应用液-液色谱联用技术在线富集测定环境水中西维因和抗蚜威的残留量

为高效液相色谱法测定环境水样中残留的西维因和抗蚜威的含量,应用了液-液色谱联用技术,实现了在线富集、分离和检测。分析中用YWG-C_(18)(10mm×4.6mm,10μm)和YWG- C_(18)(150mm×4.6mm,10μm)依次作为富集柱和分离柱,以体积比45比55混合的甲醇与水的混合溶液作为流动相,其流速为1.0mL·min^(-1)。选定紫外光度检测的波长为228nm,试样溶液在富集柱上的流速为5.0mL·min^(-1)。此外,对渗漏体积等参素也作了试验,分析信号值与两农药的质量浓度在0.04～1.0μg·L^(-1)范围内呈线性关系。测得方法的检出限(S/N=3)为0.008μg·L^(-1)(西维因)和0.014μg·L^(-1)(抗蚜威),回收试验的结果在93%～119%之间。还对日内和日间精密度进行了试验,测定西维因的日内精密度为1.3%～20.0%,日间精密度为2.3%～22.2%,测定抗蚜威的日内精密度为2.0%～10.8%,日间精密度为2.2%～5.8%。

分散液液微萃取-高效液相色谱/质谱法测定乳制品中氯霉素含量

建立了分散液液微萃取-液相色谱/质谱法(DLLME-LC/MS)测定乳制品中氯霉素(CAP)残留的分析方法。采用单次单因子法对影响DLLME富集因子的实验参数进行了优化,试验结果表明,在最优DLLME条件下,方法检测限为0.2μg/kg,当CAP质量分数在1μg/kg^20μg/kg范围内线性关系良好(R2=0.9961),样品回收率(86%～94%)、方法精密度(2.7%～3.9%)均符合要求,该方法可用于乳制品中CAP残留的检测。

高速逆流色谱在保健食品功能成分纯化中的应用

高速逆流色谱(high speed countercurrent chromatography,简称HSCCC)是一种快速、高效、连续的液-液色谱分离技术,在中药、生化、保健食品、天然产物化学、环境分析等领域有着广泛的应用,本文综述了高速逆流色谱在食品功能成分分离纯化领域的应用,并对高速逆流色谱今后的研究方向提出了一些建议。

血浆中可替宁和3-羟基可替宁的固体介质液液萃取-超高效液相色谱-串联质谱同时测定法

目的 建立血浆中可替宁和3-羟基可替宁的固体介质液液萃取-超高效液相色谱-串联质谱同时测定法。方法 在血浆样品中加入同位素内标和氢氧化钾后,使用96孔固体介质液液萃取提取盘净化,净化液经异丙醇二氯甲烷溶液洗脱、氮气吹干,纯水复溶,使用ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以0.1%氨水溶液-乙腈为流动相进行梯度洗脱,使用多反应监测模式采集数据。结果 可替宁和羟基可替宁在0.1~300 ng/ml线性范围内方法线性良好(r>0.999),可替宁方法定量下限为0.03 ng/ml,三羟基可替宁方法定量下限为0.02 ng/ml。可替宁的加标回收率为95.0%~99.4%,RSD为2.1%~3.3%。三羟基可替宁的加标回收率为95.8%~97.4%,RSD为2.1%~4.2%。结论 该方法取样量少、操作简便、方法精密度高,适合大规模样品的筛查检测工作。

血浆中可替宁的固体介质液液萃取-液相色谱-串联质谱测定法

目的建立一种快速准确测定血浆中可替宁的方法。方法将血浆样品碱化并加入内标后,通过96孔固体介质液液萃取对其进行分离纯化。经洗脱、氮气吹干、复溶后,使用Hypersil Gold C18 Selectivity色谱柱(50 mm×3 mm,1.9μm),以醋酸铵缓冲溶液-甲醇为流动相进行梯度洗脱,使用ESI源正离子扫描方式测定。结果本方法在0.50~200 ng/ml范围内线性良好(R2=0.999 5),定量检出限为0.25 ng/ml,加标回收率为84.0%~96.1%,RSDs为3.08%~7.66%(n=6)。结论该方法简单快捷,溶剂使用量小,具有高通量、高灵敏度的特点,适用于非吸烟人群及主动吸烟人群血浆中可替宁含量的测定。

Stability of Baicalin Aqueous Solution by Validated RP-HPLC

Aim In the present study a RP-HPLC method was developed and validated toinvestigate the stability of baicalin aqueous solution. Methods The influences of temperature and pHon the stability of baicalin aqueous solution were investigated by classic homoiothermicacceleration test, and the pH for the most stable solution was determined. Results The time whenbaicalin suffered 10% loss was found to be 18.1 h, and the degradation activation energy of baicalinwas 79.1 kJ·moL^(-1) . The pH at which baicalin is most stable is 4.28. Conclusion The temperatureshould be kept at a lower level and the pH should be adjusted to near that for the most stablesolution in the production of baicalin preparations.

