反相高效液相色谱法测维生素A含量

目的：建立一种用反相高效波相色谱测定维生素Ａ的方法。方法：色谱柱：Ｈｙｐｅｒｓｉｌ ＯＤＳ（１５０ｍｉｎ×４６ｍｍ）,流动相：甲醇－水（９６：４）,流速１．０ｍｌ／ｍｉｎ,紫外检测器波长３２５ｎｍ,柱温２５℃,外标法测定。结果：维生素 Ａ在 ２．５～３０．０μｇ／ｍｌ之间有良好的线性关系,最低检测浓度为 １．５μｇ／ｍｌ,平均回收率为９８．５％,精密度ＲＳＤ±ｌ、５６％（ｎ＝３）。结论：本方法准确、灵敏、适应面广。     

罗非鱼中红霉素残留量的高效液相色谱法的测定

建立了以高效液相色谱法测定罗非鱼中红霉素残留量的方法。提出了以乙腈为提取剂,经过液-液萃取、固相萃取、反萃取等步骤对样品进行分离净化的样品处理方法,该方法准确、灵敏度高。方法的检出限为400μg/kg。     

促排卵素3号的反相高效液相色谱测定法

用反相高效液相色谱法测定促排卵素３号的含量,采用Ｃ１８反相柱,通过对流动相和检测波长的筛选,取得了良好的回收率和线性关系,建立了促排卵素３号含量的测定方法。     

反相高效液相色谱法测定褐藻中岩藻黄素的含量

建立一种反相高效液相色谱法用于海藻中岩藻黄素的定量分析,并测定新鲜及干制褐藻中岩藻黄素含量,探究不同干制方式对褐藻岩藻黄素的影响。样品经甲醇提取、C18柱固相萃取净化后,采用ZORBAX SB-C18色谱柱,流动相采用水与甲醇梯度洗脱,流速0.7 ml/min、进样量10μl、柱温35℃、检测波长450 nm进行液相色谱分析。在该条件下,岩藻黄素在0.11~50 mg/L范围内有良好的线性关系(Y=38.46X+0.8899,R~2=0.9999),检测限为0.03 mg/L,回收率为92.66%~109.06%,相对标准偏差(RSD)为3.46~4.61%。采用该方法,岩藻黄素提取净化完全,杂质干扰小,回收率高,能够对海藻及其制品中岩藻黄素进行准确定量。采用该方法测得新鲜海带和马尾藻的岩藻黄素含量(干基)分别为559.2和680.4 mg/kg,而干制褐藻样品中岩藻黄素的含量明显低于新鲜褐藻。不同干制方式造成的岩藻黄素损失程度为真空冷冻干燥45℃烘干≈自然晾干。因此,在规模化分离制备海藻岩藻黄素时,宜采用新鲜或冻干海藻作为原料,从而保证岩藻黄素的得率。     

中华绒螯蟹血淋巴内盐酸环丙沙星的反相高效液相色谱法的建立

应用反相高效液相色谱法(RP-HPLC)建立了盐酸环丙沙星在中华绒螯蟹血淋巴内含量的测定方法.本法采用肌肉注射的给药方法,给药剂量为8.17±0.56mg·kg-1和6.25±0.85mg·kg-1.血淋巴从心区抽取.经各种抗凝方法的比较,认为等量的ACD与等量的血淋巴混合,可得到持久的抗凝效果.血淋巴样经甲醇去蛋白,3000r/min离心10min后,取上清液上HPLC.色谱柱,RP-ODS-5μl50×4.6mm;流动相,甲醇-磷酸缓冲液-四丁基溴化铵=25075020(v/v/v);流速,l mL·min-1;进样量,2μL;检测波长,280nm.研究表明,本法线性关系与重复性良好;雌、雄蟹平均回收率分别为100.08±3.43%和99.11±8.20%;日间精密度、平均提取精密度分别为1.19±0.92%、1.55±0.01%.用该方法对肌肉注射给药后48h内中华绒螫蟹雄蟹血淋巴中的盐酸环丙沙垡的血药浓度进行检测,揭示出由37.13±0.08μg·mL-1降至5.96±0.18μg·mL-1的代谢规律.     

