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Determination and Analysis of Alkaloid Content in Aconitum kusnezoffii Seeds from Different Areas

SHAN Dan, QU Lu-Lu, LI Xiao-Jie, MA Yi-Ming, WANG Jun-Jie

Bulletin of Botanical Research 2020, 40 (5): 768-774. DOI: 10.7525/j.issn.1673-5102.2020.05.016

Abstract （1101）

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The experiment was conducted to determine the content of alkaloid in the seed of Aconitum kusnezoffii Reichb. from different habitats, and to compare its alkaloid content with its content in root and leaf. High pressure liquid chromatography(HPLC) was used to determine alkaloid content in seed of wild A.kusnezoffii collected from Zhalute of Tongliao City, Aru Kerqin of Chifeng City, and Bahrain of Chifeng City. The HPLC column used was Supersil ODS2 5 μm(250 mm×4.6 mm, 5 μm). The HPLC mobile phase was 0.1 mol·L ^-1 methanol and triethylamine(V:V 75:25) and the flow rate was 0.8 mL·min ^-1 under the detection wavelength of 235 nm with column temperature as 35℃. Two types of alkaloid were detected in the seed of A.kusnezoffii and the content of benzoyl aconitine was the highest while the content of diester alkaloid was comparatively lower. Total alkaloid content in seed content from Zhalute of Tongliao City, Alu Kerqin of Chifeng City, and Balin of Chifeng City was 7.647 9, 8.256 3 and 7.512 6 mg·g ^-1, respectively. The total A.kusnezoffii seed alkaloid content collected from Tongliao was significantly higher than that from Chifeng City. The content of monoester alkaloid in A.kusnezoffii seed was significantly higher than diester alkaloid, and can be used for the comprehensive utilization, screening, and breeding of A.kusnezoffii germplasm with different seed alkaloid content.

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Optimization of Ultrasound-assisted Extraction of Dihydromyricetin from Seeds of Hovenia dulcis Thunb. Using Response Surface Methodology

ZHANG Xiao-Nan, ZHU Hong-Wei, ZHAO Shan-Bo, BAI Chen, SHI Ya-Nan, XU Hao-Peng, YIN Lian, HU Shu-Min, YIN Hua

Bulletin of Botanical Research 2020, 40 (5): 775-781. DOI: 10.7525/j.issn.1673-5102.2020.05.017

Abstract （1107）

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Dihydromyricetin is an important component in the seeds of Hovenia dulcis Thunb. The extraction of dihydromyricetin from seeds of H.dulcis by ultrasound-assisted method was carried out on the basis of single factor investigation of ethanol volume fraction, ultrasound irradiation power, extraction temperature, liquid-solid ratio and ultrasound irradiation time. The Box-Behnken response surface design method was used to establish a quadratic polynomial model of ultrasound irradiation power, ultrasound irradiation time and liquid-solid ratio, and optimized extraction process. The optimum conditions for the extraction were as follows:ethanol volume fraction of 60%, ultrasonic irradiation power of 140 W, ultrasonic irradiation time of 30 min, liquid-solid ratio of 20.5 mL·g ^-1, and extraction temperature of 40℃. Under these optimal conditions, the yield of dihydromyricetin was 2.14±0.09 mg·g ^-1. The extraction method is simple, rapid, and high in efficiency, which is beneficial to the comprehensive processing and utilization of the H.dulcis resources.

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Comparison of Essential Oil Components in Ajania tenuifolia(Jacq.) Tzvel. at Different Altitudes

TAN Yong-Jia, GAO Cui-Fang, CHEN Xue-Lin

Bulletin of Botanical Research 2020, 40 (5): 782-788. DOI: 10.7525/j.issn.1673-5102.2020.05.018

Abstract （936）

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The aerial parts of Ajania tenuifolia(Jacq.) Tzvel. growing in different regions were collected as raw materials, and the volatile oil was extracted by water distillation. The chemical constituents of volatile oil were analyzed by GC-MS. The correlation between common components and altitudes was studied by SPSS 22.0 statistical software. The volatile oil components of A.tenuifolia were significantly different at different altitudes. There were five common components:camphor, α-pinene, β-pinene, eucalyptol, (-)-terpinen-4-ol. The chemical substances with the highest content of volatile oil at each altitude were eucalyptol, camphor, o-cymene, camphor, and eucalyptus. The same compound with a relative content of more than 5% at each elevation is only eucalyptol. By correlation analysis the content of these five common components was not significantly correlated with altitude. The altitude has no regular influence on the content of essential oils.

