Mercury ion (Hg2+) pollution poses severe environmental and health risks, necessitating the development of highly sensitive and reliable detection methods. Although ratiometric and genetically encoded fluorescent probes each offer distinct advantages, their integration for Hg2+ sensing remains largely unexplored. Herein, we report the rational design of a novel ratiometric genetically encoded fluorescent probe through the fusion of a circularly permuted green fluorescent protein (cpEGFP), a mercury-binding domain (MerBD), and a large Stokes shift reference fluorescent protein (LSSmOrange). This probe enables self-calibrated Hg2+ quantification by measuring the fluorescence intensity ratio of the response channel (F512) to the stable reference channel (F572), which effectively minimizes interferences from probe concentration, environmental fluctuations, and instrumental variations. The probe exhibits an ultra-high affinity for Hg2+ (apparent Kd' = 2.71 × 10-13 M) and a detection limit of 4 nM, alongside long-term stability and high selectivity against most common metal ions. Competitive titration, circular dichroism, and fluorescence lifetime analyses reveal that the probe operates through a cooperative conformational-change mechanism, which translates picomolar-level binding into a nanomolar-level fluorescence response. This work not only addresses a critical gap in ratiometric genetically encoded probes for Hg2+ but also provides a robust and versatile platform for accurate Hg2+ monitoring in environmental and biological systems.

Ten 4 '- (R-phenyl) -2,2 ': 6 ', 2 ' - terpyridine ligands (R = hydrogen (L1), hydroxyl (L2), methoxyl (L3), methylsulfonyl (L4), methyl (L5), phenyl (L6), fluoro (L7), chloro (L8), bromo (L9), and iodo (L10)) were synthesized. The reaction of these ligands with copper(II) nitrate led to complexes 1-10. The characterization of 1-10 was carried out by means of mass spectrometry, elemental analysis, infrared spectroscopy and X-ray single crystal diffraction. Four cell lines including esophageal cancer cell line (Eca-109), human liver cancer cell line (Bel7402), human breast cancer cell line (SIHa) and human normal liver cell line (HL-7702) were selected to carry out antiproliferation and cytotoxicity experiments in vitro. The results showed that the complexes have strong inhibitory ability on the growth of tumor cells. In order to study the anticancer mechanism of the complexes, the binding mode and binding ability of the complexes with DNA were further determined and discussed with UV-Vis spectroscopy and circular dichroism. The effects of the lowest binding energy and hydrogen bond on the binding were studied using molecular docking calculation.

评价广西贺州市K326植烟区土壤肥力状况,为高产优质烟叶生产提供参考.以贺州市K326烟区为评价单元,以约6.67 hm2为一个采样单元对贺州市主产烟区采集187个土壤样品(富川县140个、钟山县44个、昭平县3个),测定土壤化学肥力指标,对土壤养分丰缺进行评价和适宜性分析.结果表明,贺州市K326烟区土壤养分肥力整体较好,有机质、氮营养供应充足,磷营养处于丰富水平,钾营养处于中等水平,部分区域钾营养供应不足,pH总体处于中等至偏碱水平.植烟土壤钙营养供应过多,部分区域土壤镁供应不足,而硫营养供应较合适.钾钙镁供应平衡性差,钾镁比和钙镁比大于适宜范围的样本占比大,容易导致缼镁.植烟土壤的有效铜、锌、铁、锰4种营养元素供应充足,硼、钼营养元素普遍缺乏.通过土壤养分丰缺评价和适宜性分析,为贺州市K326烟区高产优质烟叶生产的施肥管理提供参考.

Purpose To study the effects of different fertilization methods and different selenium enrichment levels on the contents of elements and active components in flue-cured tobacco. MethodsYunyan87 was used as test material, and the following treatments were applied: no application of exogenous selenium application (CK), application of selenium nanoparticles in the rhizosphere (S1), foliar spray application of selenium nanoparticles with 33.75 g/hm2 (S2) and foliar spray application of selenium nanoparticles with 67.50 g/hm2 (S3). Compositional and multiple omics differences between flue-cured tobacco under different treatments were studied using inductively coupled plasma mass spectrometer, inductively coupled optical emission spectrometer, and ultra-performance liquid chromatography-quadruple time-of-flight mass spectrometer. ResultsThere were obvious differences of element distribution among the four groups of flue-cured tobacco, with the main contributing elements being S, Mg, Mn, Al, P, and Na. The metabolomics of the four groups also displayed obvious differences, with a total of 29 differential metabolites identified, including four phytosterols and their derivatives, nine phenolic compounds, seven amino acid compounds, three aldehyde compounds, and six other compounds. The effect of foliar application of selenium fertilizer on the elements and chemically active components of flue-cured tobacco was found to be more significant than that of rhizosphere application. ConclusionThese results may serve as a reference for investigating the absorption and bioaccumulation mechanism of selenium-enriched flue-cured tobacco’s elements and chemically active components.

