Development and optimization of a supercritical fluid chromatography tandem mass spectrometry method for the high‐throughput determination of nimodipine in beagle plasma

A simple, sensitive, and efficient supercritical fluid chromatography with tandem mass spectrometry method was established for the determination of nimodipine in beagle plasma. One‐step protein precipitation with acetone was used to extract the analytes from the plasma. Nitrendipine was used as the internal standard. The chromatographic separation was achieved on an ACQUITY UPC²? BEH 2‐EP column, and a gradient elution program was applied at a flow rate of 1.5 mL/min. The detection was carried out on a triple quadrupole tandem mass spectrometer with an electrospray ionization source operating in positive ion mode. Quantification was performed using multiple reaction monitoring of the transitions of m/z 419.3→301.3 for nimodipine and m/z 361.4→315.2 for nitrendipine. A satisfactory linearity was obtained over the concentration range of 0.5–800 ng/mL (r > 0.996). The intra‐ and interday precision and accuracy results were <9.1% across the quality control levels. The peak concentration and area under concentration‐time curve (0–720 min) values of the test and reference formulations were 279.28 ± 211.46 and 265.13 ± 149.26 ng/mL, 25608.00 ± 17553.65 and 28553.67 ± 20207.92 ng·min/mL, respectively. The validated method was successfully applied to reveal the pharmacokinetic profiles of nimodipine in beagle dogs after oral administration. Moreover, the analytical method could be used for further bioequivalence studies.     

A sensitive,high‐throughput,and ecofriendly method for the determination of lumefantrine,artemether, and its active metabolite dihydroartemisinin by supercritical fluid chromatography and tandem mass spectrometry

A quick and sensitive supercritical fluid chromatography with tandem mass spectrometry method for the simultaneous determination of lumefantrine, artemether, and its active metabolite dihydroartemisinin in rat plasma was developed and validated. The chromatographic separation was performed on an ACQUITY UPC²™ BEH 2‐EP column within 2.5 min by gradient elution using compressed CO₂ and methanol containing 2 mM ammonium acetate as the mobile phases. Detection was achieved by multiple reaction monitoring using electrospray ionization in the positive ionization mode. For sample preparation, 50 μL of the sample was processed by modified high‐throughput, one‐step protein precipitation using hydrogen peroxide as a stabilizer to protect the endoperoxide‐containing artemisinin derivatives from degradation. The calibration curves were linear over the concentration range of 2.0–1000 ng/mL for both artemether and dihydroartemisinin, and 1.0–5000 ng/mL for lumefantrine. The values of selectivity, lower limit of quantification, linearity, accuracy, precision, matrix effects, stability, and recovery met the acceptable range according to the Food and Drug Administration guidelines. The developed method enables high resolution and speed as well as low cost, low solvent consumption, and short time and was successfully applied to pharmacokinetic studies through the intravenous administration of an artemether–lumefantrine lipid emulsion in rats.     

Determination of rabeprazole enantiomers in dog plasma by supercritical fluid chromatography tandem mass spectrometry and its application to a pharmacokinetic study

Rabeprazole is a novel benzimidazole proton pump inhibitor used for the treatment of gastrointestinal disorders. It is a chiral molecule that gives rise to the possibility of stereoselective pharmacokinetics. To investigate this phenomenon, a rapid and sensitive chiral assay based on supercritical fluid chromatography tandem mass spectrometry was developed and applied to the determination of (R )‐rabeprazole and (S )‐rabeprazole in dog plasma. Sample preparation involved protein precipitation with acetonitrile after the addition of (R )‐lansoprazole as internal standard. Baseline separation of enantiomers in 4.5 min was achieved on an Acquity UPC² system using an ACQUITY UPC² Trefoil CEL2 column maintained at 60°C and a mobile phase consisting of methanol/CO₂ (30:70, v/v) delivered at 2.5 mL/min. Detection was achieved by multiple reaction monitoring of the transitions at m/z 360.0→242.2 (rabeprazole) and 370.3→252.0 (internal standard) in the positive ion mode. The assay was linear in the range of 1–1000 ng/mL and free of matrix effects. Intra‐ and interday precisions were less than 10.0% with accuracy in the range of –2.6 to 3.1%. The method was successfully applied to a pharmacokinetic study of rabeprazole enantiomers after administration of a single oral dose of 10 mg racemate to beagle dogs.     

