Development of a rapid UPLC-MS/MS method for the determination of toddalolactone in mouse blood and its application in pharmacokinetics

AbstractA rapid and simple UPLC-MS/MS method was developed to determine toddalolactone in mouse blood and applied to measure the pharmacokinetics of toddalolactone in mice. Blood samples were first preprocessed by ethyl acetate liquid-liquid extraction. Oxypeucedanin hydrate (internal standard, IS) and toddalolactone were gradient eluted from a UPLC BEH C18 column using a mobile phase consisting of acetonitrile and water (0.1% formic acid). Using electrospray ionization (ESI) as the ionization source, multiple reaction monitoring was used to detect the precursor and product ions of m/z 309.2 and 205.2, respectively, for toddalolactone and of m/z 305.1 and 203.0 for IS, respectively, for quantitative detection. A calibration curve was run over the concentration range of 5–4,000 ng/mL (r > 0.995). The matrix effects ranged from 93.5 to 98.4%, and the recovery was higher than 77.3%. The precision was less than 13%, and the accuracy ranged from 90.9 to 108.4%. The developed UPLC-MS/MS method was successfully used for measuring the pharmacokinetics of toddalolactone in mice after oral (20 mg/kg) and intravenous administration (5 mg/kg), and the absolute bioavailability of toddalolactone was 22.4%.

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Determination of eugenitin in mouse blood by ultra-performance liquid chromatography-tandem mass spectrometry and its application to a pharmacokinetic study

Acta Chromatographica ◽

10.1556/1326.2021.00937 ◽

2021 ◽

Author(s):

Chongliang Lin ◽

Dezhen Song ◽

Haodong Jiang ◽

Lvqi Luo ◽

Xi Bao ◽

...

Keyword(s):

Biological Fluids ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Quantitative Detection ◽

Syzygium Aromaticum ◽

Mouse Blood ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

Abstract Eugenitin is a non-volatile chromone derivative which is always found in dried flower buds of Syzygium aromaticum L. (Merr.) & L.M. Perry. Until now, there were no reports about the pharmacokinetics of eugenitin in biological fluids. A UPLC-MS/MS method developed to determine eugenitin in mouse blood. The blood samples were prepared by protein precipitation with acetonitrile. Chrysin (internal standard, IS) and eugenitin were gradient eluted by mobile phase of acetonitrile and water (0.1% formic acid) in a BEH C18 column. The multiple reaction monitoring (MRM) of m/z 221.1→206.0 for eugenitin and m/z 255.1→152.9 for IS with an electrospray ionization (ESI) source was used for quantitative detection. The calibration curve ranged from 0.5 to 500 ng/mL (r > 0.995). The accuracy ranged from 98 to 113%, the precision was less than 12%, and the matrix effect was between 86 and 94%, the recovery was better than 81%. The developed method was successfully used for pharmacokinetics of eugenitin in mice after intravenous (5 mg/kg) and oral (20 mg/kg) administration, and the absolute availability of eugenitin was 12%.

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Simultaneous Determination of Six Uncaria Alkaloids in Mouse Blood by UPLC–MS/MS and Its Application in Pharmacokinetics and Bioavailability

BioMed Research International ◽

10.1155/2020/1030269 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

Lianguo Chen ◽

Jianshe Ma ◽

Xianqin Wang ◽

Meiling Zhang

Keyword(s):

Simultaneous Determination ◽

Oral Absorption ◽

Matrix Effects ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Mouse Blood ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

A specific ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the simultaneous determination of six Uncaria alkaloids in mouse blood with midazolam as the internal standard (IS). Only 20 μL blood was needed for sample preparation, and the protein was precipitated with acetonitrile. The UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) was used for chromatographic separation. The mobile phase consisted of 0.1% formic acid and acetonitrile with gradient elution within 5.5 min. Multiple reaction monitoring (MRM) and the positive electrospray ionization model were used for quantitative analysis. The accuracy of the UPLC-MS/MS method ranged from 86.5% to 110.4%. The precision for intraday and interday was ≤15% each. The mean recovery and the matrix effects were found to be 64.4-86.8% and 94.1-109.4%, respectively. The calibration curves in blood were linear in the range of 1-1000 ng/mL with a favorable correlation coefficient (r2) of 0.995. The pharmacokinetic results showed that six Uncaria alkaloids metabolized rapidly in mice with a half-life between 0.6 h and 4.4 h. The bioavailability of corynoxeine, isocorynoxeine, rhynchophylline, isorhynchophylline, hirsutine, and hirsuteine was 27.3%, 32.7%, 49.4%, 29.5%, 68.9%, and 51.0%, respectively, which showed satisfactory oral absorption of each alkaloid.

