氨基糖苷类抗生素在蒸发光散射检测器中响应因子的一致性考察

目的考察不同的氨基糖苷类抗生素在蒸发光散射检测器中的响应因子是否一致,进而考察ELSD在该类药的有关物质检查、多组分同类物质的相对比例测定及氨基糖苷类新药基准品的建立等方面应用的可能性。方法用HPLC-ELSD法测定了一组氨基糖苷类抗生素(阿米卡星、西索米星、奈替米星、依替米星和一类新药威替米星)的线性方程。色谱条件:Diamonsil C₁₈柱150 mm×4.6 mm,5 μm;流动相0.2 mol·L^-1三氟醋酸-甲醇(94∶6);流速0.6 mL·min^-1。检测器条件:漂移管温度110℃,气体流速2.80 L·min^-1。结果上述5种物质的线性方程间无显著性差异(P＞0.05)。结论5种不同氨基糖苷类抗生素在蒸发光散射检测器中的响应因子一致,可以尝试用HPLC-ELSD法测定该类药中的有关物质、控制多组分药物的相对比例并建立一类新药基准品。     

HPLC-ELSD法测定玉屏风口服液中黄芪甲苷的含量

目的 建立玉屏风口服液中黄芪甲苷的含量测定方法。方法 采用HPLC-ELSD法,Kromasil-C₁₈(250mm×4.6mm,5μm)色谱柱;流动相为甲醇-水(74∶26);流量为0.8ml·min^-1;柱温为30℃;ELSD检测器检测;ELSD漂移管温度为90℃;气体流量1.48L·min^-1。结果 平均加样回收率为99.1%,RSD为2.76%。结论 方法准确、可靠,可用于玉屏风口服液的质量控制。     

HPLC-ELSD法测定贝母中异甾类生物碱及糖苷类成分的含量

目的建立同时测定贝母中5种异甾类生物碱——peimissine, imperialine, sinpeinine A, imperialine-3β-glucoside和yibeinoside A含量的HPLC分析方法。方法C₁₈柱;流动相:乙腈-水(含0.1%二乙胺);梯度洗脱,流速1.0 mL·min^-1;检测器:Alltech 500蒸发光散射检测器(ELSD)。结果线性范围为peimissine 13.1～288.2 mg·L^-1(r²=0.997 5), imperialine-3β-glucoside 7.7～169.4 mg·L^-1 (r²=0.999 3), yibeinoside A 7.3～160.6 mg·L^-1 (r²=0.999 7), imperialine 16.5～363.0 mg·L^-1 (r²=0.999 2), sinpeinine A 8.7～191.4 mg·L^-1 (r²=0.994 2)。 5个化合物的精密度和重现性RSD均<5%。结论本方法简便、有效、可行,可用于贝母中5种异甾类生物碱的含量测定。     

湖北贝母的HPLC指纹图谱分析

目的建立湖北贝母的指纹图谱分析方法。方法采用高效液相色谱-蒸发光散射检测法以Hypersil ODS(250 mm×4.6 mm ID，5 μm)为色谱柱；流动相：甲醇(含0.05%三乙胺)-水梯度洗脱；流速：1.0 mL·min^-1；记录时间：60 min；以蒸发光散射检测器(ELSD)检测，检测条件：漂移管温度75 ℃，氮气流速1.9 L·min^-1。结果用梯度洗脱得到的色谱图各色谱峰分离较好，达到指纹图谱要求。结论为更好的控制湖北贝母的内在质量提供了可靠的分析方法。     

