蔬菜中邻苯二甲酸酯的残留研究

实验建立气相色谱-质谱法同时测定蔬菜（白菜、芹菜、番茄和黄瓜）中18 种邻苯二甲酸酯。初步开展对安徽省基地蔬菜中邻苯二甲酸酯残留监测,为指导消费者蔬菜安全膳食,向管理者提出风险管理建议提供理论依据。研究结果表明：18 种邻苯二甲酸酯的质量浓度在0.05~5.0 mg/L 范围内呈良好线性,相关系数为0.9972~0.9998,最小检出量(S/N=3)为0.05 ng;在0.5~5.0 mg/kg 加标水平下的该方法平均回收率为72.28%~112.57%、相对标准偏差(RSD,n=5)为0.94%~12.47%,方法可满足蔬菜中邻苯二甲酸酯的检测要求。采用该方法对蔬菜基地4 种蔬菜中邻苯二甲酸酯进行残留风险监测,样品中检出了邻苯二甲酸二异丁酯(DIBP)、邻苯二甲酸二（2-丁氧基）乙酯(DBEP)、邻苯二甲酸二（2-乙基）己酯(DEHP)和邻苯二甲酸二壬酯(DNP),其总含量为0.20~0.72 mg/kg,而DEHP在4 种蔬菜中均有检出、残留量为0.02~0.22 mg/kg。     

巴戟天胚乳诱导愈伤组织及三倍体再生

目的:利用巴戟天种子内胚乳为外植体进行离体培养获得三倍体。方法:选取未成熟巴戟天种子内胚乳,置于不同培养基上诱导愈伤组织和分化再生植株,并对植株倍性进行根尖染色体鉴定。结果:在MS+2,4-D(2,4-二氯苯氧乙酸)2.0 mg/L+6-BA(6-苄基嘌呤)1.0 mg/L培养基上诱导获得愈伤组织,诱导频率达58.9%。愈伤组织在MS+6-BA1.0~2.0 mg/L+IBA 0.1~0.5 mg/L分化培养基上分化出不定芽。不定芽在1/2 MS+IBA 0.5 mg/L上诱导生根,生根率100%。再生植株根尖细胞染色体数目为2 n=3 x=33。结论:利用巴戟天胚乳培养获得三倍体是创造多倍体新种质的有效途径。     

除草剂氯嘧磺隆在培养基中的提取和检测方法研究

试验建立了黑曲霉(TR-H)在基础培养基中对氯嘧磺隆除草剂降解的提取和检测方法。结果表明:采用二氯甲烷作为提取溶剂,氯嘧磺隆在基础培养基中的添加回收率均大于96.0%,标准偏差均小于4.0%。采用甲醇-水(pH2.5,用85.0%的磷酸调节)作为液相色谱的流动相体系,可将氯嘧磺隆与其它物质较好的分离。氯嘧磺隆的浓度在0.1~200.0mg/L范围内具有良好的线性关系,相关系数为r=0.9998(n=3)。氯嘧磺隆在基础培养基中添加的最低浓度应在0.05mg/L(S/N=3)以上方能被检出;此外,氯嘧磺隆在菌体上的吸附均低于其检出限。     

饼肥与无机肥的不同配比对白肋烟烟叶中的游离氨基酸的影响

建立了一种用于烟草中游离氨基酸测定的反相高效液相色谱法。实验采用超声波水解、邻苯二甲醛/3-巯基丙酸作为衍生剂进行柱前衍生。色谱柱为依利特C18柱(4.6mm i.d. ×250mm,5μm),流动相A为18mmol/L的醋酸钠溶液（pH7.2）含体积分数为0.002%的三乙胺和0.3%的四氢呋喃,流动相B组成为：100mmol/L的醋酸钠溶液（pH7.2）-乙腈-甲醇（体积比为1:2:2）,流速为1.0ml/min,柱温为40℃。荧光检测器,激发波长350nm,发射波长450nm。方法的回收率为95.3% ~100.7%,RSD为2.32%~9.24%(n=6)。该方法简便、准确、重现性好。测定不同肥料配比生产的白肋烟烟叶中17种游离氨基酸的含量。结果表明,不论有机肥与无机肥怎样配比,天冬氨酸的含量与各氨基酸相比都是最高的;随着饼肥的加入,大部分氨基酸的含量是先增加后逐渐降低的趋势, 15%饼肥+85%无机肥与30%饼肥+70%无机肥配比时,大部分游离氨基酸的含量接近,30%饼肥+70%无机肥配比时的游离氨基酸总量最高,综合考虑,30%饼肥+70%无机肥配比时的烟叶质量最好。     