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Optimization of the ActiveCompoundsExtraction Process from Fig Leaves by Anionic and Cationic Surfactants and Preliminary Functional Evaluation

MENG Yao, YU Lian, QIN Xiang-Yu, LIU Chang, WANG Ze-Lin, ZHAO Chun-Jian, FU Yu-Jie

Bulletin of Botanical Research 2020, 40 (5): 789-794. DOI: 10.7525/j.issn.1673-5102.2020.05.019

Abstract （945）

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The experimental conditions for formulated surfactant combined with microwave-assisted extraction of effective bioactive substances in fig leaves were designed and the process parameters were optimized. The final results were as follows:1%(w/v) formulated surfactant(sodium dodecyl sulfate(SDS):dodecyl trimethyl ammonium bromide(DTAB)), extraction temperature of 40℃, extraction time of 10 min, liquid-solid ratio of 30:1 mL·g ^-1. Under the above optimized conditions, the average extraction yields of psoralen and bergapten were 15.37 and 3.59 mg·g ^-1, respectively. The antioxidant activity of crude fig extract obtained by surfactant-assisted extraction was significantly enhanced as compared with that by conventional aqueous extraction. Meanwhile, the degradation rate of the main substance in fig leaveswas also significantly inhibited. This study provides the necessary scientific basis for the rational and effective development and utilization of fig leaf resources. Also, it is of great theoretical significance for the green and efficient extraction of natural and effective target products and functional evaluation in plants.

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Detection of Alpha-linolenic Acid Content and Antioxidant Activity in Peony Seed Oil Prepared by Squeezing Method

GE Yun-Long, ZHAO Xiu-Hua, ZU Yuan-Gang, YUAN An-Long

Bulletin of Botanical Research 2020, 40 (5): 795-800. DOI: 10.7525/j.issn.1673-5102.2020.05.020

Abstract （861）

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The peony seed oil was prepared by physical pressing method, and the content of alpha-linolenic acid(ALA) in the obtained peony seed oil and the market peony seed blending oil was determined, and the antioxidant properties of the two peony seed oil were compared and analyzed. The results showed that the content of alpha-linolenic acid in peony seed oil and peony seed blending oil was 43.12% and 29.99%, respectively. The scavenging capacity of DPPH free radical, the scavenging capacity of ABTS, the reducing capacity of Fe ²⁺ and the scavenging capacity of -OH free radical of peony seed oil were stronger than that of peony seed blending oil, which were 1.29, 1.51, 3.62 and 1.44 times, respectively. It indicated that peony seed oil has stronger antioxidant ability.

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Preparation of Resveratrol-Nanostructured Lipid Carrier and Evaluation of in vitro Antioxidant Activity

WANG Xin-Yu, LI Hong-Kun, YU Liang, ZHAO Xue-Lian, YU Xue-Ying, FU Yu-Jie

Bulletin of Botanical Research 2020, 40 (4): 623-628. DOI: 10.7525/j.issn.1673-5102.2020.04.017

Abstract （2402）

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Resveratrol is a natural functional component, with whitening, antioxidant and other effects, can be used to improve skin lesions and cell aging. Resveratrol has low solubility and poor stability in water, which impedes its application in medicine and cosmetics. The nanostructured lipid carrier loaded with resveratrol was prepared to improve the stability and functional activity of resveratrol. The resveratrol nanostructured lipid carrier(NLC) was prepared by thermal high pressure homogenization method, and the homogenization pressure was determined to be 600 bar with GMS as the solid lipid, oleic acid as the liquid lipid, with their ratio of 1:1, 4% Tween 80 as the emulsifier, and the smallest particle size of resveratrol lipid carrier was 179 nm after three cycles.The results of MTT experiment and ROS scavenging experiment also showed that resveratrol did not produce obvious toxicity to cells when the lipid carrier concentration of resveratrol was less than 100 μmol·L ^-1, and had better antioxidant activity than resveratrol API.

