HPLC法測定黃連中的鹽酸小檗堿-上禾生物 | HPLC法測定黃連中的鹽酸小檗堿-上禾生物 |
| 發(fā)布時間:2023-10-08 信息來源:admin 發(fā)布人:admin 點擊次數(shù):279 |
HPLC法測定黃連中的小檗堿-上禾生物 1)試劑 a. 乙腈（分析純） b. 重蒸水 c. 磷酸（分析純） d.十二烷基磺酸鈉（SDS） 2)儀器 高效液相色譜儀（Shimadzu） 分析天平（1/10000） 超聲波清洗儀 0.45 mm 微孔過濾器 50 ml 容量瓶 帶塞小玻璃瓶 3)色譜條件 色譜柱 Hypersil ODS C18 5 mm 4.6´200mm 波長 282nm 流動相 A:水（含0.02%mol/L磷酸、0.2%十二烷基磺酸鈉） B: 乙腈 A:B=52:48(等梯度) 流速 0.8ml/min 進樣量 10 mm 4)樣品制備 （1）待測樣品 精確稱取30mg提取物于50ml容量瓶中，用0.1mol/L磷酸超聲溶解，冷卻定容，0.45 mm微孔濾膜過濾為供試液。 （2）標樣制備 精密稱取標樣用0.1mol/L磷酸溶解，使其濃度為0.15mg/ml，過0.45mm濾膜，置冰箱貯存。 5)計算 根據(jù)各峰峰面積利用外標法計算各種成份的含量。
鹽酸小檗堿生產(chǎn)企業(yè)-上禾生物
（27） Determination of Berberine by HPLC
1) Reagents a. Acetronitrile (HPLC) b. Nanopure water c.SDS d.Phosphate acid(A.R) 2) Apparatus HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner 0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition Column: Hypersil ODS C18 5μm , 4.6×200mm Detector: UV 282nm Mobile Phase: A=water ( including 0.02ml/L phosphate acid and 0.2% SDS) B= Acetronitrile A/B=52/48 Flow Rate: 0.8ml/min Injection: 10μl 4) Procedure Samples Preparation Accurately weigh 30mg extraction into 50ml volumetric flask. Add 0.1%mol/L Phosphate acid to volume and sonicate to mix, filter through a 0.45μm PTFE filter into a screw cap vial and proceed with the analysis. Standard preparation /calibration Prepare a standard solution (approximately 0.6mg/ml in 0.1%mol/L Phosphate acid) using a commercial reference standard of berberine，Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
5) Calculation Determine the peak areas of the berberine on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the reference standard solution. Calculate the weight percent.
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