Lycium ruthenicum Murr. treatment attenuates APP_(SWE)/PS1ΔE9 mouse model-like mitochondrial dysfunction in Slc25a46 knockout mouse model

Mitochondrial dysfunction is proposed to be substantially associated with ageing and ageing-related diseases like Alzheimer's disease(AD). However, it is unclear whether different mouse models with mitochondrialrelated diseases have similar changes in mitochondrial morphology of the same tissues. Moreover, whether similarities in mitochondrial morphology can be a suitable marker for screening and/or discovering mitochondrial-protective substances remains unknown. Mitochondria morphology in different tissues of a novel mitochondrial outer membrane protein Slc25a46 knockout mouse and a traditional APP_(SWE)/PS1ΔE9 transgenic mouse were examined using transmission electron microscope(TEM). Both young Slc25a46 knockout mice and aged APP_(SWE)/PS1ΔE9 mice models showed similar mitochondrial damage in cerebellum tissues. The results indicated that different mitochondrial-related diseases shared similar alteration and defects in mitochondrial morphology. Furthermore, Lycium ruthenicum Murr. extract, a bioactive food substance with cognition-improving property, could effectively improve muscle strength and increase body weight in the Slc25a46 knockout mice. These findings suggest that mitochondrial morphology defects in mice models, particularly in the mitochondrial compartment, represent a unified and effective marker for screening and validating natural product-derived functional substances with mitochondrial protective properties. It also holds potential application in mitochondrial-impaired senile neurodegenerative diseases, especially in AD.

Effects of NaCl Stress on Seed Germination of Lycium ruthenicum Murr.

[Objective]To study the seeds germination of Lycium ruthenicum Murr.under different concentrations of NaCl,as well as to find the optimal concentration of NaCl for the germination of L.ruthenicum.[Method]The seeds of L.ruthenicum were treated with different concentrations of NaCl,and the state of seed germination was measured.[Result]With the increasing of concentration of NaCl,the seed germination rate of L.ruthenicum showed an obvious increasing trend.when the concentration was of 0.3%-0.4 %,the germination rate was the highest,and when the concentration of NaCl was greater than 0.4,the germination rate showed a decline trend.[Conclusion]After treated with appropriate concentrations of NaCl before sowing,the germination rate of seeds of L.ruthenicum would increase.

Karyotype Analysis of Lycium ruthenicum Murr.

Karyotype analysis of Lycium ruthenicum Murr. was carried out in this study. The results showed that the chromosome number was 2n=2x=24; the arm index was 48; the ratio of the longest chromosome to the shortest one was 1.31; the proportions of chromosomes with arm ratio higher than 2 was 0.08; the asymmetry index was 57.02; the karyotype type was 2A; and the karyotype formula was 2n=-24=20m＋4sm.

Study on Tissue Culture and Rapid Propagation of Lycium ruthenicum Murr.

[Objective] This study aimed to establish a tissue culture and rapid propagation system for Lycium ruthenicum Murr.[Method] Using tender stems of L.ruthenicum as explants,MS as basic culture medium,the effects of different factors on primary culture,subculture and rooting of L.ruthenicum plantlets were investigated.[Result] The most appropriate medium for primary culture of L.ruthenicum was MS ＋ ZT 0.2 mg/L ＋ IBA 0.01 mg/L,in which axillary buds grew well and were rarely vitrified with the germination rate of 88.73%.In addition,ZT exerted significantly better effects on subculture and proliferation of L.ruthenicum plantlets than 6-BA.The most appropriate medium for subculture and proliferation of L.ruthenicum plantlets was MS ＋ ZT 0.15 mg/L ＋ IBA 0.01 mg/L,in which L.ruthenicum plantlets grew rapidly and robustly without vitrification,and the proliferation multiple reached 5.83 times.The most appropriate medium for rooting of L.ruthenicum plantlets was MS ＋ IBA 1.0 mg/L,in which the rooting rate reached 100%.The most appropriate substrate for transplanting and hardening of L.ruthenicum plantlets was humus soil：perlite = 1：1,in which L.ruthenicum plantlets grew well with the survival rate of 92.37%.[Conclusion] This study provided theoretical basis for largescale production and popularization of L.ruthenicum.