Determination of Candicidin/FR-008 and Related Components in Fermentation Broth by RP-HPLC

Aim A liquid chromatographic method for the determination ofcandicidin/FR-008 and related components in fermentation broth has been developed. Methods Therewere four major components in the candicidin/FR-008 complex, which were separated by HPLC under thefollowing conditions: SB-C8 column (4.6 mm x 250 mm, 5 μm) was used, the mobile phase consisted ofacetonitrileam-monium acteate (20 mmol·L^(-1) , pH 4.0) (40:60, V/V) , with a flow rate of 1 .0mL·min^(-1) , the UV detection wavelength was 380 nm, and the whole process was performed at 25℃ .Results The linearity was obtained in the range of 6.25 - 500 μg· mL^(-1) candicidin/FR-008 withthe regression equation of Y = 20 461 x + 30 748 and the correlation coefficient of 0.999 1. Theinstrument precision was 1.84% and the method precision was 3.8%. Conclusion This method isaccurate, rapid and simple; it can be used for determination of candicidin/FR-008 and relatedcomponents in fermentation broth.

液液分配色谱及其与液相色谱联用分离天然花色苷的研究进展

液液分配色谱(liquid-liquid partition chromatography,LLC)是一种连续高效的分配色谱技术,如今已广泛应用于天然产物领域。天然花色苷是一种黄酮类天然色素,具有多种生理活性。花色苷极不稳定,LLC因其具有操作条件温和、可避免样品不可逆吸附等特点目前已广泛应用于天然花色苷的制备性分离。综述了2010—2019年LLC及其与液相色谱联用(liquid-liquid partition chromatography-liquid chromatography,LLC-LC)分离纯化天然花色苷的研究进展。

Improved determination of salicylaldoxime in water samples by liquid-liquid extraction followed by high performance liquid chromatographic analysis

For the determination of salicylaldoxime in environmental water samples,a stable and rapid method with low detection was proposed and established,based on the liquid-liquid extraction-high performance liquid chromatography with ultraviolet detector.Parameters including extraction solvent,ionic strength,solution pH,and extraction pattern were discussed for the optimal quantification of salicylaldoxime-spiked water.When the described method was applied to four spiked water samples,the obtained average extraction recovery rate was found between 87%–107%and relative standard deviation was below 6%.At the same time,good linear relationships were observed for spiked water samples from 0.01 to 10μg/mL(R2=0.9993).In addition,the detection limit of salicylaldoxime was revealed between 0.003–0.008μg/mL,which is two orders of magnitude lower than previously reported results.Thus,the presented method may be advantageous for the high-efficiency determination of salicylaldoxime in water samples.

Determination of risperidone in human plasma by HPLC-MS/MS and its application to a pharmacokinetic study in Chinese volunteers

This study presents a rapid, specific and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) assay for determination of risperidone (RIS) in human serum using paroxetine as an internal standard (IS). An Alltima-C18 column (2.1 mm×100 mm, 3 μm) and a mobile phase consisting of 0.1% formic acid-acetonitrile (40:60, v/v) were used for separation. The analysis was performed by selected reaction monitoring (SRM) method, and the peak area of the m/z 411.3→191.1 transition for RIS was measured versus that of the m/z 330.1→192.1 transition for IS to generate the standard curves. The assay linearity of RIS was confirmed over the range 0.25～50.00 ng/ml and the limit of quantitation was 0.05 ng/ml. The linear range corresponds well with the serum concentrations of the analytes obtained in clinical pharmacokinetic studies. Intraday and interday relative standard deviations were 1.85%～9.09% and 1.56%～4.38%, respectively. The recovery of RIS from serum was in the range of 70.20%～84.50%. The method was successfully applied to investigate the bioequivalence between two kinds of tablets (test versus reference products) in 18 healthy male Chinese volunteers. The result suggests that two formulations are bioequivalent.