高效液相色谱法同时测定饮料中的5种添加剂

建立了一种高效液相色谱法(HPLC)同时测定饮料中安赛蜜、糖精钠、苯甲酸、山梨酸、脱氢乙酸5种食品添加剂检测技术。采用Venusil ASB C18色谱柱(4.6 mm×250 mm,5μm),以浓度0.02 mol/L的乙酸铵-甲醇(90∶10,V∶V)为流动相,柱温35℃,用二极管阵列检测器在波长230 nm处进行检测,分离过程在10 min内完成。安赛蜜、糖精钠、苯甲酸、山梨酸、脱氢乙酸的检出限分别为1.5,0.8,0.5,0.5,1.2 mg/kg,在1.0～50.0μg/m L内具有良好的线性关系(R20.999 0)。加标回收率在80%～100%,相对标准偏差小于1.0%(n=7)。该方法以水作为溶剂并超声提取,然后再加入沉淀剂除杂,简单、快速、准确、实用性强。     

高效液相色谱法测定辣椒中的辣椒碱类成分

建立了高效液相色谱法同时测定辣椒中辣椒碱和二氢辣椒碱含量的方法。采用Ultimate AQ-C18型色谱柱(4.6 mm×250 mm,5μm),流动相为水-甲醇(40∶60,V/V),流速1.0 mL/min,柱温30℃,检测波长280 nm。辣椒碱和二氢辣椒碱在0.001~0.150 mg/m L质量浓度范围内线性关系良好(R~2=0.999 9);加样回收率均值分别为99.35%和98.26%,RSD分别为0.82%和0.73%(n=9);稳定性试验RSD为0.27%;中间精密度RSD低于3.79%(n=18)。该法准确度高、重现性好,能快速测定辣椒中辣椒碱和二氢辣椒碱的含量。     

高效液相色谱-荧光检测法测定淀粉制品中孔雀石绿隐形孔雀石绿残留量的研究

采用高效液相色谱-荧光检测的方法检测粉条等淀粉制品中孔雀石绿和隐形孔雀石绿的残留量;通过乙腈提取、氮吹浓缩、酸性氧化铝小柱净化的方法分离出淀粉制品中的目标物;通过C_(18)-ODS柱(5μm,4.6 mm×250 mm)(柱后接25%的二氧化铅氧化柱),以乙腈与0.1 mol/L的乙酸铵缓冲溶液(用冰乙酸调节p H值至4.5)体积比80∶20为缓冲液,实现孔雀石绿和隐形孔雀石绿的分离。于波长588 nm和618 nm处测定2种物质的质量浓度在0.3~8.0μg/m L线性关系良好,相对偏差(n=7)小于2.5%,检出限为0.08μg/m L,分析时间为20 min。以粉条为基体进行回收试验,回收率为77.9%~89.5%。     

容舒口服液原药材HPLC指纹图谱的建立及成品的质量评价

目的 建立容舒口服液单味药材的HPLC指纹图谱,并建立成品口服液HPLC检测方法。方法 对不同产地的口服液原药材酸枣仁、当归、五味子和玫瑰进行HPLC指纹图谱评价。建立以槲皮素和山奈素为指标的容舒口服液的HPLC检测方法,色谱条件：Welch Ultimate XB-C₁₈ (4.6×250 mm, 5 μm);甲醇：0.4%磷酸溶液(40:60),360nm; 1mL.min_-1; 40℃;10 μL。结果 不同产地的主药材的HPLC指纹图谱峰形区分度较高,共有峰相似度高于98.2%,可以显著区别不同产地药材的含量差异。成品HPLC检测方法建立成功,检测指标槲皮素(5.79-38.6 μg.mL_-1,r=0.9999)和山奈素(3.68-24.55 μg.mL_-1,r=0.9999)在规定范围内线性关系良好;平均加样回收率分别为100.89%和99.86%,RSD值分别为1.96%(n=6)和1.39%(n=6);结论 通过指纹图谱法和HPLC法控制容舒口服液的质量,结果准确,方法稳定可行。     

