应用反相高效液相色谱定性和定量浮游植物光合色素

利用本实验室建立的反相高效液相色谱实验体系分析了19 种高纯度的浮游植物光合色素标准样品的特征参数。结果显示, 该体系可将各种光合色素较好分离, 但个别色素的保留时间、洗脱顺序、最大吸收波长及响应因子与文献报道的其他实验体系有所差异。表明在利用反相高效液相色谱定性、定量光合色素时不仅要参考其保留时间, 还要参考其最大吸...     

厦门港表层沉积物对海水中铅的吸附动力学

模拟研究厦门港表层沉积物对海水中Ｐｂ的吸附动力学及水环境条件的影响,当海水温度由２０－３０℃、ＰＨ５－８和盐度３０－４变化时,吸附速率分别提高约５０％、、个数量和５倍。吸附速率与盐度之间呈负指数相关关系。吸附过程可用交换吸附动力学模式描述。测定吸附速率常数、表观活化能和平衡吸附量等参数研究表明,沉积物对Ｐｂ的吸附性质以华东作用为主。     

光合色素标记法测定不同类群浮游植物的光合速率和生长速率

尝试利用放射性同位素14C示踪方法测定浮游植物不同类群的光合速率和生长速率,并通过实验室和现场海域的培养实验来初步评估该技术的可行性.通过高效液相色谱法分离光合色素,手动收集并测定各对应光合色素峰的放射性活度,依据Redalje-Laws法并结合类胡萝卜素标记法计算出各大类群浮游植物的光合速率和生长速率.实验结果表明,在实验室纯种培养的4株藻光合速率范围为347.73~678.98 mg/（dm3.d）,最高的为绿藻的小球藻,最低的为定鞭金藻的球石藻.其生长速率范围为0.12~0.24 d-1,最高的为球石藻,最低的为小球藻.在现场海域各类群的光合速率相对较低,变化范围为0.63~68.78 mg/（dm3.d）,生长速率在0.07~1.68 d-1之间.本研究首次在我国海域通过现场原位测定得到不同类群浮游植物的光合速率和生长速率.但在测定过程中存在的样品含量和放射活度较低等问题还有待于进一步解决.     

近岸海域光合色素的生物标志作用研究Ⅰ.台湾海峡特征光合色素的分布及其对浮游植物类群结构的指示

１引言 由于高效液相色谱法（ＨＰＬＣ）的建立,利用光合色素,尤其是类胡萝卜素作为区分浮游植物类群的指示物,成为更方便和有效的化学分类法［１,２］．原因是色素能够很好地进行定量,可涵盖浮游植物的全部粒级范围;而且,适用于大批量样品的分析,甚至可实现现场的船上测定．海洋浮游植物光合色素的研究,比较多地集中在开放大洋,多为利用类胡萝卜素表征浮游植物的类群结构［３－５］;或以光合色素及其降解产物作为浮游植物参与的生物地球化学过程的生物标志物［６－８］．相比较而言,在近岸、陆架海域开展的此类研究较少,且多…     

厦门筼筜湖潟湖底栖微藻光合色素的时空变化特征及与环境因子的关系

2010年7月至2011年7月,利用高效液相色谱分离分析技术研究了厦门筼筜湖潟湖表层沉积物的底栖微藻生物量(以叶绿素a计)和光合色素组成,探讨了其与环境因子的关系.结果表明,表层沉积物中检出岩藻黄素、别藻黄素、玉米黄素、叶绿素b四种光合色素,对应的微藻类群包括硅藻、隐藻、蓝藻和绿藻.底层(表层沉积物)与水层(水体)微藻叶绿素a生物量呈显著负相关关系(p＜0.05),但底层-水层对应的光合色素不存在显著相关关系,此暗示底层光合色素主要是底栖微藻的贡献,而非来自水层浮游微藻的沉降.底栖微藻生物量随时间变化明显,夏季(6～8月)较低(1.30 mg/m2),冬季(12月至翌年2月)较高(86.16 mg/m2).各测站底栖微藻生物量存在空间差异,随湖区的水深增加而降低,与沉积物表层光辐照度呈显著的正相关关系(p＜0.05).岩藻黄素与降雨量呈显著负相关,别藻黄素与水体无机氮含量呈显著正相关,玉米黄素分布主要受温度影响,除玉米黄素外,其他光合色素含量与光辐照度呈正相关.初步估算表明,底栖微藻占该浅水潟湖生态系统水层-底层微藻叶绿素a生物量和固碳量的27.64％和12.56％.     

