Comparative in vivo constituents and pharmacokinetic study in rats after oral administration of ultrafine granular powder and traditional decoction slices of Chinese Salvia

Salvia miltiorrhiza, one of the most well‐known herbal medicines, is commonly used for the treatment of coronary heart diseases in China. Besides traditional decoction slices (TDS), another relatively new product of S. miltiorrhiza, ultrafine granular powder (UGP; D90 < 45 μm), is also increasingly being used. In this paper, a UHPLC‐LTQ‐Orbitrap MS technique was developed for a metabolite profile study after oral administration of UGP and TDS of S. miltiorrhiza. The results showed that the number of in vivo absorbed compounds from UGP was much greater than that from TDS, and different types of products from S. miltiorrhiza will have different metabolic processes in vivo. Furthermore, a UHPLC‐Q‐Trap MS/MS method for simultaneously determining four tanshinones (tanshinone IIA, dihydrotanshinone I, tanshinone I and cryptotanshinone) was established and applied to assess the pharmacokinetics of the two types of products. All of the analytes displayed significant higher area under the concentration–time curve and peak concentration after oral administration of UGP than after TDS, indicating that ultrafine powder product could improve the bioavailability and absorption of cryptotanshinon,tanshinone II A,dihydrotanshinonE I and tanshinone I in vivo. The present study provides scientific information for further exploration of the pharmacology of these two types of S. miltiorrhiza and offers a reference for clinical administration of S. miltiorrhiza.     

Evaluation of pressurized liquid extraction and pressurized hot water extraction for tanshinone I and IIA in Salvia miltiorrhiza using LC and LC-ESI-MS

Pressurized liquid extraction (PLE) and pressurized hot water extraction (PHWE) using a laboratory-made system are applied for the extraction of thermally labile components such as tanshinone I and IIA in Salvia miltiorrhiza. PLE and PHWE are carried out dynamically at a flow of 1 mL/min, temperature between 95-140 degrees C, applied pressure of 10-20 bars, and extraction times of 20 and 40 min, respectively. Effects of ethanol added into the water used in PHWE are explored. PLE is found to give comparable or higher extraction efficiencies compared with PHWE with reference to Soxhlet extraction for tanshinone I and IIA in Salvia miltiorrhiza. The tanshinone I and IIA present in the various medicinal plant extracts are determined by liquid chromatography and liquid chromatography-mass spectrometry.     

Hollow Fiber/Solvent Bar Microextraction Coupled with High Performance Liquid Chromatography for Preconcentration and Determination of Tanshinones and Salvianolic Acids in Radix Salvia miltiorrhiza

Hollow fiber solvent bar microextraction coupled with high-performance liquid chromatography was developed for the preconcentration and determination of active ingredients in Radix Salvia miltiorrhiza. These ingredients include dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone IIA, salvianolic acid B, danshensu, and protocatechuic aldehyde. To evaluate the technique, seven compounds of varying polarity were used as model analytes, and a polyvinylidene fluoride hollow fiber (1.0 cm) with octanol (2 µL) was used as microextraction bar. The extraction conditions, including the identity of the hollow fiber, organic solvent, pH, salt addition, agitation speed, extraction time, and volume, were investigated and optimized. The extraction mechanism was analyzed and verified. The two main parameters, extraction recovery and enrichment factor, were obtained. Under the most favorable conditions, the enrichment factors of the analytes were 0.7–612, the limits of detection were below 1.11 ng mL^?1, and the recoveries were between 95.4% and 101.3%. Thus, hollow fiber solvent bar microextraction is simple, rapid, and practical with a wide range of potential applications.     

On‐line ionic liquid‐based dynamic microwave‐assisted extraction‐high performance liquid chromatography for the determination of lipophilic constituents in root of Salvia miltiorrhiza Bunge

On‐line continuous sampling, ionic liquid‐based dynamic microwave‐assisted extraction high performance liquid chromatography has been developed and applied to the extraction of lipophilic constituents from root of Salvia miltiorrhiza Bunge. Several operating parameters were optimized by single‐factor and Box–Behnken design experiments. The type and concentration of ionic liquids, power of microwave irradiation, flow rate of sample suspension, amount, and particle size of sample were investigated. The limits of detection for tanshin‐one I, cryptotanshinone, and tanshinone II_A are 0.014, 0.009, and 0.009 mg/g, respectively. The RSDs of interday and intraday were lower than 2.02 and 2.16%, respectively. The recoveries for target analytes were in the range of 90.7–101.8%. The homogeneity of the suspension and stability of the analytes were investigated and the results were satisfactory. The proposed method was compared with the off‐line ionic liquid‐based dynamic microwave‐assisted extraction, off‐line ethanol‐based dynamic microwave‐assisted extraction, ionic liquid‐based ultrasonic‐assisted extraction, and ionic liquid‐based maceration extraction. The results indicated that the proposed method is effective for the extraction of the active components in Chinese herbal medicine and has some advantages over the other methods.     

