生附片化学成分的HPLC/ESI-MS~n研究

采用高效液相色谱与电喷雾质谱联用技术,对生附片的化学成分进行了系统的研究.并辅以提取离子色谱方法.发现微量的化学成分.通过保留时间,质荷比及多级串联质谱数据,共鉴定了48个成分,其中双酯型生物碱8个,单酯型生物碱7个,脂型生物碱29个.其中双酯型生物碱是生附片中的主要成分,而单酯型和脂型生物碱的含量和种类较少.

西洋参与北沙参的HPLC、ESI-MS指纹图谱识别

目的:通过HPLC、ESI-MS指纹图谱对中药西洋参与北沙参进行鉴别。方法:利用HPLC和ESI-MS技术,优化西洋参、北沙参提取物的色谱和质谱分离分析条件,建立二者HPLC、ESI-MS指纹图谱。结果:确定出西洋参与北沙参HPLC的各色谱峰相对保留时间;根据MS图中各成份的m/z值,确定相应成分的分子量。二者的液相色谱和质谱指纹图谱完全不同。结论:通过HPLC、ESI-MS指纹图谱可以完全鉴别中药西洋参与北沙参,方法简单准确,重现性好,具有实用价值。

刺五加叶的HPLC-UV和ESI-MS指纹图谱研究

对12批不同来源的刺五加叶提取物进行指纹图谱研究,并利用ESI-MS指纹图谱鉴别刺五加叶与山楂叶。分别采用高效液相色谱(HPLC-UV)和电喷雾电离质谱(ESI-MS)测定12批不同来源的刺五加叶提取物,利用ESI-MS技术测定刺五加叶与山楂叶提取物,得到了分离度、精密度和重现性均较好的刺五加叶HPLC-UV及ESI-MS指纹图谱;同时,利用刺五加叶与山楂叶ESI-MS指纹图谱的差异,成功鉴别了二者,可为刺五加叶药材的质量控制提供参考。

麻黄炮制条件的ESI-MS法优化

ESI-MS was used to optimize the Ephedra sinica refinement. The ratio of honey to drug is 20/100, and the ratio of water to honey is 1/2. The toast temperature is 80 ℃, and the toast time is 2 h. The change of Ephedra sinica after processing was given.

ESI-MS和HPLC-UV法研究大黄、黄柏、赤芍炮制前后化学成分变化

目的:考察大黄、黄柏、赤芍炮制前后化学成分在质和量方面的变化,以探讨中药的炮制机理。方法:利用电喷雾质谱法(ESI-MS)和高效液相色谱法(HPLC-UV),对大黄、黄柏、赤芍炮制前后的化学成分进行详细研究。质谱:电喷雾电离源(ESI),加热毛细管温度为200℃,喷雾电压为4.5kV;液相色谱:色谱柱为Agilent Zorbax C18柱(4.6mm×150mm,5μm),柱温均为28℃。结果:大黄经酒制和醋制后,其化学成分的含量变化较大;黄柏经酒制后巴马汀的含量略有增加,而小檗碱的含量呈下降趋势;盐制对于黄柏的化学成分几乎无影响;赤芍经酒制和炒制后,芍药苷的含量都呈下降趋势。结论:不同炮制方法对化学成分的影响不同,为进一步阐明中药材炮制入药的科学内涵提供了实验依据。

复方板蓝根颗粒水提部位的ESI-MS指纹图谱研究

Sialic acid and lysine as an internal standard substance,the ESI-MS fingerprinting of extracts of Compound Indigowoad Root Granule which from different origin was studied.The fingerprinting was obtained with better precision and reproducibility.This work can provide a reference for the quality control of Compound Indigowoad Root Granule.

ESI-MS法研究白芍炮制前后化学成分的变化

ESI-MS analytical method was employed to study extracts of unprocessed and processed Radix Paeoniae alba.The results of the experiments prove that different processed methods is making a certain different influence on the contents of chemiacl components in Radix Paeoniae alba.This method is simple and accurate.It can be used for analyse of Traditional Chinese Medicine(TCM).

乌头碱在家兔肠道内代谢产物的LC/ESI-MSn研究

采用LC/ESI-MSn的方法对家兔肠道内的乌头碱代谢产物进行研究,经与空白组比较发现,给药后家兔小肠内容物中新增加6个化合物峰(M1～M6),盲肠内容物中新增加5个化合物峰(M2～M6),粪便中新增加2个化合物峰(M3、M4). 分别测定各化合物的准分子离子及各级串联碎片离子,并与标准品的质谱断裂规律进行比较,同时参考文献,推断肠道内化合物M1为16-O-去甲基-8-O-去乙酰基乌头碱,M2为8-O-去乙酰基乌头碱,M3为16-O-去甲基乌头碱,M4为乌头碱(AC),M5为去氧乌头碱,为印乌头碱。

Studies on the content variation of chemical constituents during the combination of ginseng with veratrum nigrum by ESI-MS and HPLC-ESI-MS

To study the content variation of ginsenosides and alkaloids during combination of ginseng with veratrum nigrum, the ginsenosides and alkaloids in the decoction of ginseng with veratrum nigrum were analyzed and compared by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) and electrospray ionization-mass spectrometry (ESI-MS). In the compatible decoction, eight ginsenosides and eight alkaloids. were detected, and the contents of six ginsenosides were found to be reduced, on the contrary, the contents of six alkaloids were increased. During combination of ginseng with veratrum nigrum, the contents of ginsenosides were reduced and those of the toxic alkaloids were increased. From the chemical point of view, the traditional theory is right that ginseng and veratrum nigrum are incompatible with each other.

