高效合相色谱同时测定血浆中12种磺胺类药物

建立了一种同时分离测定血浆中12种磺胺类药物的高效合相色谱(Ultra Performance Convergence Chromatography,UPC~2)方法。血浆样品中加入乙腈,涡旋混合后离心,上清液过滤后采用UPC~2方法测定,采用Acquity UPC~(2TM)BEH色谱柱,CO_2和甲醇作为流动相,梯度洗脱,柱温箱温度40℃,系统背压14.5MPa。该法在6 min内实现了对12种磺胺类药物的基线分离,定量限为0.5μg/mL,回收率在86.2%~99.5%之间。方法适用于血浆中多种磺胺类药物的同时检测。     

高效液相色谱法测定血浆中阿魏酸的含量

建立了一种快速、稳定的高效液相色谱-二极管阵列检测法(HPLC-DAD),用于测定健康人口服阿魏酸钠片后,血浆中阿魏酸的含量。该方法采用DIONEX C18色谱柱(250×4.6mm i.d.,5μm),流动相为甲醇-磷酸缓冲溶液(0.1%),梯度洗脱,检测波长为322nm。结果显示:人体血浆中的阿魏酸提取回收率大于60%。阿魏酸的含量在0.025～8.0μg/mL范围内线性良好,相关系数r=0.9994。方法检出限(S/N=3)为0.025μg/mL。日内相对标准偏差(RSD)小于1.02%,日间RSD小于2.1%。该法可用于人体血浆中阿魏酸的药代动力学研究。     

固相萃取–高效液相色谱法同时测定饲料中4种磺胺类药物残留

建立固相萃取–高效液相色谱法同时测定饲料中的磺胺嘧啶、磺胺二甲嘧啶、磺胺甲恶唑、磺胺喹恶啉4种磺胺类药物残留的方法。样品用乙腈提取,然后用碱性氧化铝固相萃取柱净化,色谱柱为C_(18)柱(250 mm×4.6mm,5μm),以水–乙腈(体积比为75∶25,含0.3%乙酸)为流动相,流量为1.0 mL/min,检测波长为270 nm。4种磺胺类药物的质量浓度在1~10μg/mL范围内与其色谱峰面积呈良好的线性,相关系数均大于0.999,检出限为0.025~0.133μg/g。磺胺类药物测定结果的相对标准偏差为0.22%~0.30%(n=6),样品加标回收率为93.6%~106.7%。实际饲料样品中均未检出这4种磺胺组分。该方法具有干扰少,灵敏度高,重复性好的优点,可以作为饲料中的磺胺类药物残留的一种检测方法。     

超高效液相色谱-串联质谱法测定血浆与尿液中14种麻痹性贝类毒素北大核心CSCD

人体生物基质中麻痹性贝类毒素的检测对其引起的食物中毒诊断和救治具有重要意义。研究建立了超高效液相色谱-串联质谱法测定血浆、尿液中14种麻痹性贝类毒素的分析方法。实验比较了不同固相萃取柱的影响,优化了前处理条件和色谱条件,血浆样品采用0.2 mL水、0.4 mL甲醇、0.6 mL乙腈提取后直接上机测定,尿液样品采用0.2 mL水、0.4 mL甲醇、0.6 mL乙腈提取,聚酰胺(PA)固相萃取柱净化后上机测定。采用Poroshell 120 HILIC-Z色谱柱(100 mm×2.1 mm,2.7μm)对14种贝类毒素进行分离,流动相为含0.1%(v/v)甲酸的5 mmoL/L甲酸铵缓冲溶液和0.1%(v/v)甲酸乙腈溶液,流速为0.50 mL/min。在电喷雾模式(ESI)下进行正负离子扫描,采用多反应监测(MRM)模式检测,外标法定量。结果表明,对于血浆和尿液样品,14种贝类毒素分别在0.24~84.06 ng/mL范围内线性关系良好,相关系数均大于0.995。尿液检测的定量限为4.80~34.40 ng/mL,血浆检测的定量限为1.68~12.04 ng/mL。尿液和血浆样品在1、2和10倍定量限加标水平下平均回收率为70.4%~123.4%,日内精密度为2.3%~19.1%,日间精密度为4.0%~16.2%。应用建立的方法对腹腔注射14种贝类毒素小鼠血浆和尿液进行测定,20份血浆样本中检出含量分别为19.40~55.60μg/L和8.75~13.86μg/L。该方法操作简便,样品取样量少,方法灵敏度高,适用于血浆和尿液中麻痹性贝类毒素的快速检测。     