SPE of Tanshinones from Salvia miltiorrhiza Bunge by using Imprinted Functionalized Ionic Liquid-Modified Silica

Ionic liquid-modified silica, with functional groups based on imidazole as the cation, was obtained. A molecular imprinting technique was introduced to form the order of functional groups. The selectivity of the obtained ionic liquid-modified silica was successfully used as a special imprinted sorbent in the solid-phase extraction to isolate cryptotanshinone, tanshinone I and tanshinone IIA from Salvia miltiorrhiza Bunge. Several washing and elution solvents with different polarities were evaluated. The ionic liquid-modified silica as the sorbent exhibited a higher selectivity than blank ionic liquid-modified silica, traditional silica and C₁₈ cartridges. A quantitative analysis was conducted by liquid chromatography with a C₁₈ column and methanol/water (75:25, v/v, containing 0.5% acetic acid) as the mobile phase. A good linearity was obtained from 0.5 × 10^?4 to 0.1 mg mL^?1 (r ²  > 0.99) with relative standard deviations that were less than 4.6%.     

Direct purification of tanshinones from Salvia miltiorrhiza Bunge by high‐speed counter‐current chromatography without presaturation of the two‐phase solvent mixture

Tanshen, the rhizome of Salvia miltiorrhiza Bunge, is a famous Traditional Chinese Medicine for multiple therapeutic remedies. This work presents the isolation and purification of tanshinone I and tanshinone IIA from the extract of the rhizome of S. miltiorrhiza by using high‐speed counter‐current chromatography (CCC) without presaturation of the two‐phase solvent mixture. The CCC method combines the results of CCC solvent system selection and components analyses of solvent mixture by GC, and thus it is possible to add accurately each individual solvent to prepare single saturated solvent phase without presaturation. The optimum CCC solvent system is a system of hexane–ethyl acetate–ethanol–water (8:2:7:3, v/v), which has been determined by usual solvent system selection and CCC runs. As a result, over 98% pure tanshinone IIA and over 94% pure tanshinone I have been obtained by using less solvent volume. Their structures have been identified by ESI‐MS, NMR spectra.     

Pharmacokinetic comparison of five tanshinones in normal and arthritic rats after oral administration of Huo Luo Xiao Ling Dan or its single herb extract by UPLC‐MS/MS

A fast, sensitive and reliable ultra performance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS) method has been developed and validated for simultaneous quantitation and pharmacokinetic study of five tanshinones (tanshinone I, tanshinone IIA, tanshinone IIB, dihydrotanshinone I, cryptotanshinone), the bio‐active ingredients of Huo Luo Xiao Ling Dan (HLXLD) in rat plasma. After liquid–liquid extraction, chromatographic separation was accomplished on a Shim‐pack XR‐ODS column (75 × 3.0 mm, 2.2 µm particles) and eluted with a mobile phase consisting of acetonitrile–0.05% formic acid aqueous solution (80:20, v/v) at a flow rate of 0.4 mL/min, and the total run time was 7.0 min. The detection was performed on a triple quadrupole tandem mass spectrometry equipped with an electrospray ionization source in positive ionization and multiple reaction monitoring mode. The lower limits of quantification were 0.050–0.400 ng/mL for all the analytes. Linearity, precision and accuracy, the mean extraction recoveries and matrix effects all satisfied criteria for acceptance. This validated method was successfully applied to a comparative pharmacokinetic study of five bio‐active components in rat plasma after oral administration of HLXLD or Salvia miltiorrhiza extract in normal and arthritic rats. The results showed that there were different pharmacokinetic characteristics among different groups. Copyright © 2016 John Wiley & Sons, Ltd.     