HPLC-ELSD和IC-ICD检测磺丁基-β-环糊精中的杂质离子

目的 建立了高效液相色谱-蒸发光散射法（HPLC-ELSD）及离子色谱-抑制电导检测法（IC-ICD）测定磺丁基-β-环糊精中氯化钠、4-羟基-1-丁烷磺酸钠、双（4-磺丁基）醚二钠3种杂质含量的方法。方法 HPLC-ELSD采用CD-Screen-IEC柱（4 mm×150 mm，3 μm），梯度洗脱，流速为1.2 mL·min^-1；IC-ICD采用Dionex IonPac AS19（4 mm×250 mm）阴离子交换色谱柱分离，30 mmol·L^-1氢氧化钠溶液为淋洗液，等度洗脱，流速为1.0 mL·min^-1。结果 2种方法各组分分离度良好，HPLC-ELSD法中氯化钠、4-羟基-1-丁烷磺酸钠、双（4-磺丁基）醚二钠检出限（S/N=3）分别为0.230，0.270和0.340 ng，IC-ICD法检出限分别为0.041，0.027和0.014 ng。结论 HPLC-ELSD和IC-ICD在磺丁基-β-环糊精中氯化钠、4-羟基-1-丁烷磺酸钠、双（4-磺丁基）醚二钠3种杂质的含量测定结果上没有明显不同，可根据实际情况选择，只是根据2种方法的检出限及定量限值，IC-ICD法灵敏度更高。     

HPLC-ELSD同时测定丹参川芎嗪注射液中间体中7种糖类成分

目的 建立同时测定丹参川芎嗪注射液中间体中果糖、葡萄糖、蔗糖、蜜二糖、棉子糖、甘露三糖和水苏糖7种糖类成分的HPLC-ELSD方法，并研究丹参川芎嗪注射液生产过程中糖类成分的变化规律。方法 采用Prevail Carbohydrate ES色谱柱（250 mm×4.6 mm，5 μm），以乙腈-水为流动相进行梯度洗脱，柱温为25 ℃，进样量为5 μL，流速为0.6 mL·min^–1，ELSD检测器雾化器温度为70 ℃，漂移管温度为60 ℃，氮气流量1.0 L·min^–1。结果 7种糖类成分在定量范围内线性关系良好，R²均>0.998 0，进样、日内、日间和中间精密度RSD值均<5%，供试品溶液在10 h内稳定，低、中、高浓度平均加样回收率为97.00%~104.34%。结论 建立的分析方法稳定、准确、重复性好，可用于丹参川芎嗪注射液中间体中7种糖类成分含量测定。本研究推进了丹参川芎嗪注射液安全性再评价质量控制研究，也为其他中药制剂中糖类成分的定量、转移、转化规律研究提供了参考。     

日本救心丹多维指纹图谱研究

目的综合运用多维联用技术，建立日本救心丹HPLC-UV/ELSD和HPLC-UV/MS³指纹图谱研究方法，初步阐明其化学物质基础。方法Altima C₁₈色谱柱(250 mm×4.6 mm ID，5 μm)；流动相为乙腈-0.2%甲酸水体系；梯度洗脱；流速1.0 mL·min^-1；检测波长286 nm。ELSD条件为漂移管温度110 ℃，N₂流速2.5 L·min^-1。采用电喷雾离子阱质谱分析，正、负离子三级扫描。结果建立了日本救心丹HPLC-UV/ELSD多维指纹图谱，同时通过MS定性鉴别出21种成分，并推测可能存在去氧苷胆酸及其异构体。方法稳定且重现性好。结论该研究为分析汉方中药复杂体系提供了一种有效、可靠的模式。     

HPLC-CAD法与ELSD法用于测定蒺藜皂苷提取物中蒺藜皂苷K的对比研究

目的：分别建立高效液相色谱-电雾式（CAD）和蒸发光散射（ELSD）2种通用型检测器测定蒺藜皂苷提取物中蒺藜皂苷K含量的方法。方法：采用高效液相色谱法，Waters Symmetry Shield RP18为色谱柱，以乙腈-水为流动相，梯度洗脱，体积流速为1.0 mL·min^-1，柱温为25 ℃；使用CAD和ELSD检测器测定，并对样品实测结果进行比较。结果：2种方法线性关系均良好，精密度、重复性、稳定性及加样回收率皆符合方法学要求，CAD的检出限和定量限均比ELSD低。结论：2种检测方法都可用于蒺藜皂苷提取物中蒺藜皂苷K的含量测定，CAD检测器比ELSD检测器具有更高的灵敏度和精密度。     

