Supercritical CO2 Extraction of Volatile Oil in Pine Needle of Pinus yunnanensis from Different Habitats and Analysis by GC-MS / 中国实验方剂学杂志

Objective:To analyze and compare the volatile oil components of pine needle of Pinus yunnanensis from different habitats. Method:The volatile oils of pine needle of P. yunnanensis from 12 different habitats were extracted by supercritical CO2 extraction （SFE-CO2），the components were analyzed by gas chromatography-mass spectrometry （GC-MS），and relative percentage of each component was determined by peak area normalization method，and SPSS 20.0 software was used for principal component analysis. Result:The 137 compounds were isolated from the volatile oil of pine needle of P. yunnanensis from 12 different producing areas，with 1 sample identified 57 species，2 samples identified 57 species，3 samples identified 54 species，4 samples identified 48 species，5 samples to identify 58 species，6 samples identified 60 species，7 samples identified 53 species，8 samples identified 55 species，9 samples identified 56 species，10 samples to identify 58 species，11 samples identified 54 species and 12 samples to identify 50 species，composed of hydrocarbon，acid，alcohol，ester，aldehyde，ketone，phenol，ether 8 compounds，including 11 common composition including alpha pinene，beta pinene，beta caryophyllene，7C，10C，16C-hexadecatrienoic acid and palmitic acid methyl ester，palmitic acid，phthalic acid dibutyl do acid，alcohol，stearic acid，mountain，wood tar acid. Conclusion:In this study，supercritical CO2 extraction （SFE-CO2） was used to extract the volatile oil of pine needle of P. yunnanensis from different habitats. It was found that the volatile oil of pine needle of P. yunnanensis was mainly composed of hydrocarbons，acids，esters and alcohols. And the content and species of volatile oil of pine needle of P. yunnanensis from different producing areas differ greatly which lay a foundation for the further development and utilization of pine needle of P. yunnanensis resources.

Effect of Sulgidduk containing pine needle juice on lipid metabolism in high fat-cholesterol diet induced dyslipidemic rats / 한국영양학회지

PURPOSE: Dyslipidemia is a major risk factor for cardiovascular disease. Pine needles (Pinus densiflora seib et Zucc) are a traditional medicine used to treat dyslipidemia in clinical settings. This study examined the potential effects of sulgidduk, a Korean traditional rice cake containing pine needle juice to protect against dyslipidemia induced by a high-fat/sugidduk diet in a rat model. METHODS: Twenty one male Sprague-Dawley rats were divided randomly into three groups: normal control (NC), Sulgidduk diet (SD), Sulgidduk diet containing pine needle juice (PSD). The blood lipid levels, production of lipid peroxide in the plasma and liver, total cholesterol and triglyceride in the liver and feces, antioxidant enzyme activities in plasma and erythrocytes were measured to assess the effects of PSD on dyslipidemia. RESULTS: A high-fat/Sulgidduk diet induced dyslipidemia, which was characterized by significantly altered lipid profiles in the plasma and liver. The food intake was similar in the three groups, but weight gain and food efficiency ratio (FER) were reduced significantly in the PSD group compared to those in the SD group. The level of total cholesterol, LDL-cholesterol and TBARS in the plasma showed tendencies to decrease in the PSD group compared to those in the SD group. The levels of high-fat/Sulgidduk diet-induced sterol regulatory element-binding protein 2 (SREBP2) gene expression were reduced significantly in the PSD group. The supplementation of PSD reduced the hepatic triglyceride and total cholesterol levels significantly, and enhanced the fecal excretion of triglyceride and hepatic antioxidant enzyme activities compared to the SD group. CONCLUSION: These results suggest that the addition of 0.4% pine needle juice to Sulgidduk may be an alternative snack to control dyslipidemia.