Eco-Friendly pH Indicator Based on Natural Anthocyanins from Lycium ruthenicum

An eco-friendly and visual pH indicator was developed based on cotton fabrics dyed with natural Lycium ruthenicum extract(LRE),whose color changed from red to purple,blue and green at pH of 2-10.Three methods for dyeing cotton fabrics with LRE were attempted,namely pre-mordanting,simultaneous mordanting,and post-mordanting methods with different dyeing temperatures,dyeing times and dyebath pH values.The cotton fabrics exhibited the highest K/S value when they were dyed under a dyebath of pH 6 at 20℃for 90 min in the case of the simultaneous mordanting method.Meanwhile,the dyed cotton fabrics also showed reversible pH-dependent color changes.The developed flexible pH indicator based on renewable natural materials is suitable for multiple applications in environmental monitoring and smart textiles.

Comparative Analysis of Proanthocyanidins and Polysaccharides on Wild Lycium ruthenicum

[Objectives] To compare content of proanthocyanidins and polysaccharides in wild Lycium ruthenicum. [Methods] The wild L. ruthenicum collected from different regions such as Qinghai, Inner Mongolia, Ningxia and Gansu Province were taken as the research objects. The conventional indicators such as proanthocyanidins and polysaccharides of the experimental materials were determined, and the proanthocyanidins and polysaccharides of the experimental materials in different regions were compared and analyzed. The difference in content and correlation, and the cluster analysis method were used to divide clusters of the experimental materials. [Results] The absorbance of proanthocyanidins in the fruit of wild L. ruthenicum was No.4>No.1>No.5>No.6>No.3>No.2, among which the absorbance of anthocyanin(2.43) of wild L. ruthenicum variety No.4 was significantly higher than other experimental materials(P<0.05), and proanthocyanidin of No.2 had the lowest absorbance value of 1.35. There was no significant difference between No.3 and No.6(P>0.05), and there were significant differences among other experimental materials(P<0.05). The content of polysaccharides was: No.3>No.7>No.2>No.4>No.5>No.6>No.1; there was no significant difference between No.3 and No.7(P>0.05), but significantly higher than other materials(P<0.05). Besides, proanthocyanidins and polysaccharides showed significant variability, but there was no consistency in the correlation between them. [Conclusions] In terms of the absorbance of proanthocyanidins, the experimental materials No.1 and No.4 can be classified into a cluster; experimental materials No.2, No.3, No.5 and No.6 can be classified into another cluster. This can provide a theoretical basis for the introduction and breeding of fine varieties.

Structural Characteristics of Lycium ruthenicum Community & Soil Properties on Different Types of Desert Rangeland in the Lower Reaches of Shiyang River

[Objectives] To study structural characteristics of Lycium ruthenicum community and soil properties on different types of desert rangeland in the lower reaches of Shiyang River, and provide theoretical basis for ecological protection and management in the lower reaches of Shiyang River. [Methods] One-way ANOVA analysis and correlation analysis were carried out with the aid of SPSS19.0 software for different types of desert rangeland soils. [Results](i) Among the communities surveyed, there are 31 species belonging to 9 families and 28 genera, including 12 shrub plants, 8 perennial herbs and 11 annual herbs. There are more single genus, occupying 50% of total genus. L. ruthenicum occupies an important position in the community with relatively higher importance value. It plays a dominated role in the community.(ii) In the soils with depths of 0-10 cm, 10-20 cm, 20-40 cm and 40-60 cm in different types of desert rangeland, the organic carbon contents in the saline alkali soil were the highest, with the values of 1.08%, 0.98%, 0.89% and 0.972%, respectively. Compared with that in sandy land, soil organic carbon in saline alkali land(0-10 cm) had the significant difference(P<0.05), available soil potassium in saline alkali land(except the depth of 20-40 cm) and the total phosphorus content in saline alkali land(0-10 cm) had the significant difference(P<0.05), the total soil nitrogen, alkali-hydrolyzable nitrogen, available phosphorus and the total potassium content had no significant difference(P>0.05). Soil organic carbon, available soil phosphorus in saline alkali land, total soil potassium on sand dune and total soil phosphorus in saline alkali land and gravel in different soil layers of the same type of desert rangeland had distinct aggregation effect.(iii) The soil microbial biomass in different types of desert rangeland displayed from high to low as saline alkali land > sandy land > fixed and semi fixed sandy land > gravel. The discrepancy of soil microbial biomass varied in different types of desert rangeland. The four soil enzyme activities in salinization was the highest. Soil invertase activity in saline alkali land in different soil layers were the highest. For different soil layers in the same type of desert rangeland, soil microbial biomass and invertase decreased gradually with the deepening of soil layer.(iv) There was a significant positive correlation between soil organic matter and microbial biomass carbon, microbial biomass nitrogen, phosphatase and sucrase(P<0.01), and a significant positive correlation with urease and catalase(P<0.05). Phosphorus content was significant positively correlated with urease, phosphatase and invertase(P<0.01), and positively correlated with microbial biomass carbon(P<0.05). Soil available potassium content was significant positively correlated with microbial and phosphatase(P<0.01). [Conclusions] The species of L. ruthenicum community and soil properties on different types of desert rangeland in the lower reaches of Shiyang River clearly shows that most species belong to a few families, and a few species belong to most families, and many species are single genus or single family. Soil organic carbon, soil available phosphorus, total phosphorus, soil microbial biomass and enzyme activity in salinization land had distinct surface aggregation effects, so salinization land provides excellent growth environment for the growth of L. ruthenicum, which is of great significance for the protection of L. ruthenicum population.