HPLC方法分析表层海水叶绿素a含量

以ODS-2 Spherisorb C18反相HPLC分析柱,流动相A(体积比为80：20的甲醇,pH=7．2的0．5mol／L乙酸氨水溶液,0.01％BHT W：V)、流动相B(87．5：12．5的乙腈：水,0.01％BHT W：V)和流动相C(乙酸乙酯),梯度洗脱,流速为1mL／min,紫外检测波长为436nm,HPLC方法分析辽东湾表层海水中的叶绿素a含量。该方法灵敏度高,检测限为10ng／L。对HPLC、荧光法和分光光度法的叶绿素a分析结果进行了比较。     

基于光合色素化学分类和流式细胞术结合的珠江口自养微微型浮游生物种群时空分布特征

为探究珠江口海域自养微微型浮游生物种群时空分布特征及其与环境之间的关系,于2013年5～11月,运用高液相色谱(HPLC)法和流式细胞术对珠江口海域表层水体中微微型浮游生物进行测定。流式细胞计数结果显示,珠江口海域自养微微型浮游生物由聚球藻(Synechococcus, Syn)和微微型真核生物(Picoeukaryotes,PEUK)组成。聚球藻始终占据总细胞丰度的主导地位。光合色素化学分类法(Chemotaxonomy,CHEMTAX)分析表明,自养微微型浮游生物群落结构具有明显的季节性变化,春季和夏季生物量以聚球藻为主,秋季生物量以青绿藻为主。CHEMTAX分析和流式细胞计数结果的相关性分析表明,在春季和夏季Syn细胞丰度与CHEMTAX生物量(即Syn贡献chla)之间呈现极显著正相关(P<0.01),PEUK细胞丰度与CHEMTAX生物量(即PEUK贡献chla)也存在显著正相关(P<0.05);然而,在秋季则无显著性相关关系(P>0.05)。冗余分析表明,温度和营养盐浓度是影响自养微微型浮游生物群落分布与组成的重要因素。另外,盐度、透明度、悬浮颗粒物对自养...     

基于反相C8色谱柱的海洋浮游藻色素高效液相色谱分析方法有效性验证

对一种海洋浮游藻色素高效液相色谱分析方法进行了有效性验证。该方法使用反相C8色谱柱,以甲醇、乙腈和丙酮为流动相,并在流动相中添加吡啶/醋酸溶液作为修饰剂改善色素峰的分离效果。结果表明,4种色素标准在一定的浓度范围内线性关系良好,叶绿素a的线性范围为189.6~18 960μg/L,叶绿素b为89.2~8 920μg/L,β,β-胡萝卜素为7.77~777.2μg/L,叶黄素为15.28~1 528μg/L。除β,β-胡萝卜素外各色素标准的回收率在不同浓度下均在90%以上,相对标准偏差也普遍低于5%。在4个参考藻种和123个胶州湾现场样品中共检测出37种色素,一些关键的特征色素均获得了良好的分离效果。该方法操作简便,具有分离度好、灵敏度高、重复性好、回收率高等优点,适用于培养的藻种和现场样品中的海洋浮游藻色素分析。     

固相萃取-高效液相色谱-荧光检测法测定海水中的多环芳烃

建立了用固相萃取-高效液相色谱-荧光检测法测定海水中多环芳烃的方法,优化了色谱条件和萃取条件。除苊不能用荧光检测器检出外,其余15种多环芳烃的空白加标回收率在64.5%(苯并[g,h,i],茚并[1,2,3-cd]芘)～88.7%(苯并[a]蒽)之间,相对标准偏差(n=5)为4.9%(荧蒽,苯并[b]荧蒽)～11.1%(苯并[g,h,i],茚并[1,2,3-cd]芘),方法的检出限在0.72(蒽)～14.10 ng/L(荧蒽)之间,基本上达到了痕量分析的要求。利用该方法测得青岛湾表层海水中多环芳烃的浓度在0.125(苯并[k]荧蒽)～25.996 ng/L(萘)之间,但苯并[a]芘未检出。     

Summer phytoplankton pigments and community composition related to water mass properties in the Gulf of Gabes

