饲料中盐酸克伦特罗的高效液相色谱法测定

用高效液相色谱法测定饲料中盐酸克伦特罗的含量。选用ShimpackCLC－ODS分析柱（46mm×150mm），流动相：甲醇：水＝75：25，检测波长：243nm，流速：1ml／min。盐酸克伦特罗浓度在0．1－2．0μg／ml之间具有良好线性关系，相关系数0．9996。平均回收率为（101．2±9．75）％。该法快速、准确、重现性好。

国产盐酸氨溴索胶囊人体生物等效性

目的 :研究被试制剂国产盐酸氨溴索胶囊和标准参比制剂 (沐舒坦R○)的药动学和人体生物等效性。方法 :采用高效液相色谱法测定 12名健康受试者单剂量口服被试制剂或参比制剂 90mg后 ,血浆中盐酸氨溴索的浓度 ,经 3p97计算程序拟合 ,计算其药动学参数 ,比较其与标准参比制剂的人体生物等效性。结果 :经 3p97计算程序拟合 ,两者在体内的过程皆符合血管外给药一室模型 ,采用梯形法计算的两者AUC0 -t均值分别为 (2 42 1.2± 76 1.3) μg·h·L-1和 (2 485 .9± 80 3 .9) μg·h·L-1,实测Cmax均值分别为 (385 .5± 98.3) μg·L-1和 (378.9± 90 .3) μg·L-1,实测Tpeak均值分别为 (1.5± 0 .3)h和 (1.6± 0 .4)h。被试制剂的相对生物利用度为 (97.9± 6 .7) %。结论 :经统计学分析 。

HPLC Determination of Harpagoside and Cinnamic Acid in Radix Scrophulariae

A gradient HPLC method was established for the determination of harpagoside and cinnamic acid in Radix Scrophulariae (Xuanshen) and a proposition was put forward for the lowest content of the characteristic and active constituent (harpagoside) for qualified Radix Scrophulariae. The experimental conditions were as follows: Ultrasphere ODS column (250 mm 4.6 mm, 5 mm), mobile phase: acetonitrile-water (containing 1.0% acetic acid) (20:8050:50) (20 min), flow-rate 1 mLmin-1, room temperature, detection wavelength 278 nm. Twenty-eight samples of Xuan-shen (Radix Scrophulariae) from different districts of China were analyzed and the contents of harpagoside and cinnamic acid in Xuan-shen were 0.041~0.244% and 0.012~0.068% respectively. The recoveries (RSD)% were 97.13(0.80)% for harpagoside and 99.38(0.51)% for cinnamic acid. The method is simple and accurate. It can be used for the quality control of Radix Scrophulariae. We propose that the content of harpagoside in qualified Radix Scrophularia should be no less than 0.05%.

Determination of contents of eight alkaloids in fruits of Macleaya cordata (Willd) R. Br. from different habitats and antioxidant activities of extracts

A reliable ultrasound-assisted extraction （UAE） method combined with HPLC-UV for quantification of eight active alkaloids in fruits of Macleaya cordata （Willd） R. Br. was developed. The optimization conditions of UAE were obtained by using Box-Behnken design of response surface methodology. Chromatography was carried out using a Kromasil C18 column by gradient elution with 0.1% phosphoric acid aqueous solution for HPLC-UV. All calibration curves showed good linear correlation coefficients （R^2〉0.999 6） and recoveries （from 97.3% to 104.9%） were acceptable. 1,1-diphenyl-2-picrylhydrazyl （DPPH） method was employed to test the antioxidant activity of the extract from the samples. The proposed method was successfully applied to quantifying eight components in nine samples of M.cordata, and significant variations of alkaloid contents and antioxidant aetivity of the samples from different habitats were demonstrated. It presents a powerful proof for the selection of the best sources to extract eight kinds of alkaloids.

Production of Aflatoxins from Aspergillus flavus in Liquid Medium

Aflatoxins are produced mainly by Aspergillus flavus and Aspergillus parasiticus, and can be found in many grains such as peanuts, soybeans and com. This study aimed to qualitatively and quantitatively evaluate the production of aflatoxin in liquid media using strains of Aspergillus flavus obtained from peanuts marketed in the city of Fortaleza, CEo Strains of Aspergillus flavus were inoculated into a liquid medium malt extract and after 2 days inoculated into a second medium containing sucrose 5%, MgSO4·7H20 0.1%, KH2PO4 1%, ZnSO4·7H2O 0.0176 g, and cultured for 3 more days. The media were kept at room temperature ranging from 24°C to 32 °C with agitation of 130 rpm and aeration of 4.17 Llmin. Qualitative analysis was performed by thin layer chromatography and quantitatively by high performance liquid chromatography with fluorescence detection, demonstrating the production of aflatoxin B I （588 mg/L） and B2 （929 mg/L）.