The Effects of Different Fixation Methods on Quality of Lycium ruthenicum Murr Bud-tea

The effects of three fixation methods( steaming,blanching and microwave fixation) on chemical composition and sensory quality of Lycium ruthenicum bud-tea were investigated.The results showed the fixation technique( the leaf weight 3 g/cm^2,the time 1.5 min,and the temperature 100 ℃) was the best,which was beneficial to the conservation of free amino acid and soluble sugar in L.ruthenicum bud-tea.The loss of chlorophyll was limited,and the score of sensory quality review was higher.

Quality Analysis of Lycium ruthenicum Murr. from Different Producing Areas

[Objectives]The quality of Lycium ruthenicum Murr.from different producing areas was studied.[Methods]The content of proanthocyanidins(UV spectrophotometry),water-soluble vitamins(VB 2,VB 6,VB 12,Vc,niacinamide,and folic acid)),fat-soluble vitamins(VD,and VK 1)(high performance liquid chromatography),and trace elements Cu,Zn,and Fe(flame atomic absorption method)in L.ruthenicum from Ejin Banner of Inner Mongolia,Nuomuhong of Qinghai and Korla of Xinjiang were measured.[Results]The content of proanthocyanidins,VB 2,nicotinamide and VB 12 in L.ruthenicum from Ejin Banner was higher,and the content of Vc,VB 6,Cu,Zn,VD and VK 1 in L.ruthenicum from Nuomuhong was higher,while the content of folic acid and Fe in L.ruthenicum from Korla was higher.The proanthocyanidin content of L.ruthenicum from Ejin Banner was the highest,so it is an ideal plant for extracting proanthocyanidins.[Conclusions]This study provides a theoretical basis for the research,development and utilization of L.ruthenicum from Nuomuhong,Ejin Banner and Korla.

Technical Study on Extraction of Procyanidins in Lycium ruthenicum Murr. Using Sub-critical Fluid 1,1,1,2-Tetrafluoroethane( R134a)and Content Determination from Different Producing Areas

[Objectives] To study the optimal conditions for extracting procyanidins fromLycium ruthenicum Murr. with sub-critical fluid R134 a( 1,1,1,2-tetrafluoroethane) in 1 L extraction kettle. [Methods]Taking the extraction rate of procyanidins as an indicator,the influence of pressure,temperature,and extraction time on extraction rate of procyanidins fromL. Ruthenicum Murr. was studied by single factor experimental methods and orthogonal array design. [Results]The order of factors affecting extraction rate of procyanidins was extraction temperature > extraction pressure > extraction time. The optimum extraction conditions were as follows: the extraction rate of procyanidins fromL. ruthenicum Murr. was the highest with extraction pressure of 1. 2 MPa,extraction temperature of 50℃ and extraction time of 90 min. The content of procyanidins in L. ruthenicum Murr. from different producing areas was determined by vanillin-HCl method under the optimal conditions. [Conclusions] The method has the advantages of easy operation,good selectivity,low extraction temperature and high extraction efficiency,which is suitable for extraction of procyanidins in L. ruthenicum Murr.

