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A rapid, sensitive and convenient method based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the simultaneous quantification of calycosin-7-O-β-d-glucoside (CCSG), ononin, calycosin, (6aR,11aR)-9,10-dimethoxypterocarpan-3-O-β-d-glucopyanoside (DPPG), and 7,2′-dihydroxy-3′,4′-dimethoxyisoflavan-7-O-β-d-glucopyanoside (DIFG) in rat plasma after oral administration of the methanol extraction from Radix Astragali. Theophylline played the role of internal standard (IS). Preparation of plasma samples by liquid-liquid extraction method with ethyl acetate after precipitation of protein with methanol. The analytes were detected with a triple quadrupole tandem mass spectrometery (MS) in multiple reaction monitoring (MRM) mode and a positive ion electrospray ionization (ESI). The method was validated with the concentration ranges of 1.96–62.69 ng/mL for CCSG, 1.70–54.5 ng/mL for ononin, 1.85–59.06 ng/mL for calycosin, 2.14–137.24 ng/mL for DPPG and1.96–125.25 ng/mL for DIFG, respectively. The method had the lower limit of quantification (LLOQ) with 0.49, 0.21, 0.92, 1.07, and 0.98 ng/mL for CCSG, ononin, calycosin, DPPG and DIFG respectively, and the precision less than 10%. The RSD of the accuracy was in the range of −4.35–8.91%. The results may be helpful to provide more accurate references to clinical application of this herb.

A sensitive and rapid liquid chromatography with tandem mass spectrometry method has been developed for simultaneous determination of berberine (I), jateorhizine (II), palmatine (III), tetrahydropalmatine (IV), phellodendrine (V), protopine (VI) and columbamine (VII) in rat plasma after oral administration of Phellodendri chinensis cortex extraction. The plasmas were extracted by liquid-liquid extraction. The tandem mass spectrometric detection was performed in the multiple reaction monitoring mode in the positive ionization. The intra- and interday precisions and accuracies were in range from -12.18 to 13.21%. Mean absolute recoveries of all analytes and internal standard were between 78.6 and 98.9%. The seven alkaloids were proven to be stable during sample storage and analysis procedures. The established method was validated and successfully applied to pharmacokinetics study in rat plasma after oral administration of Phellodendri chinensis cortex extract. The t1/2 of palmatine, columbamine, pellodendrine, berberine, tetrahydropalmaine, jatrorrhizine, and protopine were 5.16, 5.96, 7.18, 19.84, 6.28, 7.08, 6.90 h, respectively. The seven compounds could be rapidly absorbed into blood (time for maximal concentration, 1.80-1.93 h). This study could establish a foundation for further research of Phellodendri chinensis cortex and might provide more useful information to guide the clinical usage.

Pharmacokinetic studies were conducted on rats for protopine, corynoline, 7'-(3',4'-dihydroxyphenyl)-N-[(4-methoxyphenyl)-ethyl]propenamide, acetylcorynoline, and 8-oxocorynoline, five main active components from Corydalis bungeana Turcz (C. bungeana Turcz). An ultra high performance liquid chromatography with tandem mass spectrometry method has been developed and validated for the simultaneous determination of the pharmacokinetic characteristics of these components in rat plasma. Chromatographic separation was achieved on an Agilent SB-C18 column (1.8 μm, 150 × 2.1 mm) using a gradient elution program with a mobile phase consisting of acetonitrile and water containing 0.1% acetic acid at a flow rate of 0.3 mL/min. The analytes were detected in the multiple reaction monitoring mode with positive electrospray ionization. Lower limits of quantification were >0.680 ng/mL and matrix effects ranged from 91.26 to 100.38%. The mean extraction recoveries of quality control samples were less than 79.32%, and the precision and accuracy were within the acceptable limits. All analytes were proven to be stable during sample storage and analysis procedures. The validated method was successfully applied to the pharmacokinetic study of five alkaloid components after oral administration of C. bungeana Turcz extract to rats. The obtained results may be helpful to reveal the mechanism of action and to guide the clinical application of C. bungeana Turcz.

