•Development of a sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC–MS/MS) method.•Application of UFLC–MS/MS for the determination of RA, sal D, LA and sal B in SAFI in rat plasma.•The first pharmacokinetic study of salvianolic acid for injection (SAFI).•Investigation of dose-dependent pharmacokinetics of SAFI.A simple, sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC–MS/MS) method was established for simultaneous determination and pharmacokinetic study of rosmarinic acid (RA), salvianolic acid D (Sal D), lithospermic acid (LA) and salvianolic acid B (Sal B) in rat plasma after intravenous administration of salvianolic acid for injection (SAFI). Three doses of administration, containing 14, 28 and 56 mg/kg, were investigated in this study. Plasma samples were pretreated using protein precipitation (PP) with pre-cooled acetonitrile. Chromatographic separation was achieved on a CORTECS™ UPLC C18 column (1.6 μm, 2.1 × 100 mm) with a mobile phase composed of 0.1% formic acid aqueous (V/V) and 0.1% formic acid acetonitrile (V/V). Analytes were detected using electrospray ionization (ESI) source in negative ionization mode and quantified in multiple reaction monitoring (MRM) mode. The validated method is stable and reliable. No significant difference of half lives (t1/2) of four analytes at three doses was observed. Area under the curve (AUC0-∞) and peak concentration (Cmax) of the four analytes demonstrated a linear increase in across the doses with the linear correlation r of each analyte at three doses were greater than 0.95. It indicated that the pharmacokinetic behavior of SAFI is positively related to dose at the range of 14–56 mg/kg.

The present study was designed to establish a multi-wavelength quantitative fingerprinting method for San-Huang Tablets (SHT), a widely used and commercially available herbal preparation, where high performance liquid chromatography (HPLC) with a diode array detector (DAD) was employed to obtain the fingerprint profiles. A simple linear quantitative fingerprint method (SLQFM) coupled with multi-ingredient simultaneous determination was developed to evaluate the quality consistency of the tested samples qualitatively and quantitatively. Additionally, the component–activity relationship between chromatographic fingerprints and total radical-scavenging capacity in vitro (as assessed using the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay) was investigated by partial least squares regression (PLSR) analysis to predict the antioxidant capacity of new samples from the chromatographic fingerprints and identify the main active constituents that can be used as the target markers for the quality control of SHT. In conclusion, the strategy developed in the present study was effective and reliable, which can be employed for holistic evaluation and accurate discrimination for the quality consistency of SHT preparations and other traditional Chinese medicine (TCM) and herbal preparations as well.

Separation of active components from Traditional Chinese medicine (TCM) is always difficult due to its complex matrix. The prep-HPLC-based multidimensional chromatography, which combined the characteristics of different separation techniques to improve separation capability and efficiency, is more conductive for separating and purifying complex TCMs. In this paper, we reviewed the basic principles, separation mode, key techniques of prep-HPLC-based multidimensional chromatography and its application in TCM research.In a preparative HPLC-based multidimensional chromatography system, crude drug or crude isolates that prepared by column chromatography or other technical is purified on the 1D-LC. Then a further purification is performed on the 2D-LC for those unseparated target component by off-line or on-line mode.

•Determination of two novel analogs of salvianolic acid B in rat plasma by UFLC-MS/MS.•Only 3.0 min was in need for one analytical run.•Application to the study of pharmacokinetic behaviors of the two novel compounds.•This study benefits further researches in the process of drug development.7′(Z)-(8″S, 8‴S)-epi-Salvianolic acid E (compound 1) and (7′R, 8′R, 8″S, 8‴S)-epi-salvianolic acid B (compound 2), two novel analogs of salvianolic acid B (Sal B), have been recently isolated from Salvianolic acid for injection. They both show powerful antioxidant effects, including inducing NQO1 activity and scavenging DPPH free radical, and potential protecting effects for cerebral ischemia. However, no reports have been described the pharmacokinetic study of them. In this study, an ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed and validated for the determination of compound 1, compound 2 and Sal B in rat plasma, respectively. Plasma samples were pretreated by liquid–liquid extraction with ethyl acetate. Chromatographic separation was achieved on a Waters Acquity UPLC® HSS T3 column (1.7 μm particles, 2.1 mm i.d. × 100 mm) with the mobile phase of 0.1% aqueous formic acid (A)-acetonitrile (B) (65:35, v/v). Quantification was performed on a triple quadruple tandem mass spectrometry with electrospray ionization (ESI) by multiple reaction monitoring (MRM) in the negative ion mode. Monitored transitions were set at m/z 717.0 → 519.0, 717.1 → 519.1, 717.2 → 518.9 and 320.9 → 152.1 for compound 1, compound 2, Sal B and chloramphenicol (internal standard, IS), respectively. Linear calibration curves were acquired over the concentration range of 2.0–1000 ng/mL for the three analytes in rat plasma. The extraction recoveries, matrix effects, intra- and inter-day precisions and accuracies of the three analytes were all within acceptable limits. The validated method was successfully applied to the pharmacokinetic study of compound 1, compound 2 and Sal B after intravenous administration of 6.0 mg/kg in rats, respectively. The results indicated that compound 1 and compound 2 were both eliminated more slowly than Sal B. Exposure levels of both compound 1 and Sal B were higher than compound 2 in the same dosage range. This study provided critical reference for the pharmacokinetic study of compound 1 and compound 2.

