高效液相色谱-荧光检测法测定罂粟籽和火锅汤料中的罂粟碱

用所建立的高效液相色谱-荧光检测法测定了罂粟籽和火锅汤料中的罂粟碱。采用的色谱柱为RP-C18柱(250 mm×4.6 mm i.d.,5 μm);检测激发波长为285 nm,发射波长为355 nm;流动相为甲醇-0.02 mol/L乙酸铵(体积比为70∶30),流速0.8 mL/min。实验结果表明,罂粟碱的进样量为1×10-4~0.1 μg时其质量浓度与相应峰面积有良好的线性关系,最低检测限(以信噪比大于3计)达到0.02 ng。罂粟籽中罂粟碱的回收率为99.0%～100.8%。方法快速准确,简便灵敏,分离度高,能够满足有关食品中罂粟碱的检测要求。     

超高效超临界色谱分析聚合物制品中的7种添加剂

采用超高效超临界色谱(UPSFC)-二极管阵列检测器(PDA)建立了快速分析高分子材料中7种常用聚合物添加剂的方法。在检测波长为220 nm条件下,通过考察样品的稀释溶剂、流动相添加剂、柱温、背压、柱流速对分离的影响,最终确定7种添加剂分离的最优化条件：采用正己烷/异丙醇(1/1,v/v)为稀释溶剂,流动相添加剂为甲醇/乙腈(1/1,v/v),流速2 mL/min,柱温50 ℃,背压12.41~13.79 MPa条件下,7种聚合物添加剂能够在5 min之内达到完全基线分离。利用微波辅助萃取方法对实际样品中的添加剂进行提取,经UPSFC-PDA分析,7种添加剂的回收率为69.9%~118.9%,相对标准偏差(RSD,n=9)低于10%。该方法简便、快速,选择性强,能够准确快速地分析聚合物制品中的添加剂。     

高效液相色谱法同时测定食品中7种非食用色素

建立了高效液相色谱法同时测定食品中7种非食用色素(碱性嫩黄O、碱性橙、酸性橙Ⅱ、酸性金黄、玫瑰红B、对位红、苏丹红Ⅰ)的方法。依次采用乙腈、甲醇和碱性甲醇提取豆制品和肉制品;采用乙腈和70%乙腈依次提取调味品。采用SunFireTMC18色谱柱(250 mm×4.6 mm,5μm),以甲醇-50 mmol/L乙酸铵水溶液(H3PO4调至pH 4.5)为流动相,梯度洗脱,流速1 mL/min,测定波长为450和520 nm。7种非食用色素的在各自相应浓度范围内线性相关系数均大于0.998;检出限(LOD)在0.01～0.1 mg/L之间;定量限(LOQ)在0.18～1.2 mg/kg之间。平均回收率均大于80%;相对标准偏差(RSD,n=3)在2.0%～5.7%之间。     

Evaluation of a reversed-phase column (supelcosil LC-ABZ) under isocratic and gradient elution conditions for estimating octanol-water partition coefficients

The solvation parameter model is used to identify suitable chromatographic models for estimating the octanol-water partition coefficient for neutral compounds of varied structure by reversed-phase liquid chromatography. The stationary phase Supelcosil LC-ABZ with methanol-water mobile phases affords a series of suitable correlation models for estimating the octanol-water partition coefficient (log KOW) under isocratic and gradient elution conditions. Isocratic separations with mobile phase compositions containing from about 25 to 40% (v/v) methanol provide the most accurate results for log KOW values in the range -0.1 to 4.0. Gradient separations programmed from 5 to 100% (v/v) methanol are suitable for faster separations of compounds with large log KOW values. The standard error in the estimate for the regression models of the predicted log KOW values against literature values are 0.135 log units for the 30% (v/v) methanol-water isocratic system and 0.263 log units for the methanol-water gradient system. Isocratic retention factors predicted from two gradient separations with gradient times of 15 and 45 min afford a poorer fit for the correlation models between log KOW and the estimated retention factors than that of either the above isocratic and gradient models. Plots of the retention factor (log k) as a function of mobile phase composition are generally non-linear. Values of log kw obtained by non-linear extrapolation to a volume fraction of 0% (v/v) methanol do not afford a useful model for estimating log KOW.     

