Pressurized hot water extraction of berberine, baicalein and glycyrrhizin in medicinal plants

Pressurized hot water extraction (PHWE) using a laboratory made system was applied for the extraction of thermally labile and reasonably polar components such as berberine in coptidis rhizoma, glycyrrhizin in radix glycyrrhizae/liquorice and baicalein in scutellariae radix. PHWE was carried out dynamically at a flow of 1 ml/min, temperature between 95 and 140 °C, an applied pressure of 10-20 bar and extraction time of 40 min. Extraction by PHWE was found to give efficiencies comparable to Soxhlet extraction for baicalein in scutellariae radix and sonication for berberine in coptidis rhizoma, and glycyrrhizin in radix glycyrrhizae. Effects of ethanol added into the water used in PHWE were explored. Pressurized liquid extraction (PLE) with methanol as solvent was used for extraction of baicalein in scutellariae radix. The marker compounds present in the various medicinal plant extracts were determined by gradient elution HPLC.     

Poly(methacrylic acid-ethylene glycol dimethacrylate) monolith in-tube solid-phase microextraction applied to simultaneous analysis of some amphetamine derivatives in urine by capillary zone electrophoresis

A method based on in-tube solid-phase microextraction and capillary zone electrophoresis (CZE) was proposed for simultaneously determining four amphetamines (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, and 3,4-methylenedioxymethamphetamine) in urine. A poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column, which can provide sufficient extraction efficiency, was introduced for the extraction of amphetamines from urine samples. The hydrophobic main chains and acidic pendant groups of the monolithic column make it a superior material for extraction of basic analytes from aqueous matrix. After extraction, the samples were analyzed by CZE. The best separation was achieved using a buffer composed of 0.1 M disodium hydrogen phosphate (adjusted to pH 4.5 with 1 M hydrochloric acid) and 20% methanol v/v, with a temperature and voltage of 25 degrees C and 20 kV, respectively. By applying electrokinetic injection with field-amplified sample stacking, detection limits of 25-34 microg/L were achieved. Excellent method of reproducibility was found over a linear range of 0.1-5 mg/L. Determination of these analytes from abusers' urine sample was also demonstrated.     

原子发射光谱法间接测定黄连素片剂中盐酸小檗碱的研究

本文提出了离子对-原子发射光谱间接测定黄连素片剂中盐酸小檗碱的新方法。发现用1,2-二氯乙烷作萃取剂效果最佳。用此怯测定实际样品的结果与药典法一致,相对误差为0.61％,单一测定的相对标准偏差为0.67％。本法还可在盐酸普鲁卡因共存下单独测定盐酸小檗碱。方法可靠、适用,选择性较好。     

Preparation of a sulfonated fused-silica capillary and its application in capillary electrophoresis and electrochromatography

In the present paper, two new methods, sol-gel and chemical bonding methods, were proposed for preparation of sulfonated fused-silica capillaries. In the sol-gel method, a fused-silica capillary was coated with the sol solution obtained by hydrolysis of 3-mercaptopropyltrimethoxysilane (MPTS) and tetramethoxysilane, and followed by age; while in the chemical bonding method, a capillary was chemically bonded directly with MPTS. Then, both the resulting capillaries were oxidized with an aqueous solution of hydrogen peroxide solution (H2O2) (30%, m/m) to obtain the sulfonated capillaries. The electroosmotic flow (EOF) for the sulfonated capillaries was found to remain almost constant within the studied pH range, and greater than that of the uncoated capillary. However, the coating efficiency of the capillary prepared by chemical bonding method was higher than that by sol-gel method, by comparing their magnitude of the EOF, the degree of disguise of the silanol and reproducibility of preparation procedure. The effects of the electrolyte's concentration and the content of methanol (MeOH) on the EOF were also studied. Especially, the study of the apparent pH (pH*) on the EOF in a water-MeOH system was reported. Finally, capillary electrophoretic separation of seven organic acids was achieved within 6.5 min under optimal condition using the chemically bonded sulfonated capillary. Moreover, separation of four alkaloids on the sulfonated capillary was compared with that on uncoated capillary in different conditions. Ion-exchange mechanism was found to play a key role for separation of these four basic analytes on the sulfonated capillary.     