HPLC-ELSD法测定复胃散胶囊中黄芪甲苷的含量

目的 建立高效液相色谱-蒸发光散射检测器法测定复胃散胶囊中黄芪甲苷的含量。方法 采用Agilent 1200高效液相色谱仪和Waters XSELECT C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为乙腈-水（32:68）,流速为1.0 mL·min^-1,进样量为20 μL,蒸发光散射检测器（漂移管温度为70 ℃,载气流速为2.0 L·min^-1）。结果 黄芪甲苷在0.050 5～1.011 mg·mL^-1的范围内呈良好的线性关系,标准曲线为lgA=1.3555lgC+3.277 6,（r=0.999）;精密度和重复性的RSD分别为1.2%和1.6%;平均回收率为96.1%,RSD为1.7%。结论 该方法简便准确、快速、方便、精密度高、重复性好,可用于复胃散胶囊中黄芪甲苷的含量测定。     

HPLC-ELSD法测定麻黄根药材中麻黄宁A的含量

摘 要 目的： 建立麻黄根药材中麻黄宁A的HPLC-ELSD测定方法。方法: 采用高效液相色谱法和蒸发光散射检测器;色谱柱：Alltima TM C18柱（250 mm×4.6 mm,5 μm）;柱温：30℃;流动相：乙腈 水（28∶72）;流速：0.7 ml·min^-1;ELSD条件：漂移管温度为105℃,载气流速为2.8 L·min^-1。结果: 麻黄宁A在42.56～383.04 μg·ml^-1范围内线性关系良好,r为0.999 8;平均回收率为99.9%,RSD为1.96%（n=6）。结论:该方法简便,结果准确,可用于麻黄根药材的质量控制。     

Mutagenesis induced by 12 quinolone antibacterial agents in Escherichia coli WP2uvrA/pKM101.

The mutagenic potential of 12 quinolone antibacterial agents (quinolones) was examined at concentrations of 3.91-1000 ng/plate with or without S9 mix in Escherichia coli WP2uvrA/pKM101. All quinolones showed mutagenic potential in the strain: the maximum numbers of revertant colonies were observed at 7.81 ng/plate for clinafloxacin and sitafloxacin; 15.63 ng/plate for ciprofloxacin, gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, and trovafloxacin; and 31.25-500 ng/plate for enoxacin, lomefloxacin, norfloxacin and ofloxacin. The numbers for all quinolones were comparable between the groups with and without S9 mix. In all quinolones, bactericidal effects were observed at one or two higher concentrations than their mutagenic concentrations except for enoxacin without S9 mix. From these results, the WP2uvrA/pKM101 strain is proved to be highly sensitive to quinolones.     

Structure-phototoxicity relationship in Balb/c mice treated with fluoroquinolone derivatives, followed by ultraviolet-A irradiation

We examined the structure-phototoxicity relationship for fluoroquinolone antimicrobial agents (quinolones) using female albino Balb/c mice. First of all, to obtain an optimum dosage level for induction of phototoxicity, the prototype phototoxicant sparfloxacin was intravenously administered once at 10 mg/kg, 30 mg/kg or 100 mg/kg to female mice, followed immediately by ultraviolet-A (UVA) irradiation for 4 h (21.6J/cm2). The auricular thickness was measured at pre-dose (0 h), 4, 24, 48, 72 and 96 h post-dose, and then the histopathological examination of the auricle was performed. As results, the auricular thickness increased from 30 mg/kg, in conjunction with edema, cellular infiltration, epidermal necrosis and focal loss of the auricle. On the basis of these information, ciprofloxacin, enoxacin, fleroxacin, gatifloxacin, lomefloxacin, norfloxacin and ofloxacin were given intravenously to mice at a fixed dose of 100 mg/kg to compare their potential phototoxicities. Certain quinolones caused the auricular lesions in the following rank order (from lowest to highest): vehicle control (non-phototoxicity)=gatifloxacin=ofloxacin相似文献   

Safety profile of antimicrobial agents

Many antibiotics have been developed and used for the treatment of infectious diseases. Although they have been known to have various adverse effects, most of the mechanisms remain still unknown. New quinolones are well known to induce convulsions and their convulsant activity enhanced by concurrent administration of anti-inflammatory drugs. Each new quinolone has an individual convulsant activity with individual drug-interaction with anti-inflammatory drugs. And enoxacin, lomefloxacin and gatifloxacin have been reported to decrease blood glucose levels in a dose-depend- ent manner, but ciprofloxacin and levofloxacin had no effect on the levels. It should be important to know the safety profile of antimicrobial agents before doctors administer these agents to the patients with infectious diseases.     