Variations in phytoplankton pigments and community composition were examined in the Gulf of Gabes in relationship to water mass properties, characterised by the influence of the Modified Atlantic Water and by the thermal stratification. Data were collected on board the R/V Hannibal during July 2005.Distinct water masses were identified using cluster analysis of temperature–salinity (T–S) characteristics. Three major clusters appeared based on the combined effects of temperature and salinity. The first cluster was identified as the cool and less salty bottom Modified Atlantic Water (MAW). The warmer and saltier Mediterranean Mixed Water (MMW) represented the second cluster. The third cluster was the Transition Water (TW) separating the two previous clusters.The pigment and taxonomic composition of these water masses were examined. Chlorophyll a was rather low (<200 ng l⁻¹). Chlorophyll b was generally the most abundant accessory pigment and fucoxanthin dominated the accessory pigments in the MAW. Proportions of chlorophyll a associated with different phytoplankton classes were estimated using CHEMTAX software, and did not present significant variations among water groups. The results pointed out variations in the relative contribution of each phytoplankton taxa in each station group. Chlorophytes and prasinophytes accounted for 65% of chlorophyll a in the MMW. Diatoms and chlorophytes were relatively abundant in the MAW contributing to almost 63% of chlorophyll a. An unstructured community, slightly dominated by prasinophytes, chlorophytes and cryptophytes, characterised the TW. Different trophic statuses were observed in these water masses, the MMW and the MAW being characterised by mesotrophy, while an oligotrophy was observed in the TW. Nutrient availability, particularly the P-limitation supported by the summer stratification, as revealed by the high N:P ratio (greater than 20), seems to enhance the development of small-sized phytoplankton, thereby supporting the regenerated production.     

Spatial variability of phytoplankton absorption coefficients and pigments off Baja California during November 2002

We have estimated the spatial variability of phytoplankton specific absorption coefficients (a* _ph ) in the water column of the California Current System during November 2002, taking into account the variability in pigment composition and phytoplankton community structure and size. Oligotrophic conditions (surface Chl < 0.2 mg m⁻³) dominated offshore, while mesotrophic conditions (surface Chl 0.2 to 2.0 mg m⁻³) where found inshore. The specific absorption coefficient at 440 [a* _ph (440)] ranged from 0.025–0.281 m²mg⁻¹ while at 675 nm [a* _ph (675)] it varied between 0.014 and 0.087 m²mg⁻¹. The implementation of a size index based on HPLC data showed the community structure was dominated by picoplankton. This would reduce the package effect in the variability of a* _ph (675). Normalized a _ph curves were classified in two groups according to their shape, separating all spectra with peaks between 440 and 550 nm as the second group. Most samples in the first group were from surface layers, while the second group were from the deep chlorophyll maximum or deeper. Accessory photoprotective pigments (APP) tended to decrease with depth and accessory photosynthetic pigments (APS) to increase, indicating the importance of photoprotective mechanisms in surface layers and adaptation to low light at depth. Samples with higher ratios of APP:APS (>0.4) were considered as phytoplankton adapted to high irradiances, and lower ratios (<0.26) as adapted to low irradiances. We found a good relationship between APP:APS and a* _ph (440) for the deeper layer (DCM and below), but no clear evidence of the factors causing the variability of a* _ph (440) in the upper layer.     

应用光合色素标记物研究2001年2～6月厦门西海域浮游植物群落结构

利用反相高效液相色谱（RP-HPLC）对厦门西海域2001年2～6月的浮游植物色素进行了测定,并首次运用CHEMTAX软件对该海域进行了浮游植物群落组成的分析,同时分析了环境因子对群落组成的作用.我们首先改进了RP-HPLC分离方案,分离的时间缩短了近1/3,达到了提高分离效率的目的.利用改进后的RP-HPLC方案进行色素分析显示,厦门西海域2001年2～6月期间,主要的特征色素是岩藻黄素（fucoxanthin）、19’-丁酰基氧化岩藻黄素（19’-butanoyloxyfucoxanthin）、19’-己酰基氧化岩藻黄素（19’-hexanoyloxyfucoxanthin）和多甲藻素（peridinin）,月平均含量分别在267.824～655.076、20.118～126.764、65.693～214.115、26.955～203.927μg/m3的范围变动.CHEMTAX计算的结果进一步表明：硅藻是主要优势类群,其次是定鞭金藻和甲藻,分别占总生物量的36.14%～79.27%、4.50%～37.40%和4.83%～21.37%.总生物量从2月到6月呈增长趋势,温度是主要的决定因子.硅藻和定鞭金藻所占生物量的份额呈相反趋势变化,表明种间竞争也是重要的因素之一.     