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Pharmacokinetics and bioavailability of liensinine in mouse blood by UPLC-MS/MS

Acta Chromatographica ◽

10.1556/1326.2020.00847 ◽

2020 ◽

Author(s):

Shuhua Tong ◽

Yuqi Zeng ◽

Jianshe Ma ◽

Congcong Wen

Keyword(s):

Pharmacokinetic Study ◽

Mobile Phase ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Quantitative Detection ◽

Reaction Monitoring ◽

Mouse Blood ◽

Blood Samples ◽

The Matrix ◽

Better Than

AbstractLiensinine is a bisbenzyltetrahydroisoquinoline alkaloid extracted from lotus (Nelumbo nucifera GAERTNER., Nelumbonaceae), especially in its embryo loti “Lien Tze Hsin” (green embryo of mature seed). A rapid and simple UPLC-MS/MS method was developed to determine liensinine in mouse blood and its application to a pharmacokinetic study. The blood samples were preprocessed by protein precipitation using acetonitrile. Midazolam (internal standard, IS) and liensinine were gradient eluted by mobile phase of methanol and water (0.1% formic acid) in a Waters UPLC BEH C18 column. The multiple reaction monitoring of m/z 611.3 → 206.1 for liensinine and m/z 326.2 → 291.1 for IS with an electrospray ionization (ESI) source was used for quantitative detection. The calibration curve ranged from 0.5 to 400 ng/mL (r > 0.995). The accuracy ranged from 92.2 to 108.2%, the precision of intra-day and inter-day was less than 14%, and the matrix effect was between 100.0% and 109.6%, the recovery was better than 71.0%. The developed UPLC-MS/MS method was successfully used for a pharmacokinetic study of liensinine in mice after oral (5 mg/kg) and intravenous administration (1 mg/kg), and the absolute availability of liensinine was 1.8%.

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Quantification of Lappaconitine in Mouse Blood by UPLC-MS/MS and Its Application to a Pharmacokinetic Study

BioMed Research International ◽

10.1155/2019/6262105 ◽

2019 ◽

Vol 2019 ◽

pp. 1-6

Author(s):

Fan Chen ◽

Xiuwei Shen ◽

Peng Huang ◽

Huiyan Fu ◽

Yue Jin ◽

...

Keyword(s):

Limit Of Detection ◽

Multiple Reaction Monitoring ◽

Ultrahigh Pressure ◽

Absolute Bioavailability ◽

Quantitative Detection ◽

Intragastric Administration ◽

Linear Calibration ◽

Ionization Source ◽

Mouse Blood ◽

Limit Of Quantitation

Lappaconitine is extracted from Aconitum sinomontanum Nakai, which belongs to the Ranunculaceae. Lappaconitine is as a diterpenoid alkaloid used as a nonaddictive analgesic. To assure the rational use of the drug, ultrahigh-pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was conducted to determine lappaconitine in mouse blood and its application to pharmacokinetics. In this study, khasianine was used as internet standard (IS). A UPLC BEH C18 column was used for chromatographic separation and the mobile phase consisted of acetonitrile and 10 mmol/L ammonium acetate (0.1% formic acid). The flow rate of was 0.4 mL/min. Quantitative detection was performed in a multiple reaction monitoring (MRM) mode using an electrospray ionization source in positive mode. Twenty-four mice were randomly divided into four groups, three of which received 2, 4, and 8 mg/kg lappaconitine by intragastric administration, while the other group received 1 mg/kg lappaconitine by intravenous administration. After 0.0833, 0.5, 1, 1.5, 2, 3, 4, and 8 h, blood samples were collected and acetonitrile was used for protein precipitation. A linear calibration relationship (R2 = 0.9979) in the range of 0.1-500 ng/mL in mouse blood indicated good results. The lower limit of quantitation was 0.1 ng/mL and the limit of detection was 0.04 ng/mL. The intra-day and inter-day precision were below 13% and 14%, respectively. The accuracy was 90.1-107.2%, and the recovery exceeded 81.1%. The matrix effect ranged between 102.1 and 108.8%. The absolute bioavailability of lappaconitine was 2.0%. UPLC-MS/MS achieved high sensitivity, speed, and selectivity. Methodological verification indicated this method as suitable for determination of lappaconitine in mouse blood.