高效液相色谱法同时测定饲料中VE和VD3的研究

研究了用反相高效液相色谱法同时测定饲料中V3和VD3的技术及试验溶液的皂化、提取、浓缩等试样前处理方法。选出了最佳色谱条件，流动相：甲酵＋水（95＋5），流速：1ml/min，温度：室温，进样体积：20μl，检测波长：264nm。     

高效液相色谱法测定配合饲料中呋喃唑酮含量

呋喃唑酮是人工合成抗菌药,如果过量添加,会引起畜禽中毒甚至癌症,容易引起抗药菌株。准确检测饲料中呋喃唑酮的含量能正确评估饲料质量,减少因呋喃唑酮含量过高引起的畜禽疾病,降低畜禽业风险,提高养殖效益,从而达到规范饲料行业的目的。     

高效液相色谱法在药残测定中的研究进展

通过介绍高效液相色谱法（HPLC）的原理，阐述了高效液相色谱法在实际应用中的影响因素如流动相、固定相载体、柱温、柱长及柱内径和透射光波长等等；同时也综述了国内外在中、西药成分检测和机体内、外药物残留测定中诸因素的优化筛选；最后笔者结合目前新技术的研究进展提出了HPLC在与其它先进技术联用中的应用展望及应克服的不良因素。     

高效液相色谱法在药残测定中的研究进展

通过介绍高效液相色谱法(HPLC)的原理,阐述了高效液相色谱法在实际应用中的影响因素如流动相、固定相、载体、柱温、柱长及柱内径和透射光波长等等;同时也综述了国内外在中、西药成分检测和机体内、外药物残留测定中诸因素的优化筛选;最后笔者结合目前新技术的研究进展提出了HPLC在与其它先进技术联用中的应用展望及应克服的不良因素。     

Use of ideal protein concept for precision formulation of amino acid levels in fish-meal-free diets for juvenile Nile tilapia (Oreochromis niloticus L.)

This study was undertaken in a closed system with Nile tilapia (Oreochromis niloticus) to examine the effects of total replacement of fish meal (FM) by soybean meal. Nile tilapia fingerlings with an average weight of 5.34±0.08 g were hand‐fed one of the five isoenergetic (≈13.5 MJ digestible energy kg^?1) and isoproteic (≈31% of digestible protein) experimental diets to satiation, six times a day during 85 days in eight replicate fibreglass tanks (six fish per tank). The control diet containing FM was substituted by soybean meal, with and without essential amino acids (lysine, methionine and threonine) or dicalcium phosphate supplementation. The supplemental amino acids were added at levels to simulate the reference amino acid profile of Nile tilapia carcass protein, based on the ideal protein concept. The results showed that soybean meal diet supplemented only with dicalcium phosphate was inferior to the control diet with FM and soybean meal diets supplemented with dicalcium phosphate and essential amino acids. Multiple essential amino acids and dicalcium phosphate incorporation in soybean meal diets was associated with performance, whole‐body composition and carcass yield equal to that of the fish fed with the control diet containing FM. These data suggest that a diet with all plant protein source, supplemented with essential amino acids, based on tissue amino acid profile, can totally replace FM in a diet for Nile tilapia, without adverse effects on the growth performance, carcass yield and composition.     

高效液相色谱法测定水产品中ATP关联化合物

建立了水产品中三磷酸腺苷(ATP)、二磷酸腺苷(ADP)、腺苷酸(AMP)、肌苷酸(IMP)、次黄嘌呤核苷(HxR)和次黄嘌呤(Hx)6种ATP关联化合物的高效液相色谱检测方法。样品中的ATP关联物经高氯酸提取并用氢氧化钠调节pH值至6.0-6.4后,以AQ-C18(250 mm×4.6 mm,5μm)柱为分析柱,0.02 mol/L KH2PO41-0.02 mol/L K2HPO4(V/V=1/1)的缓冲液为流动相进行洗脱,在254 nm波长下用高效液相色谱进行检测,外标法定量。研究表明,ATP、ADP、AMP、IMP和HxR线性范围为0.2-40.0μg/ml,Hx线性范围为0.1-20.0μg/ml,相关系数均大于0.999 0;ATP、ADP、AMP、IMP和HxR检出限为5.00 mg/kg,Hx检出限为2.5 mg/kg(S/N=3);对真鲷、鱼师鱼、鲑鱼、凡纳滨对虾和梭子蟹5种水产品进行3个浓度水平的加标实验,回收率在85.5%-105%之间,相对标准偏差(RSD)小于12.9%。实验结果表明,该方法简便、准确,适合于水产品中ATP关联物的测定。     

