黄精炮制品多糖多元指纹图谱的建立及化学模式识别

目的 构建黄精炮制品多糖的酶水解产物指纹图谱,结合化学计量学方法评价不同蒸晒次数黄精炮制品多糖的差异。方法 采用糖苷酶水解,结合高效薄层色谱（high thin-layer chromatography,HPTLC）、亲水作用色谱-高效液相色谱-蒸发光检测器（hydrophilic interaction liquid chromatography-high performance liquid chromatography-evaporative light scattering detector,HILIC-HPLC-ELSD）和超高效液相色谱-四级杆飞行时间串联质谱（ultra high pressure liquid chromatography-quadrupole time of flight tandem mass spectrometry,UHPLC-QTOF/MS）对相应的酶解产物进行高效分离和检测。由此产生的自下而上的指纹图谱反映了黄精炮制品多糖的变化,通过相似度和偏最小二乘-判别分析（partial least squares-discriminant analysis,PLS-DA）对指纹图谱进行综合评价。结果 HPTLC分析发现,果糖随着蒸晒次数的增加逐渐降低,结合色谱分析发现,六蒸六晒和十二蒸十二晒为黄精炮制过程中的转折点。同时根据酶水解中寡糖的变化,利用相似度和PLS-DA对不同蒸晒次数的黄精炮制品进行了清晰的聚类。结合UHPLC-QTOF/MS分析表明,不同蒸晒次数黄精炮制品中含有蔗糖、蔗果三糖、蔗果四糖、蔗果五糖、蔗果六糖、蔗果七糖及蔗果八糖。结论 基于酶解法分析不同蒸晒次数样品的质量动态变化,结合所建立的多元指纹图谱,进一步阐释了不同蒸晒次数黄精炮制品的科学内涵。     

Validated HPTLC methods for the determination of salicin in Salix sp. and of harpagoside in Harpogophytum procumbens

Preparations of Harpagophytum procumbens and of Salix species are successfully used for the treatment of degenerative rheumatism and painful arthrosis. For the quality control of both drugs, rapid methods of quantification are desirable. Here we report the development of two HPTLC methods enabling the determination of harpagoside in Harpagophyti radix and of salicin in Salicis cortex. We focused on a standardized methodology and thorough validation including two laboratories. The methods allow the analyses of up to 16 samples in parallel demonstrating the proposed methods as very rapid and cost efficient.     

The ICH guidance in practice: stress degradation studies on indinavir sulphate and development of a validated specific stability-indicating HPTLC assay method

A sensitive, selective, precise and stability-indicating high-performance thin layer chromatography (HPTLC) method for analysis of indinavir sulphate both as a bulk drug and in formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of carbon tetrachloride/chloroform/methanol/10% v/v ammonia (4:4.5:1.5:0.05, v/v/v/v). Densitometric analysis of indinavir sulphate was carried out in the absorbance mode at 260 nm. This system was found to give compact spots for indinavir sulphate (Rf value of 0.43 +/- 0.02, for six replicates). Indinavir sulphate was subjected to acid and alkali hydrolysis, oxidation, dry and wet heat treatment, and photo degradation. The drug undergoes degradation under acidic and basic conditions, oxidation, dry and wet heat treatment, and photo degradation. Also the degraded products were well resolved from the pure drug with significantly different Rf values. The method was validated for linearity, precision, robustness, limit of detection (LOD), limit of quantitation (LOQ), specificity and accuracy. Linearity was found to be in the range of 100-6000 ng/spot with significantly high value of correlation coefficient r2 = 0.997 +/- 0.64. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.999 +/- 0.002 in the working concentration range of 1000-6000 ng/spot. The LOD and LOQ were 40 and 120 ng/spot, respectively. Statistical analysis proves that the method is repeatable and specific for the estimation of the said drug. As the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating one. Moreover, the proposed HPTLC method was utilized to investigate the kinetics of acid degradation process. Arrhenius plot was constructed and activation energy was calculated.     

An HPTLC method for the determination and the purity control of ciprofloxacin HCl in coated tablets

A high-performance thin-layer chromatographic method (HPTLC) has been developed for the determination and the purity control of a synthetic fluoroquinolone antibiotic ciprofloxacin hydrochloride in coated tablets when desfluoro compound, ethylene diamine compound, by-compound A and fluoroquinolonic acid are considered as impurities. Silica gel F₂₅₄ was used as a stationary phase and a mixture of acetonitrile, ammonia 25%, methanol and methylene chloride (1:2:4:4, v/v/v/v) as a mobile phase. The method was validated in terms of linearity (range), selectivity (placebo and related compounds), precision, accuracy (Recovery), system suitability (repeatability, peak symmetry, resolution) and impurities limit of detection (LOD). The analysis of variance (ANOVA) and t-test were applied to correlate the results of ciprofloxacin hydrochloride determination in coated tablets by means of the HPTLC method and the official British Pharmacopoeia (BP 1999) high-performance liquid chromatographic (HPLC) method.     