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Determination of Trans-loxoprofen-alcohol Tromethamine in Pregnant SD rats by a Validated LC-MS/MS method：Application to a Toxicokinetics and Tissue Distribution study

Current Pharmaceutical Analysis ◽

10.2174/1573412917666201222100518 ◽

2020 ◽

Vol 17 ◽

Author(s):

Lingling Xu ◽

Wenjun Zhou ◽

Yanjuan Yuan ◽

Qing Shao ◽

Hongqun Qiao

Keyword(s):

Tissue Distribution ◽

Matrix Effects ◽

Multiple Reaction Monitoring ◽

Ion Pairs ◽

Internal Standard ◽

Quantitative Detection ◽

Ionization Source ◽

Distribution Study ◽

Sd Rats ◽

Tissue Distribution Study

Background: As an anti-inflammatory prodrug, loxoprofen is metabolized into trans-loprofenol to treat diseases related to pain and inflammation. Although loxoprofen has fewer adverse effects than other NSAIDs, the safety of its usage during pregnancy remains unclear and needs to be considered. Fortunately, the toxicokinetics and tissue distribution study of trans-loxoprofen-alcohol in pregnant rats can resolve the problem. Objective: The purpose of this study is to establish a simple, sensitive and effective LC-MS/MS analysis method for determination the concentration of trans-loxoprofen alcohol in plasma and tissues. Methods: The analytic samples were precipitated by methanol in one step and separated using a reverse-phase Poroshell 120 EC-C18 column (4.6 mm×50mm; 2.7μm). And the gradient mobile phase at a flow rate of 0.6 mL/min was composed of acetonitrile and 0.1% formic acid in water. The quantitative detection was achieved by multiple-reaction monitoring mode with positive electrospray ionization source, transitional ion pairs of m/z 265.9>184.8 for trans-loxoprofen-alcohol, and 268.8>187.9 for rac-trans-loxoprofen-D3 alcohol (Internal standard). Results: A good linearity of calibration curves for plasma and tissues were observed in the concentration range from 5.0 to 5000ng/mL, and the lower limit of quantification was detected at 5.0ng/mL. The intra-day and inter-day precision in plasma and tissues were within 8.94% and 7.26%, respectively. The mean extraction recovery and matrix effects in plasma and tissues were in the range of 89.08~109.27% and 89.00~106.80%, respectively. Precision of stability in plasma and tissues were within 8.91% and 7.08%, respectively. Conclusion: Complying with the requirements of bioanalytical guidelines by validation, this method was successfully adopted to the toxicokinetics and tissue distribution study after intravenously administrated trans-loxoprofen-alcohol into pregnant SD rats.

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Determination of narciclasine in mouse blood by UPLC-MS/MS and its application to a pharmacokinetic study

Acta Chromatographica ◽

10.1556/1326.2021.00865 ◽

2021 ◽

Author(s):

Ke Ren ◽

Tiantian Feng ◽

Hai Shi ◽

Jianshe Ma ◽

Yongxi Jin

Keyword(s):

Intravenous Administration ◽

Pharmacokinetic Study ◽

Mobile Phase ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Hydroxy Derivative ◽