快速溶剂萃取-高效液相色谱法同时测定牛肉中5种磺胺类药物残留

建立了同时测定牛肉中5种磺胺药物残留量的快速溶剂萃取-高效液相色谱法。以乙腈为萃取剂在120℃、10MPa条件下用快速溶剂萃取仪提取样品中的目标物,提取液经冷冻除脂净化后,采用UltimateXB-C18色谱柱(4.6mm×250mm×5μm)分离,乙腈-3%乙酸溶液(体积比25:75)为流动相,流速1.0mL/min,柱温为30℃,在紫外检测波长268nm条件下进样10μL。该方法在0.01～0.10g/L范围内线性关系良好(r=0.999),最低检出限为0.011mg/kg,加标回收率在89.0%～107.8%范围内,相对标准偏差小于5.3%(n=5)。该方法快速、准确、简便,能够满足检测要求。     

高效液相色谱内标法检测鸡血浆中甲砜霉素的浓度

以氟甲砜霉素作内标,乙腈作为提取溶剂,采用高效液相色谱内标法检测鸡血浆中甲砜霉素的浓度。色谱柱为Shim-pack CLC-ODS(150 mm×6 mm,5μm),流动相为乙腈-水(体积比25∶75),流速为1.0 mL/min,检测波长为225 nm,柱温为40℃。在此色谱条件下,在0.25~32.00 mg/L浓度范围内,甲砜霉素浓度与甲砜霉素和氟甲砜霉素峰面积比呈线性关系,相关系数r为0.9999,最低检测浓度为0.1 mg/L;在高、中、低3个浓度水平下日内、日间精密度均大于6.1%(n=5),提取回收率大于98.68%,方法回收率为99.20%~100.25%。建立的方法符合生物样品的分析要求,可用于临床药代动力学研究。     

固相萃取/高效液相色谱法同时测定保健食品中7种磺酰脲类降血糖药物

建立了同时测定保健食品中7种磺酰脲类降血糖药物(格列吡嗪、甲苯磺丁脲、妥拉磺脲、格列齐特、格列本脲、格列美脲和格列喹酮)的固相萃取/高效液相色谱(SPE/HPLC)分析方法。样品经甲醇超声提取,用C18固相萃取柱净化后进行HPLC分析。采用Agilent Zorbax SB-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.02%磷酸溶液(B)为流动相,梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长228 nm。结果表明,7种目标物在0.2~20μg/mL范围内线性关系良好,相关系数(r)大于0.999 9,检出限(LOD,S/N≥3)均为0.1 mg/kg,在5种不同基质中的加标回收率为81%~115%,相对标准偏差(RSD)小于9%。     

高效液相色谱-二极管阵列检测法同时测定临床中毒患者血浆中的甲基苯丙胺及苯丙胺

建立了临床中毒患者血浆中甲基苯丙胺(MA)及其代谢产物苯丙胺(AP)的高效液相色谱-二极管阵列检测器(HPLC-DAD)联用的定性定量方法。用固相萃取小柱分离、纯化、富集血浆中的MA和AP,在C8柱上测定,条件为以乙腈-甲醇-磷酸二氢钾缓冲液为流动相,流速为1.0 mL/min,二极管阵列检测波长为210 nm,检测范围为200-260 nm。利用保留时间、紫外光谱进行定性分析。结果表明,血浆中MA和AP分别为0.036-1.438 mg/L与0.047-1.500 mg/L时线性关系良好;其回收率分别为     

高效液相色谱法研究邻烯丙基苯酚在小麦植株中的向基传导和分布

采用一种快速、灵敏的高效液相色谱-二极管阵列检测器法(HPLC-DAD)研究拟银杏杀菌剂邻烯丙基苯酚在小麦幼苗中的自顶向基传导作用,并定量分析其进入小麦植株后的具体分布。HPLC-DAD检测条件为:等度流动相65%乙腈/水(0.1%醋酸),HYPERSIL BDS C18,5μm,250 mm×4.6 mm i.d色谱柱,检测波长280 nm。邻烯丙基苯酚的线性回归方程为Y=8.0266X 5.019(r=0.9999),在2~500 mg/L浓度范围内线性关系良好。日内与日间精密度均≤12.44%;平均回收率为83.55%;最低检出限为1.5 ng,最低定量限为2 mg/L。分析方法符合小麦体内药物含量测定的要求。经HPLC-DAD检测,确证了邻烯丙基苯酚具有自小麦叶部向基部传导的特性,施药4h后即可传至根部。给药处理72 h后药剂在小麦叶、茎及根部的积累水平相当,基本完成自顶向基传导过程。     