3种生物杀虫剂防治烟草烟粉虱的室内毒力及田间药效试验

为了筛选控制烟草烟粉虱的有效药剂,采用3种生物杀虫剂对烟草烟粉虱进行室内毒力测定及田间药效试验。室内毒力测定结果表明,对烟草烟粉虱的毒力顺序为：0.5%依维菌素乳油＞0.5%藜芦碱可溶液剂＞6.0%鱼藤酮微乳剂,其LC50值分别为1.396 mg/L、5.083 mg/L、12.287 mg/L,显著优于对照药剂高效氯氟氰菊酯(LC50值为30.67 mg/L)。田间药效试验结果表明,6.0%鱼藤酮微乳剂、0.5%依维菌素乳油,0.5%藜芦碱可溶液剂稀释2000倍液喷雾处理后,药后3d、7d、14d更正防效对烟草烟粉虱持效性较好,更正防效达70﹪以上。因此,3种生物杀虫剂对烟田未产生药害,烟草植株生长安全,可在生产上进一步推广应用。     

毛细管气相色谱内标法测定防霉剂中富马酸二甲酯

建立了巴豆酸为内标物,用毛细管气相色谱快速测定防霉剂中富马酸二甲酯含量的方法.样品用乙酸乙酯提取,以DM-FFAP毛细管柱分离,火焰离子化检测器检测.结果表明,在0.25～3.0 mg/mL浓度范围内,富马酸二甲酯和巴豆酸的浓度比与峰面积比的回归方程为y=-0.0009 0.7174x,相关系数r=0.9996,检出限为5.0 mg/L;样品加标回收率(n=6)为97.0%～104.0%,相对标准偏差为3.26%.该方法简单、快速、准确度好,可用于测定防霉剂中的富马酸二甲酯含量.     

伞形科植物挥发油杀螨活性筛选及成分研究

为了研究6种伞形科植物挥发油的杀螨效果,分析活性最好的植物挥发油的主要活性成分。采用水蒸气蒸馏法提取香菜、白芷、柴胡、大齿山芹、辽蒿本、川芎6种伞形科植物挥发油。以玻片浸渍法测定其挥发油杀螨活性后,使用气质联用仪(GC-MS)对具有较好杀螨活性的挥发油化学成分进行分析。结果表明香菜、白芷、柴胡、大齿山芹、辽蒿本、川芎挥发油杀螨的半数致死浓度(LC50)分别为2.9006、1.1575、3.2386、1.6314、1.8854、0.3767 mg/mL,其中以川芎挥发油的杀螨活性最强,其主要成分为4-萜烯醇(11.81%)、双环(2,2,1)庚烷-2-醇1,7,7-三甲基-2-乙酸酯(8.28%)、4-乙烯基-2-甲氧基苯酚(14.88%)、苯戊酮(7.92%)、苯己醇(9.87%)。本研究首次发现川芎挥发油对朱砂叶螨具有较好的触杀作用。毒杀效果与市售常用杀螨剂螺螨酯(LC50=356.8878 mg/L)相对持平。因此推测4-萜烯醇可能是杀螨的活性成分之一。     

催眠睡茄的离体快繁及四倍体的诱导

以催眠睡茄腋芽为外植体建立快繁体系,利用秋水仙素进行催眠睡茄四倍体诱导,并将其与二倍体进行了初步的形态学和细胞学方面的比较.结果发现:催眠睡茄二倍体(2n=2x=48)最适增殖培养基为MS 6-BA 0.5 mg/L NAA 0.05 mg/L,生根培养基为1/2 MS IBA 0.2 mg/L;四倍体(2n=4x=96)诱导率因秋水仙素浓度和处理时间的不同而异,其中用0.5%的秋水仙素对丛生芽浸泡处理48 h诱导率最高,诱导率可达13.79%.四倍体与二倍体相比:叶片明显变厚,颜色加深,叶片气孔及其保卫细胞大小增大,气孔密度减小,且保卫细胞内叶绿体数目增加.     

