| Pine pollen contains rich nutrient components, and it has the characteristics that canregulate the function of the human body with multi-components, multiple target points,and through multiple pathways. Many experts and departments have carried out a greatdeal of research on pine pollen. At present, the development of health food direction istransferring to the third generation product containing clear functional factors. Studies onthe functional factors have been paid much attention to both at home and abroad.Polyunsaturated fatty acids, saponins and trace elements are important functional factors.Based on single species of pine pollen and methodology research, we usedspectrophotometric method, GC-MS and TXRF detection, traditional Chinese medicinefingerprint similarity calculation software to analyze test results, in order to get thesimilarities and differences in the efficacy factors of different varieties of pine pollen. Inaddition, we made a preliminary study on pine pollen from different angles, such as thefour properties of Chinese traditional medicine, efficacy and material basis.This paper consists of four parts.The first part briefly introduced the research status on the composition, pharmacologyand efficacy of pine pollen; reviewed the detection means and the research significance offatty acids, flavonoids and saponins in pine pollen; described the advantage ofsimultaneous determination of multiple elements with TXRF.In the second part, a GC-MS method for the determination of methylesters of fattyacids in the pone pollen, after the soxhlet extraction process, was established. The contentsof12pieces of stress fatty acids in the pine pollen, which collected from different pinespecies, were determined and contrasted. The results showed that the GC-MS method is afast and effective one for the determination of fatty acids in pine pollen. The results alsoshowed that there was a significant difference of10pieces of fatty acids in the5pieces ofpine pollen, except the palmitic acid and heptadecanoic acid(P≤0.05), and there was a verysignificant difference of saturated fatty acid or unsaturated fatty acid in them(P≤0.01).The third part: the total saponins in pine pollen of different tree species and from different habitats was studied by the spectrophotometric method. By orthogonalexperiments and confirmatory experiments, the best conditional combination of extractingand determination of the total saponins in pine pollen was solid-to-liquid ratio(1:20),ultrasound extraction time(60min), number of ultrasonic times(twice), developingtemperature(60℃), developing time(15min). It was found from range R that factorsaffecting extracting and determination of the total saponins were number of ultrasonictimes, ultrasound extraction time, solid-to-liquid ratio, developing temperature anddeveloping time in turn. The research results showed that there were significantdifferences between total saponins in wall-broken pollen of masson pine and that inwall-unbroken pine pollen. The total saponin of the broken pine pollen was more thanthree times higher than that of unbroken pine pollen. Total saponins content is different inthe pollen of masson pine from different habitats.The results showed that there is a significant difference of the total saponins amongpollen of Pinus massoniana from different geographical areas (P≤0.01). The order of thetotal saponins contents in them from high to low is Shandong, Jiangxi, Anhui, Chongqing,Shanxi. And there is also a significant difference of it among different varieties pine pollenfrom Shandong (P≤0.01). The order of it from high to low is Pinus massoniana, Japanesered pine, oil pine, Japanese black pine.The last part：A method using microwave digestion and TXRF for the determinationof15life elements （K、Ca、Cr、Mn、Fe、Co、Ni、Cu、Zn、Rb、Sr、Sn、Ti、Cdand Pb） in the pine pollen has been developed.（All the results of Cd in this thesis wereobtained by ICP-MS, because of the limiting factor of TXRF.）The pine pollen wasdigested by microwave digestion. The counts, QXAS software and internal standardmethod were used for TXRF measurement. Under optimized conditions，the relativestandard deviations （RSDs） were lower than6.1%（reagent blank, n=5），and thestandard added recoveries were94.3%~102.4%．Quantification limits were from0.003mg/kg（Ni, Cu, Zn or Rb） to0.120mg/kg（K）．After microwave digestion,15elementsin pine pollen were simultaneously determined by TXRF. The results show that all the15elements were founded in all pine pollen. There was a significant difference among the average content of the element such as Ca, Mn, Zn, Rb and Ti in different group of pinepollen (P≤0.01). There was a difference between the average content of the element suchas K, Co, Fe, Cu, Sr, Sn and Cd in them (≤P0.05). And there was no difference betweenthe average content of the element such as Cr, Ni and Pb in them.The results also showed that pine pollen has the spectral characteristics of warmproperty or cold property drug. There were closely related to the tree species and thegrowth environment or the growth area. |
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