高效液相色谱法测定家兔血浆中棉酚含量

利用丙酮-乙腈-正庚烷-磷酸盐溶液提取家兔血浆中游离棉酚,建立家兔血浆中棉酚的高效液相色谱测定方法。色谱柱为ODS-C18(4.6mm×200mm,5μm)柱,流动相为V(甲醇)∶V(1%H3PO4溶液)=85∶15,流速1.0mL/min,检测波长235nm。棉酚在0.66～3.30μg/mL质量浓度范围内峰面积与其浓度间呈现良好的线性关系,r=0.9994;平均加样回收率为95.7%～96.3%;日内、日间RSD分别为0.7%～1.4%和3.7%～7.4%。可用于棉酚生物样品分析。     

Simultaneous determination of rifampicin and sulbactam in mouse plasma by high-performance liquid chromatography

A simple and rapid high-performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed and validated for the simultaneous determination of rifampicin and sulbactam in mouse plasma. Plasma samples were deproteinized with acetonitrile and separated by HPLC on a RP-18 (125 x 4 mm, 5 microm) column and gradient elution with potassium dihydrogen phosphate solution (pH 4.5; 50 mm) and acetonitrile at a flow-rate of 1.0 mL/min. Rifampicin and sulbactam were monitored at 230 nm and confirmed by means of their UV spectra using a diode-array detector. The method was linear at plasma levels from 1 to 100 microg/mL for rifampicin and from 5 to 200 microg/mL for sulbactam. The limits of quantification were 0.6 microg/mL for rifampicin and 4.2 microg/mL for sulbactam. The intra- and inter-day precisions of the method (RSD) were lower than 5% for both compounds. Average recoveries of rifampicin and sulbactam from mice plasma were 98.2 and 89.3%, respectively. The developed method was successfully applied to the determination of the pharmacokinetic profile of both compounds in mice.     

反相高效液相色谱法测定小鼠血浆及组织中的阿昔洛韦浓度

 建立了测定小鼠血浆、肝、肾、脾、肺等组织中阿昔洛韦 (ACV)浓度的高效液相色谱法。色谱柱为HypersilODS ,流动相为甲醇 水 冰醋酸 (体积比为 1∶99∶0 5 )混合溶液 ,流速为 1 5mL/min ,检测波长为 2 5 2nm。ACV血浆最低检测浓度为 2 0 μg/L ,各组织最低检测浓度为 5 0ng/g。血浆及组织匀浆中的ACV浓度在 0 1mg/L～ 4mg/L及 0 1μg/g～ 4μg/ g时线性关系良好 (r >0 99)。血浆及肝匀浆中的ACV回收率分别为 97 5 %～ 10 0 0 %和 10 0 0 %～ 10 6 0 % (n =5 )。该法精密度高 ,方便 ,快捷 。     

Optimizing high-performance liquid chromatography method for quantification of glucosamine using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate derivatization in rat plasma: application to a pharmacokinetic study

A sensitive and reliable HPLC method with fluorescence detection based on the precolumn derivatization of glucosamine with 6-aminoquinolyl-N-hydroxylsuccinimidyl carbamate (AQC) was established for the quantitative determination of glucosamine in rat plasma. The plasma protein was precipitated by acetonitrile, followed by vortex mixing and centrifugation. The supernatant was divided into the organic layer and aqueous layer by adding sodium chloride, and then the aqueous layer was derivatized with AQC in 0.2 M borate buffer of pH 8.8 before the HPLC analysis. An amino acid analysis column (3.9 x 150 mm, 4 microm) was applied, with 140 mM sodium acetate buffer (pH = 5.25) and acetonitrile as mobile phase at a flow rate of 1 mL/min. A linear correlation coefficient of 0.9987 was calculated within the range of 0.1-30 microg/mL of the standard curve for glucosamine. The limit of detection was 30 ng/mL. The intra- and inter-day precisions (as RSD) were less than 7.38 and 12.72%, respectively. The intra- and inter-day accuracy ranged from 91.8 to 110.0%. Extraction recoveries of glucosamine in plasma were more than 90%. The validated method was successfully applied for the quantitative determination of glucosamine in rat plasma and evaluation for pharmacokinetic study of glucosamine. It was also possible to be applied for the quantitative determination of other compounds containing amino group in biological samples.     