Binding characteristics of fluoroquinolones to synthetic levodopa melanin

To define the binding characteristics of fluoroquinolones to synthetic levodopa melanin, the binding of various drugs, including levofloxacin and ofloxacin, and positive controls (timolol and chloroquine), was investigated in-vitro. The affinity and capacity of the drug binding were calculated by Langmuir's adsorption isotherm. The affinity constant (K) and the binding capacity (r(max)) of levofloxacin were similar to those of timolol and much lower than those of chloroquine. Racemic ofloxacin and its enantiomers showed similar K and r(max), suggesting that the binding lacked stereoselectivity. The binding experiment with levofloxacin derivatives indicated that the basic nitrogen atom at position 7 of the quinolone ring, but not carboxyl group at position 3, would play a critical role in the interaction of fluoroquinolones with melanin. The melanin-drug complexes of levofloxacin and chloroquine were washed with neutral phosphate buffer, ethanol and 1 M HCl solution to explain the nature of the interaction of melanin with the drugs. Electrostatic forces mainly participate in the formation of the chloroquine-melanin complex, whereas van der Waals' and hydrophobic interactions are involved in the levofloxacin-melanin complex in addition to electrostatic forces. The interactions of various fluoroquinolones such as norfloxacin, enoxacin, sparfloxacin, ciprofloxacin and lomefloxacin with melanin were also studied. The results showed that the relative K value was: chloroquine approximately ciprofloxacin, sparfloxacin >/= lomefloxacin > timolol, levofloxacin approximately enoxacin, norfloxacin, and that the relative r(max) value was: norfloxacin, enoxacin >/= chloroquine, sparfloxacin > levofloxacin, ciprofloxacin, timolol, lomefloxacin. The fluoroquinolones vary in their affinity and capacity to bind with melanin, and ciprofloxacin and sparfloxacin showed a stronger interaction with melanin than the other fluoroquinolones studied.     

Permeability classification of representative fluoroquinolones by a cell culture method

This study was undertaken to categorize representative fluoroquinolone drug substance permeability based on the methods outlined in the Food and Drug Administration's biopharmaceutic classification system (BCS) Guidance for Industry. The permeability of ciprofloxacin, levofloxacin, lomefloxacin, and ofloxacin was measured in an in vitro Caco-2 assay with previously demonstrated method suitability. The permeability class and efflux potential were ascertained by comparing test drug results with standard compounds (metoprolol, atenolol, labetalol, and rhodamine-123). All 4 quinolones drugs demonstrated concentration-dependent permeability, indicating active drug transport. In comparing absorptive versus secretive in vitro transport, the tested fluoroquinolones were found to be subject to efflux in varying degrees (ciprofloxacin > lomefloxacin > rhodamine 123 > levofloxacin > ofloxacin). Based on comparison to labetalol, the high permeability internal standard, ciprofloxacin was classified as a low permeability drug, whereas lomefloxacin, levofloxacin, and ofloxacin were classified as high permeability drugs. The in vitro permeability results matched human in vivo data based on absolute bioavailabilities. This laboratory exercise demonstrated the applicability of an in vitro permeability method for classifying drugs as outlined in the BCS Guidance.     