Extraction technology optimization of total flavonoids in Wudang pine needle tea by orthogonal test / 国际中医中药杂志

Objective To optimize the best extraction technology of total flavonoids in Wudang pine needle tea.Methods The contents of total flavonoids were taken as index, and orthogonal design of L9(34) was applied to process ultrasonic extraction.Results The order of factors of affecting extraction technology was solid to liquid ratio > ultrasonic power > ultrasonic time. The optimized extraction technology was as follows: adding 500 times volume of purified water, ultrasonic extracting for 15 minutes with ultrasound power 500 W at 40℃. The average content of total flavonoids was 39.701 mg/g.Conclusions The optimal extraction technology is simple, efficient and feasible, and can be used for extracting total flavonoids from Wudang pine needle tea, providing basis for the formulation of quality standards.

What deters plant colonization in a tropical pine plantation? / ¿Cuáles son los factores que limitan la colonización vegetal en una plantación de pino tropical?

AbstractPine plantations in the tropics are often employed to recondition eroded slopes from mudslides, as the Pinus caribaea plantation that shields the Universidad Simón Bolívar campus in Caracas (Venezuela). However, mismanagement of this plantation has led to its rapid degradation. The best option to maintain the protective service is to restore the plantation and direct its successional trajectory towards the neighbouring montane forest. Through experimental manipulation, we aimed to determine which factors block secondary succession and to investigate their effects. Within the experimental constraints imposed by the plantation small area, we analysed the effects of light and fertility limitation, litter accumulation and access to seed on plantation restoration. Light availability was manipulated by clearing and thinning three 800 m2 main plots. Fertilization and litter removal was applied to sub-plots within the light plots. Soils were analysed, microclimate was monitored and, for four years, stem density, species richness and basal area were tallied. Our results showed that light accessibility was the main factor deterring the successional trajectory of the plots, with varying grades of interaction with the sub-treatments. By the end of the fourth year, the cleared plot showed the largest responses in all traits (triplicating stem density and basal area and >20 times higher species richness). The main colonizers were Croton megalodendron, Ocoteafendleri, and Clusia spp. all dominant trees in the nearby native forest. We concluded that the results of this pioneer study, showed that small clearings, repeated in 3-4 year cycles are appropriate for similar restoration schemes. This procedure would create a mosaic of vegetation patches at different successional stages while protecting the slopes from erosion and increasing local biodiversity. Rev. Biol. Trop. 64 (2): 461-471. Epub 2016 June 01.

ResumenEn los trópicos, las plantaciones de pino se emplean comúnmente para proteger laderas erosionadas. Este es el caso de la plantación de Pinus caribaea que resguarda el campus de la Universidad Simón Bolívar en Caracas (Venezuela) de los deslaves de lodo. Sin embargo, el inadecuado manejo de la plantación está conduciendo a su rápido deterioro. La opción más adecuada para mantener el papel protector de la plantación es la restauración dirigida hacia una sucesión similar a la del bosque montano vecino. Mediante manipulaciones experimentales nos proponemos determinar cuáles son los factores que bloquean la sucesión secundaria e investigar sus efectos específicos. Dentro de las limitaciones impuestas por el reducido tamaño de la plantación y su rol protector, aquí analizamos los efectos de la reducida radiación solar y fertilidad del suelo, la acumulación de acículas de pino y el acceso a la fuente de semillas para la restauración. La disponibilidad de radiación solar se varió cortando y entresacando los pinos de parcelas de 800 m2. Los tratamientos de fertilidad y la remoción de acículas (por fuego y manualmente) se realizaron en subparcelas replicadas dentro de las parcelas principales. Los suelos se analizaron fisicoquímicamente, se monitoreo el microclima y, durante 4 años, se censó la densidad, el área basal y la riqueza de especies de los elementos leñosos en las parcelas. Los resultados muestran que el acceso a la radiación solar fue el factor principal que influyó sobre la colonización y crecimiento de nuevos individuos en las parcelas con diversos grados de interacción con los sub-tratamientos. Al finalizar el cuarto año, la parcela totalmente deforestada mostró las respuestas más elevadas (tres veces superior en cuanto a densidad y área basal y hasta veinte veces mayor en cuanto a riqueza de especies) comparada con las parcelas control y parcialmente deforestada. Las principales especies leñosas colonizadoras fueron: Croton megalodendron, Ocotea fendleri y Clusia spp., todas ellas dominantes en el bosque montano vecino. Concluimos que este estudio pionero muestra que el aclareo total de parcelas pequeñas, repetido en ciclos de 3-4 años es apropiado para proyectos de restauración similares. Así se obtendría un mosaico de vegetación en diferentes estadios sucesionales que mantienen el papel protector de la cubierta vegetal e incrementando la biodiversidad local.