高效液相色谱-程序波长紫外检测法同时测定血浆中的色氨酸及其代谢物

建立了一种高效液相色谱-程序波长紫外检测法同时测定血浆中色氨酸(Trp)及其主要代谢产物犬尿氨酸(Kyn)和5-羟色胺(5-HT)。以茶碱为内标(IS),采用BDS-Hypersil-C8柱(150 mm×4.6 mm, 5 μm)分离。流动相为10 mmol/L醋酸钠缓冲液(pH 4.5)-乙腈(94:6, v/v),流速为0.6 mL/min;柱温为25 ℃;紫外检测波长设定: Kyn和IS为360 nm, 5-HT为220 nm, Trp为302 nm。3种物质的平均回收率为87%～113%;线性范围分别为3.97～400 μmol/L(Trp), 0.421～20.2 μmol/L(Kyn), 4.36～980 nmol/L(5-HT);检出限分别为0.134 μmol/L(Trp), 0.0160 μmol/L(Kyn), 2.03 nmol/L(5-HT)。利用该方法对15例抑郁症患者和15例健康志愿者的血浆进行测定,结果表明两组间Trp的代谢存在显著的差异。     

High performance liquid chromatography for the determination of glucosamine sulfate in human plasma after derivatization with 9-fluorenylmethyl chloroformate

In this study, we developed a simple, rapid, sensitive, and reliable method for the determination of glucosamine sulfate in human plasma, which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by reverse-phase HPLC-FLD. For the first time, FMOC-Cl was introduced into derivatization of glucosamine sulfate in human plasma. The amino groups of glucosamine sulfate and vertilmicin sulfate (the internal standard) were trapped with FMOC-Cl to form glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts, which can be very suitable for HPLC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C18 column (DIAMONSIL 150 x 4 mm id, 5 microm) with a mobile phase gradient consisting of acetonitrile and water at a flow-rate of 1 mL/min. The retention times of glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts were 8.9 and 21.2 min, respectively. This method was shown to be selective and sensitive for glucosamine sulfate. The limit of detection was 15 ng/mL for glucosamine sulfate in plasma and the linear range was 0.1-10 mg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 5.2-8.1% and 6.1- 8.5%, respectively. Extraction recoveries of glucosamine sulfate in plasma were greater than 90%. The validated method was successfully applied to the determination of glucosamine sulfate in human plasma samples.     

高效液相色谱法同时测定血清中的犬尿氨酸和色氨酸

建立了一种能同时检测血清中的犬尿氨酸(kynurenine,Kyn)和色氨酸(tryptophan,Trp)的高效液相色谱-紫外检测法。采用的色谱柱为Symmetry Shield RP-C18柱(150 mm×3.9 mm i.d.,5 μm),流动相为15 mmol/L乙酸钠-乙酸溶液(含2.7%乙腈,pH 3.6),流速为1.0 mL/min,紫外检测波长为225 nm。血清标本经5.0%(体积分数)高氯酸溶液去除蛋白质后取上清液直接进样分析测定。研究结果表明,Kyn保留时间为3.5 min,线性范围为0.098~49 μmol/L,最低检出浓度为0.02 μmol/L,回收率为90.82%~93.45%;Trp保留时间为8.1 min,线性范围为4.9~490 μmol/L,最低检出浓度为0.20 μmol/L,回收率为95.51%~98.67%。Kyn和Trp日内、日间测定的相对标准偏差均小于4%,苯丙氨酸、酪氨酸、5-羟色胺和犬尿喹啉酸等物质对该法均无干扰。该方法简便、快速、稳定、可行,可应用于临床和科研工作。     

固相萃取/高效液相色谱法对肝癌细胞中多西紫杉醇的测定

建立了固相萃取/高效液相色谱法测定肝癌细胞中多西紫杉醇浓度的方法。细胞样品经三氯乙酸沉淀蛋白,Bond-elut C18固相萃取柱提取,采用Agilent TC-C18(5μm,4.6 mm×150 mm)色谱柱分析,以乙腈-0.02 mol/L醋酸铵缓冲液(体积比50∶50,醋酸调至pH5.0)为流动相,流速为1 mL/min,检测波长为230nm,进样体积为20μL。多西紫杉醇的质量浓度在0.05~2.25 mg/L范围内线性关系良好,相关系数r=0.999 3,检出限为0.03 mg/L,提取回收率为93%~95%,其日内、日间精密度(n=5)不大于6.9%,且稳定性良好。方法简便、快速、灵敏度高、干扰小,可用于生物样品中低浓度多西紫杉醇的测定。     