超高效液相色谱法同时测定纺织品中18种荧光增白剂

建立了同时测定纺织品中18种荧光增白剂（FWAs）的超高效液相色谱-荧光检测（UPLC-FLR）法。试样由三氯甲烷-乙醇（6：4,v/v）超声提取,以ACQUITY UPLC HSS T3柱（100 mm×2.1 mm,1.8 μm）为分离色谱柱进行分析,以5 mmol/L乙酸铵水溶液和甲醇为流动相,梯度洗脱,流速为0.4 mL/min,荧光激发波长为350 nm,发射波长为430 nm,外标法定量。18种FWAs在各自范围内呈良好的线性关系,相关系数（R²）均≥ 0.9992;方法的定量限（LOQs,S/N=10）为0.002~0.1 mg/L。样品的平均加标回收率为88.3%~104.5%,相对标准偏差（RSD）为2.0%~5.5%（n=6）。该方法灵敏度高,精密度好,准确度高,适用于各种纺织品中FWAs的测定。     

高效液相色谱–光电二极管阵列法测定虾青素的含量

建立虾青素含量测定的高效液相色谱–光电二极管阵列法。采用Purospher STAR RP 18(4.6 mm×250mm,5μm)色谱柱,以甲醇–水(体积比为95∶5)为流动相,流速1.0 mL/min,检测波长为482 nm,柱温为30℃,进样量为20μL。在所选定的液相色谱条件下,虾青素主峰与其它杂质峰分离良好,虾青素在0.2~16μg/mL范围内线性良好,线性相关系数r=0.999 9,检出限为0.01μg/mL,测定结果的相对标准偏差为0.42%(n=6),平均回收率为100.4%。该法分析快速准确、灵敏度高、重现性好。     

Identification of degradation impurities in aripiprazole oral solution using LC–MS and development of validated stability indicating method for assay and content of two preservatives by RP-HPLC

A simple and simultaneous reverse phase high-performance liquid chromatographic method was developed for the quantification of aripiprazole (ARI) and two preservatives, namely, methyl paraben and propyl paraben in ARI oral solution. The method was developed on ACE C18 (4.6?×?250?mm, 5?µm) column using gradient elution of 0.1% v/v trifluoroacetic acid in water and acetonitrile as mobile phase components. Flow rate of 1.0?mL/min and 30°C column temperature were used for the method at quantification wavelength of 254?nm. The developed method was validated in accordance with International Conference on Harmonization guideline for various parameters. Forced degradation study was conducted in acid, base, peroxide, heat, and light stress conditions. ARI was found to degrade in oxidation, acid hydrolysis, and heat while it was stable under the remaining conditions. Specificity of the method was verified using Photo Diode Array (PDA) detector by evaluating purity of peaks from degradation samples. Major degradation impurities formed during stress study were identified using liquid chromatography–mass spectrometry method. The present method was useful for determining the content of all the three main analytes present in the oral solution without interference from degradation impurities. The method was robust under the deliberately modified conditions.     

Determination and locational variations in the quantity of hydroxyanthraquinones and their glycosides in rhizomes of Rheum emodi using high-performance liquid chromatography

Locational variations in the quantity of five hydroxyanthraquinone derivatives (emodin glycoside (1), chrysophanol glycoside (2), emodin (3), chrysophanol (4) and physcion (5)) in the rhizomes of Rheum emodi are described. A simple and reliable method was developed for quantitation of compounds (1-5) in the methanolic extract of rhizomes of R. emodi using reverse-phase high-performance liquid chromatography (HPLC) with photo-diode array detector (PDA). The separation was carried out using a Purospher((R))-Star RP-18 e column (4.6mm i.d.x 250 mm, 5 microm) under the following conditions: acetonitrile:methanol (95:5, v/v) (solvent A) and water:acetic acid (99.9:0.1, v/v) (solvent B) as mobile phase with a linear gradient elution at a flow rate of 0.8 mL/min. The detection wavelength was set at 290 nm. Regression equation revealed a linear relationship (r(2)>0.9901) between the mass of hydroxyanthraquinone derivatives injected and the peak areas. The detection limits (S/N=3) ranged from 0.56 to 3.50 ng/mL and the recoveries ranged from 95.7 to 103.5% for five hydroxyanthraquinone derivatives. Compound 2 was found in maximum quantity (up to 2.23%) in the rhizomes from all the three locations (L(1), L(2) and L(3)) while compound 5 was found in the least quantity (up to 0.19%).     