氮磷添加对黑果枸杞(Lycium ruthenicum)营养器官非结构性碳水化合物特征的影响

非结构性碳水化合物(NSC)在植物生长、繁殖、防御和生存等方面发挥着基础性作用。为探究黑果枸杞(Lycium ruthenicum)NSC积累的器官间规律及其对土壤氮磷供应水平的响应,以3年生黑果枸杞为对象,测定了3个氮磷比例和供应量条件下根茎叶NSC含量。结果表明:(1)黑果枸杞根、茎、叶NSC组成均以可溶性糖为主,淀粉约占37.3%;器官NSC含量粗根和细根>叶片>新梢>一二年生枝条,各器官可溶性糖含量粗根和细根>叶片>新梢>一年生枝>二年生枝;淀粉含量细根>粗根>二年生枝>一年生枝>叶片和新梢;糖淀比叶片>新梢>粗根>一年生枝>二年生枝>细根。(2)氮磷比和供应量对黑果枸杞根系可溶性糖含量无显著影响,对茎叶NSC含量的影响存在显著交互效应。氮磷比5∶1时,茎叶NSC含量随氮磷供应量升高而增加;氮磷比15∶1时,茎叶NSC含量随氮磷供应量增加显著降低;氮磷比45∶1时,茎叶NSC含量随供应量的增加而显著升高。(3)一年生枝NSC含量于萌芽前达到最大值(5.05%DW),于春梢旺盛生长期降至最低值(3.79%DW);二年生枝NSC含量在萌芽前最低(4.23%DW),于秋季落叶期达到最大值(4.33%DW)。(4)粗根和叶片NSC含量与一二年生枝NSC含量负相关;新梢NSC含量与叶片NSC、二年生枝淀粉含量正相关;一年生枝可溶性糖含量与二年生枝可溶性糖含量负相关;二年生枝淀粉含量与根系淀粉和NSC含量负相关。黑果枸杞通过调整各器官NSC积累及分配应对生长季各器官能量需求和环境氮磷供应变化,在氮磷限制环境中将更多NSC储存在根系,可平衡当前碳同化与未来能量需求的矛盾,增强个体生存能力,也为黑果枸杞应对非生物逆境胁迫提供更大的灵活性。

黑果枸杞(Lycium ruthenicum)种子萌发及幼苗生长对盐胁迫的响应

黑果枸杞(Lycium ruthenicum)是民勤绿洲-荒漠过渡区主要的盐生植物,近年来由于对其营养价值和药用价值的认识,新疆、青海和甘肃等地拟在荒漠-绿洲过渡带开展黑果枸杞种植以改善盐碱土和增加当地农民经济收入。主要通过插枝和种子繁殖黑果枸杞,而盐胁迫对黑果枸杞种子在实验室和大田的萌发以及对黑果枸杞幼苗生长的影响还未知。于2014年在民勤开展了不同盐度溶液对黑果枸杞萌发和幼苗生长的影响研究。结果表明:淡水条件下,黑果枸杞种子的萌发及幼苗生长优于盐水条件;在室内培养试验中,黑果枸杞种子累积萌发数和萌发率随浇灌水盐度的增加而减少;室外大田试验淡水处理下黑果枸杞种子累积萌发数和萌发率高于其他盐度处理;黑果枸杞种子萌发和幼苗生长对盐胁迫较敏感,其形态和生理均显示出抑制特征,不同处理的幼苗根芽比随着培养液盐度的增加而增加。大田种植时,降低表层土壤盐度是黑果枸杞成功建殖的关键。