A rapid, simple and sensitive ultra-performance liquid chromatography-electrospray-ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) method was developed and validated for the simultaneous determination of aesculin, aesculetin, fraxetin, fraxin and polydatin in beagle dog plasma for the first time. Plasma samples were pretreated by protein precipitation with methanol. Chromatographic separation was performed on an Acquity UPLC HSS T3 C18 column (2.1 mm &times, 100 mm, 1.8 &mu, m) with gradient elution at a flow rate of 0.4 mL/min, using a mobile phase consisting of 0.1% formic acid (A) and acetonitrile (B). The analytes and IS were detected by multiple reaction monitoring (MRM) via negative ion mode with ion transitions of m/z 339.1&ndash, m/z 176.8 for aesculin, m/z 176.8&ndash, m/z 88.9 for aesculetin, m/z 206.8&ndash, m/z 192.1 for fraxetin, m/z 369.1&ndash, m/z 206.9 for fraxin, m/z 389.1&ndash, m/z 227.0 for polydatin and m/z 415.2&ndash, m/z 295.1 for puerarin. This method was validated according to the FDA guidelines and the results met the requirements of analysis. The calibration curves of analytes were linear with correlation coefficients more than 0.9980. The intra- and inter-day precisions were less than 15% and the accuracy was within &plusmn, 15%. The maximum plasma concentration (Cmax) of aesculin, aesculetin, fraxetin, fraxin and polydatin was 46.75 &plusmn, 7.46, 209.9 &plusmn, 57.65, 369.7 &plusmn, 48.87, 67.04 &plusmn, 12.09 and 47.14 &plusmn, 12.04 ng/mL, respectively. The time to reach the maximum plasma concentration (Tmax) was 1.32 &plusmn, 0.38 h for aesculin, 1.03 &plusmn, 0.27 h for aesculetin, 0.94 &plusmn, 0.23 h for fraxetin, 0.83 &plusmn, 0.18 h for fraxin and 1.15 &plusmn, 0.15 h for polydatin. The results indicated that the absorption of aesculin might be slow in beagle dog plasma. This method was successfully applied for pharmacokinetics in beagle dog plasma after oral administration of the extracts of Ledum palustre L. at a dosage of 0.27 g/kg.

A rapid and sensitive Ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the simultaneous determination of fourteen alkaloids in beagle dog plasma after a single oral dose of the Corydalis bungeana Turcz (C. bungeana) extract selected bifendate as the internal standard (IS). The plasma samples were preprocessed by liquid-liquid extraction (LLE) with aether before separation on an Agilent SB-C18 column (1.8 &micro, m, 150 &times, 2.1 mm) using a gradient elution program. The mobile phase consists of 0.2% acetic acid and acetonitrile at the flow rate of 0.3 mL/min. In the positive ion mode, the analytes were detected by multiple reaction monitoring (MRM). The results indicated that calibration curves for fourteen analytes have good linearity (R2 = 0.9904). The lower limits of quantification (LLOQ) of fourteen alkaloids and IS were all over 4.87 ng/mL and the matrix effects ranged from 94.08% to 102.76%. The mean extraction recoveries of Quality control samples at low (LQC), medium (MQC) and high (HQC) and IS were all more than 78.03%. The intra- and inter-day precision (R.S.D.%) also met the criterion, at the same time the deviation of assay accuracies (R.E) ranged from &minus, 13.70% to 14.40%. The Tmax values of fourteen alkaloids were no more than 1 h. The range of Cmax was from 74.16 &plusmn, 8.71 to 2256 &plusmn, 255.9 ng/mL. The assay was validated in the light of the regulatory bioanalytical guidelines and proved acceptable, which was successfully applied to a pharmacokinetic study of these compounds in beagle dogs after oral administration of Corydalis bungeana Turcz extract.

