长柱十大功劳中药根碱、巴马汀和小檗碱的高效液相色谱法同时测定

建立了高效液相色谱法同时测定长柱十大功劳中药根碱、巴马汀、小檗碱含量的方法.样品用盐酸-甲醇(体积比1:100)混合溶液超声提取,C18柱色谱分离,以乙腈-0.05 mol/L磷酸二氢钾缓冲液(磷酸调Ph值至3.0)(体积比28:72)为流动相,检测波长:265 nm.方法线性良好,相关系数均大于0.999,盐酸药根碱、盐酸巴马汀、盐酸小檗碱的加标回收率及相对标准偏差分别为99%,1.43%; 100%,1.93%; 99%,2.82%.     

胶束薄层扫描法测定黄连及其制剂中的小檗碱、巴马汀和药根碱

以2％CPC-醋酸乙酯(9：1)胶束溶液为展开剂,在聚酰胺薄膜上成功地分离了小檗碱、巴马汀和药根碱。以345nm为测定波长,550nm为参比波长进行扫描测定,建立了一种新的同时测定黄连及其制剂中小檗碱、巴马汀和药根碱的胶束薄层扫描法。小檗碱、巴马汀和药根碱的线性范围分别为0．2—2．4、0．1—1．0和0．1—1．0μg;回收率分别为97．7％-99．4％、101．7％～102．6％和96．7％-97．8％;相对标准偏差分别为1．4％-1．7％、1．9％-2．7％和1．8％-2．2％。     

黄连生物碱的反相高效液相-电喷雾离子阱串联质谱的研究

采用反相高效液相-电喷雾离子阱串联质谱法对由乙醇提取的黄连生物碱进行了研究.优化出了反相高效液相色谱分离黄连生物碱的条件:流动相为V(乙腈):V(H2O)(三乙胺2 mmol/L)=30:70;柱温为30℃;流速为0.5 mL/min,并结合电喷雾串联质谱检测出了黄连生物碱中的小檗碱、药根碱、巴马汀、黄连碱以及微量的表...     

毛细管电泳安培法检测黄连中的生物碱

本文建立了一种简单快速的毛细管电泳安培检测法分离检测黄连中的黄连碱、盐酸小檗碱、巴马汀、和药根碱.以150 μm的铂电极为工作电极,考察并优化了影响分离和检测的条件.在80 mmol/L磷酸盐缓冲液中添加50%甲醇(pH 6.0),分离电压15 kV,检测电位1.2 V (vs.Ag/AgCl)的条件下,黄连碱、盐酸小檗碱、巴马汀和药根碱在8min内获得良好分离.黄连碱、盐酸小檗碱、巴马汀和药根碱的峰电流面积和浓度分别在1.0×10-5～2.0×10-7,1.0×10-5 ～8.0×10-8 mol/L,1.0×10-5～1.0×10-7 mol/L和1.0×10-5～2.0×10-7mol/L范围内呈良好的线性关系.检出限(S/N=3)低达10-8mmol/L.方法应用于微波辅助溶剂提取黄连中生物碱的测定,回收率在97.0%～104%,RSDs≤3.8%,结果满意.     

离子液体作为流动相添加剂高效液相色谱法同时测定黄连中5种生物碱

建立了高效液相色谱法(HPLC)同时测定黄连中5种生物碱含量的方法。黄连药材经甲醇超声提取后,用Spherigel C_(18)色谱柱(250×4.6 mm,5μm)进行HPLC测定,流动相为含有1-己基-3-甲基咪唑四氟硼酸盐添加剂的甲醇-水(25:75,V/V),流动相流速为1.0 m L/min,检测波长为345 nm,同时测定了黄连药材中药根碱、表小檗碱、黄连碱、巴马汀和小檗碱含量。在1~200μg/m L浓度范围内,5种生物碱的线性相关系数均大于0.9990,药根碱、表小檗碱、黄连碱、巴马汀和小檗碱的检测限(LOD)分别为0.19,0.13,0.11,0.18,0.15 mg/L。测定了3种不同产地的黄连生物碱的含量并进行加标回收,回收率在98%~102%之间。     