反相高效液相色谱法测定饲料添加剂阿散酸中的阿散酸及掺假物质对氨基苯磺酸

建立了用于饲料添加剂阿散酸质量控制的反相高效液相色谱法。采用的色谱条件: Waters Bondapak C18柱(150 mm×4 mm, 5 μm)分离,以甲醇-水(用稀磷酸调节pH至2.9) (1:4, v/v)作流动相,流速1.0 mL/min,紫外检测波长为244 nm。在优化的色谱条件下,阿散酸和掺假物质对氨基苯磺酸在3 min内实现了基线分离。阿散酸和对氨基苯磺酸的线性范围均为5～200 mg/L,检出限(S/N=3)分别为0.20 mg/L和0.15 mg/L。该方法简便快速,适合饲料添加剂阿散酸的分析以及监测对氨基苯磺酸掺假。     

高效液相色谱-紫外法快速测定塑料类食品接触材料及制品中7种对苯二甲酸酯或苯甲酸酯的特定迁移量

建立了高效液相色谱-紫外检测（HPLC-UV）法快速测定从塑料类食品接触材料及制品迁移至10%（v/v）乙醇、3%（m/v,即3 g/100 mL）乙酸、4%（v/v）乙酸、20%（v/v）乙醇、50%（v/v）乙醇、95%（v/v）乙醇和橄榄油7种食品模拟物中对苯二甲酸二甲酯、对苯二甲酸二辛酯、苯甲酸甲酯、苯甲酸乙酯、苯甲酸丙酯、苯甲酸丁酯和新戊二醇二苯甲酸酯的特定迁移量。考察了多种提取溶剂、QuEChERS dSPE EMR-Lipid试剂盒和Captiva EMR-Lipid试剂盒对橄榄油食品模拟物中7种对苯二甲酸酯或苯甲酸酯的提取或净化效果。以甲醇和水为流动相进行梯度洗脱,7种对苯二甲酸酯或苯甲酸酯在苯基柱上于17 min内达到基线分离。检测波长为237 nm,进样量为10 μL。7种对苯二甲酸酯或苯甲酸酯在7种食品模拟物中的定量限为0.2~8.1 mg/kg、1~80 mg/L或8~160 mg/kg,相关系数r≥0.9998。在2或8、60、80或160 mg/kg 3个加标水平的回收率为91.7%~106%,相对标准偏差为0.1%~3.1%。该方法样品前处理简便,色谱分离和线性关系好,回收率和重复性较好,已应用于实际样品的检测。     

Quantitative determination of porphyrins, their precursors and zinc protoporphyrin in whole blood and dried blood by high-performance liquid chromatography with fluorimetric detection

Methods for the determination of porphyrins, delta-aminolevulinic acid (ALA), porphobilinogen (PBG) and zinc protoporphyrin of heme biosynthesis in whole blood and dried blood are described. Erythrocyte porphyrins and the precursors ALA and PBG were extracted from whole blood (50 microliter) with 0.3 ml of methanol and 1.5 M hydrochloric acid (2:1, v/v). Zinc protoporphyrin was extracted with an acetone-pyridine-Sterox solution. Other major interfering metabolites were removed by centrifugation. An aliquot of the supernatant was injected onto the reversed-phase C18 column for detection of porphyrins with excitation wavelength at 405 nm and emission wavelength at 630 nm. The mobile phase was 0.1 M phosphate-methanol-tetrahydrofuran (18:30:16, v/v/v), pH 5.38. The ALA and PBG were derivatized with o-phthalaldehyde before injection. The detection excitation wavelength occurred at 330 nm and the emission wavelength at 418 nm. The mobile phase was 0.1 M phosphate-methanol (7.5:5), pH 3.38. For the dried blood specimen of filter paper, two 0.64-cm discs punched out from the blood-impregnated filter paper were placed in a test tube containing 200 microliter of 0.9% saline for 60 min or longer at room temperature and then treated as whole blood.     

基质固相分散-高效液相色谱法测定鲫鱼肌肉中残留的辛硫磷

建立了鲫鱼肌肉中残留的辛硫磷的基质固相分散-高效液相色谱-二极管阵列检测(MSPD-HPLC-DAD)的分析方法。通过优化样品处理条件,确定选取0.50 g鲫鱼肌肉样品与1.5 g弗罗里硅土、0.5 g无水硫酸钠混合研磨,并采用丙酮-正己烷溶液(体积比为40:60)为洗脱剂,洗脱剂用量为25 mL。优选的最佳色谱条件为：ODS色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇-水(体积比为50:50),流速0.6 mL/min,检测波长270 nm,进样量为20 μL。在上述条件下,辛硫磷质量浓度在0.01～10 mg/L范围内与响应信号呈良好的线性关系(r20.9994),检出限为3.3 μg/kg;相对标准偏差为1.1%～6.3%(n7);3个添加水平(0.05,0.1,1 mg/kg)下得到的回收率为88%～112%。该方法操作简单,耗时少,精密度高,符合农残分析的要求。     