Recovery of Astaxanthin from Discharged Wastewater During the Production of Chitin

In this paper, studies were carried out to extract astaxanthin from discharged wastewater during the production of chitin and to reveal the scavenging effect of the obtained pigment on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical. Different ratios of dichloromethane/methanol (V/V) were used to extract astaxanthin. When the ratio of dichloromethane/methanol was 2:8 and the ratio between the mixed organic solvent (dichloromethane/methanol, 2:8, V/V) and wastewater was 1:1, the highest yield of pigment was obtained (8.4 mg/ 50 mL). The concentration of free astaxanthin in the obtained pigment analyzed by HPLC was 30.02%. The obtained pigment possessed strong scavenging ability on DPPH radical and IC50 was 0.84mg/ml.

Persistence of malachite green and leucomalachite green in perch (Lateolabrax japonicus)

The persistence of malachite green (MG), and its metabolite leucomalachite green (LMG), in fish tissues is still unclear, leading to many trade disputes. In this research, we established and evaluated an HPLC method that could detect MG and LMG simultaneously, and then investigated the persistence of these two toxins in the tissues of juvenile perch (Lateolabrax japonicus) post sub-chronic MG exposure at 1.0 mg/L. Exposure lasted for 2 h everyday and was repeated six times. The perch were then placed in MG-free seawater for 100 d to eliminate the toxins. Results show that MG accumulated in the tissues, including the gills, liver, muscle, blood and viscera, and then was metabolized rapidly to LMG. The concentrations of these two toxins increased significantly with the accumulation process. In general, the highest concentrations of MG and LMG in all tissue exceeded 1 000 μg/kg, except for MG in the muscle. The order of accumulation levels (highest to lowest) of MG was gill>blood>liver>viscera>muscle, while that of LMG was liver>blood>gill>viscera>muscle. High levels of MG or LMG could persist for several hours but decreased rapidly during the elimination process. The concentration of LMG was much higher than that of MG during the experiment, especially in the gill, liver and blood. Therefore, the three tissues play important roles in toxin accumulation, biotransformation, and elimination. Although the MG and LMG concentrations in muscle were much lower than in other tissues, the content still exceeded the European minimum required performance limit (MRPL), even after 2 400 h (100 d) of elimination. This demonstrates that it is extremely difficult to eliminate MG and LMG from tissues of perch, and therefore use of these toxins is of concern to public health.

Alkaloids Production from Callus of Hyoscyamus niger L, in Vitro

Callus cultures of Hyoscyamus niger L. were initiated from leaf segments cultured on Murashige and Skoog （MS） medium supplemented with 0.5 mg/L Benzyl Adenine （BA） and 0, 1, 2 and 3 mg/L Naphthalene acetic acid （ NAA ）. Half of cultures were incubated under light of 16 hr/day, while the other half was incubated under complete darkness. The incubation temperature was 25 ± 1 ℃in both incubation conditions. The fresh and dry weight of the produced callus was obtained after six weeks of incubation. Callus produced were recultured on medium that gave the highest production of callus. Constant weight （300 mg） of callus was cultured in each of these medium supplemented with abiotic elicitor of 50 g/L sucrose, 200 mg/L NaCI, 50 or 100 mg/L proline and 2 mg/L BA each one added separately and incubated under complete darkness. The fresh and dry weights of callus were measured after six weeks. HPLC was used to determine the tropane alkaloids （Hyoscyamine and Scopolamine）. The results showed that the significant highest average of fresh and dry weight of callus （112 and 89.6 mg） achieved using the medium contained 0.5 mg/L BA and 2 mg/L NAA under dark condition. The amount of fresh and dry weight of callus produced under dark condition was significantly higher than that produced under light condition, with increase in percentage of 51.3 and 37.62% respectively. The addition of abiotic elicitors caused reduction in both fresh and dry weight of callus, therefore the highest fresh weight average was 1,727 mg using 100 mg/L proline. The results indicated that addition of 50 or 100 mg/L proline led to increase in Hyoscyamine concentration of 58.03 and 21.37% respectively compared with the control. While other abiotic elicitors were found to cause reduction in Hyoscyamine concentration. Percentage of Scopolamine concentration were increased to 129.03, 166.94, 205.51 and 149.20% after addition of sucrose （50 g/L）, NaC1 （200 mg/L） and proline （50 or 100 mg/L） respectively compared with the control.

Optimization of Yield and Chemical Properties of Agar Extracted from Melanothamnus Somalensis from Oman Sea

The red seaweed Melanothamnus somalensis was investigated as potential economic source of agar. The effect of different conditions of alkali pre-treatment on chemical properties of agar was evaluated. Agar was extracted by various concentrations of NaOH （4%, 6% and 8%） and heated at different temperatures （70 ℃, 75 ℃ and 80 ℃） for different durations （2 h, 2.75 h and 3.5 h）. The yields-molecular weight （Mw） and sulfate contents of extracted agar were analysed and characterized by FTIR spectroscopy. The yield was significantly increased at these treatments from 23.29% to 30.86%. Mw studied by HPLC ranged from （.12.45 ± 0.21） × 10^5 to （8.60 ± 2.40） × 10^5 Da. FTIR bands show sulfate groups in C4 and C6 ofgalactose and no sulfate group were found on both C2 of galactose and C2 of 3,6-anhydrogalactose. All treatments showed a high sulfate content that ranged from 5.4% to 10.1%. These properties were found to be significantly affected by the alkali pre-treatment concentration （p 〈 0.05）. In conclusion, agar extracted in this study was considered acceptable for industrial application and the optimal conditions for extraction were found to be at 6% NaOH at 70 ℃ for 2 hours.