Spectroscopic and Binding Properties of Berberine to DNA and Its Application to DNA Detection

Berberine(BER) binds to the double helical DNA with a high affinity There is only a much smaller hypochromism and no shifts in the absorption spectra when BER binds to calf thymus DNA(CT DNA) The fluorescence yields increase dramatically when BER binds to DNA, with no shifts in the emission maximum. These spectral changes are in contrast to the behavior observed with many fluorescent intercalates Groove binding rather than intercalation was suggested to be the cause of these spectral changes. Consistent with groove binding, for polyamide anion quenching studies showed that the magnitude of K_sv of the bound BER was higher than that of the free BER. The addition of salt to the solution releases the DNA-bound drug action from the groove and causes a decrease in the fluorescence yield. The results of all above studies proved the groove binding of BER to DNA. The large fluorescence enhancements observed when BER binds to DNA and the poor fluorescence yield of BER in the absence of DNA can be used for sensitive detection of DNA The linear concentration range was 0–20μg/ml The limit of detection for CT DNA was 12 ng/ml     

Inclusion Modes of Berberine with β-Cyclodextrin in Aqueous Solution

The possible inclusion modes of berberine(Berb)with β-cyclodextrin(BCD)in aqueous solution were predicted by molecular docking,molecular dynamics(MD)simulation and binding free energy calculations.Firs...     

Isolation of berberine tannate using a chromatography activity-guided fractionation from root bark of Berberis hispanica Boiss. & Reut.

Root bark of Berberis hispanica Boiss. & Reut. constitutes an important source of alkaloids and is traditionally used in Algeria and Morocco in many herbal formulations particularly for the treatment of stomach infections and colon cancer. Activity-guided fractionation of Berberis extract is developed by step-gradient elution on column chromatography followed by preparative HPLC to isolate the most antistaphylococcal compound. Solvent extraction-activity screening indicates that ethanol should be used as the extracting solvent as it has exhibited the highest activity against Staphylococcus aureus(Rosenbach ATCC6538) followed by water extract, whereas no activity is recorded for acetone and hexane extracts. Screening of collected fractions from chromatographic separations indicates that the fraction presenting the highest antistaphylococcal activity is identified as berberine tannate by UV spectrum, melting point measurement and confirmed by liquid chromatography coupled to mass spectrometry (LC/MS). The minimal inhibition concentration value of berberine tannate against S. aureus is evaluated at 5?µg?mL^?1.     

The auxiliary determination of the binding site of berberine binding to human serum albumin by surface-enhanced Raman scattering

We report on the joint application of fluorescence, ultraviolet-visible (UV-Vis) and Raman spectroscopy to the study of berberine with human serum albumin (HSA). We propose the surface-enhanced Raman scattering (SERS) technique to improve the understanding of the quenching interaction caused by berberine which could be applied in recognition process of fluorescent drugs with large biomolecules. The fluorescence and UV-Vis spectroscopic results show that the fluorescence intensity of HSA is significantly decreased in the presence of berberine, and the quenching mechanism is static. The SERS technique demonstrates clear advantages over direct measurements in physiological conditions. By means of this method, we are able to deduce important information concerning the binding property of berberine when interacting with HSA. We show the nitrogen atom is free but the dioxolane is involved in the spontaneously electrostatic inducement and subsequently hydrophobic binding.     

微孔板发光二极管诱导荧光-表面活性剂增敏法测定盐酸小檗碱

研制了微孔板发光二极管诱导荧光分析仪,以发光二极管为激发光源,Y形光纤束传导激发光和荧光,采用平面型光敏二极管接收荧光.采用二维定位器移动微孔板,可实现对多个样品快速连续的测定.利用弱碱性介质中阴离子表面活性剂十二烷基苯磺酸钠(SDBS)对盐酸小檗碱荧光的增敏作用,在pH 9.0的Britton-Robinson缓冲溶...     

HPD-100树脂吸附小檗碱的热力学及动力学研究

研究了小檗碱在HPD-100树脂上的吸附热力学和动力学行为。考察了小檗碱在HPD-100树脂上的吸附等温曲线和吸附动力学曲线,确定了吸附过程的热力学及动力学参数。结果表明,HPD-100树脂对小檗碱的等温吸附可采用Langmuir方程描述、吸附为自发进行的放热型物理吸附过程;Dunwald–Wagnen内扩散方程能较好地拟合吸附动力学数据,吸附过程以颗粒内扩散为主要控制步骤,该吸附过程的表观活化能为29.11kJ/mol。本文为HPD-100树脂分离富集小檗碱类生物碱的实际应用提供基础数据及指导。