A New Method For Detemination Of 1-Deoxynojirimycin

The mulberry leaf is the mulberry branch plant mulberry's dry leaf. In all previous dynasties Chinese medicine books records the mulberry leaf to be able to treat any disease with thirst and excessive urination sickness. The modern pharmacology research indicated that 1-Deoxynojirimycin (DNJ) is one characteristic value ingredient (one kind of glycosidase inhibitor), which falls the blood sugar as the main active constituent. Due to its remarkable effect of the cure of the diabetes, antivirus and anticancer, it arouses the attention of many researchers in the world.GC–MS has been the general method for the determination of DNJ by virtue of the high resolution of GC and the additional structural information obtained by MS. Because DNJ was lacks of the chromophore in its molecular structure, so it is very difficult to use the HPLC to determinate DNJ immediately. In order to establish one kind of accurate and reliable determination method for DNJ, this experiment has done the following three work:1: In the research work, we used piperidine and 2,4-Dinitro-1-fluorobenzene in the reaction, thus this study we obtained good results. The result were about the following conclusion: (1) This reaction is easy to carry on, and then completely finished in the normal temperature in 30 minutes. (2) Piperidine-DNP demonstrated that it has intense absorption in 374nm, and its molar extinction coefficientεwas17000, the melting point is 91.5℃-92℃.2: DNJ has extracted from the silkworm.3: A rapid and reliable method suitable for assays of a large number of Morus alba leaves for 1-deoxynojirimycin (DNJ) has been developed. DNJ in 0.2 g of dried leaves was double-extracted in 10 mL of 0.1 % methanol acetic acid, by ultrasound for 5 min at 50°C, derivatized with 2,4-Dinitro-1-fluorobenzene (DNFB). Then DNJ-DNP analyzed by high performance liquid chromatography equipped with ultraviolet detector. Mobile phase consisted of acetonitrile-0. 1% methanol acetic acid (25∶75 V /V )with a flow rate of 1.0 mL/min. The calibration curve was linear over the concentration range from 0.16μg/mL to 8.14μg/ mL, the correlation coefficient was 0. 9994, the limit of detection was 0. 03μg/mL (3σ). The results showed that the content of DNJ in leaves of Morus alba L. was 0. 30 %, the average recovery was 96.8%, RSD was 2.0% ( n = 6). This determination method is simple, fast, and accurate for DNJ. And it has provided the important value for determination of DNJ.