为了明确靖西市植烟区土壤交换性钙、镁含量及其比值的特征,阐明其与土壤酸碱的相关性,以靖西市主要植烟区土壤为研究对象,2022年6月采集植烟区具有代表性的土壤,测定土壤交换性钙、镁含量及计算其钙镁比值,并分析其与土壤酸碱的相关性.结果表明,靖西市主要植烟区土壤交换性钙、镁含量丰富,平均值分别为5 114.59、268.57 mg/kg,土壤交换性钙镁比值大于10,占71.01%.钙镁比值表现为强变异,且交换性钙的变异性大于交换性镁.植烟区土壤pH普遍偏高,pH与土壤交换性钙含量呈极显著正相关,与交换镁含量呈显著负相关,pH对交换性钙含量的影响大于交换性镁,植烟土壤为微碱性时,交换性钙含量处于高及以上水平的占比达100%.综上所述,靖西市主要植烟区土壤交换性钙、镁含量总体处于偏高水平,钙镁比变异强度大,易出现钙、镁平衡失调.因此,今后在生产中需注意酸碱性钙镁肥料的使用以调节土壤酸碱性,并适时培肥土壤,调节土壤钙、镁平衡,提高烤烟产量及品质.

[目的]探索水稻中不同营养元素响应稻瘿蚊取食的差异,为稻瘿蚊防治及水稻抗稻瘿蚊机理阐释提供新思路.[方法]以稻瘿蚊抗性近等基因系43W-NIL和感性品系9311为材料,利用电感耦合等离子体质谱(ICP-MS)测定抗感水稻材料在接种稻瘿蚊后第0、5、7和9 d时植株体内的营养元素含量;利用水培法验证关键元素受稻瘿蚊取食诱导的变化;通过qRT-PCR测定与营养元素代谢相关基因在抗感材料中的表达量.[结果]离子组分析共鉴定25种营养元素,大部分元素在抗感材料及稻瘿蚊取食前后差异明显.接种稻瘿蚊后,与9311的第0 d相比,43W-NIL中Ti、K、P和S元素含量较高,而Pb、Sr和Ca元素含量较低.离子组的主成分分析发现,PC1和PC2分量分别解释40.0％和24.7％的变异.主成分载荷分析结果表明,Ca、Sr、Co、Pb、Be、Sb、Fe、Ti、P、S和K的含量变化可以区分抗性和感性材料,且影响稻瘿蚊抗性最重要的5种元素依次为Ca、Sr、P、K、Co.水培植株的接虫试验发现,高浓度的外源Ca处理可促进稻瘿蚊幼虫在抗性水稻中生长,而低浓度P、K处理会抑制稻瘿蚊幼虫在感性材料中生长.qRT-PCR结果显示,钙调蛋白基因(OsCML20)表达量与Ca2+含量的变化趋势相同;钾转运蛋白基因(OsHAK12)的表达量在抗感材料中均表现为先下降后上升.[结论]Ca、P和K等3种元素可能在水稻响应稻瘿蚊取食中发挥关键作用.OsHAK12和OsCML20基因可能是稻瘿蚊抗性表达机制中的重要基因.离子组学可从元素、营养的角度发现水稻在被稻瘿蚊取食时的差异,为植物抗虫机理研究提供新思路.

为探明接种丛枝菌根真菌对烤烟湿润育苗烟苗生长及其产质量的影响,并与常规漂浮育苗进行比较,以烟草"K326"品种为试验材料,采用湿润育苗和大田试验方法,设置育苗基质配制10％缓释肥并接种低、中、高水平丛枝菌根真菌(CQ1、CQ2、CQ3)处理和漂浮育苗对照,同时培育成苗后移栽大田试验,分析各处理对烟苗素质、烤烟生长及烟叶产质量的影响.结果表明:与漂浮育苗对照相比,接种中、高、低水平丛枝菌根真菌均能促进烟苗生长,提高烟苗根长、根数量、根系体积和烟苗生物量.接种丛枝菌根真菌处理的烟叶产量均显著高于对照处理,其中CQ3处理烤烟产量提高16.27％;CQ2处理烟叶产值和上等烟叶产量最高,且与对照相比差异显著,其中产值增加11.42％;CQ2处理的烟叶上等烟比例也比对照处理高,达到85.71％.综上所述,接种丛枝菌根真菌处理可提高烤烟湿润育苗的烟苗素质,并能显著提高烤烟烟叶的产量、产值和品质,可替代常规漂浮育苗.

