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.     

Levels and spatial distribution of polycyclic aromatic hydrocarbons (PAHs) in superficial sediment from 15 Italian marine protected areas (MPA)

Surface sediment from 15 Italian marine protected areas (MPA) were analysed for polycyclic aromatic hydrocarbons (PAHs). The organic carbon percentage was also determined. Total PAH concentrations (ng g⁻¹ d.w.) ranged from 0.71 (Penisola del Sinis) to 1550 (Miramare). Individual PAH analysis showed that three and four rings PAHs were the most frequently detected isomers and accounted for 60-70% of the PAH total concentrations. PAH ratio analysis showed a prevalence of pyrolytic PAH origin at most of the MPAs with exception of Porto Cesareo and Ustica where a petrogenic origin was detected. Results for organic carbon percentages ranged from 0.3% (Capo Rizzuto) to 2% (Punta Campanella). These results are comparable to other Mediterranean marine environments. However, our results shows that some MPAs, such as Miramare, Porto Cesareo, Isola Capo Rizzuto and Punta Campanella, are subject to strong pressure from urban and industrial activities where high PAH levels were detected.     

微管径高效液相色谱及其应用

介绍了微管径高效液相色谱法 (microboreHPLC ,μHPLC)及其应用 ,包括 μHPLC的结构特点、优点及其在制药、生物产品、环境监测和临床药物试验中的应用     

Early chlorin diagenesis in a recent aquatic sediment

Chlorophylls a and b, pheophytins a and b, pheophorbides a and b and pyropheophorbide a were identified in a Recent sediment (Priest Pot from Cumbria, U.K.) by laboratory preparation and structure elucidation by NMR and FAB-MS, and HPLC coelution with sedimentary extracts. Quantitative depth profiles revealed a steady decrease of the ratios of chlorophylls to pheophytins, and also a different but fairly constant a to b ratio for the pheophytins and the chlorins.     

Stability and changes in astaxanthin ester composition from Haematococcus pluvialis during storage

In this paper, we investigated the effects of temperature, oxygen, antioxidants, and corn germ oil on the stability of astaxanthin from Haematococcus pluvialis under different storage conditions, and changes in the composition of astaxanthin esters during storage using high performance liquid chromatography and spectrophotometry. Oxygen and high temperatures （22-25~C） significantly reduced the stability of astaxanthin esters. Corn germ oil and antioxidants （ascorbic acid and vitamin E） failed to protect astaxanthin from oxidation, and actually significantly increased the instability of astaxanthin. A change in the relative composition of astaxanthin esters was observed after 96 weeks of long-term storage. During storage, the relative amounts of free astaxanthin and astaxanthin monoesters declined, while the relative amount of astaxanthin diesters increased. Thus, the ratio of astaxanthin diester to monoester increased, and this ratio could be used to indicate if astaxanthin esters have been properly preserved. If the ratio is greater than 0.2, it suggests that the decrease in astaxanthin content could be higher than 20%. Our results show that storing algal powder from H. pluvialis or other natural astaxanthin products under vacuum and in the dark below 4~C is the most economical and applicable storage method for the large-scale production of astaxanthin from H. pluvialis. This storage method can produce an astaxanthin preservation rate of at least 80% after 96 weeks of storage.     

高效液相色谱法检测水产品中的孔雀石绿和结晶紫

样品中的孔雀石绿、结晶紫经试剂盒提取,浓缩后用经固相萃取柱净化、硼氢化钾还原、反向色谱柱分离,使用荧光检测器检测,孔雀石绿、结晶紫的加标回收率在76.1%～92.5%之间,相对标准偏差1.9%～5.8%,检出限均为0.5μg/kg。结果表明:该方法检测孔雀石绿(MG)、结晶紫(GV)处理简单,灵敏度高,节省时间,可用于大量样品的快速分析。     

Day–night and depth differences in haemolymph melatonin of the Norway lobster, Nephrops norvegicus (L.)