Pre-column derivatization of amines with 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate followed by LC-fluorescence and LC-APCI-MS

A pre-column derivatization method for the sensitive determination of amines using the labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate (BCIC-Cl) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives is carried out by high performance liquid chromatography/atmospheric pressure chemical ionization (LC-APCl-MS-MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent is replaced by 1,2-benzo-3,4-dihydrocarbazole-9-isopropyl functional group, which results in a sensitive fluorescence derivatizing reagent BCIC-Cl. BCIC-Cl can easily and quickly label amines. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography and show an intense protonated molecular ion corresponding m/z [MH](+) under APCl in positive-ion mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 260 corresponding to the cleavage of CH2-OCO bond. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3 to 4-fold molar reagent excess. In addition, the detection responses for BCIC derivatives are compared with those obtained using CEOC and FMOC as derivatization reagents. The ratios of l(BCIC)/l(CEOC) and l(BCIC)/l(FMOC) are, respectively, 1.23-3.14 and 1.25-3.08 for fluorescent (FL) responses (here, l is relative fluorescence intensity). Separation of the derivatized amines had been optimized on reversed-phase Eclipse XDB-C-8 column. Detection limits are calculated from 1.0 pmol injection, at a signal-to-noise ratio of 3, are 10.6-37.8 fmol. The mean interday accuracy ranges from 94 to 105% for fluorescence detection with the largest mean %CV < 7.5. The mean interday precision for all standards is < 6.0% of the expected concentration. Excellent linear responses are observed with coefficients of > 0.9997.

HPLC-ESI-MS method development for the elucidation of nicardipine degradation products

Purpose: Nicardipine is a member of a family of calcium channel blockers named dihydropiridines that are known to be photolabile and may cause phototoxicity. It is therefore vital to develop analytical method which can study the photodegradation of nicardipine. Method: Forced acid degradation of nicardipine was conducted by heating 12 ml of 1 mg/ml nicardipine with 3 ml of 2.5 M HCl for two hours. A gradient HPLC medthod was developed using Agilent Technologies 1200 series quaternary system. Separation was achieved with a Hichrome (250 x 4.6 mm) 5 μm C18 reversed phase column and mobile phase composition of 70% A(100%v/v water) and 30% B(99%v/v acetonitrile + 1%v/v formic acid) at time zero, composition of A and B was then charged to 60%v/v A;40%v/v B at 10minutes, 50%v/v A; 50%v/v B at 30minutes and 70%v/v A; 30%v/v B at 35minutes. 20μl of 0.8mg/ml of nicardipine degradation was injected at room temperature (25oC). The gradient method was transferred onto a HPLC-ESI-MS system (HP 1050 series - AQUAMAX mass detector) and analysis conducted with an acid degradation concentration of 0.25mg/ml and 20μl injection volume. ESI spectra were acquired in positive ionisation mode with MRM 0-600 m/z. Results: Eleven nicardipine degradation products were detected in the HPLC analysis and the resolution (RS) between the respective degradants where 1.0, 1.2, 6.0, 0.4, 1.7, 3.7, 1.8, 1.0, and 1.7 respectively. Nine degradation products were identified in the ESI spectra with the respective m/z ratio; 171.0, 166.1, 441.2, 423.2, 455.2, 455.2, 331.1, 273.1, and 290.1. The possible molecular formulae for each degradants were ambiguously determined. Conclusion: A sensitive and specific method was developed for the analysis of nicardipine degradants. Method enables detection and quantification of nicardipine degradation products that can be used for the study of the kinetics of nicardipine degradation processes.

Development and validation of a dried blood spot-LC-APCI-MS assay for estimation of canrenone in paediatric samples

A selective and sensitive liquid chromatography (LC)-atmospheric pressure chemical ionisation (APCI)-mass spectroscopic (MS) assay of canrenone has been developed and validated employing Dried Blood Spots (DBS) as the sample collection medium. DBS samples were prepared by applying 30 mu l of spiked whole blood onto Guthrie cards. A 6 mm disc was punched from the each DBS and extracted with 2 ml of methanolic solution of 17 alpha-methyltestosterone (Internal Standard). The methanolic extract was evaporated to dryness and reconstituted in acetonitrile:water (1:9, v/v). The reconstituted solution was further subjected to solid phase extraction using HLB cartridges. Chromatographic separation was achieved using Waters Sunfire C18 reversed-phase column using isocratic elution, followed by a high organic wash to clear late eluting/highly retained components. The mobile phase consisted of methanol:water (60:40, v/v) pumped at a flow rate of 0.3 ml/min. LC-APCI-MS detection was performed in the selected-ion monitoring (SIM) mode using target ions at m/z 341.1 and 303.3 for canrenone and internal standard respectively. The selectivity of the method was established by analysing DBS samples from 6 different sources (individuals). The calibration curve for canrenone was found to be linear over 25-1000 ng/ml (r >0.994). Accuracy (% RE) and precision (% CV) values for within and between day were

Development and validation of a method for the confirmation of nicarbazin in chicken liver and eggs using LC-electrospray MS-MS according to the revised EU criteria for veterinary drug residue analysis

A method is described for the quantitative confirmation of 4,4'-dinitrocarbanilide (DNC), the marker residue for nicarbazin in chicken liver and eggs. The method is based on LC coupled to negative ion electrospray MS-MS of tissue extracts prepared by liquid-liquid extraction. The [M-H](-) ion at m/z 301 is monitored along with two transition ions at m/z 137 and 107 for DNC and the [M-H](-) ion at m/z 309 for the internal standard, d(8)-DNC. The method has been validated according to the new EU criteria for the analysis of veterinary drug residues at 100, 200 and 300 mug kg(-1) in liver and at 10, 30 and 100 mug kg(-1) in eggs. Difficulties concerning the application of the new analytical limits, namely the decision limit (CC) and the detection capability (CC) to the determination of DNC in both liver and eggs are discussed.