Baphicacanthus cusia (B. cusia) was traditionally used as raw material to prepare indigo naturalis; however, many factories have been shut down by local governments on account of their serious pollution contributions. In this study, a green and sustainable production of B. cusia leaf extracts is proposed. Namely, new indigo extract (NIE) and phenolic acid extract (PAE) were produced in an integrated fashion by enzyme transformation coupled membrane separation and freeze-drying technologies. The small-scale process was optimized, and its feasibility was verified by a pilot experiment. Moreover, the physicochemical properties, dyeing effect, and wound-healing ability of the two extracts were systematically evaluated. The NIE could be obtained within 1 day and exhibited more than 20 times the indigo content and 1.8 times the pure indigo yield than indigo naturalis. Moreover, the NIE showed superior dye-uptake rate and washing and rubbing fastness on pure cotton poplin compared with natural indigo from India. The PAE exhibited excellent broad spectrum antibacterial activities, especially S. aureus (IC50 = 32 mu g/mL). Significantly, the PAE had no toxicity and presented over 98% of the wound-healing rate on mice by increasing their collagen fiber content. All of above work is of great significance for the extension and sustained healthy development of a B. cusia industry chain.

In this study, Ginkgo biloba leave polyprenols (GBP) and polypyridine metal complex were individually utilized as functional ligand and main ligand, four kinds of novel GBP-based polypyridine metal complexes were successfully synthesized and their cell absorption capacity, light-dark stability, photodissociation efficiency, ROS production capacity, light-controlled antibacterial and anti-tumor activities as well as mechanisms were systematically investigated by ultraviolet visible spectrophotometer (UV–vis), confocal laser scanning microscope (CLSM), gel electrophoresis (GE), scanning electron microscope (SEM), oxford cup method, MTT method etc. The lipid water distribution coefficients of complex 1, 2 and 4 were all within the range of 0∼3, demonstrating their better cell absorption capacity and more competitive bioavailability potentiality compared with GBP. All of the synthesized complexes possessed excellent stability in a dark environment, and could conduct ligand dissociation under the condition of visible light irradiation except complex 1. In which, complex 2 and complex 4 were able to achieve degradation rates of 37.9 % and 54.4 % within 5 min, separately. In addition, complex 2 and complex 4 exhibited superior inhibitory activities on the HN-3 tumor cells on account of their stronger ROS production capacity. Moreover, the constricted expression of BCL-2 and NF-kB p-p65, especially the promoted expression of BAX may be one of the root cause. The four synthesized complexes had preferable inhibition effects against S. aureus under the condition of visible light irradiation in contrast to darkness, in which complex 4 was the best and its MIC and MBC values were 6.25 and 12.5 μg/mL, respectively. The antibacterial mechanism of the complex 4 may be in relation to the synergistic effect of multiple factors, including leakage of bacterial inclusion, change of cell membrane permeability and disruption of cell wall etc. All of the above generalized researches will pave a way for the high-value development and application of GBP-based functional products.

The effect of acidolysis time, hydrochloric acid concentration, ozonization time and pH on the yield of indigo were investigated by single factor experiment using dried leaves of Baphicacanthus cusia (Nees) Bremek (DLB) as raw material. On this basis, the technological condition for preparing new indigo extract (NIE) by directional chemical conversion method was optimized by orthogonal experiment. Then, the physiochemical properties of NIE were analyzed by combustion assay, nitric acid assay, thin layer chromatography identification assay, moisture detection and water-soluble pigment observation assay. Moreover, UPLC-Q-TOF-MS/MS technology was utilized to compare chemical composition difference between the NIE and traditional indigo naturalis (TIN). Finally, NIE emulsion was prepared by ultrasound assisted high pressure homogenization method and its stability, antibacterial activity as well as antibacterial mechanism were evaluated. The experimental results showed that optimum process for preparing NIE were hydrochloric acid concentration of 2 %, ozonization time of 3 min, pH of 7 and acidolysis time of 1 h; Under this condition, the content and yield of indigo in the NIE were significantly improved, which could reach up to 42.6 ± 0.28 % and 2.26 ± 0.04 %, separately. NIE possessed similar physiochemical properties with TIN described in Chinese pharmacopoeia 2020, and had more diverse active ingredients than TIN. The prepared NIE emulsion demonstrated excellent stability, whether at dark or bright place, and no matter at low temperature (4 ℃) or room temperature (25 ℃), the particle size, PDI, pH and indigo content of which displayed little variation. In addition, the NIE emulsion owned good inhibitory activity against S. aureus, and its MIC and MBC values were 40 and 80 μg/mL, respectively. Moreover, NIE emulsion exhibited antibacterial effect by influencing cell membrane integrity, Ca2+-Mg2+-ATPase activity on the cell membrane and intracellular Ca2+ levels of S. aureus. All of above results will lay the foundation for the green and sustainable utilization of Baphicacanthus cusia (Nees) Bremek.