Efficient separation of tanshinones by polyvinylpyrrolidone‐stabilized graphene‐modified micellar electrokinetic chromatography

In this work, a PVP‐stabilized graphene was used in MEKC for the separation of tanshinones. Seven structurally similar tanshinones were studied, that is, tanshinone IIB, dihydrotanshinone I, tanshinone I, cryptotanshinone, 1,2‐dihydrotanshinone I, miltirone, and tanshinone IIA. To achieve optimal conditions, graphene concentration, sample solvent composition, SDS concentration, 2‐propanolconcentration, and buffer pH were investigated. At a separation voltage of 30 kV and a 41.5 cm effective length fused‐silica capillary, good resolution within 12 min was performed using 10 mM borate buffer (pH 9.3) containing 30 mM SDS, 10% v/v 2‐propanol and 6 μg/mL graphene. The method was validated in terms of linearity (r² > 0.9970), intra‐ and inter‐day precision were less than 3.56 and 4.83%, respectively. The proposed method was then successfully applied to Danshentong capsule, an herbal preparation from Salvia miltiorrhiza. Our results indicated the high separation efficiency of PVP‐stabilized graphene provided new opportunities for the analysis of complex samples.     

Identification of Salvia species using high‐performance liquid chromatography combined with chemical pattern recognition analysis

Salvia miltiorrhiza, also known as Danshen, is a widely used traditional Chinese medicine for the treatment of cardiovascular diseases and hematological abnormalities. The root and rhizome of Salvia przewalskii and Salvia yunnanensis have been found as substitutes for Salvia miltiorrhiza in the market. In this study, the chemical information of 14 major compounds in Salvia miltiorrhiza and its substitutes were determined using a high‐performance liquid chromatography method. Stepwise discriminant analysis was adopted to select the characteristic variables. Partial least squares discriminant and hierarchical cluster analysis were performed to classify Salvia miltiorrhiza and its substitutes. The results showed that all of the samples were correctly classified both in partial least squares discriminant analysis and hierarchical cluster analysis based on the four compounds (caffeic acid, rosmarinic acid, salvianolic acid B, and salvianolic acid A). This method can not only distinguish Salvia miltiorrhiza and its substitutes, but also classify Salvia przewalskii and Salvia yunnanensis. The method can be applied for the quality assessment of Salvia miltiorrhiza and identification of unknown samples.     

Determination of Three Tanshinones from Radix Salvia Miltiorrhiza by Molecularly Imprinted Solid-phase Extraction

A selective molecularly imprinted solid‐phase extraction procedure was developed for the selective separation of tanshinone I, tanshinone IIA, and cryptotanshinone from Radix Salvia Miltiorrhiza samples. Tanshinone IIA imprinted polymers (MIP) synthesized in ethanol‐dodecanol system show high affinity to tanshinone IIA and its structure analogs in aqueous environment and the affinity can be controlled by adjusting the intensity of the eluents. By using 60% water‐40% methanol (volume ratio) and 99.5% methanol‐0.5% trifluoroacetic acid (volume ratio) as washing and eluting solvents, most interferences originating from the salvia matrix were eliminated. The extracts were sufficiently clean enough to be directly injected into HPLC for further chromatographic analysis. Good linearity was obtained from 0.4 to 500.0 mg·L^?1 (r²=0.999) with the relative standard deviations less than 4.2%. The mean recoveries of tanshinone IIA in Radix Salvia Miltiorrhiza were more than 85.6% at three different concentrations and the limits of detection were 0.06–0.09 mg·L^?1. This method is a viable alternative tool to the existing HPLC methods for analyzing the content of the three tanshinones in Radix Salvia Miltiorrhiza samples.     

Ultrasonication-assisted extraction and preconcentration of medicinal products from herb by ionic liquids

Ionic liquid-based extraction of medicinal or useful compounds from plants was investigated as an alternative to supercritical fluid, cloud point and conventional organic solvent extractions. The method integrated extraction and preconcentration. Medicinal products were first extracted by an ionic liquid solution, part of which was then converted to a hydrophobic form by anion metathesis for preconcentration. The remaining soluble ionic liquid acted as a dispersive agent to enhance the efficiency of preconcentration. Protein in the extract was precipitated spontaneously without addition of further solvents. Ultrasonication assisted this method for extraction and preconcentration of cryptotanshinone, tanshinone I and tanshinone II A from Salvia Miltiorrhiza Bunge. 0.233 mg g⁻¹, 0.695 mg g⁻¹ and 0.682 mg g⁻¹ of each, respectively, were extracted using [OMIM][Cl], and preconcentrated in a [OMIM][PF₆] phase at respective concentrations of 148.1, 507.1 and 486.1 μg mL⁻¹. The method exhibited potential applicability with other medicinal products.     