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Development of Microsatellite Primers of Quercus championii with RAD-seq Data

NING Xin, JIANG Xiao-Long, DENG Min, XU Gang-Biao

Bulletin of Botanical Research 2020, 40 (4): 629-634. DOI: 10.7525/j.issn.1673-5102.2020.04.018

Abstract （2565）

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Microsatellite(simple sequence repeat, SSR) is a co-dominant molecular marker commonly found in plant genome with high polymorphism. This genotyping method is widely used in the study of spatial genetic structure of populations. With the blooming of the new sequencing technologies, the development method of SSR is also more diverse. Quercus championii is a precious wood species in the evergreen broad-leaved forest of South China. The development of its molecular markers can promote the species breeding and germplasm conservation. We used the reads obtained from RAD-seq(restriction-site associated DNA sequence) of four individuals of Q.championii to developed SSR primers. The pyRAD analysis showed that:(1)the reads containings 46 000-84 000 repeats sequence in each individual; (2)within the individual ranged clustering, 5 500-24 000 loci were detected; (3)1 158 concordance loci were obtained after clustering between individuals. Finally, 186 loci with flanking sequences were not mutated and repeating base was in the middle were obtained. A total of 25 primers pairs were designed using premier 5.0 and validated in 36 individuals from 3 populations. The results showed that 17 primer sets were successfully amplified and 106 alleleswere obtained. The number of alleles per primer between 2-12, and the average number is 6.2. The expected heterozygosity and observed heterozygosity of the primers were 0.19-0.88 and 0.11-0.76, respectively. The rapid, effective, and low cost in this method can be applied to the development of molecular markers of population genetics.

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Response Surface Optimized Enzymatic Hydrolysis-microwave Assisted Extraction Process of Mulberry Leaf Polysaccharide

ZHU Ya-Wei, WANG Li-Tao, AN Juan-Yan, Lü Mu-Jie, SUN Jian-Hang, GUO Na, FU Yu-Jie

Bulletin of Botanical Research 2020, 40 (4): 635-640. DOI: 10.7525/j.issn.1673-5102.2020.04.019

Abstract （2804）

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The response surface analysis was used to optimize the process of extracting polysaccharide from mulberry leaves by enzymatic hydrolysis-microwave assisted method.Based on the single factor test, the Box-Behnken method was used to study the effects of liquid-solid ratio, extraction time and extraction temperature on the extraction rate of mulberry leaf polysaccharides. The fitting equation was significant, and the optimal extraction conditions of mulberry leaf polysaccharides were finally determined as follows:enzyme content 2%, enzymolysis pH6, enzymolysis temperature 50℃, enzymolysis time 20 min, liquid-solid ratio 15 mL·g ^-1, extraction time 13 min, and extraction temperature 76℃.Under these conditions, the actual extraction rate of mulberry leaf polysaccharide was 15.23%, which was closed to the theoretical simulation value of 15.12%. Therefore, the established model was true and reliable. The method can be used for extracting polysaccharide components in mulberry leaves with the advantages of simple process, low cost and high application value.

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Root CO ₂ Efflux Variations of Betula platyphylla Among Sites and Root Diameter Classes in 12 Different Provenances

SUN Shuo, WANG Xiu-Wei, DU Meng-Tian, LI Jing-Hang, WANG Bo-Yi, LIU Gui-Feng

Bulletin of Botanical Research 2020, 40 (3): 476-480. DOI: 10.7525/j.issn.1673-5102.2020.03.020