Lacking of substantial physiological activity and low utilization remains a problem for most conventional drug carriers. Polyprenol with beneficial medical effects and high availability could be an ideal candidate for solving this issue. Here, Ginkgo biloba leaves polyprenol (GBP)-based derivative was prepared by Michael addition reaction of poly (β-amino esters) (PBAE) with GBP and galactose (Gal). The intervention of poly (β-amino ester) and galactose promoted GBP-PBAE-Gal to depict as micellar carrier, enhancing the loading of hydrophobic DOX and the sensitivity to the specific tumor microenvironment, with the largest DOX loading of 28.62 ± 1.49 % and the efficient DOX release rate of 90.30 %. In the meantime, GBP-PBAE-Gal exhibited enhanced colloidal stability at 640-folds of dilution and in the presence of serum and realized the possibility of long-term storage at room temperature. Additionally, GBP-PBAE-Gal was safe for human red blood cells and human normal liver cells HL-7702. When applied for DOX delivery to HepG2 cells, GBP-PBAE-Gal increased the targeting of DOX to intensify its inhibition on HepG2 cells. Compared to free DOX, the DOX loaded into GBP-PBAE-Gal presented stronger anticancer activity, with IC50 of 0.56 μg/mL at 72 h. Besides, the anticancer mechanism study revealed that GBP-PBAE-Gal arrested the cell cycle in HepG2 cells, suggesting the potential of GBP-based carrier for intensive treatment. This research evidenced the feasibility and high availability of the GBP to use as a drug carrier, providing a novel candidate for drug delivery systems.

The effects of treatment with edible coatings containing decolorizing shellac incorporated with natural antibacterial juglone from walnut green husk extract (WGHE) on the postharvest quality of dried Wichita pecans during 180 days at 4℃ were investigated. WGHE with a high yield of juglone was obtained by Soxhlet extraction in orthogonal test and mixed with main film former of decolorizing shellac to prepare edible coatings. The present results showed that the edible coating preserver NO. 4 (CP4) group had better waterproof performance and could coat pecan nuts in a natural color, decrease the pure kernel yield and the fat content, but the acid value, the peroxide value, and the soluble protein content were increased compared with the blank (BL) group. Moreover, the changing trends of the pure kernel yield were consistent with the fat content but had an opposite relationship to the acid value, as the heat map and correlation coefficients of different indexes after treatment of CP4. Besides, WGHE enhanced the antibacterial activity of CP4, meanwhile, Wichita pecans had a good overall taste in the determination of sensory evaluation, indicating CP4 minimized the hydrolysis and oxidation of internal fats, especially their auto-oxidation, maximized the stability of their functional unsaturated fatty acids and antioxidant functional components. In conclusion, CP4 treatment is a potential method in delaying significantly the oxidation process of pecans for maintaining the pecan nutrition and quality under low temperature during long-term storage.

以海南生物结香的沉香木为原料,建立了沉香精油的基质固相分散-气相色谱/质谱联用(MSPD-GC/MS)分析方法.采用基质固相分散技术进行沉香精油的提取,采用单因素试验得到优化工艺条件为:吸附基质为硅酸镁,吸附基质与样品的质量比为4:1,分散溶剂为异丙醇,分散溶剂与样品的比例为4:1(mL:g),洗脱溶剂为体积比9:1的正己烷和乙酸乙酯混合液,洗脱溶剂用量6 mL,在此条件下,沉香精油的提取得率为(9.19±0.15)％,明显高于乙醚超声波提取方法的(6.65±0.22)％.采用GC/MS分析沉香精油成分,并与乙醚超声波提取方法进行比较.基质固相分散提取的沉香精油成分组成与乙醚超声波提取相似,主要为2.13％的芳香族类、25.67％的倍半萜类、26.85％的色酮类和7.5％的脂肪族.相比乙醚超声波提取,基质固相分散提取的沉香精油中倍半萜类化合物的成分更多,主要有 γ-芹子烯、β-檀香醇、缬草-4,7(11)二烯、α-愈创木烯、白菖醇和1,9-马兜铃二烯.MSPD-GC/MS分析方法能够真实地反映沉香精油的化学成分,可作为沉香品质评价的一种快速鉴别方法.

