Study On The Extraction,Separation And Analysis Of Phospholipids In Radix Polygoni Multiflori

Radix Polygoni Multiflori (RPM) is dry earthnut of Polygonum Multiflorum Thunb. It is a Chinese herbal restorative medicine, which is used for cure the graying of beard and hair, strengthening of bones, bowel movement etc. It has highly effective for people suffering from high cholesterol and constipation. In recent years of studies, Radix Polygoni Multiflori is found to contain many chemical constituents such as anthraquinones, stilbenes, phospholipids and etc. Phospholipid is one of the many efficiency components in RPM. It can prevent liver loss of function and aids in cardiovascular system. It has many hygienical and medicinal functions. So study on the extraction, separation and analysis method of Phospholipids in RPM has important applied values and academic senses in exploitation and using this resource. Referred to a lot of literatures, in this paper using Ultrasonicator Extraction and Microwave Assisted Extraction techniques with Inductively Coupled Plasma Atomic Emission Spectrum (ICP-AES) to study the extraction conditions of Phospholipids and found a new analysis method of Phospholipids in RPM. In this experiment, we have done a research on the Phospholipids in RPM. On a base of Thin Layer Chromatography, it established the structure of fatty acids by gas chromatography-mass spectrum. The main results of study are the following:1. The method of Ultrasonic extraction was applied to extract the Phospholipids from Chinese traditional medicine RPM. The effects of solvent volume, extraction time, power of Ultrasonic Extraction on Phospholipids extraction were investigated. The content of Phospholipids in extracted compounds was determined by ICP-AES. The result indicated that the optimal extraction conditions for 1.5g RPM were 20ml Folch solvent, time 35min and power 24 kHz.2. Found a new Microwave assisted extraction method for the Phospholipids of RPM. The effects of solvent, proportion of solid to liquid, extraction time and the temperature were investigated. The experiments select chloroform-methanol (1:2,V/V) as the extraction solvent, the proportion of solid to liquid was 1:20, extraction time was 15min and the temperature was set at 50℃.3. A high performance liquid chromatography method was found to determine the Phospholipids in RPM. Separation was performed on a Nuclesil 100-5 column (250mm×4.6mm i.d., 5μm), using methanol as mobile phase at a flow rate of 0.8 mL／min. The carrier gas flow rate and drift tube temperature of evaporative light scattering detection were set at 1.8 L/min and 66℃, separately. The injecting volume was 15μL. The linear ranges for the concentration of Phospholipids were Phosphatidylglycerol (PG) 0.0388-0.388mg/mL (r=0.9991), Phosphatidylethanolamine (PE) 0.0500-0.250mg/mL (r=0.9996), Phosphatidylcholine (PC) 0.0103-0.165mg/mL (r=0.9995), Lysophosphatidylcholine (LPC) 0.0103-0.343mg/mL (r=0.9995). The proposed method was successfully applied to the determination of Phospholipids in RPM samples.4. Studied the separation conditions of Phospholipid on Thin Layer Chromatography. For Phosphatidylglycerol (PG), Phosphatidylethanolamine (PE), Phosphatidylcholine (PC), Lysophosphatidylcholine (LPC), Phosphatidylinositol (PI), Phosphatidylserine (PS), it selects chloroform-methanol-acetic acid (5:2:1.5, V/V/V) to spread 8cm on silica gel G board for once. It found PE in RPM by separation conditions. Using GC-MS to determine the structure of PE, the result indicated that the side chains of PE contain kittul acid, oleic acid, suboleic acid and stearic acid in RPM.