Liquid Chromatography-tandem Mass Spectrometry for Analysis of Acylcarnitines in Dried Blood Specimens Collected at Autopsy from Neonatal Intensive Care Unit

Objective To investigate the feasibility of analyzing acylcarnitine in dry filter-paper blood spots by liquid chromatography-tandem mass spectrometry(LC-MS/MS) which could be applied to detect inborn errors of metabolism in neonates.Methods We obtained filter-paper blood from 26 dead infants from a neonatal intensive care unit(NICU) between October 1,2008 and September 30,2009.Acylcarnitine and amino acid profiles were obtained with LC-MS/MS.Four infants underwent routine autopsy.The postmortem blood specimens were compared with newborn blood specimens,and with specimens obtained from older infants with metabolic disorders.Results Of all the 26 patients,5(19.2%) were diagnosed as having different kinds of diseases:3 with methylmalonic acidemia(the concentration of C3,and the ratio of C3/C16,C3/C2 increased),1 with maple syrup urine disease(the concentration of leucine and isoleucine increased),and 1 with isovaleric aci-demia(the concentration of C5 increased).Conclusions Postmortem metabolic test can explain infant deaths and provide estimates of deaths attributable to inborn errors of metabolism in NICU.LC-MS/MS is suitable for analysis of postmortem specimens and can be considered for routine application in NICU autopsy.

A highly sensitive SPE-liquid/liquid extraction-RPLC analytical method for the determination of 6β-hydroxycortisol and cortisol in cancer patients' urine

A highly sensitive SPE-liquid/liquid extraction RPLC method has been developed for the analysis of 6β-hydroxycortisol and cortisol in the urine of cancer patients. Methods： After SPE column purification and liquid-liquid extraction, the sample test solutions were analyzed with RPLC using a C18 analytical column. This improved analytical method has been validated for linearity, accuracy （recovery from urine）, repeatability （within-day and between-day precision）, specificity, sensitivity, and stability. This SPE-liquid/liquid extraction-RPLC is rapid, simple, accurate and reproducible. The technique is particularly useful for monitoring the CYP3A activity of cancer patients in clinical settings. The results are expressed as the ratio of 6β-hydroxycortisol to cortisol. Results： The CYP3A activity from a total of 153 samples was measured using this improved method. Considerable variation in the CYP3A activity of different cancer patients has been documented. Thus, personalized medical treatment based on the individual metabolic enzyme activity level is necessary. Conclusion： This new analytical method facilitates such individualized medical treatments.

Reaction Kinetics of Trans-Sobrerol and 8-p-Menthen-1,2-diol with Hydroxyl Radical in Aqueous Solution： A Combined Experimental and Theoretical Study

Trans-sobrerol （Sob） and 8-p-menthen-1,2-diol （Limo-diol） are the primary products in the atmospheric oxidation of α-pinene and limonene, respectively. Because of their low volatility, they associate more likely to the liquid particles in the atmosphere, where they are subject to the aqueous phase oxidation by the atmospheric oxidants. In this work, through experimental and theoretical study, we first provide the rate constants of Sob and Limo-diol reacting with hydroxyl radical （.OH） in aqueous solution at room temperature of 3044-3 K and 1 atm pressure, which are （3.05±0.5）×10 9 and （4.57±0.2）×10 9 L/（mol.s）, respectively. Quantum chemistry calculations have also been employed to demonstrate the solvent effect on the rate constants in aqueous phase and the calculated results agree well with the measurements. Some reaction products have been identified based on liquid chromatography combined with mass spectroscopy and theoretical calculations.

Changes of Metabolites and Macro- and Micro-elements in Hungarian Potatoes under Organic and Conventional Farming

To study the effect of cultivation system （conventional and organic） on potato tuber components, 42 parameters （anti-nutritives, micro- and macro-elements and 23 metabolites） were studied in three multi-resistant Hungarian potato varieties for three years in 2007-2009. Discriminant analysis of data proved that all investigated factors （farming technology, genotype and season/year） had significant effect on metabolites. Reversed phase high performance liquid chromatographic （HPLC-DAD, HPLC-MS） and elemental analysis methods have been adapted to quantify the major components, i.e., steroidal alkaloids, nitrite, nitrate, sugars, amino acids and micro-elements, in potato tubers. The absolute amount and changes of tuber components were influenced differentially by the technology, genotype and season in a complex manner. Any examined component, except nitrate content, was found to be significantly higher or lower consistently in relation to the production practice （organic or conventional） during the three years trials. Under the examined circumstances, no consequent positive effect of organic fanning on the total amount of anti-nutritive components, vitamins or micro- and macro-elements of potato tubers could be proven.