Amphidiploids between Cyclamen persicum Mill. and C. purpurascens Mill. induced by treating ovules with colchicine in vitro and sesquidiploids between the amphidiploid and the parental species induced by conventional crosses

Summary We cultured colchicine-treated hybrid ovules in vitro to produce fertile amphidiploids of C. persicum (2n=2x=48. referred to as AA) × C. purpurascens (2n=2x=34, referred to as BB). Seedlings and mature plants were obtained from the ovules without colchicine and those exposed to 50 mg/l colchicine for 5, 10 and 15 days, whereas they were not obtained from the ovules exposed to 50 mg/l colchicine for 20 days and 500 mg/l for 5, 10, 15 and 20 days. Although 8 mature hybrids derived from the ovules without colchicine produced a few fertile pollen grains, they failed to produce viable seeds by self-fertilization. The hybrids had 41 somatic chromosomes. Four and 3 mature plants were derived from ovules exposed to 50 mg/l colchicine for 10 and 15 days, respectively. One each among 4 and 3 mature plants showed a high frequency of pollen grain fertility, produced several seeds by self-fertilization, and had 82 somatic chromosomes which is twice the number of hybrid chromosomes (2n=41, AB). These findings indicated that these plants are amphidiploids (2n=82, AABB) between C. persicum and C. purpurascens. Three and 2 viable seeds were derived by the conventional crosses of diploid C. persicum × the amphidiploid and the amphidiploid × C. purpurascens, respectively. Flowering plants that developed from the seeds of diploid C. persicum × the amphidiploid were barely fertile and had 65 somatic chromosomes (2n=65, AAB), whereas those that developed from the seeds of the amphidiploid × C. purpurascens were barely fertile and had 58 somatic chromosomes (2n=58, ABB). The somatic chromosomes indicated that these plants are probably sesquidiploids between the amphidiploid and either C. persicum or C. purpurascens. The interspecific cross-breeding of cyclamen using the amphidiploids and the sesquidiploids is discussed.     

莲蓬壳中挥发油成分的定性和定量分析

为研究莲蓬壳中挥发油的成分及含量,以莲蓬壳为试材,采用索氏提取法从莲蓬壳中提取出挥发油,并通过气相色谱-质谱联用技术（GC-MS）结合计算机检索系统对其挥发油的主要化学成分进行定性和定量分析。结果表明,莲蓬壳挥发油共鉴定出32种化学成分,占总挥发性物质含量的99.83%,主要分为烃类、醇类、酮类、酯类、酚类、酸类等7类化合物。通过面积归一化法对其含量进行分析,研究发现莲蓬壳挥发油的主要成分为?琢-香树精、?茁-香树精、12-齐墩果烯-3-酮、?茁-谷甾醇油酸酯、谷甾醇、乙酸环阿屯酯、羽扇烯酮等,其中,?琢-香树精含量最高,为27.93%。本研究确定了莲蓬壳中挥发油的主要化合物组成,为合理开发和利用莲蓬壳提供理论依据。     

Somatic cell genetic studies inBrassica species. II. Production of androgenetic haploid plants inBrassica nigra (L.)Koch

Summary Callus growth and its subsequent regeneration into complete plantlets was achieved from in vitro cultured anthers ofBrassica nigra (L.)Koch. Callus was induced on a modified N6 medium containing trace elements, organics of B5 medium and 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Morphogenesis of callus in the form of shoots on MS medium containing indole-3-acetic acid (IAA) and N⁶-benzyladenine (BA) 0.5 mg/l each and embryoids on MS medium containing 0.5–1.0 mg/l IAA and 3.0–5.0 mg/l BA could be accomplished. Chromosomal analysis revealed presence of 41% haploids (n=8) amongst the regenerated plants.     