The in vivo absorbed constituents and metabolites of Danshen decoction in rats identified by HPLC with electrospray ionization tandem ion trap and time‐of‐flight mass spectrometry

Danshen, the dried root and rhizome of Salvia miltiorrhiza Bunge, is widely used for the treatment of cardiovascular and cerebrovascular diseases. This research focuses on the in vivo metabolism of Danshen decoction (DSD) in rats. After oral administration of DSD, the absorptive constituents and their metabolites in urine and plasma were analyzed by HPLC coupled with a photodiode array detector and electrospray ionization hybrid ion trap and time‐of‐flight mass spectrometry. Samples were separated on a C₁₈ column by gradient elution using 0.1% (v/v) aqueous formic acid and acetonitrile. As a result, 93 compounds from urine and 38 compounds from plasma were identified. Among them, lipo‐soluble diterpenoids (24 in urine and 15 in plasma) were reported for the first time as in vivo metabolites of DSD. According to the quantities and contents of the identified compounds, tanshinone IIA, cryptotanshinone and tanshinone I were deduced to be the major absorptive diterpenoids of DSD. Moreover, nine water‐soluble phenolics (caffeic acid, ferulic acid, danshensu, etc.) were proved to be the major absorptive constituents as reported. Most of the absorbed constituents underwent sulfation, glucuronidation, hydrogenation and hydroxylation in vivo. This investigation provided scientific evidence to obtain a more comprehensive metabolic profile of DSD. Copyright © 2014 John Wiley & Sons, Ltd.     

Thermal characterization of dried extract of medicinal plant by DSC and analytical techniques

The increased search for herbal products has generated an increasing interest in improving the quality control of dried extracts by pharmaceutical industry since these are raw materials of great importance by their quality and versatility. This work aimed at the application of various analytical techniques (thermal analysis, X-ray diffraction, scanning electron microscopy, and infrared Fourier transform spectroscopy) in the characterization of dried extracts of two plants from the Brazilian semiarid region with medicinal properties. The DSC curves for the dried extracts of Ximenia americana L. and Schinopsis brasiliensis Engl. showed that thermal processes occur between 33.50 and 118.58 °C and between 39.17 and 126.14 °C. The X-ray powder diffraction revealed high degree of amorphization, but the dried extract of X. americana L. showed some diffraction peaks of high intensity. The IR spectra showed high variety of metabolites in the extracts dried. Through this study it was possible to verify the feasibility of applying these techniques in the characterization of raw materials from medicinal plants for use in the herbal medicines production.     

Characterization of Phenolics in Lavandula angustifolia

ABSTRACT

The lavender flowers and their essential oil are widely used in therapy in Romania and the European Community. Since the European Pharmacopoeia only allows the use of Lavandula sp flowers for medicinal purposes, the objective of this study was to investigate the chemical composition of Lavandula angustifolia extracts obtained by ultrasound-assisted extraction, rapid pressurized extraction at 6.7?bar, and subcritical fluid extraction. The solvents used for the first two methods were mixtures of water and alcohol, glycerin, and propylene glycol. These extracts were analyzed by high-performance thin-layer chromatography, infrared spectroscopy with attenuated total reflectance, and Raman spectroscopy. The total phenolics were evaluated using a modified Folin–Ciocalteu method. The primary phenolics were chlorogenic acid, gallic acid, umbelliferone, luteolin 7-O-glucoside, vitexin, and isoquercitroside. The extracts were variable in composition, with the highest yield by subcritical fluid extraction, followed by extraction at 6.7?bar. The infrared and Raman spectroscopy results confirmed the chromatography measurements.     