高效液相色谱法测定香连丸及组方药材中5种活性组分的含量

建立了香连丸及组方药材中药根碱、巴马汀、小檗碱、木香烃内酯和去氢木香内酯的含量测定方法．采用反相高效液相色谱,Hypersil ODS（5μm,4．6mm×250mm）色谱柱,流动相为乙腈：水（70：30,V：V）,流速1．0mL／min,柱温30℃,检测波长为210n／n测定木香烃内酯和去氢木香内酯的含量;采用高效液相胶束色谱,Kromasil ODS（5la,m,4．6mm×150mm）色谱柱,流动相为0．2mol／LNaH2P04水溶液：7．00mmol／L十二烷基硫酸钠：乙腈（35：35：30,V：V：V）,流速1．0mL／min,柱温30℃,检测波长为350nm测定药根碱、巴马汀、小檗碱的含量．结果显示5种活性成分在适当的线性范围内均具有良好的线性关系（r大于0．9993）,平均回收率在87．67％～102．37％之间,RSD小于3．13％;结果表明方法简便、灵敏、准确,重现性好．     

反相高效液相色谱法同时测定黄连、左金丸及反左金丸中三种生物碱的含量

用反相高效液相色谱(RP-HPLC)法,以CH3CN-0.1%,H3PO4的0.5%SDS溶液(50:50)为流动相体系,有效分离了黄连、左金丸和反左金丸样品中的小檗碱型生物碱组分.该方法线性关系良好,相关系数均在0.9999以上,回收率高.同时测定了黄连、左金丸(黄连-吴茱萸6:1)及反左金丸(黄连-吴茱萸1:6)中药根碱、巴马汀以及小檗碱等3种生物碱的含量,并比较了其间存在的差别.本方法分离度高,准确、简便、快捷.     

SiO2@松香基高分子(Ru131)色谱柱分离测定岩黄连中脱氢卡维丁、盐酸巴马汀和盐酸小檗碱

建立测定岩黄连中脱氢卡维丁、盐酸巴马汀、盐酸小檗碱含量的液相色谱分析方法.确定了脱氢卡维丁、盐酸巴马汀和盐酸小檗碱的色谱分离条件为:SiO2@松香基高分子(Ru131)色谱柱(250 mm×4.6mm,5μm),以乙腈-0.0050 mol·L-1磷酸盐缓冲溶液(pH=3.0)=20∶80(V/V)为流动相,紫外检测波...     

甘肃产三颗针植物中生物碱的测定及分布状态的研究

建立梯度洗脱双波长HPLC法同时测定小檗属植物中小檗胺、药根碱、巴马汀、小檗碱含量的方法,并应用该法分析采集于甘肃不同产地、不同品种小檗属植物的茎木、根木、茎皮、根皮样品,研究小檗属植物中生物碱的分布状态,考察其药用价值。小檗胺、药根碱、巴马汀、小檗碱线性范围分别为：0．015～2．56μg(r=0．9998);0．012～2．0μg(r=0．9996);0．010～0．52μg(r=0．9999);0．028～4．74μg(r=0．9998)。结果表明,甘肃不同产地、不同品种,不同部位三颗针植物中生物碱的含量分布有明显的差异。     

基于表面静电排斥/反相混合模式色谱的黄连生物碱分析方法

建立了基于表面静电排斥/反相混合模式色谱的黄连生物碱分析方法。选用实验室自制的C18HCE柱,以乙腈和水为流动相,考察了甲酸、乙酸两种流动相添加剂及其在流动相中的体积分数对黄连生物碱的保留、峰形及选择性的影响。最终确定0.1%（v/v）乙酸作为添加剂能实现黄连生物碱的良好分离,结合质谱和文献对其主要色谱峰进行了识别,分别为黄连碱、表小檗碱、非洲防己碱、药根碱、小檗碱、巴马汀。参考2015版《中国药典》对黄连生物碱的含量测定方法,以盐酸小檗碱进行方法学考察,结果表明,在0.5~100 mg/L范围内线性关系良好,相关系数为0.9996,平均加标回收率为93.74%。利用所建立的方法测定了湖北和重庆两个产地不同批次的黄连样品中各生物碱的含量。该方法简便,重复性好,精密度高,可为其他碱性化合物的分离分析提供参考。     