为研究叶面喷施氨基酸螯合硒对水稻籽粒硒含量及形态的影响,以水稻品种百香139为材料,通过大田试验,在水稻破口期叶面喷施不同用量的氨基酸螯合硒,并在水稻拔节孕穗期、始穗期、扬花授粉期、灌浆期、蜡熟期5个不同生育期叶面喷施13.5 g/hm2(以硒计)的氨基酸螯合硒,在水稻成熟期测定、分析各处理组水稻地上部不同组织器官硒含量和籽粒硒形态.结果表明,喷施外源氨基酸螯合硒可提高水稻各器官硒含量,在适量范围内水稻植株各器官硒含量随着喷硒浓度的增加而提高;水稻籽粒中硒主要以硒代蛋氨酸存在,水稻籽粒中硒代蛋氨酸含量与总硒含量的变化规律相一致.喷施氨基酸螯合硒用量为6.75、13.5、27.0 g/hm2时水稻籽粒中硒含量分别为0.168、0.431、0.439 mg/kg,均达到《富硒农产品硒含量分类要求》(DB45/T 1061-2014)的要求.拔节孕穗期至灌浆期喷硒处理水稻籽粒硒含量呈现增高趋势,灌浆期喷硒处理的水稻籽粒总硒含量最高,达到0.320 mg/kg;水稻籽粒中硒代蛋氨酸含量占总硒含量的比例为43％～91％,硒代蛋氨酸含量占总硒含量的比例由大到小依次为:拔节孕穗期>始穗期>扬花授粉期=灌浆期>蜡熟期,硒代蛋氨酸比例随着施硒时期越靠近水稻成熟期越低,表明水稻叶片中硒的转化时间对稻米中硒形态有显著影响,提早喷施硒有利于有机态硒的形成,提高稻米中有机态硒的比例.在硒的生物强化生产富硒水稻中,水稻破口期叶面喷施氨基酸螯合硒的最佳用量为13.5 g/hm2,在5个不同生育期喷施13.5 g/hm2的氨基酸螯合硒均能生产出富硒大米,且最佳喷施外源硒的时期为灌浆期.

本研究通过对木薯根系分泌物进行提取、分离与鉴定,探究不同木薯品种根系分泌物的差异,为筛选抗化感耐连作木薯新种质提供参考.以木薯组培苗琼脂培养基为试验材料,采用正交试验设计,结合GC-MS技术考察不同萃取材料和洗脱剂对木薯根系分泌物的提取和分离效果,选择最优方案测定'新选048'南植199'和'华南205'的根系分泌物.结果表明:(1)木薯根系分泌物水溶性物质提取的最优方案为:用去离子水超声提取捣碎的琼脂培养基30 min,固液分离后用XAD-2萃取、无水乙醇洗脱,浓缩后用GC-MS检测,成功鉴定出包括有机酸类、醇类、酯类、酮类、醛类等26种有机化合物.(2)醇溶性物质提取的最优方案为:用50％乙醇超声提取捣碎的琼脂培养基30 min,固液分离后用XAD-4萃取、无水乙醇洗脱,浓缩后用GC-MS检测,鉴定出包括有机酸类、醚类、酯类、酮类、醛类等15种有机化合物.(3)不同品种根系分泌物的水溶性和醇溶性成分均有差异.'南植199'根系分泌物的主要水溶性物质有羟乙酸甲酯(相对含量为3.72％)、羟基丙酮(2.40％)、甲肼(1.79％)等,主要醇溶性物质有乙醇醛(18.89％)、羟基丙酮(2.47％)、甲酸(2.25％)等;'新选048'的主要水溶性物质有甲酸(2.68％)、1,5-戊二醇(2.39％)、丙烯酸羟乙酯(2.01％)等,主要醇溶性物质有乙醇醛(17.00％)、甲酸(2.62％)、羟基丙酮(2.46％)等;'华南205'的主要水溶性物质有甲酸(2.23％)、羟基丙酮(1.80％)、羟乙酸甲酯(1.43％)等,主要醇溶性物质有乙醇醛(16.58％)、甲酸(3.06％)、八氟戊醇(2.98％)等.不同木薯品种的根系分泌物种类和含量均有差异,从而导致其抗化感耐连作能力的差异,为筛选耐连作品种缓解木薯连作障碍成为可能.