专项整治前后某院喹诺酮类抗菌药应用情况对比分析

目的:分析评价南京某三甲医院抗菌药专项整治前后喹诺酮类抗菌药使用情况和发展趋势,为抗菌药临床应用专项整治活动及管理提供依据。方法:调取该院2010年(整治前)和2011~2013年(整治后)所用喹诺酮类抗菌药的使用记录,采用回顾性方法,对该类药物的销售金额、用药频度(DDDs)、限定日费用(DDC)、细菌耐药情况及药品不良反应进行统计分析。结果:整治后(2011~2013年)喹诺酮类抗菌药销售金额及占抗菌药总销售金额比例较整治前(2010年)有明显下降;整治前注射用门冬氨酸洛美沙星的销售金额和DDDs最高,整治后的2012~2013年,莫西沙星注射剂和盐酸左氧氟沙星注射液的销售金额最高,分别位于第1、2位;盐酸左氧氟沙星片和盐酸左氧氟沙星注射液的DDDs最高,分别位于第1、2位;莫西沙星注射剂DDC最高。主要耐药菌葡萄球菌、肠杆菌科细菌、非发酵菌对左氧氟沙星、诺氟沙星、环丙沙星的耐药率有所下降;整治后该院抗菌药所致不良反应例数及喹诺酮类抗菌药所致不良反应占抗菌药不良反应的比例低于整治前。结论:该院通过抗菌药专项整治,喹诺酮类抗菌药的各项应用指标明显改善,喹诺酮类抗菌药的合理应用也取得成效,专项整治的效果比较明显。但该院还应加强对第4代喹诺酮类抗菌药的临床应用加以督导。     

In vitro method for prediction of the phototoxic potentials of fluoroquinolones

The phototoxic potential of eight fluoroquinolones (norfloxacin, ofloxacin, enoxacin, ciprofloxacin, lomefloxacin, tosufloxacin, sparfloxacin and gatifloxacin) was evaluated by using three in vitro methods of cytotoxicity against mammalian cells, erythrocyte lysis and DNA strand breakage. All fluoroquinolones tested with the exception of gatifloxacin, an 8-methoxy quinolone, showed DNA strand breaking activities under UV-A irradiation. Their cytotoxicity against HeLa cells was also enhanced by UV-A irradiation. In particular, the phototoxic potential of sparfloxacin, enoxacin and lomefloxacin was high in both methods. Ofloxacin is very photocytotoxic against HeLa cells, while it has low potential to cause DNA strand breakage. Norfloxacin, ciprofloxacin and enoxacin were very photohemolytic, but sparfloxacin was not, indicating that the in vivo phototoxic potencies of fluoroquinolones might not be predictable by the photohemolysis study. Gatifloxacin, a non-phototoxic quinolone, showed no phototoxic potential in any of these three in vitro tests. These results suggest that determination of DNA strand breaking activity, combined with cytotoxicity against mammalian cells, is available to predict the phototoxic potential of fluoroquinolones without laboratory animals.     

Celecoxib does not induce convulsions nor does it affect GABAA receptor binding activity in the presence of new quinolones in mice

We sought to determine whether celecoxib would induce convulsions when coadministered with new quinolone antimicrobial agents in mice. The oral administration of celecoxib (500 mg/kg) alone or in combination with enoxacin (500 mg/kg), lomefloxacin (1000 mg/kg), ciprofloxacin (1000 mg/kg), or levofloxacin (1000 mg/kg) induced no convulsions in mice. In contrast, some nonsteroidal anti-inflammatory drugs (NSAIDs), fenbufen (200 mg/kg), indomethacin (500 mg/kg), and naproxen (500 mg/kg) induced convulsions in combination with the majority of the new quinolones tested. gamma-Aminobutyric acid (GABA)(A) receptor blockade-mediated neuronal excitation is assumed to be involved in these toxic convulsions. Enoxacin (100 microM) and lomefloxacin (100 microM) only slightly reduced [3H]muscimol binding to GABA(A) receptors in mouse whole brain membrane. However, these reductions were markedly enhanced by the addition of fenbufen (100 microM), indomethacin (100 microM), or naproxen (100 microM). Conversely, celecoxib (100 microM) had no apparent effect on [3H]muscimol binding when applied alone or in combination with enoxacin or lomefloxacin. These results suggest that celecoxib may be a more desirable anti-inflammatory agent with respect to drug interactions with new quinolones compared with some conventional NSAIDs.