Extraction and Powder Product of Fructo-oligosaccharide from Jerusalem Artichoke

Fructo-oligosaccharide （FOS） is a type of dietary fiber. It can not be digested by enzymes in the body. It also helps to improve a digestion in the intestine, an excretion system and prevent the cancer. Jerusalem artichoke is scanned and analyzed by High Performance Liquid Chromatography （HPLC） for quantity of FOS. They are selected for extraction and drying process. Studies on the extraction for a powder making, the conditions are optimum for maximum yield. Experiments manage using a factorial real 2 × 4 × 6 in 2 blocks of Randomized Complete Block Design （RCBD） models. The first factor studies on two types of solvent. The second factor is the four levels of temperature in the extraction with water at 25 ℃, 35 ℃, 50℃ and 60℃ for 30 min. The third factor is the ratio of sample per solvent as 5, 6, 10, 15, 20 and 30 times. The data and comparison of average is analyzed by Duncan＇s New Mutiple Range Test at the significant level 0.05. The concentrated extracts are processed to powder by freeze drying, a hot air and vacuum drying. Optimal conditions control the temperature and time by heating in both vacuum and conventional oven. The results of temperature on the extraction are significantly different at a = 0.05. In drying process with high temperature, the color of product is more yellow （b） and less white （L）. The difference of color value is statistically significant at level a = 0.05. The sensory evaluation of food products are added the extracted powder and attribute by panelist. High level acceptance of product is aspectual about texture and flavor. FOS is the trend of functional foods for health. It is focus on low energy and on obesity including increasing the absorption of calcium.

Biodegradation of Acid Anthraquinone Dye in a Facultative-aerobic Process:Kinetics and Products

The transformation of an anthraquinone dye blue 324 in a facultative-aerobic(F-A) system was investigated.Kinetic parameter study showed that higher Vmax coupled with more recalcitrant chemical oxygen demand(COD) were found in the facultative biofilm reactor(FBR) than in the aerobic reactor(AR).Results of the product analyses indicated that most of dye molecular could be facultatively broken down into simple intermediates,which would be further degraded under subsequent aerobic condition.The main metabolites in each reactor were detected by infrared(FT-IR) and high performance liquid chromatography and mass spectrometry(HPLC-MS).Comparison of the toxicities among the dye and its metabolites was conducted,surprisingly,the colorless intermediates from FBR possessed less inhibitory than original dye and the median effective luminescence concentration(EC50) in 15 min for aerobic effluent could not be detected,showing that hardly toxic products existed in the aerobic process effluent.

A rapid and simple ultraperformance liquid chromatography coupled to tandem mass spectrometry(UPLC-MS/MS) method for the detection of glucuronic acid-conjugated steroid metabolites

In the present study, we effectively detected 10 steroids and glucuronic acid-conjugated steroid metabolites in 12 min by ultraperformance liquid chromatography coupled to tandem mass spectrometry （UPLC-MS/MS）. Steroids testosterone （T）, 5ct-dihydrotestosterone （DHT）, androsterone （ADT）, etiocholanolone （ETIO）, estradiol （E2） and their glucuronide conjugates were well-separated on an Eclipse Plus C18 column （2.1 mm×50 ram, RRHD 1.8μm）. The mobile phase consisted of a mixture of methanol and ultrapure water （containing I mM ammonium formate） at a ratio of 60：40 （v/v）, and the flow rate was set at 0.25 mL/min. The LC eluate was detected by electrospray ionization （ESI） source in both positive and negative ion modes. Neutral loss （NL of 176, 194, 211 and 229 Da in positive mode） and precursor ion （PI ofm/z 141,159 and 177 in positive mode and 75, 85 and 133 in negative mode） methods were applied for the detection of steroid glucuronides. The multiple reaction monitoring （MRM） transitions were m/z 289.3→97.1,291.3→105, 291.3→199.2, 273.2→145.4 and 255.2→159.1 for T, DHT, ADT, ETIO and E2 in positive mode, respectively; as well as m/z 463.3→85 for T glucuronide （T-G）, m/z 465.3→75 for DHT glucuronide （DHT-G）, ADT glucuronide （ADT-G）, ETIO glucuronide （ETIO-G） and m/z 447.3→271 for E2 glucuronide （Ez-G） in negative mode. In addition, the analytical method was also applied for the detection of steroid glucuronides in pooled human liver microsomes （HLM）, which might serve as a basis for further investigation of steroid metabolism in vivo and in vitro.