This study has developed a sensitive and simple ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry method for the simultaneous determination of corydaline, dehydrocorydaline, tetrahydropalmatine, protopine, palmatine, tetrahydroberberine, columbamine, berberine, coptisine and berberrubine in beagle dog plasma after the oral administration of the Corydalis yanhusuo W.T. Wang and Yuanhuzhitong tablets. Chromatographic separation was achieved on an Agilent Eclipse Plus C18 RRHD column (1.8 &micro, m, 50 &times, 2.1 mm) using a gradient elution program with a mobile phase consisting of acetonitrile and water containing 0.1% formic acid at a flow rate of 0.3 mL/min. A tandem mass spectrometric detection was conducted by multiple reaction monitoring (MRM) mode via an electrospray ionization source in the positive mode. The calibration curves of all analytes showed good linear (r2 &gt, 0.9800). The intra-day and inter-day precisions were less than 15% and the accuracies were within &plusmn, 15%. The extraction recoveries conformed to the acceptable range. And there was no interference of endogenous substances in the sensitive assay method. All analytes were proven to be stable during sample storage and analysis procedures. The pharmacokinetic study indicated that the Yuanhuzhitong tablets could get a better absorption than Corydalis yanhusuo W.T. Wang.

A simple and high sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of peimine and peiminine in beagle dog plasma after the oral administration of Fritillariae ussuriensis Maxim and Fritillariae thunbergii Miq powder. Chromatographic separation was achieved on an ACQUIT UPLC&reg, BEH C18 column (1.7 &mu, m, 2.1 &times, 100 mm) in a gradient elution way with a mobile phase consisting of acetonitrile and water containing 0.1% formic acid at a flow rate of 0.4 mL/min. The plasma samples were prepared by a liquid&ndash, liquid extraction (LLE) method with ethyl acetate. The analytes were detected with a triple quadrupole tandem mass spectrometry (MS) in multiple reaction monitoring (MRM) mode and a positive ion electrospray ionization (ESI) of the transitions at m/z 432.4&rarr, 414.4 for peimine and m/z 430.3&rarr, 412.3 for peiminine. The method was linear for two analytes over the investigated range with all determined correlation coefficients exceeding 0.9900. The lower limit of quantification (LLOQ) was 0.988 ng/mL for peimine and 0.980 ng/mL for peiminine. The mean extraction recoveries of peimine and peiminine at three quality control samples (QC) levels were ranged from 82.56 to 88.71%, and matrix effects ranged from 92.06 to 101.2%. The intra-day and inter-day precision and accuracy were within the acceptable limits at LLOQ and QC levels. The method was effectively and successfully applied to the pharmacokinetics of peimine and peiminine after oral administration of powder to beagle dogs. The obtained results may be help to guide the clinical application of Fritillaria ussuriensis Maxim and Fritillaria thunbergii Miq.

A rapid and sensitive Ultra High Performance Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (UHPLC-ESI-MS/MS) method was developed and validated to simultaneously determine the concentration of seven phenolic acids (syringic acid, ferulic acid, caffeic acid, vanillic acid, p-coumaric acid, 3,4-dihydroxybenzoic acid and 4-hydroxybenzoic acid) in rat plasma after oral administration of Echinacea purpurea extract. After mixing with the internal standard (IS), butylparaben, plasma samples were prepared by liquid–liquid extraction with ethyl acetate. The separation was performed using the Agilent Eclipse Plus C18 column (1.8 μm, 2.1 mm × 50 mm) with a gradient system consisting of solution A (0.1% acetic acid in water) and solution B (methanol) at a flow rate of 0.3 mL/min. The detection was accomplished by a multiple reaction monitoring (MRM) mode with electrospray ionization (ESI). The method was validated in terms of linearity, precision, accuracy, extraction recovery, matrix effect and stability. This method was successfully applied to study the pharmacokinetic properties of the seven compounds after oral administration of Echinacea purpurea extract in rats.