Antioxidant activity and analysis of proanthocyanidins from pine (Pinus densiflora) needles

In this study, we evaluated the antioxidant activity of pine needle extracts prepared with hot water, ethanol, hexane, hot water-hexane (HWH), and hot water-ethanol (HWE), using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical method. The hot water extract possessed superior antioxidant activity than the other extracts. We also compared the antioxidant activity of pine needle extracts through ROS inhibition activity in a cellular system using MC3T3 E-1 cells. The hot water extract exhibited the lowest ROS production. The pattern of HPLC analysis of each extract indicated that the hot water extract contained the highest proanthocyanidin level. The pine needle hot-water extract was then isolated and fractionated with Sephadex LH-20 column chromatography to determine the major contributor to its antioxidant activity. The No.7 and 12 fractions had high antioxidant activities, that is, the highest contents of proanthocyanidins and catechins, respectively. These results indicate that the antioxidant activity of procyanidins from the hot water extract of pine needles is positively related to not only polymeric proanthocyanidins but also to monomeric catechins. Moreover, the antioxidant activity of the pine needle hot water extract was similar to well-known antioxidants, such as vitamin C. This suggests that pine needle proanthocyanidins and catechins might be of interest for use as alternative antioxidants.

Comparison of methods for proanthocyanidin extraction from pine (Pinus densiflora) needles and biological activities of the extracts

Flavonoids are known to be effective scavengers of free radicals. In particular, proanthocyanidins are flavonoids that possess cardiovascular protection, antioxidative activities, and immunomodulatory activities. Here, we evaluated proanthocyanidin contents in the total polyphenolic compounds of pine needle extracts prepared by hot water, ethanol, hexane, hot water-hexane (HWH), and hot water-ethanol (HWE). Analysis of each extract indicated that the ethanol extract contained the highest proanthocyanidin concentration. The HWH and hexane extracts also contained relatively high concentrations of proanthocyanidin. On the other hand, proanthocyanidin content analyses out of the total polyphenolic compounds indicated that the HWH extract contained the highest content. These results suggest that HWH extraction is a suitable method to obtain an extract with a high level of pure proanthocyanidins and a relatively high yield. The HWH extract possessed superior activity in diverse antioxidative analyses such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferrous ion chelating (FIC), and ferric-ion reducing power (FRAP) assays. In addition, upon assessing the effects of the pine needle extracts on macrophages (Raw 264.7 cell), the HWH extract exhibited the highest activity. In this study, we discerned an efficient extraction method to achieve relatively pure proanthocyanidins from pine needles and evaluated the biological functions of the resulting extract, which could potentially be used for its efficacious components in functional food products.

Furfural from Pine Needle Extract Inhibits the Growth of a Plant Pathogenic Fungus, Alternaria mali

The antifungal effect of pine needle extract prepared by a distinguishable extraction method and the dry distillation method, was examined. The effect of this extract itself was insignificant. The chemical components of pine needle extract were then investigated by gas chromatographic analysis, and four chemical components, acetol, furfural, 5-methyl furfural, and terpine-4-ol, were identified. The antifungal effects of those four chemical components against Alternaria mali (A. mali), an agent of Alternaria blotch of apple, were then examined. It was observed that the minimum inhibitory concentrations (MICs) were 6.25, 0.78, 0.78, and 12.5 (mg/ml) of acetol, furfural, 5-methyl furfural, and terpine-4-ol, respectively. MICs of furfural and 5-methyl furfural had the same order of magnitude as that of an antifungal agrochemical, chlorothalonil. Although furfural itself can not be completely substituted for an antifungal agrochemical, a partial mixture of furfural and antifungal agrochemical may be used as a substitute. The use of agrochemicals for the prevention of plant disease caused by pathogenic fungus such as A. mali could be partially reduced by the application of this mixture.