Analysis of protoberberine alkaloids in several herbal drugs and related medicinal preparations by non-aqueous capillary electrophoresis

A simple, rapid, reproducible, and universal non-aqueous capillary electrophoresis method has been developed for the separation and determination of three major active protoberberine alkaloids including berberine, palmatine, and jatrorrhizine within 7 min. The effects of the concentrations of acetic acid and electrolyte, the ratio of organic solvent, and the applied voltage on the separation were investigated. The optimum running buffer was composed of 50 mM ammonium acetate, 0.5% (v/v) acetic acid, and 10% (v/v) acetonitrile in methanol. The applied voltage was 18 kV. The analytes were detected by UV at 214 nm. The linearities between peak areas and the concentrations of the analytes were also investigated, and they exhibit excellent linear behavior over the concentration ranges (correlation coefficients: 0.9975-0.9986). The method was successfully applied to determine the three alkaloids in several families of herbal drugs (Rhizoma Coptidis, Cortex Berberidis, Cortex Phellodendri, Herba Chelidonii, Caulis Mahoniae) and their relevant medicinal preparations for the first time, and the recoveries of the three constituents ranged between 95.6-103.2% for berberine, 97.5-103.3% for palmatine, and 96.1 -103.6% for jatrorrhizine.     

强阳离子交换毛细管液相色谱-反相加压毛细管电色谱二维系统的构建及其在中药黄柏提取物分离中的应用

加压毛细管电色谱(pCEC)具有电泳和液相色谱的双重分离机理,其柱效高、选择性强、分辨率高和分离速度快并可进行梯度洗脱。我们在此基础上加入离子交换色谱模式,构建了强阳离子交换-反相加压毛细管液相色谱(micro strong cation exchange liquid chromatography/reversed phase pressurized capillary electrochromatography, μ-SCXLC/RP-pCEC)二维系统,并对中药黄柏的提取物进行了优化分离。第一维μ-SCXLC采用线性盐梯度分离,样品被切割成11个馏分洗脱收集后进入第二维,第二维脱盐后,采用RP-pCEC进行分离分析,梯度洗脱。以中药黄柏提取物为样品,此二维系统的分辨率和峰容量都较一维系统有很大提高,理论峰容量可达900左右,证明构建的二维体系非常适合复杂样品的分离分析。     

Chemical fingerprint analysis of Phellodendri Amurensis Cortex by ultra performance LC/Q-TOF-MS methods combined with chemometrics

Ultra performance LC with quadrupole TOF MS (UPLC/Q‐TOF‐MS) fingerprinting is first developed for the identification of the major components of Phellodendri Amurensis Cortex (PAC). The PAC samples are separated using a Waters ACQUITY UPLC BEH C18 (2.1×50 mm, 1.7 μm) by linear gradient elution using water (containing 0.2% formic acid) and acetonitrile (containing 0.2% formic acid) as the mobile phase. Ten batches of PAC are selected to construct the UPLC/Q‐TOF‐MS fingerprint. Sixteen common peaks in the fingerprint are obtained, ten of which are tentatively identified, with reference to the literature data, as phellodendrine, magnoflorine, tetrahydropjatrorrhizine, menisperine, tetrahydropalmatine, jatrorrhizine, palmatine, berberine, obacunone, and limonin. Chemometric methods are also employed to evaluate the variation of herbal drugs and other closely related herbs based on the characteristics of peaks in the UPLC/Q‐TOF‐MS profiles. The developed fingerprint assay is a powerful method that may be used to conduct quality control of PAC.     

