健脾补血片中阿魏酸的质量浓度测定

目的建立健脾补血片中阿魏酸的高效液相色谱(HPLC)测定方法。方法采用HPLC法。色谱柱Lichrospher C18(4.6 mm×250 mm,5μm),C18保护柱;流动相:乙腈-0.378 mol/L冰醋酸水溶液(30∶70);检测波长321 nm;流速0.8 mL/min;柱温:室温;进样量10μL。结果阿魏酸质量浓度在1.0-10.0 mg/L范围内,与峰面积呈良好的线性关系(r=0.9996),平均回收率为99.8%,RSD为0.26%。结论HPLC可用于健脾补血片中阿魏酸的质量浓度测定。     

引阳素胶囊中淫羊藿苷的含量测定

目的建立引阳素胶囊中淫羊藿苷的高效液相色谱(HPLC)含量测定方法。方法采用HPLC法。色谱柱Li-chrospher C18(4.6 mm×250 mm,5μm),C18保护柱;流动相为乙腈-20 g/L冰醋酸水溶液(30∶70);检测波长270 nm;流速1.0 mL/min;柱温为室温;进样量10μL。样品加50%(体积分数)乙醇超声提取,浸提液过0.45μm滤膜后测定。结果淫羊藿苷线性范围为10.0-60.0 mg/L,回归方程为C=2.042×10-5A+0.554,r=0.9993,平均回收率为99.8%,RSD为0.65%。结论本法用于引阳素胶囊中淫羊藿苷的含量测定时简便快捷、准确。     

治带片中苦参碱及氧化苦参碱的HPLC法测定

目的建立治带片中苦参碱及氧化苦参碱的高效液相色谱(HPLC)测定方法。方法色谱柱:Lichrospher-NH2(4.6 mm×250 mm,5μm),C18保护柱;流动相:乙腈-无水乙醇-0.5 mol/L磷酸水溶液(80∶10∶10);检测波长212 nm;流速1.0 mL/min;柱温:室温;进样量20μL。结果苦参碱和氧化苦参碱线性范围均为1.0-10.0μg/mL,回归方程苦参碱为:C=1.201×10-4A+0.161,r=0.9992;氧化苦参碱为:C=1.366×10-4A+0.221,r=0.9996,平均回收率分别为99.9%和99.4%,RSD分别为1.48%和4.33%。结论本法简便快捷,结果准确,可用于该制剂的质量控制。     

丹皮药材指纹图谱定性和有效成分定量分析方法研究

目的　建立丹皮药材的反相高效液相色谱定性和定量分析方法。方法　RP HPLCPlanetsilC18分析柱 (15 0mm× 4 .6mm ,5 μm) ,流动相为甲醇 水 冰醋酸 (44∶5 6∶0 .1) ,流速为 1.0mL·min-1,检测波长为 2 4 0nm ,柱温为室温 ,对丹皮药材进行指纹图谱定性分析 ,并对丹皮中的有效成分丹皮酚进行定量测定。结果　在此色谱条件下 ,13份不同丹皮药材的RP HPLC指纹图谱中可检出 11个相对位置稳定的色谱峰 ,通过色谱峰间峰面积的相关性分析 ,可以确定 10个峰可作为定性鉴别的指标峰 ;丹皮药材中丹皮酚含量为 1.32 %～ 2 .78%。结论　丹皮RP HPLC指纹图谱定性和有效成分定量分析方法具有较好的针对性和准确性 ,可用于丹皮药材的质量控制。     

液相色谱-离子阱质谱联用法测定西洋参药材中p-F_(11)含量

目的　建立液相色谱 离子阱质谱联用法 ,测定西洋参特有指标成分拟人参皂苷 p F11含量。方法　通过固相萃取小柱预处理样品 ,电喷雾质谱正离子模式全扫描法检测 ,选择离子法定量 ,用液相色谱 蒸发光散射法对该定量方法进行了佐证。结果　线性范围为 0 .4 5～ 7.2 0 μg ,回收率为 98.7% ,精密度和重现性实验RSD值分别为 2 .5 %和2 .8% ,样品在 4 8h内稳定性良好。该方法测得 p F11质量浓度为 3.0 1mg·g-1。结论　该方法有良好的精密度和重现性 ,能准确分析、测定西洋参药材中 p F11含量     

HPLC-MS/MS法测定兔血浆中黄芪甲苷的含量

目的 建立兔含药血浆中黄芪甲苷含量快速测定的新方法.方法 固相萃取法富集血浆中的黄芪甲苷;Agilent SB-C18反相色谱进行分离;采用离子阱质谱多级反应模式,以m/z 807.5→627.1为选择离子对,内标法测定含量.结果 血浆中黄芪甲苷的检测限为0.1ng/mL,线性范围为0.5～50.0μg/mL,平均方法回收率在96.3%～104.7%范围内.结论 该方法具有灵敏度高、稳定性好和快速的特点,符合生物样品的分析要求,有望用于黄芪甲苷的药代动力学研究.     