种植黑果枸杞(Lycium ruthenicum)对盐渍土盐分分布的影响

针对近年来民勤绿洲土壤盐渍化发展现状,通过田间试验初步研究了黑果枸杞(Lycium ruthenicum)在盐渍化土壤改良中的作用。本试验开始前,试验地已经过两年的咸水灌溉,灌溉水盐分浓度分别为0.8g·L-1(低盐)、2g·L-1(中盐)和5g·L-1(高盐)。在试验地移植黑果枸杞后连续两年监测土壤盐分在垂直和水平方向的分布。结果表明:(1)移植黑果枸杞后土壤电导率有明显的季节变化规律。这种季节变化受降水和参考作物蒸散量的影响;(2)种植黑果枸杞后土壤盐分向根际和冠幅下运移导致根际和冠幅下土壤盐分含量升高;(3)种植黑果枸杞可以降低冠幅外土壤盐分浓度。种植前灌溉水的盐分含量越低,黑果枸杞使土壤盐分向冠幅下运移的幅度越大。

骆驼蓬(Peganum harmala)茎叶水浸液对黑果枸杞(Lycium ruthenicum)种子萌发与幼苗生长的化感作用

采用生物测定的方法研究了荒漠植物骆驼蓬(Peganum harmala)对受体植物黑果枸杞(Lycium ruthenicum)种子发芽与幼苗生长的化感作用。结果表明:(1)骆驼蓬茎叶水浸液对黑果枸杞种子萌发和幼苗生长均有抑制作用,且随处理浓度的升高而增强;(2)苗干重和苗鲜重对水浸液作用表现最为敏感,其次为根鲜重和苗高,根长和根干重表现较不敏感;(3)丙二醛含量的综合化感效应指数(IR=34.22)明显大于其他指标,相对电导率也表现出较为敏感的抑制效应,而水浸液对受试植物可溶性蛋白含量的影响最小(IR=0.52)。由此推断,骆驼蓬茎叶水浸液中化感物质多种效应的生物起点可能是对膜系统的伤害。

Physiological characterization,transcriptomic profiling,and microsatellite marker mining of Lycium ruthenicum

Lycium ruthenicum is a perennial shrub species that has attracted considerable interest in recent years owing to its nutritional value and ability to thrive in a harsh environment. However, only extremely limited transcriptomic and genomic data related to this species can be found in public databases, thereby limiting breeding research and molecular function analysis. In this study, we characterized the physiological and biochemical responses to saline- alkaline mixed stress by measuring photochemical efficiency, chlorophyll content, and protective enzyme activity. We performed global transcriptomic profiling analysis using the IIlumina platform. After optimizing the assembly, a total of 68063 unique transcript sequences with an average length of 877 bp were obtained. Among these sequences, 4096 unigenes were upregulated and 4381 unigenes were down-regulated after saline-alkaline mixed treatment. The most abundant transcripts and over-represented items were assigned to gene ontology （GO） terms or Kyoto Encyclopedia of Genes and the Genomes （KEGG） categories for overall unigenes, and differentially expressed unigenes were analyzed in detail. Based on this set of RNA-sequencing data, a total of 9216 perfect potential simple sequence re- peats （SSRs） were identified within 7940 unigenes with a frequency of 1/6.48 kb. A total of 77 primer pairs were synthesized and examined in wet-laboratory experiments, of which 68 loci （88.3%） were successfully amplified with specific products. Eleven pairs of polymorphic primers were verified in 225 individuals from nine populations. The inbreeding coefficient and the polymorphism information content value ranged from 0.011 to 0.179 and from 0.1112 to 0.6750, respectively. The observed and expected heterozygosities ranged from 0.064 to 0.840 and from 0.115 to 0.726, respectively. Nine populations were clustered into three groups based on a genetic diversity study using these novel markers. Our data will be useful for functional genomic investigations of L. ruthenicum and could be used as a basis for further research on the genetic diversity, genetic differentiation, and gene flow of L. ruthenicum and other closely related species.