Effect of dietary conjugated linoleic acid levels on growth performance,muscle fatty acid profile,hepatic intermediary metabolism and antioxidant responses in genetically improved farmed Tilapia strain of Nile tilapia Oreochromis niloticus

An 8‐week feeding trial was conducted to determine the effect of dietary conjugated linoleic acid (CLA) on growth performance, muscle fatty acid profile, hepatic intermediary metabolism and antioxidant responses in genetically improved farmed Tilapia (GIFT) strain of Oreochromis niloticus (initial body weight: 42.6 ± 0.4 g, mean ± standard deviation). Three replicated groups of GIFT strain of Nile tilapia were hand‐fed to satiation, twice a day, with the diets in which CLA oil, containing mainly the bioactive cis‐9, trans‐11 and trans‐10, cis‐12 isomers, was included at 0 (control), 0.5%, 1%, 1.5%, 2% and 2.5%, respectively, at the expense of fish oil to maintain the constant lipid and energy levels. Growth performance and feed utilization showed no significant differences among the treatments (P > 0.05). The dietary inclusion of CLA modified total percentages of the main groups of fatty acids. Increasing saturated fatty acid content and reduced mono‐unsaturated fatty acid contents in muscle were observed with increasing dietary CLA inclusion (P < 0.05). Total n‐3 fatty acids and total polyunsaturated fatty acids tended to decline with increasing dietary CLA levels (P < 0.05), but n‐6 fatty acids showed no significant differences among the treatments (P > 0.05). Dietary CLA supplementation resulted in the significant increase in the trans‐10, cis‐12 and cis‐9, trans‐11 CLA isomers in muscle (P < 0.05) and also significantly influenced several hepatic enzymatic activities, such as succinate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, lipoprotein lipase and hepatic lipase activities (P < 0.05). Reduced superoxide dismutase and glutathione peroxidase activities, and the decline in malondialdehyde levels were observed in fish fed the CLA‐supplemented diets (P < 0.05), indicating that dietary CLA supplementation showed a powerful antioxidant effect for this fish species. Our study was the first report involved in the effect of dietary CLA inclusion on hepatic intermediary metabolism and antioxidant responses in fish, which could be used as indicators of nutritional and physiological status of the fish species.     

饲料牛磺酸对尼罗罗非鱼生长、体成分及组织游离氨基酸含量的影响

本文探讨了饲料牛磺酸水平对尼罗罗非鱼幼鱼生长性能、体成分及游离氨基酸含量的影响。选用体重为5.89±0.03g的罗非鱼300尾,随机分为5个处理组,每个处理3个重复,每个重复20尾。在基础饲料中分别添加0、0.4%、0.8%、1.2%、1.6%的牛磺酸,配制成5种试验料,分别饲喂不同处理组的罗非鱼,饲养周期为56d。结果表明： 0.8%牛磺酸组罗非鱼增重率最高,且显著高于0%和1.6%组(P<0.05);饲料牛磺酸水平(0.4%-1.2%)显著提高了罗非鱼摄食率、肝脏指数和内脏指数,显著降低了饲料系数(P<0.05)。随着饲料牛磺酸水平的提高,鱼体粗蛋白质和粗脂肪含量随之升高,鱼体水分和粗灰分含量随之降低(P<0.05);以增重率为指标,通过二次曲线回归分析得出罗非鱼饲料牛磺酸最适需要量为0.75%。罗非鱼血清、肝脏、肌肉和全鱼中牛磺酸含量与饲料牛磺酸含量存在正相关关系,且牛磺酸添加组血清、肝脏、肌肉和全鱼牛磺酸含量显著高于对照组相(P<0.05)。罗非鱼肝脏和肌肉中游离氨基酸含量随饲料牛磺酸含量的增加均呈逐渐下降的趋势,且牛磺酸添加组肝脏和肌肉中游离氨基酸含量显著低于对照组(P<0.05)。     

Effect of potential probiotic bacteria on growth and survival of tilapia Oreochromis niloticus L., cultured in the laboratory under high density and suboptimum temperature

This study examined the effect of potential probiotic bacteria on growth and survival of the tilapia Oreochromis niloticus , under high density and suboptimum temperature. Presumptive Bacillus and lactic acid bacteria (LAB) were isolated from tilapia and from their culture system and were characterized for haemolytic and enzymatic activity, and antagonism against Vibrio . Selected strains were included in the diet of juvenile tilapia and evaluated during a 134-day assay. The experiment was conducted with four treatments: (1) fish fed with commercial feed plus Dry Oil^®; (2) fish fed with commercial feed plus LAB; (3) fish with bacilli in water; (4) fish with a mixture of treatments 2 and 3. Tilapias in all treatments, including bacteria, grew significantly better than fish fed with commercial feed plus Dry Oil^® (control group). Survival was similar in all treatments. The physicochemical parameters of the culture system were maintained within the optimal ranges for the species, with the exception of temperature (19.9–24.82 °C). Animals fed diet supplemented with bacilli and LAB had good survival and the best growth performance, suggesting that bacteria are appropriate growth-stimulating additives in tilapia cultivation.     