Hempseed as a nutritional resource: An overview

Summary The seed of Cannabis sativa L. has been an important source of nutrition for thousands of years in Old World cultures. Non-drug varieties of Cannabis, commonly referred to as hemp, have not been studied extensively for their nutritional potential in recent years, nor has hempseed been utilized to any great extent by the industrial processes and food markets that have developed during the 20th century. Technically a nut, hempseed typically contains over 30% oil and about 25% protein, with considerable amounts of dietary fiber, vitamins and minerals. Hempseed oil is over 80% in polyunsaturated fatty acids (PUFAs), and is an exceptionally rich source of the two essential fatty acids (EFAs) linoleic acid (18:2 omega-6) and alpha-linolenic acid (18:3 omega-3). The omega-6 to omega-3 ratio (n6/n3) in hempseed oil is normally between 2:1 and 3:1, which is considered to be optimal for human health. In addition, the biological metabolites of the two EFAs, gamma-linolenic acid (18:3 omega-6; GLA) and stearidonic acid (18:4 omega-3; SDA), are also present in hempseed oil. The two main proteins in hempseed are edestin and albumin. Both of these high-quality storage proteins are easily digested and contain nutritionally significant amounts of all essential amino acids. In addition, hempseed has exceptionally high levels of the amino acid arginine. Hempseed has been used to treat various disorders for thousands of years in traditional oriental medicine. Recent clinical trials have identified hempseed oil as a functional food, and animal feeding studies demonstrate the long-standing utility of hempseed as an important food resource.     

In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.)

A protocol for in vitro induction of tetraploids via colchicine-treated somatic embryos from immature zygotic embryos of diploid grapevine (Vitis vinifera L.) is reported. Embryogenic callus was initiated from immature zygotic embryos cultured on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). The callus was transferred to NN medium containing 1.0 mg/l α-naphthalene acetic acid (NAA) and 0.5 mg/l benzyladenine (BA) to establish somatic embryogenesis. The vigorously growing globular embryos were selected and treated by 0, 10 or 20 mg/l colchicine for 1, 2 or 3 days, and then immediately transferred to NN medium supplemented with 0.03 mg/l NAA and 0.5 mg/l BA, for somatic embryo conversion and plant regeneration. The number of surviving embryos and regenerated plantlets following colchicine treatment decreased with increasing colchicine concentration and treatment time. Among 29 randomly investigated plantlets regenerated from colchicine-treated somatic embryos, five solid tetraploids (2n = 4× = 76) were identified by chromosome counting analysis; all others were diploid (2n = 2× = 38). Ploidy level of plant regenerated was also determined from leaves using flow cytometry. No chimeras with both 2C and 4C nuclei was produced from colchicine-treated somatic embryos. Significant differences in leaf stomata parameters were observed between diploid and induced tetraploid plantlets.     

Optimization of mature embryo-based high frequency callus induction and plant regeneration from elite wheat cultivars grown in China

We have developed an efficient procedure for plant regeneration of elite wheat cultivars using mature embryos. Firstly, we established the optimal combination of basal media, inoculation method and pretreatment method using biostatistical methods. The results indicated that the combination of MS medium and longitudinally bisected mature embryos showed the highest culture efficiency, whereas the pretreatment method had no significant effects on callus induction or plant regeneration. A 70% primary callus induction rate was achieved on MS medium containing 2 mg/l 2,4‐d for all tested cultivars. Primary calli were then transferred onto the subculture medium to initiate embryogenic calli. Supplementation of the subculture medium with the appropriate combination of phytohormones (2.0 mg/l 2,4‐d , 0.5 mg/l BA and 0.1 mg/l NAA) significantly enhanced embryogenic callus production. The addition of AgNO₃ (10 mg/l) in regeneration medium promoted plant regeneration, whereas CuSO₄ stimulated root formation. The use of this protocol achieved successful plant regeneration in eight tested cultivars. The culture efficiency ranged from 15.3% to 36.8%, suggesting this regeneration system may be an effective alternative for wheat genetic transformation.     

普通小麦(Triticum aestivum)和毛穗赖草(Leymus paboanus)的杂交,杂种细胞无性系的建立及植株再生

以3个普通小麦品种富可(Fuhuko)、中国春(Chinese Spring)及小偃759和毛穗赖草杂交,发现三个品种都可与毛穗赖草杂交,其中Fuhuko×L.paboanus平均结实率高达17.6％,杂种只有发育不全的幼胚而无胚乳。对杂种幼胚在N_6+1—2mg/11BA+0.5mgGA_3或MS(其中NH_4NO_3含量降低一半)附加1mg/1IBA的培养基上进行保姆培养,部分幼胚发育成