Analysis of berberine and total alkaloid content in cortex phellodendri by near infrared spectroscopy (NIRS) compared with high-performance liquid chromatography coupled with ultra-visible spectrometric detection

This paper developed a rapid method using near infrared spectroscopy (NIRS) to differentiate two species of Cortex Phellodendri (CP), Cortex Phellodendri Chinensis (PCS) and Cortex Phellodendri Amurensis (PAR), and to predict quantitatively the content of berberine and total alkaloid content in all Cortex Phellodendri samples. Three alkaloids, berberine, jatrorrhizine and palmatine were analyzed simultaneously with a Thermo ODS Hypersil column by gradient elution with a new mobile phase under high-performance liquid chromatography-diode array detection (HPLC-DAD). Berberine content determined by HPLC-DAD was exploited as a critical parameter for successful discrimination between them. Multiplicative scatter correction (MSC), second derivative and Savitsky-Golay (S.G.) were utilized together to correct the scattering effect and eliminate the baseline shift in all near infrared diffuse reflectance spectra as well as to enhance spectral features in order to give a better correlation with the results obtained by HPLC-DAD. With the use of principal component analysis (PCA), samples datasets were separated successfully into two different clusters corresponding to two species. Furthermore, a partial least squares (PLS) regression method was built on the correlation model. The results showed that the correlation coefficients of the prediction models were R = 0.996 for the berberine and R = 0.994 for total alkaloid content. The influences of water absorption bands present in the NIR spectra on the models were also investigated in order to explore the practicability of NIRS in routine use. The outcome showed that NIRS possibly acts as routine screening in the quality control of Chinese herbal medicine.     

乙肝解毒胶囊中5种有效成分的胶束电动色谱-质谱联用法测定

建立了胶束电动色谱-电喷雾电离质谱联用法同时测定乙肝解毒胶囊中的小檗碱、巴马汀、药根碱、黄芩苷、大黄素5种药效成分含量的分析方法。使用未涂层石英毛细管,以30 mmol/L月桂酸-70mmol/L氨水溶液(含20%乙腈,pH10.0)为缓冲液、70%异丙醇(含3 mmol/L乙酸)为鞘液。结果表明,在18 min内各组分达到基线分离,小檗碱、巴马汀、药根碱、黄芩苷、大黄素的线性范围分别为0.02~20、0.05~15、0.05~10、2.0~500、0.02~15 mg/L,检出限分别为0.007、0.02、0.02、0.60、0.006 mg/L。样品的加标回收率为95%~104%,相对标准偏差均小于3.9%。方法简便、快速、灵敏,已成功用于乙肝解毒胶囊中小檗碱、巴马汀、药根碱、黄芩苷、大黄素含量的同时测定。     

移液枪头式固相微萃取-高效液相色谱法测定细胞培养液中的4种生物碱

建立了移液枪头式固相微萃取(SPME)-高效液相色谱检测细胞培养液中4种生物碱(黄柏碱、药根碱、巴马丁和小檗碱)的分析方法。以甲基丙烯酸为反应单体、乙二醇二甲基丙烯酸酯为交联剂、偶氮二异丁腈为引发剂,在移液枪头内进行原位聚合反应,制备含有弱阳离子交换整体固定相的SPME枪头。样品经SPME净化后,用高效液相色谱-紫外检测法进行分析,流动相为乙腈-磷酸二氢钾溶液(0.05 mol/L,pH 4),紫外检测波长为270 nm,外标法定量。4种生物碱的检出限(S/N=3)为0.16~0.39μg/L;以空白细胞培养液进行加标回收率试验,加标回收率为92.73%~97.91%,相对标准偏差(RSD)为0.14%~3.31%。该方法简单、灵敏、准确,能够用于细胞培养液中微量生物碱的分析研究。     

非水毛细管电泳测定黄连饮片中5种生物碱

建立了一种非水毛细管电泳(NACE)同时测定黄连饮片生品与炮制品中小檗碱、巴马汀、药根碱、木兰碱和黄连碱含量的方法。分别考察了非水溶剂、缓冲液体系及其浓度和pH、运行电压、运行温度和检测波长等条件对实验结果的影响。在优化的实验条件下,选择非水毛细管电泳分离模式,以40 mmol/L乙酸钠-40 mmol/L乙酸铵的无水甲醇缓冲溶液(pH 5.8)为电泳介质,未涂渍标准熔融石英毛细管(64.5 cm×75 μm,有效长度56 cm)为分离通道,检测波长为254 nm,分离电压为25 kV,压力进样(5 kPa×6 s),柱温为20 ℃。结果显示,5种生物碱在20 min内可实现基线分离,加标回收率为98.37%～101.03%。该方法简单、准确,重现性较好,可用于黄连饮片内在质量的评价和控制。     

