柳酚咖敏片含量及有关物质的UFLC测定

目的建立UFLC法测定柳酚咖敏片中盐酸去氧肾上腺素、对乙酰氨基酚、无水咖啡因、水杨酰胺和马来酸溴苯那敏的含量及有关物质。方法采用Kinetex C18色谱柱,以0.5%三乙胺溶液(磷酸调节pH至3.0)和甲醇为流动相,梯度洗脱,流速为1mL.min-1。结果盐酸去氧肾上腺素、马来酸溴苯那敏、咖啡因对乙酰氨基酚和水杨酰胺分别在0.10~10.33,0.05~4.82,0.04~4.10,0.30~30.50和0.30~30.44μg范围内线性关系良好。各组分平均回收率为99.7%~100.7%,RSD为0.24%~0.67%(n=9)。结论该方法简单、快速、准确,可用于柳酚咖敏片的含量及有关物质检测。     

An ultra-fast LC method for the determination of iodiconazole in microdialysis samples and its application in the calibration of laboratory-made linear probes

Iodiconazole is a very potent antifungal agent used to treat serious fungal infections. After transdermal administration, several factors affect the exposure of iodiconazole, resulting in large variability and demanding further elucidation of drug distribution. For determination of iodiconazole in dermal microdialysate, a new, efficient, reliable and robust ultra-fast liquid chromatography (UFLC™, Shimadzu) assay using UV detection at 230 nm has been developed and validated. Iodiconazole was separated on a Shimadzu Prominence UFLC™ C18 column (2.2 μm, 50 mm × 2.0 mm i.d.) using acetonitrile–0.025% triethylamine solution, adjusted to pH 3.6 with phosphoric acid (65:35, v/v), at a flow rate of 0.5 ml/min. The retention time was 1.37 min for iodiconazole and 1.78 min for the internal standard, an isomeric compound of iodiconazole. Intra- and inter-day precision ranged from 5.3% to 7.8% and 3.7% to 8.4%, respectively. The UFLC method was used to measure iodiconazole concentrations in microdialysis samples obtained during the calibration of laboratory-made linear probes. The validation and sample analysis results show that the method is precise, accurate and well suited to support the dermal microdialysis experiments.     

Investigation of 6-O-methyl-scutellarein metabolites in rats by ultra-flow liquid chromatography/quadrupole-time-of-flight mass spectrometry

Context Scutellarin (1) has been widely used in China to treat acute cerebral infarction and paralysis induced by cerebrovascular diseases. However, scutellarin (1) has unstable metabolic characteristics.

Objective The metabolic profile of 6-O-scutellarein was studied to determine its metabolic stability in vivo.

Materials and methods In this study, a method of UFLC/Q-TOF MS was used to study the 6-O-methyl-scutellarein metabolites in rat plasma, urine, bile and faeces after oral administration of 6-O-methyl-scutellarein (3). One hour after oral administration of 6-O-methyl-scutellarein (3) (34?mg/kg), approximately 1?mL blood samples were collected in EP tubes from all groups. Bile, urine and faeces samples were collected from eight SD rats during 0–24?h after oral administration. The mass defect filtering, dynamic background subtraction and information dependent acquisition techniques were also used to identify the 6-O-methyl-scutellarein metabolites.

Results The parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces. The glucuronide conjugate of 6-O-methyl-scutellarein (M1, M2), diglucuronide conjugate of 6-O-methyl-scutellarein (M3), sulphate conjugate of 6-O-methyl-scutellarein (M4), glucuronide and sulphate conjugate of 6-O-methyl-scutellarein (M5), methylated conjugate of 6-O-methyl-scutellarein (M6) were detected in rat urine. M1, M2 and M3 were detected in rat bile. M1 was found in rat plasma and M7 was detected in faeces.

Discussion and conclusion Because the parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces, we speculate that 6-O-methyl-scutellarein (3) had good metabolic stability in vivo. This warrants further study to develop it as a promising candidate for the treatment of ischemic cerebrovascular disease.     

RP-UFLC-DAD法同时测定有柄石韦中7个咖啡酰奎宁酸类成分和3个黄酮类成分的含量

在本研究中,建立了高效、灵敏、准确的超快速液相色谱法同时快速分析测定有柄石韦中10个指标成分（新绿原酸、绿原酸、隐绿原酸、1-咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、紫云英苷、圣草酚-7-O-β-D-吡喃葡萄糖醛酸苷、山奈酚-3,7-双-O-葡萄糖苷）的含量。采用了Kromasil 100-2.5C18（100 mm×2.1 mm,2.5μm）C18快速色谱柱,以0.1%甲酸水和0.1%甲酸甲醇溶液为洗脱剂洗脱,检测波长为326 nm,流速为0.4 mL/min。10个成分在此色谱条件下能达到基线分离,标准曲线线性关系良好（r 2〉0.9998）,日内与日间精密度、重复性、稳定性均符合实验要求,加样回收率范围为99.1%–104.5%。对20批不同来源的有柄石韦样品进行了含量测定。此研究为有柄石韦中多成分含量测定提供了高效、灵敏、准确的快速分析方法。此外,还对20份样品进行了聚类分析和主成份分析,结果显示此方法可以很好地区分20份样品的来源及资源分布情况。     