Comparative Phytochemical Investigation of the Sources of Ayurvedic Drug Patha: A Chromatographic Fingerprinting Analysis

Standardization of herbal drugs based on their chemical and biological activity profile is an important prerequisite for acquiring the herbal market. The main problem encountered in standardization of Ayurvedic drugs is proper identification of the source plant. The present study was aimed to establish identification characters, quality control parameters, chemical and biological parameters for roots of three plants Cissampelos pareira, Cyclea peltata and Stephania japonica (Fam. Menispermaceae) which are being used as source of Patha, in the market. All the three plant were subjected for evaluation of quality control parameters as per WHO guidelines and root extracts and total alkaloidal fractions were subjected for HPTLC and HPLC fingerprinting analysis using a marker compound Bebeerine isolated from roots of Cissampelos pareira. The parameters studied clearly indicated the significant differences among the three plant materials. The roots of Cissampelos pareira can be distinguished from other two plants by presence of high concentration of alkaloids especially the presence of high concentration of pharmacologically active alkaloid bebeerine, which was found to be present in very low concentration in Stephania japonica and absent in roots of Cyclea peltata. The roots of Cyclea peltata were found to contain high concentration of saponins and comparatively in low concentration in Cissampelos pareira where as it was found to be absent in roots of Stephania japonica.     

Validated HPTLC Method for Simultaneous Determination of Quinapril Hydrochloride and Hydrochlorothiazide in a Tablet Dosage Form

Quinapril hydrochloride and hydrochlorothiazide were simultaneously determined by HPTLC in pharmaceutical formulations. The drugs were separated on silica gel 60 F₂₅₄ plates using suitable combination of solvents as mobile phase. The validation parameters, tested in accordance with the requirements of ICH guidelines, prove the suitability of methods.     

Development and validation of an analytical method based on high performance thin layer chromatography for the simultaneous determination of lamotrigine, zonisamide and levetiracetam in human plasma

Methods based on HPLC technology are the most frequently adopted for monitoring blood levels of novel antiepileptics. Here a rapid method based on HPTLC was developed for quantitative determination of lamotrigine (LTG), zonisamide (ZNS) and levetiracetam (LVT) in human plasma and compared with HPLC and LC-MS/MS methods. Chromatographic separation was achieved on silical gel 60F₂₅₄ plates using ethylacetate:methanol:ammonia (91:10:15 v/v/v) as mobile phase. Quantitative analysis was carried out by densitometry at a wavelength of 312, 240 and 210 nm for LTG, ZNS and LVT, respectively. Calibration curves were linear over range of 0-200 ng for LTG and ZNS and 0-400 ng for and LVT. The limit of quantification of LTG, ZNS and LTV was found to be 3.69, 3.7 and 6.85 μg/ml, respectively. Intra and inter-assay precision provided relative standard deviations lower than 10% for all three analytes. Correlation and Bland-Altman plot showed general agreement between HPTLC and LC-MS/MS quantification, with a mean bias of −0.25, −0.46 and 0.5 μg/ml for LTG ZNS and LVT, respectively. Likewise, comparison between HPLC-UV and LC-MS/MS showed good agreement for all the three compounds analyzed. In conclusion, the proposed HPTLC method is simple, rapid, precise and accurate. It therefore is appropriate for the routine quantification of therapeutic levels of LTG, ZNS and LVT in human plasma.     

甘薯长喙壳菌毒素的HPTLC实验方法研究

在 3 0℃条件下在石油醚 -醋酸乙酯 (2 :1,v/ v)和苯 -甲醇 (9:1,v/ v)两种溶剂系统中 ,对甘薯酮 (C)、甘薯酮醇 (A)等毒素有较好的分离效果 ,采用高效薄层色谱法检测甘薯酮 ,最低检测限量纯品为 0 .0 0 2μg,粗品为 0 .0 1μg。     

高效薄层层析法测定罗布麻烟中罗布麻浸膏的含量

利用高效薄层层析法,以罗布麻浸膏中的槲皮素为标准,测定罗布麻烟中罗布麻浸膏的含量。以甲苯:甲酸:乙酸乙酯:正己烷(2:0.2:1.5:0.2)作展开剂。可使得药烟中槲皮素与其它成分很好分离。分离后,用薄层扫描仪于波长410nm,参比波长750 nm扫描定量。本法需样品少,快速简便、灵敏准确。可适用于药烟生产厂家化验室检测使用。     

高效薄层扫描法测定四物汤中阿魏酸含量

用高效薄层扫描法（ＨＰＴＬＣ）测定了中药复方四物汤群煎液、分煎混合液、当归和川芎单煎液中阿魏酸的含量。以高效硅胶薄层板，苯－氯仿－乙酸乙酯－甲酸（８∶２∶２∶０．５）为展开剂，在紫外灯３６５ｎｍ波长下定位，用ＣＳ－９３０型薄层扫描仪进行测定，λｓ＝３２０ｎｍ，λＲ＝２４０ｎｍ，狭缝１．２ｍｍ×１．２ｍｍ，线形化参数：ＳＸ＝７，反射法锯齿形扫描，二点外标法定量测定