Novel molecularly imprinted magnetic nanoparticles for the selective extraction of protoberberine alkaloids in herbs and rat plasma

In this work, a novel magnetic nanomaterial functionalized with a molecularly imprinted polymer was prepared for the extraction of protoberberine alkaloids. Molecularly imprinted polymers were made on the surface of Fe₃O₄ nanoparticles by using berberine as template, acetonitrile/water as porogen, acrylamide as functional monomer and ethylene glycol dimethacrylate as cross‐linker. The optimized molar ratio of template/functional monomer was 1:7. The polymeric magnetic nanoparticles were characterized by transmission electron microscopy and Fourier transform infrared spectroscopy. The stability and adsorption capacity of the molecularly imprinted polymers were investigated. The molecularly imprinted polymers were used as a selective sorbent for the magnetic molecularly imprinted solid‐phase extraction and determination of jatrorrhizine, palmatine, and berberine. Extraction parameters were studied including loading pH, sample volume, stirring speed, and extraction time. Finally, a magnetic molecularly imprinted solid‐phase extraction coupled to high‐performance liquid chromatography method was developed. Under the optimized conditions, the method showed good linear range of 0.1–150 ng/mL for berberine and 0.1–100 ng/mL for jatrorrhizine and palmatine. The limit of detection was 0.01 ng/mL for berberine and 0.02 ng/mL for jatrorrhizine and palmatine. The proposed method has been applied to determine protoberberine alkaloids in Cortex phellodendri and rat plasma samples. The recoveries ranged from 87.33–102.43%, with relative standard deviation less than 4.54% in Cortex phellodendri and from 102.22–111.15% with relative standard deviation less than 4.59% in plasma.     

Selective and sensitive determination of protoberberines by capillary electrophoresis coupled with molecularly imprinted microextraction

In this work, we developed a novel molecularly imprinted solid‐phase microextraction with capillary electrophoresis method for the selective extraction and determination of protoberberines in complicated samples. The imprinted monolith was prepared in a micropipette tip‐based device by using acrylamide as the functional monomer, ethyleneglyoldimethacrylate as the cross‐linker and dimethylsulfoxide as the porogen, and exhibited an imprinting factor of 2.41 to berberine, 2.36 to palmatine and 2.38 to jatrorrhizine. Good capillary electrophoresis separation was achieved by using 20 mM phosphate buffer at pH 7 as running buffer with the addition of organic modifier of 10% methanol. Parameters such as sample pH value, sample flow rate and sample volume were investigated for imprinted monolith‐based solid‐phase microextraction. An imprinted solid‐phase microextraction with capillary electrophoresis method was developed, the method showed a wide linear range (0.3–50 μg/mL), good linearity (R² ≥ 0.9947) and good reproducibility (relative standard deviations ≤ 0.73%), the limit of detection was as low as 0.1 μg/mL, which was lower than some reported methods based on capillary electrophoresis for protoberberines. The method has been applied for determination of three common protoberberines in Cortex Phellodendri Chinensis, by using a molecularly imprinted monolith as the selective sorbent, most of the matrices in the Cortex Phellodendri Chinensis sample were removed and three protoberberines were selectively enriched and well determined.     

多肽的反相梯度加压毛细管电色谱分离

以C18为固定相，采用电压和压力联合驱动流动相，研究反相加压毛细管电色谱分离多肽；考察了加压电色谱中，电压对带电和中性物质迁移的影响，实现了梯度加压毛细管电色谱分离6种多肽；结果表明，加压电色谱可以很好地抑制气泡形成，实验结果准确，重复性好；梯度加压毛细管电色谱在复杂样品的分离分析中，具有很大的潜力。