沙棘油中α-生育酚含量的反相高效液相层析分析法

为了控制沙棘油的质量,建立了一种反相高效液相层析法(RP—HPLC)测定沙棘油中α—生育酚的含量。沙棘油样品皂化后,用乙醚提取,浓缩乙醚提取液,残渣溶解于无水乙醇中注入液相层析仪。层析条件:ZORBAX柱(4.6×150mm)甲醇作流动相(1ml/min),紫外检测波长,280nm。测定了陕西永寿产沙棘种子油和果油各3批,α—生育酚的平均含量分别为989mg/kg,1754mg/kg。方法回收率98.6%(种子油),102.2%(果油),变异系数小于5.44%。     

β-榄香烯增强柔红霉素杀伤人类白血病细胞的作用及机制

目的探讨β-榄香烯(β-elemene)对白血病细胞的耐药逆转及其对聚腺苷二磷酸聚合酶(PARP)和P-糖蛋白(P-gp)表达的影响。方法取对数生长期人类髓系白血病K562细胞的柔红霉素(DNR)耐药株K562/DNR及其敏感株K562细胞,采用台盼蓝拒染法测定其对β-榄香烯的药物敏感性及耐药逆转,采用流式细胞术PI染色检测细胞凋亡,采用流式细胞术检测细胞内药物累积,Western blot检测蛋白PARP和P-gp的表达,应用SPSS(13.0软件包)进行统计学分析。结果β-榄香烯明显抑制K562和K562/DNR细胞的生长,呈剂量依赖关系。低毒剂量(5μg/mL)β-榄香烯作用K562/DNR细胞,显著提高了柔红霉素的细胞毒作用,IC50降低至(0.68±0.43,P<0.05),逆转倍数为1.91倍。与柔红霉素单药组相比,加入低毒剂量β-榄香烯后,细胞药物蓄积明显增加(P<0.05)。β-榄香烯作用后诱导PARP裂解,同时下调了P-gp蛋白表达。结论β-榄香烯部分逆转K562/DNR细胞对柔红霉素的耐药性,其逆转机制与增加细胞内药物的蓄积、诱导PARP裂解及降低P-gp的表达有关。     

杨木粉无胶温压成形复合材料环境友好性评价

在运用热脱附-气相色谱/质谱(TD-GC/MS)联机技术分析杨木粉末无胶温压成形复合材料在4,60,90和160℃等4个环境温度点的挥发物总离子流图的基础上,采用色谱峰面积归一化法鉴定出杨木粉末基复合材料在各温度下的保留时间及峰面积,进而评价杨木粉末基复合材料的环境友好性。结果表明:在不同温度下,杨木粉末基复合材料挥发物中的有益成分含量明显多于有害成分,在40,60和160℃时,有益成分含量为有害成分含量的6倍以上,90℃时有益成分为有害成分的29倍,环境友好性良好;杨木粉末基复合材料总挥发物种类及有益成分种类、有害成分种类与杨木原粉相应种类大致相当,但有益成分明显增多,有害成分减少,增减幅度较大。说明这类杨木粉末基复合材料不仅无胶温压成形工艺具有环境友好性,且其制品的制备和使用过程都具有环境友好性。     

高效液相色谱法测定舒肤胶囊中芍药苷的含量

目的采用HPLC法对处方中主要成分芍药苷进行含量测定。方法采用高效液相色谱法,以KromasilC18(250 mm×4.6 mm,5μm)为分析柱,流动相为乙腈-1.5 mL/L磷酸(12∶88),流速1 mL/min,检测波长230 nm,柱温为室温。结果芍药苷回归方程:Y=83 168X+72 569(r=0.999 5,n=6),芍药苷在0.06~0.18μg范围内线性关系良好;方法的平均回收率为100.21%,精密度RSD的值为1.31%。结论本方法操作简便、快速、准确、灵敏,适用于舒肤胶囊的质量控制。     