Reaction Monitoring ◽

Mouse Blood ◽

The Matrix

AbstractNarciclasine is a 7-hydroxy derivative of lycorisidine. It was the first alkaloid isolated from the stem of narcissus (Amaryllidaceae) in 1967. Six mice were given narciclasine (5 mg/kg) by intravenous administration. A UPLC-MS/MS method was developed to determine narciclasine in mouse blood. Tectorigenin (internal standard, IS) and narciclasine were gradient eluted by mobile phase of methanol and 0.1% formic acid in a BEH C18 column. The multiple reaction monitoring (MRM) of m/z 308.1→248.1 for narciclasine and m/z 301.1→286.0 for IS with an electrospray ionization (ESI) source was used for quantitative determination. The calibration curve ranged from 1 to 6,000 ng/mL. The accuracy was from 92.5 to 107.3%, and the matrix effect was between 103.6 and 107.4%. The developed UPLC-MS/MS method was successfully applicated to a pharmacokinetic study of narciclasine in mice after intravenous administration (5 mg/kg).

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Determination and pharmacokinetics of calycanthine in rat plasma by UPLC-MS/MS

Acta Chromatographica ◽

10.1556/1326.2020.00772 ◽

2020 ◽

Author(s):

Meifei Lu ◽

Xiaojie Lu ◽

Zheng Yu ◽

Congcong Wen

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Rat Plasma ◽

Precipitation Method ◽

Absolute Bioavailability ◽

Limit Of Quantification ◽

The Matrix ◽

Acid Aqueous Solution

AbstractCalycanthine is an important class of alkaloids extracted and isolated from the roots, leaves, flowers and fruits of Chimonanthus praecox. In this work, the UPLC-MS/MS method was used for determination of calycanthine in rat plasma, and the pharmacokinetics in rats were investigated. Midazolam was used as an internal standard (IS), and methanol precipitation method was used to pretreatment the rat plasma samples. Chromatographic separation was achieved on a UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) column with the mobile phase of methanol- 0.1% formic acid aqueous solution with gradient elution. Multiple reaction monitoring (MRM) mode with positive ionization was applied for quantitative analysis, m/z 347.3 → 246.7 and 326.2 → 291.4 for calycanthine and IS, respectively. The results indicated that within the range of 1–200 ng/mL, linearity of calycanthine in rat plasma was good (r > 0.995), and the lower limit of quantification (LLOQ) was 1 ng/mL. Accuracy range was between 90.6 and 109.4%, precision (RSD) of calycanthine was less than 14%. The matrix effect was between 97.9% and 105.4%, the recovery was better than 85.6%. The developed UPLC-MS/MS method was successfully applied in the pharmacokinetics of calycanthine in rats after oral and intravenous administration. The absolute bioavailability of the calycanthine was 37.5% in rats.

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UPLC-MS/MS Method for Determination of Khasianine in Mouse Blood: Application for Its Pharmacokinetic Study

Current Pharmaceutical Analysis ◽

10.2174/1573412915666190220101658 ◽

2020 ◽

Vol 16 (6) ◽

pp. 705-711

Author(s):

Lianguo Chen ◽

Qinghua Weng ◽

Yijing Lin ◽

Xiaojie Lu ◽

Zuoquan Zhong ◽

...

Keyword(s):

Lower Limit ◽

Pharmacokinetic Study ◽

Oral Absorption ◽

Matrix Effects ◽

Limit Of Detection ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Quantitative Detection ◽

Mouse Blood ◽

Limit Of Quantitation

Background: The aim of this study was to determine the concentrations of khasianine in mouse whole blood sample and its application for the pharmacokinetics by a rapid, selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Methods: The blood samples were preprocessed by one-step protein precipitation with acetonitrile. The study was performed on an ACQUITY I-Class UPLC system with a UPLC BEH column. Lannaconitine (internal standard, IS) and khasianine were gradient eluted by a mixture of acetonitrile and water with 0.1% formic acid at a flow rate of 0.4 mL/min. The mass spectrometer was equipped with an Electrospray Ionization (ESI) source in positive mode. The quantitative detection was performed in a multiple reaction monitoring modes at transitions m/z 722.4→70.7 for khasianine and m/z 585.3→119.9 for the corresponding IS. Results: The calibration curve was of good linearity ranging from 0.5 to 1000 ng/mL (r > 0.995). The Lower Limit of Detection (LLOD) and Lower Limit of Quantitation (LLOQ) were 0.2 and 0.5 ng/mL, respectively. The inter-day and intra-day precision (RSD%) were both less than 14%, and the accuracy ranged from 86.6% to 108.3%. The matrix effects were between 98.0% and 103.7%, and the average recovery was better than 67.4%. Conclusion: This assay established a sensitive, rapid, selective UPLC-MS/MS method which was successfully used for the pharmacokinetic study of khasianine in mouse blood, and the absolute availability of khasianine was 0.78% which exhibited a poor oral absorption.