A signal-amplified mercury sensing biosensor with desired sensitivity was developed through firstly using the GFP mutant with fluorescence increasing response towards Hg2+ as the reporter module. The developed biosensor showed response for Hg2+ in a relatively wide range of 1-10,000 nmol/L, and the detection limit was improved one or two orders of magnitude in comparison with most metal-sensing biosensors in similar constructs. In addition, the biosensor could distinguish Hg2+ easily from multiple metal ions and displayed strong adaptability to extensive pH conditions (pH 4.0-10.0). More importantly, the developed biosensor was able to provide an initial assessment of Hg2+ spiked in the environmental water with the recoveries between 85.70% and 112.50%. The signal-amplified strategy performed by the modified reporter module will be widely applicable to many other whole-cell biosensors, meeting the practical requirements with sufficient sensing performance.

为研究不同营养水平下薏苡人工湿地对Cr(Ⅵ)的净化与形态转变化的影响,以生活污水(W)、1/2(体积比)Hoagland营养液(Y)、1/2 Hoagland营养液+生活污水(Y+W)为灌溉水源,构建微型模拟垂直流薏苡人工湿地,采取间歇进水方式,进水分别以0、10、30 mg/L左右Cr(Ⅵ)(以K2Cr2O7形式加入)进行处理.结果 表明:(1)根、茎、叶中总铬均随处理时间的延长和Cr(Ⅵ)浓度的提高而提高;不同器官之间相比较,总铬依次为根＞叶＞茎.(2)在不同营养条件下,基质中可交换态、铁锰氧化物结合态、有机物结合态、残渣态铬含量均随着Cr(Ⅵ)浓度的增加而提高,而碳酸盐结合态铬则先升高后降低.(3)Y处理出水Cr(Ⅵ)较高,W、(Y+W)处理均能提高人工湿地对含铬污水的处理效果.

This study reports the fabrication of orange peels (OP) extract (water:ethanol:acetonitrile) modified organic-inorganic hybrid bentonite (OP-bentonite). The synthesized composite was employed for the adsorptive removal of carcinogenic mycotoxins (aflatoxin B1 (AFB1), fumonisin B1(FB1) and ochratoxin (OTA)). The product was texturally characterized by BET, FTIR, XRD, TGA, SEM, EDX and XPS techniques while quantification of toxins was performed using HPLC/LC-MS. The effect of reaction temperature, pH, adsorbent dosage, coexisting mycotoxins and gastro-intestinal fluids on the adsorption performance of OP-bentonite was systematically investigated. Isothermal data (pH 2.5 and 7.0) for toxins adsorption agreed well with the Langmuir isotherm model revealing maximum adsorption capacity (qmax) (mg/g) of 166 and 121 for AFB1, 3.87 and 1.78 for FB1 and 1.97 and 3.19 for OTA respectively. The adsorption kinetics followed pseudo-second-order kinetic model while its adsorption capacity surpassed many adsorbents reported for AFB1, FB1 and OTA with orders of magnitude. Therefore, attributed to the advantages of being efficient, green and simplified synthesis, OP-bentonite could be envisaged as a promising adsorbent for the enhanced sequestration of mycotoxins.

Mercury pollution has always been a huge threat to human health due to its significant toxicity. Thus, it's the continuing goal to obtain new mercury detection techniques that are cost-effective, operational stable, performance efficient, and applicable to the environmental and biological milieus. In this research, the soluble pigment pyocyanin with anti-bacterial and anti-fungal activities, the biosynthesis pathway of which was engineered under the regulation of Hg2+-dependent transcriptional activator MerR, was firstly used as the visual detection signal in the whole-cell biosensor. The engineered biosensor displayed optical sensing window and a good linearity for Hg2+ in the range of 25-1000 nM, and the detection limit could reach as loW as 10 nM. It permitted on-site detection of bioavailable Hg2+ with extraordinary selectivity and could resist the interferences of extra metal ions. What's more, the developed biosensor performed function well in a wide pH range (pH 4-10) as well as the environmental water. By fully imitating and utilizing the biosystems from nature, the engineered colorimetric biosensor has great economic and performance advantages over most chemosensors as well as whole-cell biosensors in the practical application of detecting Hg2+ in the contaminated aquatic systems. (C) 2019, The Society for Biotechnology, Japan. All rights reserved.