Aralia elata (Miq.) Seems (A. elata) grow in Northeast China and the total saponins of A. elata is used to auxiliary treatment for the acute hepatitis, chronic hepatitis and the transaminase on the high side. Aralia-saponinV and Aralia-saponinVI are the major bioactive saponins in A. elata leaves. A selective and sensitive UHPLC-MS/MS method was developed and validated for the determination and pharmacokinetic study of Aralia-saponinV and Aralia-saponinVI indwelling the extract in rat plasma in this article. The sample pretreatment involved a one-step extraction of 0.2mL plasma with methanol. Shengmaxinside C was used as internal standard (I.S.). The separation was carried out on an Agilent SB-C18 column (1.8μm, 50mm×2.1mm) at 30°C with a mobile phase of acetonitrile-5mM ammonium acetate (90:10, v/v) at a flow rate of 0.2mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source operating in the negative ionization mode. The optimized mass transition ion-pairs (m/z) monitored for Aralia-saponinV, Aralia-saponinVI and I.S. were 1103.2/941.2, 1119.2/957.0 and 707.0/647.1, respectively. The current UHPLC-MS/MS assay method was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability, and it was suitable for the pharmacokinetic studies of the two saponins after oral administration of extract of A. elata leaves. The lower limits of quantification were 5.70ng/mL for Aralia-saponinV and 6.15ng/mL for Aralia-saponinVI. Intra-day and inter-day precisions were less than 7.4% and the accuracy range was from 1.19% to 8.60%. The mean extraction recoveries of analytes and I.S. from rat plasma were all more than 89.5%. This paper described a simple, sensitive and validated UHPLC-MS/MS method for simultaneous determination of Aralia-saponinV and Aralia-saponinVI in rat plasma after oral administration of the extract of A. elata leaves, and investigated on their pharmacokinetic studies as well.

Rosmarinus officinalis L. is commonly used as a spice and flavoring agent. Diterpenes are the main active compounds of R. officinalis. An Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-ESI-MS/MS) method was developed for the determination of carnosol, rosmanol, and carnosic acid isolated from R. officinalis in rat plasma, and applied to a pharmacokinetic study after oral administration of R. officinalis extract. Sample preparation involved a liquid-liquid extraction of the analytes with ethyl acetate. Butylparaben was employed as an internal standard (I.S.). Chromatographic separation was carried out on a C18 column (ACQUITY UPLC® HSS T3, 1.8 μm, 2.1 mm × 100 mm) with a gradient system consisting of the mobile phase solution A (0.1% formic acid in water) and solution B (acetonitrile) at the flow rate of 0.3 mL/min. The quantification was obtained using multiple reaction monitoring (MRM) mode with electrospray ionization (ESI). The UHPLC-MS/MS assay was validated for linearity, accuracy, precision, extraction recovery, matrix effect and stability. This study described a simple, sensitive and validated UHPLC-MS/MS method for the simultaneous determination of three diterpene compounds in rat plasma after oral administration of R. officinalis extract, and investigated on their pharmacokinetic studies as well.

Cucurbitacin B (CuB) and cucurbitacin E (CuE) are tetracyclic triterpene compounds from Cucurbitaceae, and the main bioactive compounds of cucurbitacins tablets that used to treatment of chronic hepatitis. Pharmacological research has been very comprehensive, and there are few studies on pharmacokinetics, especially about CuE. An Ultra High Performance Liquid Chromatography-tandem Mass Spectrometry (UHPLC-MS/MS) method with high selectivity, simplicity and sensitivity has been used for quantitative analysis of Cucurbitacin B (CuB) and cucurbitacin E (CuE). Plasma samples were pretreatment by Liquid-liquid extraction (LLE) method with dichloromethane. The chromatographic separation was achieved on a C18 column (Agilent Eclipse Plus, 1.8μm, 50×2.1mm) using gradient elution with water - methanol at a flow rate of 0.3mL/min and the column temperature was set at 30°C. The method was validated according to FDA guidelines. Lower limit of quantification (LLOQ) was 1.60ng/mL for CuB and 1.58ng/mL for CuE. Correlation coefficients of CuB and CuE were more than 0.99 in rat plasma. All values of intra-day and inter-day precision (RSD%) were not exceeded 15%, the accuracy (RE%) were within -5.57 to 5.20% for CuB and -3.33 to 7.37% for CuE. The mean extraction recoveries were more than 80%. Pharmacokinetic parameters were also evaluated by UHPLC-MS/MS method. The results suggestion that this method was successfully applied to pharmacokinetic study of CuB and CuE in rat plasma after oral administration cucurbitacin tablets.