A Simple Isocratic High-Performance Liquid Chromatographic (HPLC) Determination of Naproxen in Human Plasma using a Microbore Column Technique

Abstract

A simple and sensitive HPLC method was developed for the determination of naproxen in human plasma. The assay employs a microbore column packed with a C18 reversed-phase material (5 μm ODS Hypersil) with an isocratic mixture of acetonitrile and 10 mM phosphate buffer, pH 2.5 (40:60, v/v) as the mobile phase. The mobile phase was pumped at a flow rate of 0.5 ml/min. For sample analysis 200 μl of acetonitrile containing internal standard (flurbiprofen) was added to 100 μl of plasma. After centrifugation 10 mM phosphate buffer, pH 7.4 (200 μl) was added to the tube, then vortexed and centrifuged. The supernatant (20 μl) was injected onto the HPLC column. The chromatographic separation was monitored by a fluorescence detector at an emission wavelength of 350 nm with an excitation wavelength of 225 nm. The direct precipitation of plasma protein using acetonitrile gave a good recovery for both naproxen and the internal standard. The detection limit was 0.1 μg/ml for naproxen. The intra- and inter-assay coefficients of variation at different concentrations evaluated were less than 10%.     

Improved HPLC method for doxorubicin quantification in rat plasma to study the pharmacokinetics of micelle-encapsulated and liposome-encapsulated doxorubicin formulations

An improved simple, rapid and accurate HPLC method for quantification of doxorubicin derived from micelle-encapsulated or liposome-encapsulated doxorubicin formulation in rat plasma was described. The mobile phase consisting of a mixture of methanol-water [containing 0.1% formic acid anhydrous and 0.1% ammonia solution (25%), pH 3.0], 60:40, was delivered at a flow rate of 1.0 mL/min. Sample preparation for micelle- or liposome-encapsulated doxorubicin in rat plasma were achieved directly by protein precipitation with acetonitrile. Doxorubicin and daunorubicin (internal standard, IS) were separated on a C(18) reversed-phase HPLC column and quantified by a fluoresence detection with an excitation wavelength of 475 nm and an emission wavelength of 580 nm. The linearity was obtained over the range of 5.0-1000.0 ng/mL and 1.0-200.0 microg/mL for doxorubicin and the lower limit of quantitation was 5.0 ng/mL. For each level of quality control samples, inter- and intra-assay precision was less than 9.6 and 5.1% (relative standard deviation), respectively, and percentage error was within +/-2.6%. The extraction recoveries of doxorubicin in the range of 10 ng/mL to 100 microg/mL in rat plasma were between 94.1 and 105.6%. This method was successfully applied to the pharmacokinetic study of doxorubicin formulations after i.v. administration to rats.     

高效液相色谱分析化妆品中的防腐剂氯苯甘醚

建立了化妆品中防腐剂氯苯甘醚的高效液相色谱检测方法。采用C₁₈色谱柱(250 mm×4.6 mm,5 μm)和二极管阵列检测器,以1.0 mL/min甲醇-水(55：45,v/v)为流动相,检测波长为280 nm,柱温为25 ℃。方法的检出限(以3倍信噪比计)可达到3 ng;在1~500 mg/L范围内,峰面积与质量浓度呈良好的线性关系,相关系数为1.0000。高、中、低不同水平的加标回收率为99.0%~103%,相对标准偏差≤1.2%。同时进行了干扰试验和实际样品的测定,并组织3家实验室进行了方法验证。该方法操作简便,准确度、灵敏度和稳定性高,可用于化妆品中防腐剂氯苯甘醚的检测。     

Liquid chromatographic determination of honokiol and magnolol in hou po (Magnolia officinalis) as the raw herb and dried aqueous extract

A validated analytical method is described for the determination of honokiol and magnolol in Hou Po (Magnolia officinalis) as the dried raw herb and the commercially prepared dried aqueous extract. The samples were extracted with methanol by the Soxhlet method, and the extract was analyzed by liquid chromatography with photodiode array (LC/PDA) detection with confirmation of analyte identity by negative-ion electrospray ionization tandem mass spectrometry (ESI-MS/MS). A C18 column was used with a menthanol--0.1% aqueous acetic acid gradient mobile phase. Honokiol and magnolol were quantified at 288 nm. With the MS detector, the honokiol precursor ion at m/z 265 was shown to produce ions at m/z 222 and 224. For magnolol, the precursor ion at m/z 265 produced the ions at m/z 247 and 245. Comparable results were obtained for the LC/PDA and LC/ESI-MS/MS methods of quantitation. Six commercially prepared dried aqueous extracts were analyzed. The levels of honokiol and magnolol found in the raw herb were 17.0 and 21.3 mg/g, respectively. The limits of detection for honokiol and magnolol in the raw herb were 0.45 and 0.58 mg/g, respectively, and in the dried aqueous extract, 0.04 and 0.30 mg/g, respectively.     