黑果枸杞(Lycium ruthenicum)分布对气候变化的响应及其种植适宜性

基于生态位理论与MaxEnt模型,分析影响黑果枸杞(Lycium ruthenicum)分布的主要气候因子,划分其适宜种植区,并探究气候变化对黑果枸杞种植适宜性的影响,进而为引种驯化黑果枸杞提供理论依据和技术支持。结果表明:(1)影响黑果枸杞种植分布的主要因子为最热季降水量、温度年较差、年平均气温、最冷季平均温度、降水量的季节性变化、最湿月降水量、最干季平均温度、年降水量及海拔。(2)在当前气候情景下,黑果枸杞的适宜种植区面积约为207 382.8km2,主要分布在河西走廊及其周边、柴达木盆地、塔里木盆地、准格尔盆地、吐鲁番盆地。(3)在未来不同的温室气体排放情景下,黑果枸杞的适宜种植面积均有不同幅度的扩大,但不受气候变化影响的黑果枸杞适宜种植区面积将逐渐减小。

黑果枸杞(Lycium ruthenicum)组织培养与再生体系的建立

为了探索野生黑果枸杞组织培养与再生体系建立的适宜环境条件,以采自青海省格尔木地区野生黑果枸杞浆果的种子为供试外植体材料,通过制备一系列不同激素与浓度的培养基组,以诱导黑果枸杞组织分化与再生植株的建立。结果表明:(1)黑果枸杞浆果的种子经由8%的次氯酸钠浸种7 min,无菌水泡洗2次,再用浓度为75%的消毒酒精浸种2 min,无菌水泡洗2~3次后浸种后,接种在M3(6-BA 0.0 mg/L+2,4-D 0.0 mg/L+MS)培养基中出苗率为85%。(2)无菌苗茎和叶剪短至3~5 cm后,经过杀菌消毒处理后接种到Y4(6-BA 0.3 mg/L+IAA 5.0 mg/L+MS)最适愈伤组织培养基上,出愈率为86.7%。(3)愈伤组织丛生芽诱导的最适宜培养基为C3(6-BA 0.3 mg/L+NAA 0.05 mg/L+MS),不定芽分化率为80%;不定芽剪段后继续在C3上进行壮苗扩繁培养,成活率为92%。(4)壮苗扩繁后组培苗最适宜生根培养基为G3(NAA 0.2 mg/L+IBA 0.2 mg/L+1/2MS),植株生根率为41.6%。上述研究结果可为野生黑果枸杞组织苗的工厂化繁育提供参考依据。

基于高遗传转化率及低幼苗玻璃化率的黑果枸杞叶片组合培养体系

【目的】建立高效稳定的黑果枸杞遗传转化体系,有效降低其再生幼苗玻璃化率,促进其基因功能研究和提高遗传改良效率。【方法】以黑果枸杞叶片作为外植体,利用农杆菌(LBA4404、EHA105)介导的遗传转化法,通过调整基础培养基类型并添加相应浓度的植物激素,筛选出最适愈伤组织诱导培养基、分化和选择培养基、生根诱导培养基,将黑果枸杞遗传转化率提高到65%以上,同时降低幼苗玻璃化率至10%以下。【结果】(1)黑果枸杞叶片高效组合培养体系中最佳农杆菌侵染浓度(OD_(600))为0.6,侵染时间为25 min,在此条件下侵染叶片抗性愈伤诱导率达78.2%~96%;(2)黑果枸杞遗传转化中最适分化和选择培养基为:MS+肌醇50 mg/L+烟酸0.25 mg/L+维生素B60.25 mg/L+铁盐母液1 mL/L+甘氨酸1.0 mg/L+维生素B10.05 mg/L+6-BA 0.25 mg/L+蔗糖30 g/L+琼脂6 g/L+卡那霉素30 mg/L+特美汀300 mg/L(pH=6.0);最适生根诱导培养基为:WPM+IBA 0.25 mg/L+蔗糖30 g/L+琼脂6 g/L+卡那霉素30 mg/L+特美汀300 mg/L(pH=6.0);(3)在最适分化和选择培养基中,农杆菌LBA4404-pBI121侵染叶片外植体产生的幼苗玻璃化率约为65%,而农杆菌EHA105-pBI121侵染处理则低于10%;(4)采用木本植物低盐WPM培养基可使黑果枸杞再生幼苗生根效率达到81.2%左右。(5)将阳性愈伤数量与总接种叶片数量比值作为遗传转化效率的评价指标,在最适遗传转化体系中,农杆菌LBA4404-pBI121和EHA105-pBI121侵染后遗传转化率分别为51%和65.2%。【结论】黑果枸杞叶片高效组合培养体系能够显著提高其遗传转化率,降低玻璃化幼苗的发生率。