Corynoline and acetycorynoline are active compounds of Corydalis bungeana Turcz. with various pharmacological effects such as sedation, anti-leptospira and liver injury protection effects. A specific, simple and sensitive UHPLC–ESI–MS/MS method was developed and validated for the pharmacokinetic study of corynoline and acetycorynoline in rat plasma. Corynoline and acetycorynoline were extracted from plasma samples by liquid–liquid extraction. The analysis was carried out on an Agilent SB-C18 column (1.8 μm, 50 mm × 2.1 mm) with the mobile phase of acetonitrile–0.1% formic acid (80:20, v/v). The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source in positive mode. The current UHPLC–ESI–MS/MS assay was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability. The lower limits of quantification were 0.10 ng/mL for corynoline and 0.353 ng/mL for acetycorynoline. Intra-day and inter-day precision were less than 12.3% and accuracy ranged from 0.18% to 14.9%. The mean extraction recoveries of analytes and internal standard (IS) from rat plasma were all more than 76.2%. This validated method was successfully applied to pharmacokinetic study in rats after oral administration of corynoline, acetycorynoline and the extract of Corydalis bungeana Turcz.

Hernandezine, a novel anticancer AMPK activator, is a major active constituent of Thalictrum Ranunculaceae. A simple, specific and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of hernandezine in rat plasma and tissues after intravenous administration. Sample preparation was carried out through a protein-precipitation extraction with acetonitrile using tetrandrine as internal standard (IS). The chromatographic separation was achieved by using an Agilent ZORBAX Eclipse Plus C18 column with a mobile phase of acetonitrile and water (containing 10mM ammonium acetate) in an isocratic elution way. The mass spectrometry (MS) analysis was conducted in positive ionization mode with multiple reaction monitoring (MRM) transitions at m/z 653.4→411.2 for hernandezine and m/z 623.3→381.3 for tetrandrine (IS). Calibration curves were linear over the ranges of 20.0-4000ng/ml f or both plasma samples and tissue samples (r>0.991). The lower limit of quantification (LLOQ) was 20.0ng/ml. The intra-day and inter-day precision (RSD%) were less than 14.0%, while the accuracy was ranged from 85.2% to 114.9%. Finally, this developed method was successfully applied in the pharmacokinetics and tissue distribution study of hernandezine after intravenous administration.

A rapid and sensitive LC-MS/MS method based on the Triple Quad system has been developed and validated for the determination and pharmacokinetics of taxifolin and its nanodispersion in rat plasma. Taxifolin plasma samples along with butylparaben (internal standard) were pre-treated by liquid-liquid extraction with ethyl acetate, and then separated on a SB-C18 RRHD column (150 mm × 2.1 mm × 1.8 μm) using isocratic elution with a run time of 3.0 min. The mobile phase was acetonitrile–water (90:10, v/v) containing 5 mM ammonium acetate at a flow rate of 0.4 mL/min. Quantification of taxifolin was performed by the electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode with negative atmospheric ionization at m/z 303.0→285.0 for taxifolin and 193.1→92.0 for I.S., respectively. The calibration curve of taxifolin showed good linearity over a concentration range of 5.0–4280 ng/mL with a correlation coefficient of 0.9995. The limit of quantification (LLOQ) was 5.0 ng/mL. Intra-day, inter-day precision and accuracy (percent relative to standard deviation) were all within 8% at three concentration levels. A total recovery of taxifolin and I.S. was beyond 75%. The present LC-MS/MS method was successfully applied to pharmacokinetic studies of taxifolin after intravenous administration of taxifolin, oral administration of its physical mixture and nanodispersion. The absolute bioavailability of taxifolin was calculated as 0.75% for taxifolin nanodispersion and 0.49% for taxifolin, respectively.