西赤道太平洋暖池区光合色素分布及其对浮游植物群落的指示作用

利用光合色素的生物标志性可以在"纲"水平上表征浮游植物群落结构。依托大洋科学考察第20航次和21航次,通过对西赤道太平洋不同区域5个站位的HPLC藻类色素分析及CHEMTAX程序因子分析,获取了暖池区光合色素及浮游植物群落的垂直分布信息。结果显示在寡营养的暖池区,玉米黄素(Zeaxanthin)及乙二烯叶绿素a(DV Chl a)与叶绿素a浓度呈显著的正相关,浮游植物群落结构以蓝细菌、原绿球藻及定鞭金藻为优势藻纲,按对生物量的贡献率原绿球藻大于蓝细菌大于定鞭金藻的。蓝细菌和原绿球藻分布在真光层不同深度,而在营养盐丰富的次表层优势浮游藻类为定鞭金藻。     

不同品系条斑紫菜光合效率比较研究

用叶绿素荧光仪测试了7个品系紫菜叶状体在不同光照和温度条件下的光合效率,分析了受强光抑制后光合效率恢复的情况,并测试了这些品系的光合色素含量。结果显示,随着光照强度从20μmol.m-2.s-1提高到1 200μmol.m-2.s-1,总的趋势是光合效率逐渐降低,但下降幅度在不同品系间又有差异:相比20μmol.m-2.s-1光照下条件下光合效率,Yjs在1 200μmol.m-2.s-1时的光合效率下降了27%,Yw下降了33%,Gm下降了53%,Yqd下降了50%。在120μmol.m-2.s-1光照条件,5~20℃实验范围内,15~20℃条件下光合效率最高,除Yh2和Wjs,其它品系叶状体均在15℃达到顶峰,Yh2和Wjs则在20℃达到顶峰。在受到高光抑制(200μmol.m-2.s-1)后,处于光照60μmol.m-2.s-1,温度12℃条件下紫菜光合效率可在30~60 min内恢复。未发现光合色素含量与光合效率的相关性。文章还对紫菜光合效率与生态条件的关系进行了分析和讨论。     

Ecological features of phytoplankton in red tide outburst area in the Xiamen Harbor

ＥｃｏｌｏｇｉｃａｌｆｅａｔｕｒｅｓｏｆｐｈｙｔｏｐｌａｎｋｔｏｎｉｎｒｅｄｔｉｄｅｏｕｔｂｕｒｓｔａｒｅａｉｎｔｈｅＸｉａｍｅｎＨａｒｂｏｒ￥ＺｈａｎｇＳｈｕｉｊｉｎ（ＲｅｃｅｉｖｅｄＪｕｎｅ２０，１９９４；ａｃｃｅｐｔｅｄＳｅｐｔｅｍｂｅｒ２，１...     

Structure and photosynthetic properties of phytoplankton assemblages in a highly dynamic system, the Northern Adriatic Sea

The photosynthetic properties of phytoplankton populations as related to physical–chemical variations on small temporal and spatial scales and to phytoplankton size structure and pigment spectra were investigated in the Northern Adriatic Sea off the Po River delta in late winter 1997. Large diatoms (fucoxanthin) dominated the phytoplankton in the coastal area whereas small phytoflagellates (mainly 19′-hexanoyloxyfucoxanthin, chlorophyll b, 19′-butanoyloxyfucoxanthin) occurred outside the front. The front was defined by the steep gradient in density in the surface layer separating low-salinity coastal waters from the offshore waters.Physical features of the area strongly influenced phytoplankton biomass distributions, composition and size structure. After high volumes of Po River discharge several gyres and meanders occurred in the area off the river delta in February. Decreasing river discharge and the subsequent disappearance of the gyres and the spreading dilution of the river plume was observed in March. The dynamic circulation of February resulted in high photosynthetic capacity of the abundant phytoplankton population (>3.40 mg m⁻³). In March, the slow circulation and an upper low-salinity water layer, segregated from the deeper layers, resulted in lack of renewal of this water mass. The huge phytoplankton biomass, up to 15.77 mg chl a m⁻³, became nutrient depleted and showed low photosynthetic capacity. In February, an exceptionally high P_max^B, 20.11 mg C (mg chl a)⁻¹ h⁻¹ was recorded in the Po River plume area and average P_max^B was three-fold in February as compared to the March recordings, 10.50 mg C (mg chl a)⁻¹ h⁻¹ and 3.22 mg C (mg chl a)⁻¹ h⁻¹, respectively.The extreme variability and values of phytoplankton biomass in the innermost plume area was not always reflected in primary production. Modeling of circulation patterns and water mass resilience in the area will help to predict phytoplankton response and biomass distributions. In the frontal area, despite a considerable variability in environmental conditions, our findings have shown that the phytoplankton assemblages will compensate for nutrient depression and hydrographic constraints, by means of size and taxonomic composition and, as a result, the variability in the photosynthetic capacity was much less pronounced than that observed for other parameters.