以漆籽油为原料,酪蛋白酸钠为乳化剂,采用高压均质法制备漆籽油纳米乳液,通过单因素试验优化制备工艺条件,考察漆籽油纳米乳液在不同温度条件下的贮藏稳定性,并评价其抗氧化活性,采用体外细胞法考察其抗紫外辐射活性.实验结果表明:漆籽油纳米乳液较佳制备工艺条件为乳化剂质量分数为2.5％,漆籽油质量分数为6％,均质压力为100 MPa,均质4次,制备的纳米乳液平均粒径163.0 nm,Zeta电位-41.01 mV,多分散指数(PDI)为0.225;漆籽油纳米乳液具有很好的贮藏稳定性,在4、25和60℃条件下贮藏30 d,平均粒径保持在200 nm以内;对DPPH、ABTS及羟基自由基的有效中浓度(EC50)分别为24.26、23.77和21.10 mg/L,抗氧化活性优于阳性对照VC(38.48、43.25和40.56 mg/L);其对紫外线辐射损伤的NIH 3T3细胞有显著的修复作用,漆籽油纳米乳液质量浓度大于5 mg/L时,辐照30 min,长波紫外线(UVA)损伤的NIH 3T3细胞存活率高于100％,具有很好的抗紫外辐射活性.

以6种不同品种的松木材为原料,通过溶剂抽提得到不同品种松木的石油醚和甲醇提取物,并采用GC-MS和LC-MS分析其化学成分,同时采用牛津杯法评价不同品种松木提取物对密粘褶菌的抑菌效果.结果表明:6种松木石油醚提取物中共鉴定出45种化合物,主要为萜类、酯类和脂肪酸等物质;6种松木甲醇提取物中共鉴定出59种化合物,主要为酚类、有机酸类和黄酮类等物质;松木石油醚和甲醇提取物对密粘褶菌的生长均具有一定的抑制作用,且松木石油醚提取物的抑菌效果要优于甲醇提取物,表明松木中萜类、酚类等次生代谢产物对松木材防腐性能具有重要影响.

以江苏泗洪市所产的薄壳山核桃为原料,分别涂抹不同浓度的以脱色紫胶、大豆分离蛋白、乳化漆蜡为主要成膜剂的9组复合保鲜剂,以4℃、无氧及纳米聚乙烯包装组作为空白对照,考察不同保鲜处理对薄壳山核桃贮藏期间纯仁率、脂肪含量、酸值、过氧化值和蛋白质含量的影响.结果表明,效果最好的是复合保鲜剂4:脱色紫胶10 g、大豆分离蛋白10 g、沸水200 mL、乳化漆蜡2 g、青龙衣提取液3 mL、EDTA-Na2 2 g和维生素E 2 g;相比对照组,其纯仁率下降程度减少了5％,脂肪含量下降程度减少了6.60％,酸值增加程度减少了0.43倍,过氧化值增加程度减少了0.73倍,可溶性蛋白质含量增加程度提高了0.47％.因此,保鲜剂4结合低温、除氧剂及纳米聚乙烯包装对薄壳山核桃具有较好的保鲜效果,能够显著延缓薄壳山核桃的氧化过程,保持较高的贮藏品质.