Evaluation of Multiplexed CE with UV Detection for Rapid pK a Estimation of Active Pharmaceutical Ingredients

The acid dissociation constant (pK _a) is a key physicochemical parameter for characterizing active pharmaceutical ingredients (APIs). Early determination of pK _a values is highly desirable in drug discovery, pharmaceutical process research and formulation design. To overcome the challenges of limited sample availability and potential low purity of API samples at early stages of drug development, as well as to increase sample analysis throughput, a multiplexed 96-channel capillary electrophoresis with UV detection was evaluated as a practical approach for high throughput pK _a estimation of proprietary APIs in support of pharmaceutical research. Proprietary APIs with diverse structures were examined using the approach. The pK _a values were successfully determined with good accuracy and precision. System robustness was demonstrated and analysis of at least eight samples can be completed within 1 h. A rapid pK _a estimation procedure for marginally soluble APIs was proposed by performing single-point multiplexed CE–UV measurement without extrapolation using 10 or 20% methanol as co-solvent. Direct pK _a estimation of APIs using DMSO solution samples and crude reaction samples containing a large amount of solvents and reagents and high level of impurities was also demonstrated using the multiplexed CE–UV approach.     

Comparison of the chromatographic fingerprint,multicomponent quantitation and antioxidant activity of Salvia miltiorrhiza Bge. between sweating and nonsweating

Salvia miltiorrhiza Bge. is a traditional Chinese medicine applied in the treatment of various diseases in clinical practice. In the course of its processing, S. miltiorrhiza Bge. is usually processed by sweating. This study employed 10‐component contents determination coupled with high‐performance liquid chromatography (HPLC) fingerprint and antioxidant activity to investigate the effect of sweating on S. miltiorrhiza Bge. so as to evaluate the quality of S. miltiorrhiza Bge. The HPLC method was performed using C₁₈ and 0.05% phosphoric acid aqueous solution–acetonitrile with a gradient elution system. It was validated for linearity, precision, repeatability, stability and recovery. Similarity analysis, principal components analysis and antioxidant activity assays were used to compare sweated S. miltiorrhiza Bge. (SSM) and nonsweated S. miltiorrhiza Bge. (NSSM). SSM and NSSM showed good similarities in HPLC fingerprint (>0.9), but principal components analysis could classify the HPLC fingerprint and 10‐component quantitation analysis. Meanwhile, the antioxidant activity of SSM was significantly higher than that of NSSM (p < 0.01). The results of this study indicated that sweating could alter the content of chemical constituents in S. miltiorrhiza Bge., and could also improve its antioxidant activity. In addition, the method not only affords a viable strategy for comparing SSM and NSSM and assessing the quality of S. miltiorrhiza Bge., but also provides a reference for other herbal medicine that suffers from sweating.     

Identification of chemical ingredients of peanut stems and leaves extracts using UPLC‐QTOF‐MS coupled with novel informatics UNIFI platform

Peanut stems and leaves have been used traditionally as both herbal medicines and special food in Asia. In this study, the main functional compounds of peanut stems and leaves extracts were identified using UPLC separation coupled to high resolution mass spectrometry (QTOF‐MS), and a traditional medicine library. Three different extraction solvents (ethyl acetate, petroleum ether and n‐butanol) were evaluated to prepare the extracts of peanut stems and leaves. A total of 283 chemical compounds were identified in peanut stems and leaves extracts, of which 207 compounds are tentatively new identifications in Genus Arachis. The integration of data acquisition and processing with the traditional medicine library provides a simple, efficient process to effectively facilitate the identification of chemical ingredients in complex natural product extracts. The integrated workflow for separation, detection and identification of functional compounds in natural products using UPLC/QTOF‐MS greatly improves productivity for development of traditional herbal medicines. Copyright © 2016 John Wiley & Sons, Ltd.     

微粉化技术提高水不溶性药物溶解度

药物的微粉化可以改善颗粒的润湿性,进而提高水不溶性药物的溶解度和溶解速率。目前普遍采用的药物微粒化技术主要包括机械研磨、超临界流体过程、低温喷淋和溶剂蒸发沉积过程。本文介绍了这些微粉化制备技术的基本原理以及该类技术的应用进展。     

加速溶剂萃取技术在中药有效成分分析中的应用

以两种药材为研究实例,对加速溶剂萃取法（ASE）在中药材有效成分提取研究中的应用进行了简要介绍。采用正交试验法考察了提取丹参中丹酚酸B的提取条件（萃取温度、静态萃取时间、萃取溶剂以及料液比）,确定了较好的实验条件。比较了ASE、水蒸气蒸馏法、超声波提取法及索氏提取法对木香挥发油的提取效果,结果表明ASE对木香挥发油的提取效果最好。