A selective and sensitive ultra-high performance liquid chromatography with tandem mass spectrometry method was developed and validated for the determination and pharmacokinetic study of (+)-8-hydroxypinoresinol-4'-O-β -D-glucopyranoside, prinsepiol-4-O-β-D-glucopyranoside, (+)-pinoresinol-4,4'-di-O-β-D-glucopyranoside, and (-)-massoniresinol 3α-O-β-D-glucopyranoside in rat plasma after the oral administration of a Valeriana amurensis extract. The analytes and ethyl 4-hydroxybenzoate (internal standard) were separated on a Waters ACQUITY UPLC HSS T3 chromatographic column. The detection was performed on a triple quadrupole tandem mass spectrometer in multiple reaction monitoring mode using an electrospray ionization source operating in negative ionization mode. The linear ranges (ng/mL) of the standard curves were 0.39-154.00, 0.62-244.70, 0.50-198.60, and 0.34-134.50 for (+)-8-hydroxypinoresinol-4'-O-β-D-glucopyranoside, prinsepiol-4-O-β-D-glucopyranoside, (+)-pinoresinol-4,4'-di-O-β-D-glucopyranoside, and (-)-massoniresinol 3α-O-β-D-glucopyranoside, respectively. The inter- and intra-day precisions were less than 11.0%, the accuracies were between -5.9 and 7.7%, and the extraction recoveries of the four analytes were > 81.2% from rat plasma. The method was successfully applied to a pharmacokinetic study of the four analytes after oral administration of a Valeriana amurensis extract to rats. The developed method has the potential for pharmacokinetic analysis and to provide additional information in the clinical application of Valeriana amurensis.

Ethnopharmacological relevance Gentianella acuta (Michx.) Hulten belonging to the family of Gentianaceae is an annual plant mainly distributed in north of China, Mongolia plateau, Siberia and Far East areas of Russia. The whole herb was used as folk medicine to treat hepatitis, jaundice, headache and fever in Mongolia native medicine. Xanthones are the main active compounds of G. acuta and possess a lot of pharmacological and biological activities Aim of the study A selective and sensitive UHPLC–MS/MS method was developed and validated for the determination and pharmacokinetic study of swertianolin, norswertianolin, bellidifolin and demethylbellidifolin (DMB) in rat plasma after oral administration of G. acuta extract. Materials and methods Sample preparation involved a liquid–liquid extraction of the analytes with ethyl acetate. Butylparaben was employed as an internal standard. LC separation was achieved on an Agilent SB-C18 RRHD column (1.8 μm, 150 mm×2.1 mm) at 30 °C with an isocratic mobile phase consisting of acetonitrile–water (0.1% formic acid) (90:10, v/v ). The detection was accomplished by multiple-reaction monitoring (MRM) scanning with electrospray ionization (ESI) source operating in the negative ionization mode. The optimized mass transition ion-pairs ( m/z ) monitored for swertianolin, norswertianolin, bellidifolin, DMB and I.S. were 435.1/272.0, 420.8/258.9, 273.0/258.0, 258.9/214.9 and 193.0/92.0, respectively. Results The current UHPLC–MS/MS assay was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability and was suitable for pharmacokinetic studies of the four xanthones after oral administration of G. acuta extract. The time to reach the maximum plasma concentration ( T max ) was 0.40±0.12 h for swertianolin, 0.27±0.07 h for norswertianolin, 1.00±0.18 h for bellidifolin and 0.94±0.15 h for demethylbellidifolin. The elimination half-time ( t 1/2 ) of swertianolin, norswertianolin, bellidifolin and DMB, was 19.7±9.64 h, 11.3±4.51 h, 19.9±8.11 h and 24.9±8.19 h, respectively. Conclusion This study described a simple, sensitive and validated UHPLC–MS/MS method for simultaneous determination of four xanthones in rat plasma after oral administration of G. acuta extract, and investigated on their pharmacokinetic studies as well.