以秦巴山区漆籽为原料,采用溶剂(石油醚)浸提法从其种仁中提取漆籽油,在单因素试验的基础上,采用响应面法优化漆籽油提取工艺,对漆籽油的理化性质进行分析,采用气质联用分析漆籽油的脂肪酸组成,并评价其抗氧化活性.结果 表明:漆籽油的最优提取工艺条件为提取温度70℃、提取时间60 min、液料比10∶1、提取次数2次,在此条件下漆籽油提取率为96.14％;漆籽油相对密度为0.9258,折光指数(20℃)为1.4672,碘值(I)为146.83 g/l00 g,酸值(KOH)为2.58 mg/g,过氧化值为3.62 mmol/kg,皂化值(KOH)为185.36 mg/g;漆籽油主要含有油酸、亚油酸、棕榈酸、硬脂酸、亚麻酸和棕榈油酸6种脂肪酸,不饱和脂肪酸含量达86.10％,以油酸和亚油酸为主,二者含量达到82.17％;漆籽油对DPPH、ABTS及羟自由基的半数清除浓度(IC50)分别为35.38、41.29、38.52 mg/L,具有与维生素C相当的抗氧化活性.

The aim of this work was to construct a new type of system for the green extraction and efficient separation of Ginkgo biloba leaf polyprenols (GBP). First, cell disruption technologies (enzymolysis, ultrasound, microwave and ball milling) coupled with deep eutectic solvents (DES) were used to extract polyprenyl acetates (PA) from Ginkgo biloba leaves. Results showed that ball milling was the best assisted extraction method, and the amount of PA was 89.88 mg/g. Then, a novel silver-loaded material with selectivity for PA was synthesized, characterized and utilized to absorb PA from the DES extraction solution. The purity of the PA quickly reached 71.28 % after being separated by the silver-loaded material. Finally, saponification and molecular distillation were adopted to remove non-GBP ingredients, and a high-purity GBP (99.80 %) was obtained by optimizing its processing conditions. This study provides a technical support for the development of a green and efficient GBP separation method and lays the foundation for the fabrication of GBP-based drugs, health products and cosmetics.

Polyprenols extracted from Ginkgo biloba leaves is a kinds of unsaturated compound containing double bonds. Traditionally, the separation methods for the polyprenols are lack of selectivity and their separation efficiency are low. We synthesized two kinds of functional nano-silica containing silver ions materials (AgTCM and AgTCN) which have selectivity for unsaturated compounds to separate Ginkgo biloba leaves polyprenols for the first time. AgTCN displays exceptionally high selectivity for polyprenols and high stability under extended heat and light exposure, while silver is virtually immobile during solvent elution. Importantly, the exceptional stability of AgTCN gives rise to much higher polyprenols recovery than conventional silica gel during the chromatographic elution. In addition, we found that the adsorption of polyprenols onto the AgTCN conforms to pseudo-second-order kinetic model and AgTCN has strong affinity with polyprenols by analyzing Langmuir, Freundlich, Temkin-Pyzhev, and Dubinin-Radushkevich isotherms. The calculation results of thermodynamic parameters demonstrate that decrease of temperature in favor of increasing the adsorbing capacity of polyprenols onto the AgTCN, and the adsorption process of which is exothermic reaction. Our results pave the way for the novel separation methods of polyprenols from Ginkgo biloba leaves.

Hepatocellular carcinoma (HCC) is one of the most lethal diseases worldwide. Ginkgo Biloba Leaves polyprenol (GBP) is considered potential efficacy in the treatment of HCC. This study involved oil-in-water type nanoemulsion (NE) loading GBP and Fullerene C60 (C60F) were prepared by dissolving GBP and C60F in NE using inversed phase emulsification (EIP) with ultrasonic-assisted emulsification (UAE) method. Folic acid (FA) coupled Chitosan (CS) and Graphene oxide (GO) nanocomposites (NCs): FA-CS-GO-NCs were fabricated by ionic cross-linking of positively charged FA-CS-GO solution conjugates and negative charged NE with TPP. We combined these materials for preparing NCs loading GBP and C60F (GBP-C60F-FA-CS-GO-NCs) based GBP-C60F-NE. GBP-C60F-FA-CS-GO-NCs in the NE were the mean size was 44.9 nm, PdI was 0.267 and the mean zeta potential was 47.8 mV. And they had good drug loading efficiency, encapsulation efficiency, drug release property and storage stability. In this cytotoxic study, it demonstrated the GBP-C60F-FA-CS-GO-NCs were greater inhibition capacity on MHCC97H cells compared with GBP-NE, C60F-NE, GBP-C60F-NE, GBP-FA-CS-GO-NCs and C60F-FA-CS-GO-NCs. In this genotoxic study, GBP-C60F-FA-CS-GO-NCs at low C60F concentration showed low genotoxicity on MHCC97H and L02 cells. By comparison, GBP-C60F-FA-CS-GO-NCs had higher cytotoxicity on MHCC97H than L02 cells. By the cell apoptosis analysis, the results revealed that GBP-C60F-NE and GBP-C60F-FA-CS-GO-NCs could obviously induce MHCC97H cell apoptosis, especially high concentration GBP-C60F-FA-CS-GO-NCs had the strongest apoptosis inducing ability. The treatment against MHCC97H cells with moderate C60F concentrations of GBP-C60F-FA-CS-GO-NCs effectively inhibited the overexpressions of Akt1 and Akt2, and significantly increased the expression of PTEN. Our results suggest that PTEN, Akt1 and Akt2 might play an important role in the aspect of GBP-C60F-FA-CS-GO-NCs inhibiting against HCC cells. It provides further evidence to speculation that up-regulated PTEN expression and down-regulated Akt1 and Akt2 expression might be one of the important mechanisms for GBP-C60F-FA-CS-GO-NCs inhibiting HCC cells. All these results suggest that GBP-C60F-FA-CS-GO-NCs effectively inhibited the HCC malignant development and has potentially important value in the HCC treatment.