Few studies have been conducted to quantify and understand the role of melatonin in invertebrates, and particularly in crustaceans and in deep-sea animals. In this study, we examined day–night differences in haemolymph melatonin of the burrowing decapod crustacean Nephrops norvegicus (L.) during exposure to cycles of monochromatic blue light (480 nm) and darkness cycles of 10 and 0.1 lx. These differential intensity conditions simulate illumination at the depth of the shelf (80–100 m) and of the slope (300–400 m), where these lobster populations are chiefly found in the Western Mediterranean Sea. Our objectives were: (a) to verify the presence of melatonin in the haemolymph of this species using liquid chromatography/tandem mass spectrometry (LC–MS/MS) and fluorescence HPLC (HPLC); and (b) to study the relationship between diel variations in melatonin concentration and locomotor rhythms, in order to examine whether the former influences behaviour. Melatonin was identified in LC–MS/MS by Q1 and Q3 mass peaks at an elution time of 3.7 min, and it was also detected by HPLC. Melatonin concentration was found to be two orders of magnitude higher at 10 lx (4.8±5.3 ng ml⁻¹) than at 0.1 lx (0.06±0.03 ng ml⁻¹). Also, the increase at daytime in 10 lx was absent in 0.1 lx. When the locomotor rhythm of animals exposed to both photoperiod regimes was compared, the diel periodicity was found to be preserved, but the timing of activity shifted from night to day. Extrapolating these data to the field, we interpret our results to mean that locomotor activity preserves its diel character, but not its phase and amplitude, in a bathymetric range where haemolymph melatonin reduces its concentration and rhythmic fluctuation.     

广东省近岸海域麻痹性贝类毒素HPLC分析

于2001年和2002年,对广东省近岸海域的18个地点进行了贝类生物采样。采用Oshima麻痹性贝类毒素高效液相色谱法对贝类的主要毒素成分进行分析。结果显示:染毒贝类含有的高毒性毒素STX,GTX1检出率较低,而低毒性毒素C1,C2,GTX5及中等毒性毒素GTX3,GTX4检出率较高;与2001年相比,2002年染毒贝类毒素成分的检出率和检出的毒素含量有所降低,贝类消化腺在这方面变化尤其明显;主要染毒贝类是华贵栉孔扇贝、翡翠贻贝和长牡坜等;不论消化腺或壳内全部软组织,贝类毒素含量存在明显的地理分布差异,大亚湾和大鹏湾海域>深圳湾、珠海海域和粤东海域>粤西海域;大亚湾和大鹏湾的翡翠贻贝和华贵栉孔扇贝样品中,毒性效能较高的STX,GTX1和GTX4等成分均占相当的比例。     

雪卡毒素的提取纯化方法初步研究

雪卡毒素(ciguatoxin)是一类具有神经毒性的海洋藻类毒素,高纯度雪卡毒素是开展其相关研究的物质基础。以石斑鱼肝脏为原料,对其脂溶性粗提物经Florisil柱和Sephadex LH-20柱纯化后,用高效液相色谱/质谱(HPLC/MS)和细胞毒性方法对其进行鉴定,证明为太平洋雪卡毒素(P-CTX-1)。HPLC鉴定分析提取物与太平洋雪卡毒素(P-CTX-1)具有相同的保留时间。HPLC/MS检测表明纯化样品在m/z=1 111.3和m/z=1 133.0处出现雪卡毒素[M+H]+峰和[M+Na]+峰,与已有的P-CTX-1分子离子峰完全吻合。四甲基偶氮唑蓝法(MTT)细胞毒性试验证明了雪卡毒素样品对人神经母细胞瘤(SK-N-SH)具有毒性作用,毒素剂量与细胞生长抑制率成线性关系。细胞毒性试验进一步确认该提取物为雪卡毒素,且纯度较高。     

Development and validation of a method for the simultaneous extraction and separate measurement of oxytetracycline,florfenicol, oxolinic acid and flumequine from marine sediments

A simple and rapid method for the detection and extraction of oxolinic acid, flumequine, florfenicol and oxytetracycline from marine sediments was developed and validated. The analytes were extracted from the marine sediment using a solution of oxalic acid diluted in methanol with sonication before detection by HPLC using a diode-array detector (florfenicol and oxytetracycline) and fluorescence (oxolinic acid and flumequine). The quantification limits (QL) were 100 ng/g for oxytetracycline and florfenicol and 5 ng/g for oxolinic acid and flumequine. The coefficients of variation of the repeatability and intermediate precision were less than 10% in all of the analytes. The calibration curves were linear between 50 and 500 ng/ml for oxytetracycline and florfenicol and 1 and 20 ng/ml for oxolinic acid and flumequine. The recuperation rate for the analytes was above 86%.