Analysis of ganciclovir and its related substances using high performance liquid chromatography and liquid chromatography-mass spectrometry methods

Objective High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC/MS) methods were developed for the determination of ganciclovir and its related substances. Methods A Hypersil ODS2 column (4.6mm×250mm, 5μm) was used with a mobile phase of 0.02M potassium 1.0mL/min, and UV detector set at 254nm was used for monitoring the eluents. Results The method was simple, rapid, selective and capable of separating all related substances at trace level with a detection limit of 0.04μg/mL. It has been validated with respect to accuracy, precision, linearity, and limits of detection and quantification. The linearity range was 10.2-153.0μg/mL with r=0.9998. The percentage recoveries ranged from 96.7% to 101.6%, and RSD was 1.24%-1.96% (n=5). Conclusion The method was found to be suitable not only for monitoring the reactions during the process development but also for quality control of ganciclovir. For identification of related substances, LC/MS was used. The mainly related substances of ganciclovir active pharmaceutical ingredients (API) were determined as guanine, (1, 3-dioxolan-4-yl) methyl acetate, and diacetyl guanine.     

~(99m)Tc-HL91心肌乏氧组织显像的初步实验研究

目的　验证乏氧组织显像剂99mTc HL91能选择性浓集于缺血的心肌中 ,为临床显像提供依据。方法　制备兔心肌缺血模型 ,观察99mTc HL91在缺血心肌中的分布情况。按常规方法观察99mTc HL91在正常兔体内的分布情况。结果　电生理改变及NBT染色均证实兔左心室LCX区心肌缺血。左心室切片显像见LCX区心肌较LAD区放射性明显增浓。LCX/LAD组织放射性比 :12 0min ,2 .5 7;180min ,3.4 5。99mTc HL91主要分布在肝脏和肾脏 ,其次为胃肠道 ,排泄以泌尿系、肠道为主 ,正常心肌及肺脏中分布少 ,且清除较快。血液清除快。结论　缺血心肌组织较正常组织滞留99mTc HL91明显增高 ,180min时较显著。提示99mTc HL91可用于心肌乏氧显像     

肝细胞肝癌患者的血清蛋白质组学研究

目的分析肝细胞肝癌患者的血清蛋白质谱,寻找诊断肝癌和检测治疗的特异性标志物。方法收集了50例肝癌患者血清,在去除血清中6种高丰度蛋白后进行双向凝胶电泳分离及银染,将凝胶扫描图像与正常人血清比较分析,然后经质谱鉴定20个差异表达蛋白质点。结果共鉴定了11种差异表达蛋白质,其中7种蛋白质在肝癌患者血清中表达水平上调,其余4种蛋白质表达水平下调。有7个点被同时鉴定为α1-抗胰蛋白酶,均在肝癌患者血清中表达上调,提示α1-抗胰蛋白酶在肝癌中降解加快。Western blotting检测进一步证实多数肝癌血清中的α1-抗胰蛋白酶小片段增多。结论血清是诊断肝癌和检测治疗非常重要的临床标本,本实验中鉴定的蛋白质虽不能作为临床诊断标志物,但对分析肝癌血清蛋白谱的改变有一定的参考价值。     

川芎药材活性部位的高效液相色谱指纹图谱定性分析方法

目的　建立川芎药材活性部位的指纹图谱 ,为其定性鉴别提供依据。方法　川芎药材经乙醇超声提取后 ,用醋酸乙酯萃取 ,采用ODS柱为分析柱 ,以甲醇 水 冰醋酸 (30∶70∶0 .5 )为流动相 ,在 2 76nm条件下进行分析 ,建立其指纹图谱。并对 7种不同来源的川芎药材进行高效液相色谱指纹图谱定性分析比较。结果　本研究建立的分析方法的精密度、重现性较好 ,以阿魏酸峰计 ,RSD分别为 1.6 8%和 1.0 4 % ;不同川芎药材指纹图谱中主要峰群的整体图貌基本一致 ,但各成分含量的相对比值有所不同 ,不同来源的川芎药材共有峰面积的比值及共有峰面积和均有一定的差异。结论　HPLC指纹图谱分析法可简便、快速地鉴别区分不同来源的川芎药材     

