GH/T 1280-2019 in English
VALIDDetermination of cafferic acid, p-coumaric acid, fernlic acid, pinobanksin,pinocembrin, 3-O-acetylpinobanksin, chrysin and galangin inpropolis-Reversed-phase high performance liquid chromatography method
- Issued on:2019-11-28
- Implemented on:2020-03-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
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| Standard No: | GH/T 1280-2019 |
| Document status: | VALID |
| Title in English: | Determination of cafferic acid, p-coumaric acid, fernlic acid, pinobanksin,pinocembrin, 3-O-acetylpinobanksin, chrysin and galangin inpropolis-Reversed-phase high performance liquid chromatography method |
| Title in Chinese: | 蜂胶中咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素3-乙酸酯、白杨素和高良姜素含量的测定反相高效液相色谱法 |
| Language: | English |
| File Format: | Electronic (PDF) |
| Delivery: | Via email within 1~3 business days |
| Issued on: | 2019-11-28 |
| Implemented on: | 2020-03-01 |
| ICS Classification: | 65.140-Beekeeping |
| Chinese Classification: | B47-Beekeeping, silkworm keeping |
| Related Topics: | caffeic acid
white Galangin Prime+ Determination of Coumarin by HPLC GH 2 gh/t Folic Acid Biphasic liquid acid liquid + acid gh/t1247-2019 The content of hydrocortisone acetate was determined by high performance liquid chromatography. Folic acid liquid phase detection method Liquid phase method for determination of tetracycline hydrochloride Method for measuring salicylic acid content in leaves gh/t1243-2019 gh/t 1161 |
本标准规定了蜂胶中咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素 3-乙酸酯、白杨素和高良姜素的反相高效液相色谱检测方法。
本标准适用于蜂胶及蜂胶乙醇提取物中咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素-3-O-乙酸酯、白杨素和高良姜素的测定。
本标准液相色谱的检出限：咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素 3-乙酸酯、白杨素和高良姜素分别为 0.41、0.39、1.20、0.29、1.11、1.49、0.86、0.91μg/mL。
本标准液相色谱的定量限：咖啡酸、p-香豆酸、阿魏酸、短叶松素、松属素、短叶松素 3-乙酸酯、白杨素和高良姜素分别为 1.35、2.16、4.00、0.98、3.69、4.97、2.88、3.03μg/mL。
Introduction
Technical background and significance of the standard
This standard was proposed by the All-China Federation of Supply and Marketing Cooperatives and managed by the National Bee Products Standardization Working Group. It established the quantitative detection method for 8 characteristic active ingredients in propolis. As the core indicators for the quality evaluation of propolis, the content of these phenolic acids and flavonoids directly reflects the antioxidant, antibacterial and other biological activities of propolis.
Principle and technical characteristics of the method
A reverse phase high performance liquid chromatography (RP-HPLC) combined with a UV detector was used for detection at a wavelength of 280nm. The method innovatively achieved baseline separation of 8 compounds with large polarity differences through a gradient elution program (see Table 1).
| Compound Type | Representative Substances | Detection Limit (μg/mL) | Quantification Limit (μg/mL) |
|---|---|---|---|
| Phenolic Acids | Caffeic Acid, Ferulic Acid | 0.39-1.20 | 1.35-4.00 |
| Flavonoids | Pinusin, Galangin | 0.29-1.49 | 0.98-4.97 |
Key Operation Points
Sample pretreatment
Use three-stage ethanol ultrasonic extraction (15min/time) combined with rotary evaporation concentration technology to ensure the complete extraction of active ingredients. Special attention:
- Fixed volume (V1/V2/V3) needs to be precisely controlled
- Use 0.45μm organic filter membrane for filtration
Chromatographic condition optimization
It is recommended to use Sepax HP-C18 chromatographic column (150×4.6mm, 5μm), and the column temperature is controlled at 33±1℃. The mobile phase uses methanol-1% acetic acid system, and the gradient program in Appendix A is used for elution.
Implementation suggestions
Quality control measures
- Standard working solution must be prepared and used immediately
- Each batch of samples should be calibrated with standard samples
- Relative deviation of parallel samples ≤2.0%
Extended application of the method
This method can be applied to the following after appropriate adjustment:
- Quality control of propolis ethanol extract
- Identification of propolis from different plant sources
- Monitoring of composition changes of propolis during storage
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