Abstract （587）

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The 13 years old Betula platyphylla from 12 provenances in Caohekou and Maoershan were used as experimental materials. In the growing season, three diameter classes roots <0.5 mm, 2.0-5.0 mm and >5.0 mm were taken, respectively. The CO ₂ efflux rate of root system was measured by infrared gas analysis in vitro, in order to study the effect of root diameter, experiment site and provenance on CO ₂ efflux of B.platyphylla root. The root diameter and environment had significant( P<0.01) effects on root CO ₂ efflux rate, while provenance had no significant( P>0.05) effects on root CO ₂ efflux rate. Root CO ₂ efflux rate decreased significantly( P<0.01) with the increase of diameter class. All diameter class root CO ₂ efflux rate in Caohekou was greater than that in Maoershan( P<0.01). The root CO ₂ efflux rates of B.platyphylla in Caohekou and Maoershan with diameters <0.5 mm were 104.21 and 81.07 mol·g ^-1·s ^-1, the root CO ₂ efflux rates with diameters 2.0-5.0 mm were 41.08 and 30.91 mol·g ^-1·s ^-1, and the root CO ₂ efflux rates with diameters >5.0 mm were 24.87 and 11.37 mol·g ^-1·s ^-1, respectively. There were significant( P<0.05) differences in tree height and DBH among 12 provenances of B.platyphylla, and both tree height and DBH were negatively correlated with root CO ₂ efflux. The results of relevant studies will provide an important basis for more accurate assessment of CO ₂ efflux fluxes from tree roots.

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Optimization of Dry Enzymatic Extraction Process and Physicochemical Properties of Pine Seed Oil

ZU Shu-Chong

Bulletin of Botanical Research 2020, 40 (2): 308-313. DOI: 10.7525/j.issn.1673-5102.2020.02.019

Abstract （1082）

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Used a single factor experiment to optimize a dry enzyme extraction process of pine seed oil. The best preparation process for dry enzymatic extraction of pine seed oil was:0.2% of amylase, the ratio of material to liquid of 6:1, the hydrolysis temperature of 55℃, and the hydrolysis time of 8 h, and the maximum oil yield 90.2% with the residual oil rate of 6.2%. The content of unsaturated fatty acids in pine seed oil extracted after testing for the best preparation conditions was 90.09%, of which oleic acid content was 26.84%, mediating acid content was 2.41%, linoleic acid content was 46.25%, and pinolacol acid was 14.59%. The content of palmitic acid in saturated fatty acids was 6.5%, and the stearic acid was 3.41%. The pine oil had an acid value of 2.61 mg·g ^-1, a peroxide value of 1.56 mmol·kg ^-1, and a malondialdehyde content of 0.41 mg·kg ^-1. All the test results are higher than the requirements of the quality standards in pine seed oil in the "Food Industry Standard of the People's Republic of China LS/T 3242-2014".

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Preparation Process and Bioavailability Evaluation of β-cyclodextrin Inclusion Korean Pine Seed Oil

ZU Shu-Chong

Bulletin of Botanical Research 2020, 40 (2): 314-320. DOI: 10.7525/j.issn.1673-5102.2020.02.020

Abstract （1082）

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The single-factor method was used to optimize the preparation process of the Korean pine seed oil inclusion compound. Under the optimal conditions, the solidification rate of the Korean pine seed oil inclusion compound was 70.95%, and the oil content of the inclusion compound was 26.88%. The fatty acid composition, particle size, potential, and bioavailability of Korean pine seed oil and Korean pine seed oil inclusion complex were compared. There was no significant difference between the contents of each component of the Korean pine seed oil in the inclusion compound and that of the Korean pine seed oil, among which the content of pinolenic acid was in 14% to 16%. The average particle size of the Korean pine seed oil inclusion complex was 177.3±2.6 nm, and the potential was -33.01±1.4 mV. The blood drug concentration(CMAX), t1/2A, AUC and MRT were 1.27, 1.42, 1.56 and 1.04 times of the Korean pine oil inclusion complex group, respectively. The inclusion complex of Korean pine seed oil and aqueous solution formed an emulsion, the particle size decreased, the water solubility increased, the inclusion complex in vivo t1/2A and the average retention time(MRT) increased, and the absorption of pinolenic acid in vivo increased.

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