改进和优化黑果枸杞及其制品中花青素测定的pH示差法

建立一种基于美国官方分析化学师协会(Association of Official Analytical Chemists,AOAC)方法检测黑果枸杞及其制品中花青素含量的改进pH示差法。考察了黑果枸杞及其制品中花青素的最佳提取和检测条件,通过液相色谱-三重四级杆串联质谱法鉴别出黑果枸杞中花青素的具体化学结构,并计算出混合花青素的平均摩尔质量。通过分光光度法测得混合花青素的平均摩尔消光系数,对改进后的pH示差法进行方法学验证和花青素的含量测定。结果显示,最佳提取和检测条件如下:黑果枸杞花青素提取溶剂为盐酸-80%(体积分数)乙醇(3∶97,体积比),料液比为1∶100(g∶mL),提取温度为50℃,提取时间为30 min,缓冲溶液稀释5倍后静置平衡20 min。液相色谱-三重四级杆串联质谱法鉴别黑果枸杞中主要以矮牵牛素类花青素为主(占97.96%),黑果枸杞特有的混合花青素平均摩尔质量为912.7 g/mol,平均摩尔消光系数为29591 L/(mol·cm)。pH示差法改进后能够满足方法学验证要求,固体样品和液体样品最低检出限分别为28.2 mg/100 g、0.282 mg/100 mL。方法改进后花青素提取增长率均大于20%,静置平衡20 min后单次检测结果精密度小于0.3%。以矮牵牛素类花青素代替矢车菊素-3-O-葡萄糖苷计算花青素含量平均提高了2.41倍,能真实地反映黑果枸杞及其制品中花青素的含量。

黑果枸杞中矿物元素溶出特性研究

黑果枸杞,茄科枸杞属,富含多种矿物质元素,这对人体生长发育有重要的生物学作用。黑果枸杞作为一种药食同源的物质,为了解其营养价值、指导健康食用及进一步功能开发,有必要分析矿物质溶出特性,并进行相关性分析。黑果枸杞样品制备以硝酸和过氧化氢为消解酸,微波消解仪制备消解液,以不同温度纯水浸提制备浸泡液,试验建立了电感耦合等离子体光谱法测定消解液和浸泡液中K、Na、Ca、Mg、Zn、Fe、Mn、Cu、Li、Sr十种矿物元素含量的实验方法,并进行了矿物元素溶出特性研究及相关性分析。在优化的光谱分析条件下,十种元素线性关系好(r>0.9995)、检出限低(<0.01 mg·L^(-1))、准确度高、精密度好。加标回收率在97.2%~101.8%之间,相对标准偏差小于1.99%。实验发现,黑果枸杞中矿物元素含量丰富,含量最高的是K,其次是Na、Ca、Mg,然后是Fe、Li和Sr,含量较低的是Mn、Zn、Cu。黑果枸杞中K/Na值大于8,属于典型的高钾低钠食品。Ca、Mg、Fe、Zn、Li、Sr、Mn、Cu也是人体中多种酶、激素和维生素中不可缺少的活性因子,是一种理想的食品。黑果枸杞浸泡液中,除Li与Ca、Zn、Fe之间,Mg与Zn之间关系不明显外,其余元素间呈显著正相关或极显著正相关。表明,各矿物质元素中不存在明显的拮抗作用。矿物质元素的溶出率与浸泡时间及提取温度有关。实验采用沸水和40℃纯水浸泡黑果枸杞样品,选取浸提时间5、10和20 min三个时间点研究提取温度及时间对溶出率的影响。十种元素中K、Na、Zn溶出率最高,可能是因为三种元素多以离子形式存在,容易提取。Ca、Mg、Sr、Mn、Cu含量差异大,溶出率相近,可能是价态相近,提取效率相近。Li溶出率受提取温度影响很大,沸水浸泡溶出率是40℃温水的的1.5~2倍,Fe在微量元素中含量最高,溶出率却最低,主要是由于Fe与蛋白质结合,不易萃取。试验数据可为健康食用黑果枸杞及进一步开发利用提供理论参考。