双水杨醛邻苯二胺柱前衍生-高效液相色谱法测定茶叶中的Pb~(2+)

建立了利用双水杨醛邻苯二胺希夫碱(SALOPHEN)作为衍生剂结合高效液相色谱(HPLC)测定茶叶中Pb~(2+)含量的方法。Hypersil ODS2 C18反相色谱柱(250 mm×4.6 mm,5μm)为固定相,甲醇-水(80∶20,v/v)为流动相,检测波长为226 nm,反应体系pH为10.0。结果表明,样品中Pb~(2+)的线性范围为0.1~30 mg/L,线性相关系数为0.998 8,检出限为0.01 mg/L,加标回收率为91.87%~96.96%。该方法具有较高的灵敏度和稳定性,适用于茶叶中Pb~(2+)的检测。     

高效液相色谱法同时测定忍冬中7种成分

建立了在不同时间段内转换使用不同波长同时测定忍冬花和叶中7种化学成分(绿原酸、咖啡酸、芦丁、木犀草苷、异绿原酸A、异绿原酸B、异绿原酸C)的高效液相色谱分析方法,同时应用该方法分析了忍冬花、忍冬老叶和新叶中成分含量的差异。色谱柱为Agilent Eclipse Plus C18(250 mm×4.6 mm, 5 μm);流动相为0.3%甲酸水溶液(A)和乙腈(B),梯度洗脱,流速1 mL/min;采用VWD紫外检测器转换波长(330 nm、350 nm)检测。应用所建立的方法测定忍冬新叶中绿原酸、木犀草苷含量分别为2.572%、1.498‰,均比药典中规定的含量高,有必要进一步的研究和开发利用。该方法准确、简便、灵敏度高,适用于忍冬中7种化学成分含量的同时测定和忍冬的质量控制及综合评价。     

免疫亲和柱净化/柱前衍生化-高效液相色谱荧光检测法测定粮谷中的T-2毒素

建立了免疫亲和柱净化/柱前衍生化-高效液相色谱荧光检测器测定粮谷中T-2毒素含量的方法。样品经甲醇-水(体积比为80∶20)混合溶剂提取,通过免疫亲和柱(IAC)净化,以氰酸蒽(1-AN)为衍生化试剂、4-二甲基氨基吡啶(DMAP)为催化剂进行衍生,以ZORBAX Eclipse XDB-C18 柱为分离柱,乙腈-水(体积比为80∶20)为流动相进行高效液相色谱分离及荧光检测，荧光检测的激发波长为381 nm，发射波长为470 nm。T-2毒素的质量浓度为0.01~1.5 mg/L时与峰高呈良好的线性,相关系数为0.9985。在0.01~1.5 μg/g添加水平下，回收率为79.7%~94.5%,相对标准偏差小于7%；检出限（S/N＝3）为0.01 μg/g。该方法净化效果好,灵敏度高,操作简便快速。     

Comparison of HPLC and MEEKC for Miconazole Nitrate Determination in Pharmaceutical Formulation

A simple solid phase extraction (SPE) method coupled with high performance liquid chromatography (HPLC) using UV detector and microemulsion electrokinetic chromatography (MEEKC) has been developed and compared for the quantitative determination of miconazole nitrate in pharmaceutical formulation. For HPLC method, two parameters were optimized, namely, the wavelength and the mobile phases. The optimized condition was at the 225 nm wavelength and the mobile phase of ACN:MeOH (90:10 v/v). There are seven MEEKC parameters that were optimized, in this research, which were applied to voltage, temperature, wavelength, sodium dodecyl sulfate (SDS) concentration, buffer pH, buffer concentration and butan-1-ol concentration. The optimum MEEKC condition was obtained using 86.35 % (w/w) 2.5 mM borate buffer pH 9, 0.25 % (w/w) SDS, 0.8 % (w/w) ethyl acetate, 6.6 % w/w butan-1-ol and 6.0 % (w/w) acetonitrile. The combination of SPE using a diol column with HPLC–UV and the MEEKC methods were successfully applied for the determination of miconazole nitrate in a pharmaceutical formulation with the recovery percentage of 98.35 and 92.50 %, respectively.