A HPLC Method for the Determination of Salmon Calcitonin in Injection by Gradient Elution

It was concluded that the described HPLC method could be used for the assayof salmon calcitonin in injection, as it offers qualified selectivity, accuracy and precision ofanalysis.

Identification of Qishen Oral Solution with TLC and HPLC

[Objective] This study aimed to establish the standards for quality control of Qishen oral solution. [Method] Astragalus membranaceus, Atractylodes macro- cephala Koidz. and Glycyrrhiza uralensis in the preparation were identified by using thin layer chromatography （TLC）, and Codonopsis pilosula was identified by using high-performance liquid chromatography （HPLC）. [Result] By using the developed TLC system, experimental solution and reference solution showed clear spots, while negative control presented no interference. By using the developed HPLC system, the chromatographic peak of Iobetyolin was detected in experimental solution. [Con- clusion] The developed TLC and HPLC systems presented high specificity and good repeatability for identification of these four components and could be used in the quality control of Qishen oral solution.

Determination of Cordyceps Sinensis/Betaine Compound Feed Nutriment by HPLC

[Objective] The aim was to conduct HPLC analysis on Cordyceps Sinensis/Betaine compound feed nutriment. [Method] Cordyceps Sinensis/Betaine compound feed nutriment was under HPLC analysis to determine separation of Cordyceps Sinensis effluent and betaine. [Result] Different compositions in Cordyceps Sinensis/etaine compound feed nutriment would be well separated by the method. [Conclusion] The method provides a suitable platform of separation and analysis for Cordyceps Sinensis /Betaine compound feed nutriment.

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.

Economic and Qualitative Traits of Italian Alps Saffron

Saffron, obtained from the flower stigmas of Crocus sativus L., is one of the most expensive food spices. The introduction of saffron in alpine areas could help to broaden and diversify the activities of mountain multifunctional farms, with a positive impact on economy and land management. According to ISO 3632(2010/2011), saffron can be classified into three categories of quality(I, II, III) depending on the concentration of the three main metabolites responsible for its characteristic colour, flavor and aroma: Crocin, Picrocrocin and Safranal. This study represents the first investigation of the quality of saffron produced in the Italian Alps evaluated with spectrophotometry, HPLC, solid-phase microextraction(SPME), and gas chromatographic analysis combined with mass spectrometry(GC/MS). The experiments used Crocus sativus stigmas produced in 2012-2013 in different areas of the Central Italian Alps were located at an altitude between 720 and 1200 m a.s.l.. Results obtained were compared to commercial saffron. The analyses confirmed that all samples can be classified in the first quality category according to the ISO classification. This high quality is also confirmed by HPLC analysis. Moreover, the SPME-GC/MS analysis identified some differences in the aromatic profile of saffron samples, in particular regarding safranal concentration. A preliminary assessment of the economic viability of high quality saffron production for local markets was also performed. Our study provides valid information regarding the quality and economic sustainability of saffron production in the alpine area confirming this crop as a good candidate for a new source of income for multifunctional farms in mountain areas.

Ultrafiltration of Shiitake Polyphenol with Ultrasound Enhancement

Ultrafiltration （UF） experiments were subjected to the concentration of Shi- itake polyphenol（SP） with ultrasound （US） enhancement, by which the effects of the main parameters such as ultrasonic power, transmembrane pressure, temperature and axial flow-rate on the membrane fluxes were analyzed, and then the SP of the ultrafiltrate after follow-up resin adsorption by HPLC was investigated. The results in- dicated that US could significantly enhance the membrane fluxes. The optimal con- ditions obtained was as follows： 0.60 W/cm2 US power, axial flow-rate of 80 L/h, operation temperature 30 ~C, transmembrane pressure 0.10 MPa. US increased the absorption capacity of the absorbent resin during the following stage. The HPLC analysis also showed that catechins were purified to some extent as the ultrafiltrate was treated by US and macroporous resin, indicating the technology of US en- hancement coupled with UF showed the potential for concentration and purification of SP by absorbent resins.