Yizhi Granule (YZG) is a health food containing six traditional Chinese medicines (TCMs). It improves memory barriers in rat experiments. Here, we describe the first fast and sensitive ultraperformance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC/ESI-Q-TOF MS) method for analyzing YZG in plasma. We used this technique for studies in cynomolgus monkey plasma. By comparing retention time, MS, and MS/MS data of reference compounds, 70 compounds were detected in YZG. Of these, 63 were identified including 60 saponins, 2 flavones, and 1 methyl ester. There were 33 saponins, 1 flavone, and 1 methyl ester in the plasma. Next, to study the therapeutic properties of YZG, the neuroprotective effect of some of the absorbed components was evaluated using PC12 cell damage caused by the Aβ25–35 model. The results showed that 9 compounds protect PC12 cells from Aβ25–35 with cell viability (%) of 111.00 ± 8.12 (G-Rb1), 102.20 ± 4.22 (G-Rb2), 100.34 ± 6.47 (G-Rd), 102.83 ± 2.10 (G-Re), 101.68 ± 7.64 (NG-Fa), 101.19 ± 7.83 (NG-R1), 102.53 ± 0.55 (NG-R2), 106.88 ± 4.95 (gypenoside A), and 103.95 ± 4.11 (gypenoside XLIX), respectively, versus the control group (87.51 ± 6.59). These results can reveal the real pharmacodynamic basis of YZG and provide a theoretical basis for subsequent studies. It can also provide some references for the research of Alzheimer’s disease.

[目的]拟解决由于土壤有效硒含量不均导致的广西区内富硒水稻生产中出现的硒肥施用量难以把控的问题.[方法]通过大田试验,以百香139水稻品种为试材,在不同土壤有效硒条件下叶面喷施硒肥,研究不同土壤有效硒水平下的水稻外源硒生物强化适用量.[结果]叶面喷施适量外源硒可以增加水稻籽粒中硒含量,同时对水稻有一定的增产效果,且土壤有效硒含量越低,增产越显著;若不考虑土壤其他相关条件的影响,水稻籽粒中硒含量与土壤总硒含量相关性为0.573*,与土壤有效硒含量的相关性为0.691**;富硒大米硒含量为0.15～0.50mg/kg(DB45/T1061-2014)范围又基本能达到增产效果,土壤有效硒含量<20.00μg/kg、20.00～30.00μg/kg、30.00～40.00μg/kg、40.00～50.00μg/kg、>50.00μg/kg的5个不同土壤有效硒水平区外源硒用量分别为0.54～2.83g/667m2、0.60～2.50g/667m2、0.25～1.61g/667m2、0.23～1.16g/667m2、0.29～0.78g/667m2.[结论]叶面喷施适量硒肥可以提高水稻产量;提出不同土壤有效硒水平下富硒水稻外源硒叶面强化的适用量范围.

Mercury pollution has been one of the most serious environmental problems today. The detection and remediation of mercury contamination are of special concern due to its nondegradable nature, bioaccumulation effect, and significant toxicity. In this research, the whole-cell based detection and remediation systems for Hg2+ were engineered based on the cell surface engineering technique and transcriptional activation mechanism of metalloprotein MerR. The engineered biodetection system showed high selectivity for Hg2+ over other metal ions with the detection limit as low as 0.1 nM. It also permitted a rapid, visual, and on-site detection of bioavailable Hg2+ for large-scale water samples with simple processes. The engineered bioremediation system, which was more efficient than the most current biosystems with limited adsorption capacities, could continuously reduce mercury contamination in wide concentration by transforming highly toxic Hg2+ to volatile and much less deleterious Hg2+ with extraordinary selectivity. In addition, the bioremediation system could perform function in the absence of extra inductive agent and showed an increased protection of cells against Hg2+ biotoxicity. By fully imitating and utilizing the microorganisms, the engineered biochemical systems have great potential in the practical application of detecting and remediating Hg2+ in the contaminated aquatic systems.

A method for simultaneously determining indaziflam and its five metabolites in soil, water, and fruits using ultraperformance liquid chromatography/tandem mass spectrometry was established. The analytes were eluted in <4.5 min. Positive electrospray ionization mode was used. The analytes were extracted using acetonitrile containing 1% ammonium hydroxide, and then the extracts were purified using octadecylsilane and PRiME HLB cartridges. The quantification limits were 0.01-1.01 mu g kg(-1). The linearities of the calibrations for all analytes were excellent (R-2 > 0.9952). The recoveries at spike concentrations of 0.01, 0.1, and 1 mg kg(-1) were 81.3-112.1%. The intraday and interday relative standard deviations were <13.5% and <12.3%, respectively. The method was successfully used to determine indaziflam and its five metabolites in samples from markets and fields. The results confirmed that the method is an effective and robust procedure for routinely determining indaziflam and its metabolites in soil, water, and fruit samples.