以脂肪酶为催化剂,催化漆蜡与异丙醇进行酯交换反应合成漆蜡基异丙酯.首先采用傅里叶红外光谱对合成产物进行结构表征,然后采用单因素试验和正交试验对酯交换反应条件进行了优化,并对最佳条件下合成的漆蜡基异丙酯的组成进行了GC-MS分析.结果 表明:在脂肪酶催化下,漆蜡可与异丙醇反应生成目标产物;Novozyme 435脂肪酶催化酯交换反应的最佳工艺条件为醇蜡比4∶1、脂肪酶加量10 g/L、反应温度55℃、反应时间36 h,在此条件下酯交换率为86.7％,漆蜡基异丙酯的酸价(KOH)为3.28 mg/g;漆蜡基异丙酯主要由棕榈酸异丙酯(73.64％)、油酸异丙酯(19.39％)、硬脂酸异丙酯(6.15％)和亚油酸异丙酯(0.82％)组成.

The analytical methodology and method performance for the determination of polyprenol in Ginkgo biloba leaves were investigated for the first time. The results showed that of the high performance liquid chromatography with C18 as chromatographic column, isopropanol methanol (volume ratio, 32∶18) as mobile phase, 210 nm as detection wavelength showed high precision (RSD=0.89％) and good stability (RSD=3.26％) . In addition, the average standard recovery and RSD value of polyprenol were 97.4％ and 1.75％, respectively. Then, polyprenol with the purity of 98.6％ was obtained by extraction with n-hexane, saponification, de-waxing with ethanol and acetone, and silica gel column chromatography (V (diethyl ether)∶V (n-hexane) =3∶97) . On this basis, the terminal hydroxyl group of high-purity polyprenol was modified to synthesize aminopolyprenol. Firstly, by mitsunobu reaction with phthalimide, followed by reduction reaction with hydrazine hydrate, to obtain aminopolyprenol. The structure of amino polyprenol was characterized by infrared and nuclear magnetic resonance spectroscopy, which confirmed the aminopolyprenol was successfully synthesized.

为探究先后提取枸杞多糖及枸杞色素时对各自得率的影响.本研究通过分别考察这两种成分在提取过程中的提取溶剂、提取温度、料液比、提取次数及提取时间等因素对枸杞色素和枸杞多糖的得率影响,确定枸杞色素和枸杞多糖在提取次序不同时,两者的最佳提取工艺以及对DPPH·和·OH自由基清除率.结果 表明,首先提取枸杞多糖后,枸杞色素的最佳提取工艺为采用正己烷,80℃时,料液比1∶10,提取2次,每次1h,枸杞色素得率为2.48％,此时枸杞多糖得率为7.45％;而首先提取枸杞色素后,采用了超声辅助提取的方式提取枸杞多糖,发现超声效率为25％,料液比1∶10,提取20 min,枸杞多糖得率为5.23％,此时枸杞色素得率为3.93％.因此,首先提取枸杞多糖,使其平均得率为7.45％,而后提取枸杞色素,其平均得率为2.48％;总体上,枸杞色素1和枸杞多糖1对DPPH·自由基清除率都较高,枸杞多糖1对·OH自由基清除率较高,其抗氧化活性都接近Vc.