A high performance liquid chromatography (HPLC) method with diode array detector (DAD) was developed and validated for multi-wavelength fingerprint profiling to evaluate the quality consistency of a Traditional Chinese Medicine, Fangfeng Pill. The validation results demonstrate that the optimized HPLC method achieves the desired linearity, precision, accuracy, LOQ and LOD. The multi-wavelength fingerprints were developed and qualitatively and quantitatively evaluated using the Ratio Fingerprint Quantification Method (RFQM). The fingerprint analysis of the marker compounds (baicalin, wogonin, glycyrrhizic acid, baicalein, paeoniflorin, gardenoside, and ephedrine) was found to reflect the content of the marker compounds in the Fangfeng Pill samples determined using the external standard method. The classical Fenton reaction was performed to assess the antioxidant activity of the Fangfeng Pill samples in vitro, which was correlated to the fingerprint components. This study demonstrates that multi-wavelength fingerprint profiling correlated the antioxidant activity offering a reliable and efficient approach to quantitatively evaluate the quality consistency of the TCM and herbal preparations.

•DMRM is introduced into multiple components quantitation for DFF for the first time.•The issue of the comprehensive and quantitative control for the quality of DFF is solved.•A standard operating procedure for building a new DMRM method is recommended.It is a challenging task to simultaneously and quantitatively analyze multiple components in DFF [Da-Fu-Fang, namely, complex traditional Chinese medicine (TCM) preparations containing more than ten TCMs] due to their numerous and extreme complex chemical compositions possessing a wide variety of chemical and physical features, and their very low content. Rather than using a conventional mass spectrometry (MS) method with multiple reaction monitoring (MRM), in the current study, this challenge was addressed by using dynamic multiple reaction monitoring (DMRM). Using a DFF, Niuhuang Shangqing pill, which is composed of 19 TCMs, as a model, a rapid (one run in 20 min), sensitive [lower limit of detection (LOD) and limit of quantitation (LOQ) were achieved comparable with MRM] and accessible (a standard HPLC/MS/MS instrumentation was employed) MS method was successfully developed for the simultaneous quantification of 41 bioactive components which represented 15 of the 19 medicinal plants. A comparison of LOD and LOQ using MRM and DMRM was made to quantitatively reveal that the latter demonstrated advantages over the former. Meanwhile, a standard operating procedure concerning the development of a new DMRM method was recommended. The MS data were obtained in the positive ion mode with electrospray ionization as the ion source, acetonitrile and water as mobile phase and a Kinetex C18 core–shell column (100 mm × 2.10 mm, 2.6 μm, Phenomenex Inc.) as the analytical column. This method was then applied to 32 batches of samples. It transpired, through principal component analysis and orthogonal partial least squares discriminant analysis, that the consistency of the products was relatively good within one company, but poor among different companies among the 32 samples; one failed to qualify in terms of the Chinese Pharmacopeia. This work illustrated that the proposed DMRM method was particularly suitable for quantifying the trace components in DFF and capable of ensuring the quality of DFF.

A high performance liquid chromatography (HPLC) method with diode array detector (DAD) was developed and validated for multi-wavelength fingerprint profiling to evaluate the quality consistency of a Traditional Chinese Medicine, Fangfeng Pill. The validation results demonstrate that the optimized HPLC method achieves the desired linearity, precision, accuracy, LOQ and LOD. The multi-wavelength fingerprints were developed and qualitatively and quantitatively evaluated using the Ratio Fingerprint Quantification Method (RFQM). The fingerprint analysis of the marker compounds (baicalin, wogonin, glycyrrhizic acid, baicalein, paeoniflorin, gardenoside, and ephedrine) was found to reflect the content of the marker compounds in the Fangfeng Pill samples determined using the external standard method. The classical Fenton reaction was performed to assess the antioxidant activity of the Fangfeng Pill samples in vitro, which was correlated to the fingerprint components. This study demonstrates that multi-wavelength fingerprint profiling correlated the antioxidant activity offering a reliable and efficient approach to quantitatively evaluate the quality consistency of the TCM and herbal preparations.