脑肿瘤~(99m)Tc-MIBI脑SPECT显像的影像特征及应用价值

为探讨 99m Tc-甲氧基异丁基异腈 ( MIBI)脑 SPECT显像在脑肿瘤中的影像特征及应用价值 ,对 1 0例正常对照者和 30例脑内病变患者进行99m Tc- MIBI脑SPECT显像。结果 :对照组脑实质内均无核素浓聚 ,脑肿瘤组表现为阳性结果者 1 9例 ,显示出脑实质内有局限性核素浓聚 ;表现为阴性结果者 6例 ,均为脑肿瘤术后无复发者 ;5例非肿瘤组良性病变患者均表现为阴性结果。本组术前诊断脑肿瘤的灵敏度与特异性均为 1 0 0 %。表明该显像方法可用于脑肿瘤的诊断 ,尤其是发现并鉴别脑肿瘤术后残留、复发及软化灶形成。     

Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS

A global quality control method based on high performance liquid chromatography (HPLC) coupled with diode array detection (DAD), single quadrupole mass spectrometry (MS) and time-of-flight mass spectrometry (TOFMS) was developed for simultaneous determination of seven major components (mangiferin, neomangiferin, timosaponin E1, timosaponin E, timosaponin BⅡ, timosaponin BⅢ, and timosaponin AⅢ) and identification of most components in extracts of Rhizoma Anemarrhenae (RA). HPLC analysis was performed on an Agilent SB-C18 column (4.6 mm×150 mm, 5 μm) by gradient elution using acetonitrile and water-acetic acid(100∶0.05, v/v) as the mobile phase. Seven major components in RA were successfully separated. This quantitative method was fully validated in respect of the following performance criteria: linearity, precision, repeatability, stability, accuracy, limits of detection (LOD) and quantification (LOQ). A formula database of known compounds in RA was established, against which, most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS. This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China. This global quality control method, which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components, is suitable for routine quantification and comprehensive quality control of RA.     

Simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture by high performance liquid chromatography

A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.     

乌头碱在家兔检材中的降解动力学

用高效液相色谱法(High performance liquid chromatography,HPLC)分离测定生物性样品中的乌头碱。对乌头碱在离体生物检材中的降解动力学进行了研究。乌头碱在经不同处理的生物检材(凝血块、肝脏)中以表观二级动力学过程被降解,其降解速率常数K以4%甲醛溶液固定组中为最大(1.5619,0.7728μg/d),其次为95%酒精固定组(0.4891,0.7556μg/d),4℃冰箱放置组中最小(0.06372,0.0393μg/d),亦即中毒检材中乌头碱在冰箱中分解较慢,其次为在酒精固定剂中。从死亡或中毒后一段时间取检材检测,可用建立的动力学方程推算体內乌头碱是否已达中毒量或致死量。     

Study on chromatographic fingerprint of sarcandra glabra （Thunb.） by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction (MAE) was used for extraction of effective components of sarcandra glabra (Thunb.), and then chromatographic fingerprint of sarcandra glabra (Thunb.) was studied by high performance liquid chromatography/diode array detector (HPLC/DAD). The conditions of MAE were optimized by an orthogonal experiment, and then the authentication and validation of the chromatographic fingerprint were conducted. Nine peaks were identified as common peaks in the fingerprint chromatograms, and isofraxidin was considered as a reference compound and quantified. Relative standard deviations of retention time and peak area of each component were less than 3% and 8%, respectively. Similarity and difference analysis were conducted by use of PCA and relation coefficient. Twenty batches of sarcandra glabra (Thunb.) samples from two different producing areas could be classified into two different groups in the PCA model. The results showed that MAE-HPLC/DAD method was simple, efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra (Thunb.), which could provide more reliable and precise information for quality evaluation.     

大骨节病病因研究——病区天堂村饮水中有机物的测定(Ⅰ)

本研究应用XAD树脂富集,色谱/质谱联机分离鉴定的方法。对大骨节病病区(麟游县天堂村)三种饮水与对照(西安医科大学校园)深井水作了痕量有机化合物的测定分析。从天堂河水、深机井水、窑水与对照水样中分别检出了40种、31种、19种和10种有机化合物,且对每种检出物的结构式、分子量和峰面积均作了分析测定。其中芳核化合物分别为25种、19种、8种和4种。病区三种水样中均检出了烯类、烯酮类、酮类和萘类、苯并噻唑等稠环芳烃化合物;仅在河水和深井水样中检出了茚类化合物;在对照水样中未发现上述有机物。此结果未见文献报导。上述诸有机化合物可能是值得重视的可疑病因因素,但其真正的病因流行病学意义尚需进一步研究探讨。