高效液相色谱法测定降糖中成药中添加的高极性化学降糖药物

建立了测定添加在降糖中成药中高极性的盐酸二甲双胍、盐酸苯乙双胍、阿卡波糖、伏格列波糖等4种化学降糖药物的高效液相色谱方法。采用Thermo氨基色谱柱(4.6 mm×250 mm,5 μm)分离,流动相组成为磷酸盐缓冲溶液(0.06%磷酸二氢钾和0.028%磷酸氢二钠溶液)-乙腈(体积比为30∶70),流速为1 mL/min,紫外检测波长为195 nm,柱温为30 ℃,进样量为20 μL。结果表明,4种化学降糖药能完全分离,中成药基质不干扰测定;检出限为0.1～3 mg/L;4种药物具有宽的线性范围和良好的线性关系(r2≥0.9981);日内、日间测定的相对标准偏差(RSD)分别为0.10%～5.07%和0.19%～6.41%;在中成药中的加标回收率除了添加低浓度的伏格列波糖的回收率较低外,均高于80%,RSD为1.14%～4.82%。该方法用于中成药中高极性的化学降糖药的检测具有特异、快速、简便、高效的特点。     

柱前衍生高效液相色谱-荧光检测法测定血浆中同型半胱氨酸

建立了一种柱前衍生高效液相色谱-荧光检测法用于测定血浆中同型半胱氨酸(Hcy)。使用三(2-羧乙基)膦盐酸盐(TCEP)为还原剂,N-(1-芘)马来酰亚胺(NPM)为衍生剂进行样品预处理,Agilent Hypersil C-18柱(250 mm×4.0 mm, 5 μm)进行分离,流动相为15 mmol/L醋酸钠-乙腈-混合酸(300 mL水中含1 mL醋酸和1 mL磷酸)混合溶液,采用梯度洗脱,荧光检测激发波长为330 nm,发射波长为380 nm。Hcy的回收率为(102.08±4.94)%。线性范围为0.500～100 μmol/L,检出限（以信噪比为3计）为0.016 μmol/L。日内与日间相对标准偏差均小于5%。利用该方法对7例高血压患者和7例健康志愿者的血浆进行了测定,结果表明两组间的Hcy含量存在显著的差异(p＜0.05)。本方法简单、快速、灵敏、特异,适用于血浆Hcy的临床定量测定。     

Evaluation and differentiation of the Betulaceae birch bark species and their bioactive triterpene content using analytical FT-vibrational spectroscopy and GC-MS

A new simple, rapid and sensitive reversed-phase liquid chromatographic method was developed and validated for the simultaneous determination of sulpiride (SUL) and mebeverine Hydrochloride (MEB) in the presence of their impurities and degradation products. The separation of these compounds was achieved within 6 min on a 250 mm, 4.6 mm i.d., 5 m particle size Waters^?-C18 column using isocractic mobile phase containing a mixture of acetonitrile and 0.01 M dihydrogenphosphate buffer (45:55) at pH = 4.0. The analysis was performed at a flow rate of 1.0 mL/min with fluorescence-detection at excitation 300 nm and emission at 365 nm. The concentration-response relationship was linear over a concentration range of 10- 100 ng/mL for both MEB and SUL with a limit of detection 0.73 ng/mL and 0.85 ng/mL for MEB and SUL respectively. The proposed method was successfully applied for the analysis of both MEB and SUL in bulk with average recoveries of 100.22 ± 0.757% and 99.96 ± 0.625% respectively, and in commercial tablets with average recoveries of 100.04 ± 0.93% and 100.03 ± 0.376% for MEB and SUL respectively. The proposed method was successfully applied to the determination of MEB metabolite (veratic acid) in real plasma simultaneously with SUL. The mean% recoveries (n = 3) for both MEB metabolite (veratic acid) and SUL were 100.36 ± 2.92 and 99.06 ± 2.11 for spiked human plasma respectively. For real human plasma, the mean% recoveries (n = 3) were and respectively.