Effect of dietary betaine levels on growth performance and hepatic intermediary metabolism of GIFT strain of Nile tilapia Oreochromis niloticus reared in freshwater

An 8‐week feeding experiment was conducted to determine the effect of dietary betaine levels on the growth performance and hepatic intermediary metabolism of genetically improved farmed tilapia (GIFT) strain of Nile tilapia Oreochromis niloticus (mean initial body weight: 78.3 ± 1.3 g, means ± SD). Six practical diets were formulated with the incorporation of betaine at the levels of 0 (control), 5, 10, 15, 20 and 25 g kg⁻¹. Survival showed no significant differences among the treatments (P > 0.05). The highest and lowest weight gain (WG) and specific growth rate (SGR) were observed for fish fed the diets containing 5 and 0 g kg⁻¹ (control) betaine, respectively. Feed intake showed similar trend with WG and SGR. In contrast, feed conversion ratio was the lowest when dietary betaine level was 5 g kg⁻¹. In general, dietary betaine supplementation showed no significant effect on hepatic composition of tilapia. Condition factor and viscerosomatic index tended to increase with increasing dietary betaine levels from 0 to 5 g kg⁻¹ and then decline when dietary betaine levels further increased from 5 to 25 g kg⁻¹. In contrast, hepatosomatic index declined with increasing dietary betaine levels (P < 0.05). Dietary betaine levels significantly influenced several hepatic enzymatic activities, including succinate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, lipoprotein lipase and hepatic lipase, suggesting that dietary betaine addition had significant effects on nutrient metabolism in the liver. Based on the second‐order polynomial regression analysis of WG, 12.5 g kg⁻¹ of dietary betaine level seemed optimal for genetically improved farmed tilapia strain of O. niloticus.     

高效液相色谱法测定淡水渔业用水中诺氟沙星、环丙沙星、恩诺沙星残留

该文采用高效液相色谱—荧光法检测淡水渔业用水中诺氟沙星、环丙沙星和恩诺沙星3种喹诺酮类抗生素药物的残留,建立了检测淡水渔业用水中喹诺酮类抗生素药物的方法。结果显示,诺氟沙星、环丙沙星和恩诺沙星的检出限(按S/N=3计)分别为2.5、1.0、2.5μg/L,定量限(按S/N=10计)分别为5、1、5μg/L。按1倍、5倍、10倍定量限进行加标回收,3种抗生素的平均回收率为76.6%～89.9%,相对标准偏差(RSD)在1.72%～8.43%。     

基于高效液相色谱-四级杆-飞行时间质谱技术筛查渔业水域环境中抗生素残留方法的建立与应用

应用高效液相色谱—四级杆—飞行时间质谱(HPLC-Q-TOF-MS)技术建立水中常用的42种抗生素残留快速筛查方法。采用固相萃取富集净化的方法进行样品预处理,目标药物经液相色谱仪C18反相柱分离,1%甲酸乙酸铵水溶液—甲醇溶液为流动相梯度洗脱,利用正离子源全扫描模式进行检测,并将建立的方法应用于浙江省内主要渔业环境水质样品抗生素残留的筛查工作。渔业水域环境中42种抗生素定量限(LOQ,S/N=10)为0.010~0.120μg/L。3个水平加标的平均回收率为60.7%~97.1%,相对标准偏差(RSD)为0.58%~8.01%。该方法快速简便,灵敏度高,对抽取的60批次样品进行检测,共检出磺胺甲鰁唑、磺胺间甲氧基嘧啶、磺胺吡啶、磺胺氯哒嗪、恩诺沙星5种药物,含量为25.6~106.7 ng/L。本实验建立的渔业水质中抗生素药物的高效液相色谱—四级杆串联飞行质谱分析方法,灵敏、准确、定性能力强,可以成功应用于渔业环境水质样品抗生素残留的筛查工作。