HPLC法测定厚朴中的厚朴酚(Magnolol)和和厚朴酚(Honokiol)含量--上禾生物

1)试剂

   a. 重蒸水                 b. 甲醇（色谱纯）   

2)仪器

   高效液相色谱仪（Shimadzu）                 分析天平（1/10000）

   超声波清洗仪                               0.45mm微孔过滤器

   50ml容量瓶                                带塞小玻璃瓶

3)色谱条件

   色谱柱   Kromasil   C18  5mm  4.6×200mm

   波长     294nm

   流动相   A: 水 (20%)        B: 甲醇(80%)

   进样量   10ml

   流速     1.0ml/min

4)样品的制备

（1）待测样品：精确称取30mg提取物于50ml容量瓶中，用甲醇溶解，超声定容，0.45mm微孔滤膜过滤即为供试液。

（2）标准样品:  精确称取厚朴酚、和厚朴酚标准品用甲醇溶解,使其浓度为0.2mg/ml, 0.45mm微孔滤膜过滤。

5)计算

根据各峰峰面积利用外标法计算各种成份的含量。

Determination of Magnolol and Honokiol by HPLC

1) Reagents

  a. Nanopure water          b. Methanol (HPLC)

2) Apparatus

HPLC (Shimadzu)           Analytical balance (1/10000)            Ultrasonic cleaner

0.45μm PTFE filter          50ml volumetric flask                  Screw cap vials

3) HPLC condition

Column: Kromasil   C18  5mm  4.6×200mm

Detector:  UV 294nm

Mobile Phase: A= water  B= MeOH

A:B=20:80(Isocratic)     

Flow Rate: 1.0ml/min

Injection: 10μl

4) Procedure

Samples Preparation

  Accurately weigh 30mg extraction into 50ml volumetric flask. Add MeOH 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 0.2mg/ml in MeOH) using a standard of Magnolol，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 Maglolol on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard solution. Calculate the weight percent.