Using caffeic acid and icariin as internal standards, a method for the simultaneous determination of protocatechuic acid, protocatechuic aldehyde, chlorogenic acid, scutellarin, isochlorogenic acid C, baicalin, luteolin, apigenin, atractylenolide III and atractylenolide I in Fufangxingxiangtu' erfeng capsules were established by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The separation was performed on a ZORBAX RRHD Eclipse Plus C18 column (50 mm x 2.1 mm, 1.8 microm) by using water containing 0.3% formic acid and methanol as mobile phases with the gradient elution at a flow rate of 0.3 mL/min. The analytes were detected by a tandem mass spectrometer in the multiple reaction monitoring (MRM) mode via the switching of positive electrospray ionization (ESI(+)) and negative electrospray ionization (ESI(-)). Under optimum conditions, the calibration curves were linear in the range of 0.003 00-24.0 mg/L for protocatechuic acid, 0.017 0-2.00 mg/L for protocatechuic aldehyde, 0.015 0-30.0 mg/L for chlorogenic acid, 0.004 00-30.0 mg/L for scutellarin, 0.010 5-24.0 mg/L for isochlorogenic acid C, 0.003 00-30.0 mg/L for baicalin, 0.003 00-5.0 mg/L for luteolin, 0.006 00-1.50 mg/L for apigenin, 0.001 50-4.00 mg/L for atractylenolide III, and 0.000 600-0.900 mg/L for atractylenolide I with the detection limits of 1.0, 11, 5.0, 1.5, 3.5, 1.0, 1.0, 2.0, 0.50, 0.20 microg/L, respectively. The average recoveries of the ten effective components were between 92.5% and 106% with all relative standard deviations not more than 3.2%. The developed method was rapid, simple, accurate, reproducible, and suitable for the quality control of the Fufangxingxiangtu' erfeng capsules.

Objective: A method for simultaneous determination of matrine and oxymatrine in Xianning Linimentum was established by ultra performance liquid chromatography with tandem mass spectrometry( UPLCMS-MS). Method: The UPLC separation was performed on C18column( 2. 1 mm × 50 mm,1. 8 μm) by using methanol and water containing 0. 1% formic acid as mobile phase with the gradient elution at a rate of 0. 30 mL· min- 1. The analytes were detected by tandem mass spectrometry under the positive ion mode with the ESI source and carried out in the multiple reaction monitoring( MRM) mode. Result: Under the optimum conditions,the calibration curves were linear in the range of 0. 000 5-0. 6 mg ·L- 1for matrine,0. 000 4-0. 5 mg ·L- 1for oxymatrine. The detection limits were 0. 2,0. 2 μg·L- 1respectively. The average recoveries of the three effective components were between 95. 8% and 98. 6% with all relative standard deviations not more than 1. 9%. Conculsion: The developed method is simple,rapid and accurate,and suitable for the quality control of the two components in Xianning Linimentum.

AIM A method for simultaneous determination of ephedrine hydrochloride,pseudoephedrine hydrochloride and codeine phosphate in Kesuting Syrupy was established by ultra performance liquid chromatography with tandem mass spectrometry( UPLC-MS / MS). METHODS The UPLC separation was performed on a Zorbax RRHD Eclipse Plus C18column( 2. 1 mm × 50 mm,1. 8 μm) by using methanol and water which contained 0. 1% formic acid as mobile phase with the gradient elution at a flow rate of 0. 30 mL / min. The analytes were detected by tandem mass spectrometry under the positive ion mode with the ESI source and carried out in the multiple reaction monitoring( MRM) mode. RESULTS Under the optimum conditions,the calibration curves were linear in the range of 0. 001 6- 4. 5 mg / L for ephedrine hydrochloride,0. 000 8- 4. 8 mg / L for pseudoephedrine hydrochloride,0. 004- 2. 0 mg / L for codeine phosphate. The detection limits were 0. 8,0. 4,1. 2 μg / L,respectively. The average recoveries of the three effective components were between 91. 1% and 104% with all relative standard deviations not more than 2. 6%. CONCLUSION The developed method is simple,rapid and accurate,and suitable for the quality control of the three components in Kesuting Syrupy.