四季抗病毒合剂制备中连翘酯苷A和连翘酯苷B转移率的研究

目的建立连翘与四季抗病毒合剂中连翘酯苷A和连翘酯苷B的含量测定方法,阐明其在连翘药材至四季抗病毒合剂制剂间的转移率。方法采用超快速液相色谱(UFLC)法,选取3批连翘药材,每批对应5批制剂。结果 1 3批连翘药材中连翘酯苷A的含量分别为12.57,20.26和18.64mg·g~(-1),3批连翘药材中连翘酯苷B的含量依次分别为2.22,3.76和3.07mg·g~(-1);2每批连翘药材对应5批制剂中连翘酯苷A的平均含量分别为1 129.40,1 828.30和1 666.30μg·mL~(-1),每批连翘药材对应5批制剂中连翘酯苷B的平均含量依次分别为224.80,382.60和315.70μg·mL~(-1);3 15批制剂中连翘酯苷A和连翘酯苷B的转移率分别为74.86%和84.95%,RSD分别为0.88%和1.43%。结论该含量测定方法具有简便、稳定、可靠、准确等优点,可作为连翘与四季抗病毒合剂中连翘酯苷A和连翘酯苷B的定量方法;连翘酯苷A和连翘酯苷B的转移率高,表明制剂制备工艺有利于2种有效成分的溶出,为保证制剂疗效提供了理论依据。     

四季抗病毒合剂的超快速液相色谱法指纹图谱研究及4个成分的含量测定

目的:建立四季抗病毒合剂的超快速液相色谱(UFLC)法指纹图谱,并测定连翘酯苷A、连翘苷、甘草苷、甘草酸的含量。方法:采用UFLC法,色谱条件:Kromasil C₁₈柱(250 mm×4.6 mm,5 μm);流动相为乙腈-0.6%磷酸水溶液,梯度洗脱;流速为0.8 ml·min^-1;检测波长为260 nm;柱温为30℃。结果:通过中药色谱指纹图谱相似度评价系统(2004A),确定了四季抗病毒合剂的UFLC指纹图谱共有模式,标定了42个峰;并测定了12批制剂中连翘酯苷A、甘草苷、连翘苷、甘草酸的含量(mg·ml^-1)分别为1.85,0.06,0.07和0.25 mg·ml^-1,RSD分别为1.10%,1.21%,0.98%和0.84%。结论:该方法简便、准确、重复性好,为四季抗病毒合剂的质量评价与质量控制提供了科学依据。     

UPLC、RRLC及UFLC在中药质量控制中的研究进展

文章综述了近几年来超高效液相色谱(UPLC)、高分离度快速液相色谱(RRLC)和超快速液相色谱(UFLC)在中药成分定性鉴别、含量测定、指纹图谱及中药限量成分或违禁添加成分检测中的研究进展。     

Identification of metabolites of meisoindigo in rat, pig and human liver microsomes by UFLC-MS/MS

3-(1,2-Dihydro-2-oxo-3H-indol-3-ylidene)-1,3-dihydro-1-methyl-2H-indol-2-one, abbreviated as meisoindigo, has been a routine therapeutic agent in the clinical treatment of chronic myelogenous leukemia in China since the 1980s. To gain an understanding of the interspecies differences in the metabolism of meisoindigo, the relevant metabolism studies were carried out for the first time in rat, pig and human liver microsomes of different genders by ultra fast liquid chromatography/tandem mass spectrometry (UFLC-MS/MS). The qualitative metabolite identification was accomplished by multiple reaction monitoring (MRM) in combination with Enhanced Product Ion (EPI). The semi-quantitative metabolic stability and metabolite formation were simultaneously measured by MRM. The in vitro metabolic pathways of meisoindigo in three species were proposed as 3,3′ double bond reduction, followed by N-demethylation, and reduction followed by phenyl mono-oxidation. Two novel metabolic pathways involving direct phenyl mono-oxidation without reduction in the three species, and direct N-demethylation without reduction in only pig and human, were also proposed. It may be noted that the two metabolites formed after reduction followed by phenyl mono-oxidation at positions 4, 5, 6 or 7, as well as one metabolite formed from direct phenyl mono-oxidation at either of the two phenyl rings without reduction were found to be uniquely present in human. The in vitro t_1/2 and in vitro CL_int values of meisoindigo were calculated. Statistical analysis showed there were no significant differences in the metabolic stability profiles of meisoindigo among three species, and gender effect on the metabolic stability of meisoindigo was negligible. Formation profiles of the most significant reductive metabolites were obtained in the three species.     

UFLC法同时测定参芎葡萄糖注射液中6种主要成分

目的建立超快速液相色谱(UFLC)同时测定参芎葡萄糖注射液(丹参、盐酸川芎嗪)中丹参素、盐酸川芎嗪、原儿茶醛、迷迭香酸、丹酚酸B、丹酚酸A的方法。方法采用Diamonsil C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.05%磷酸为流动相进行梯度洗脱,体积流量为1 mL/min,检测波长220 nm,柱温40℃。结果 6种被测成分的线性关系良好(r>0.999),精密度、重复性、稳定性的RSD都低于3%,平均加样回收率在95.19%～101.02%,RSD在0.92%～2.70%(n=6)。结论本方法简便、快速、准确,能有效控制参芎葡萄糖注射液的质量。