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Pharmacokinetics of ebeiedinone in mouse blood by UPLC–MS/MS

Acta Chromatographica ◽

10.1556/1326.2019.00680 ◽

2020 ◽

Vol 32 (3) ◽

pp. 194-198

Author(s):

Yongxi Jin ◽

Yuyan Chen ◽

Jiawen Liu ◽

Xi Bao ◽

Yinghao Zhi ◽

...

Keyword(s):

Pharmacokinetic Study ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Absolute Bioavailability ◽

Mouse Blood ◽

Limit Of Quantification ◽

Chromatography Tandem Mass Spectrometry ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was established to determine ebeiedinone in mouse blood, and the pharmacokinetics of ebeiedinone after intravenous (0.5 mg/kg) and oral (2, 4, and 8 mg/kg) administration was studied. Twenty-four mice were randomly divided into 4 groups, 1 group was for intravenous administration (0.5 mg/kg), and other 3 groups were for oral administration (2, 4, and 8 mg/kg), with 6 rats in each group. Yubeinine was used as an internal standard. Multiple reaction monitoring (MRM) mode was used to quantitatively analyzed ebeiedinone m/z 414.4 → 91.1 and the internal standard m/z 430.4 → 412.3 in the electrospray ionization (ESI) positive interface. In the concentration range of 1–2000 ng/mL, the ebeiedinone in the mouse blood was linear (r2 > 0.995), and the lower limit of quantification was 1.0 ng/mL. In the mouse blood, the intra-day precision coefficient of variation (CV) was less than 15%, and the inter-day precision CV was less than 15%. The accuracy ranged from 85.4% to 114.6%, and the average recovery was higher than 61.3%. The matrix effect was between 87.0% and 106.5%. These data met the pharmacokinetic study requirements of ebeiedinone. The UPLC–MS/MS method was sensitive, rapid, and selective and was successfully applied to the pharmacokinetic study of ebeiedinone in mice. The absolute bioavailability of ebeiedinone was 30.6%.

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Determination of Oxadixyl in Wines by Liquid Chromatography-Tandem Mass Spectrometry: Single-Laboratory and Interlaboratory Validation Study

Journal of AOAC International ◽

10.5740/jaoacint.13-359 ◽

2014 ◽

Vol 97 (6) ◽

pp. 1701-1706

Author(s):

Armen Mirzoian ◽

Jeffrey R Ammann

Keyword(s):

Validation Study ◽

High Throughput Screening ◽

Multiple Reaction Monitoring ◽

Direct Injection ◽

Ionization Source ◽

Minimal Sample ◽

The Matrix ◽

Liquid Chromatography Tandem Mass ◽

Interlaboratory Validation

Abstract A direct injection LC/MS/MS method for the determination of the pesticide oxadixyl in wines was developed and validated. A sample divert valve was used to deliver the fraction that contained oxadixyl to the mass spectrometer's electrospray ionization source. Oxadixyl was monitored and quantitated using two transitions in multiple reaction monitoring mode. The method demonstrated recoveries of 99.2 ± 2.0 and 96.7 ± 5.2% for red and white wines, respectively, a linearity range of 2–20 μg/L, LOD at 0.7 μg/L, LOQ of 2.0 μg/L, and precision values of 8.2% (RSDr) and 6.2% (RSDR). Direct injection of the wine onto a C18 ultra-performance LC column allowed automation and high throughput screening. Benefits of this approach include minimal sample preparation, short (3 min) run times, and the use of matrix-matched calibration standards, which minimize the matrix effect due to interferences from wine phenolics, sugars, and various other components. The method's performance characteristics were not statistically different for white and red wines. An additional interlaboratory validation study involved 12 laboratories and demonstrated good data agreement with HorRat values ranging from 0.23 to 0.52.

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