.2 Chromatographic conditions Column. Sepax HP-C18, column length 150 mm, inner diameter 4.6 mm, particle size 5 µm; or equivalent column. Mobile phase flow rate. 1.0 mL/min. Gradient elution. See Appendix A, Table A.1 for elution procedures. Detection wavelength. 280 nm. Column temperature. 33 ° C. Injection volume. 5 µL. 8.3 Determination 5 μL of sample solution (8.1) and standard working solution (5.7) were injected and analyzed separately, a standard curve was drawn and quantified by external standard method. The chromatogram is shown in Appendix A, Figure A.1.9 Results calculation and representation9.1 Calculation of results In the sample, caffeic acid, p-coumaric acid, ferulic acid, phloxanthin, pinus The amount (X) is expressed in milligrams per gram (mg/g) and calculated according to formula (1). C V 3V 2 mV 1 (1) In the formula. The concentration of the compound in the sample solution calculated on the CS-standard working curve, the unit is milligram per milliliter (mg/mL); V1-Constant volume of sample solution in milliliters (mL); V2-the volume to be measured (mL) taken from the sample solution; V3-The final constant volume of the sample to be tested, in milliliters (mL); m-sample mass in grams (g). 9.2 Results Take the arithmetic mean of two independent measured values that meet the repeatability requirements as the measurement result (mg/g), and retain two significant figures. Parallel test The relative deviation of inspection shall not exceed 2.0%. X Appendix A(Informative appendix) HPLC gradient elution procedure and chromatogram Table A.1 High-performance liquid chromatography gradient elution procedure time min Flow rate mL/min Methanol 1% acetic acid Figure A.1 Mixing standards of caffeic acid, p-coumaric acid, ferulic acid, luteolin, pinusin, luteolin 3-acetate, chrysin and galangin And propolis chromatograms Figure 1 HPLC spectrum of mixed standards and propolis A. mixed standard; B. propolis 1.caffeic acid, 2.p-coumaric acid, 3.ferulic acid, 4.porphyrin, 5.pinusin, 6.porphyrin 3-acetate, 7.astaxanthin, 8.galangin ......Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of GHT1280-2019_English be delivered?Answer: Upon your order, we will start to translate GHT1280-2019_English as soon as possible, and keep you informed of the progress. The lead time is typically 1 ~ 3 working days. 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