以漆脂为原料,经过硫酸催化水解,生成漆脂脂肪酸,再与氯化亚砜反应生成漆脂脂肪酰氯,然后采用肖顿-鲍曼(Schotten-Baumann)缩合法,在弱碱性条件下,漆脂脂肪酰氯与肌氨酸钠盐进行缩合反应,再经过酸化、萃取等过程,制得漆脂酰肌氨酸.采用红外光谱和核磁共振碳氢谱对其结构进行表征,并对产物钠盐的表面性能进行分析.结果表明,合成产物为漆脂酰肌氨酸,漆脂酰肌氨酸钠盐的临界胶束浓度为6.733×10-4mol/L,表面张力为22.845 mN/m,具有较好的表面活性.当漆脂酰肌氨酸钠盐浓度为0.01 mol/L时,从乳化的液体石蜡中分出10 mL水的时间为90 s,对低极性溶剂的乳化效果较差,但漆脂酰肌氨酸钠盐的起泡和稳泡性能较好,起泡高度为130 mm,5 min后降至115 mm.

针对漆蜡的加氢工艺和复配漆蜡的复配加氢工艺进行研究.结果表明,在钯碳催化剂用量0.5％、氢气压力0.3 MPa、反应时间2h、反应温度150℃、转速500 r/min的条件下,加氢后漆蜡中硬脂酸相对含量提高到23.89％,熔点由49.6℃上升到50.9～52.6℃.在相同的加氢条件下,将加氢后的漆蜡与LH蜡(精制乌桕蜡)按复配比1:1进行复配加氢,得到硬脂酸含量为17.48％的新型复配漆蜡,接近日本精制白蜡中硬脂酸含量17.60％.由于LH蜡的市场价格远低于漆蜡的市场价格,能够降低生产成本,因而这种新型复配漆蜡有很好的发展前景和市场竞争力.

利用热重-微商热重(TG-DTG)分析漆树提取物(RWE)在氮气氛围中的热分解曲线,运用Kissinger、Flynn-Wall-Ozawa(FWO)、Friedman、Coats-Redfern和Achar法对第一步热分解过程进行动力学分析,计算热分解的表观活化能(Ea)和指前因子(A),并根据Ea和A计算热力学参数和推算漆树提取物的贮存期.研究结果表明:随着升温速率的增大,漆树提取物的热分解温度逐渐升高;漆树提取物的失重分为2个阶段(10 K/min):第一阶段189.09～266.59℃,第二阶段266.59～377.79℃,这两步热分解对应DTG曲线有2个主要的失重峰,最大热失重速率对应的温度分别为248.3和306.2℃,总失重率为57.94％.漆树提取物第一阶段热分解的机制函数为Avrami-Erofeev方程(随机成核和随后生长,n=3/4),积分形式g(α)=[-ln(1-α)]3/4,微分形式f(α)=4/3(1-α)[-ln(1-α)]1/4.计算得到Ea=101.353 kJ/mol,lnA=25.0928,A=7.9×1010 min-1;ΔG=77.799 kJ/mol,ΔH=96.978 kJ/mol,ΔS=36.446 J/mol;可以推断漆树提取物在室温(25℃)氮气氛围下贮存的话,贮存期为1.5～2年.

Gingko starch microspheres were prepared by inverse emulsion polymerization treating with gingko starch as raw material.The effects of starch concentration,volume ratio of oil and water,dosage of crosslinking agent,emulsifier concentration,dosage of initiator on adsorbing capacity were investigated.The performance of the gingko starch microspheres was observed with particle size analyzer,IR,SEM,XRD,DSC and TG.Results showed that the optimal conditions for the starch microspheres production obtained are as follows:starch concentration 10％,volume ratio of oil and water 8∶1,dosage of crosslinking agent 25％,emulsifier concentration 12％,dosage of initiator 20％,reaction temperature 60 ℃ and reaction time 2 h.There were many holes on the surface of gingko starch microspheres,the size distribution of microspheres was uniform,the average particle size was 19.6 μm,and the adsorption of methylene blue was 39.2 mg/g.Comparing with raw gingko starch,the adsorption performance and thermal stability of gingko starch microspheres were significantly improved.