Simultaneous determination of 17 bisphenols in polycarbonate by ultra‐high performance supercritical fluid chromatography with tandem mass spectrometry

A novel method was developed for the first time for the determination of 17 bisphenols by ultra‐high performance supercritical fluid chromatography with tandem mass spectrometry. Under the optimal conditions, 17 bisphenols were separated successfully on a high density diol column in 9 min using methanol and carbon dioxide as mobile phase. 0.02% ammonium hydroxide/methanol v/v was used as the post‐column compensation solvent to improve response of mass spectrometry. Linear relations of matrix‐matched calibration curve were favorable over the selected concentration range of 1–100 μg/kg with correlation coefficients greater than 0.9981. The method limit of detection and limit of quantitation were 0.1–0.5 μg/kg and 0.5–2.5 μg/kg, respectively. The average recoveries at three spiked levels in polycarbonate were in the range of 81.8–114.5%. Intra‐day and inter‐day precisions for six replicates were below 15.0%. This method was successfully applied to determine bisphenols in polycarbonate.     

Supercritical fluid chromatography coupled with tandem mass spectrometry: A high‐efficiency detection technique to quantify Taxane drugs in whole‐blood samples

We present a technique to rapid determine taxane in blood samples by supercritical fluid chromatography together with mass spectrometry. The aim of this study was to develop a supercritical fluid chromatography with mass spectrometry method for the analysis of paclitaxel, cabazitaxel, and docetaxel in whole‐blood samples of rats. Liquid‐dry matrix spot extraction was selected in sample preparation procedure. Supercritical fluid chromatography separation of paclitaxel, cabazitaxel, docetaxel, and glyburide (internal standard) was accomplished within 3 min by using the gradient mobile phase consisted of methanol as the compensation solvent and carbon dioxide at a flow rate of 1.0 mL/min. The method was validated regarding specificity, the lower limit of quantification, repeatability, and reproducibility of quantification, extraction recovery, and matrix effects. The lower limit of quantification was found to be 10 ng/mL since it exhibited acceptable precision and accuracy at the corresponding level. All interday accuracies and precisions were within the accepted criteria of ±15% of the nominal value and within ±20% at the lower limit of quantification, implying that the method was reliable and reproducible. In conclusion, this method is a promising tool to support and improve preclinical or clinical pharmacokinetic studies with the taxanes anticancer drugs.     

Assessment of tandem mass spectrometry and high‐resolution mass spectrometry for the analysis of bupivacaine in plasma

Triple quadrupole mass spectrometers coupled with high performance liquid chromatography are workhorses in quantitative bioanalyses. They provide substantial benefits including reproducibility, sensitivity and selectivity for trace analysis. Selected reaction monitoring allows targeted assay development but datasets generated contain very limited information. Data mining and analysis of nontargeted high‐resolution mass spectrometry profiles of biological samples offer the opportunity to perform more exhaustive assessments, including quantitative and qualitative analysis. The objectives of this study were to test method precision and accuracy, to statistically compare bupivacaine drug concentration in real study samples and to verify if high‐resolution and accurate mass data collected in scan mode can actually permit retrospective data analysis, more specifically, extract metabolite related information. The precision and accuracy data presented using both instruments provided equivalent results. Overall, the accuracy ranged from 106.2 to 113.2% and the precision observed was from 1.0 to 3.7%. Statistical comparisons using a linear regression between both methods revealed a coefficient of determination (R²) of 0.9996 and a slope of 1.02, demonstrating a very strong correlation between the two methods. Individual sample comparison showed differences from ?4.5 to 1.6%, well within the accepted analytical error. Moreover, post‐acquisition extracted ion chromatograms at m/z 233.1648 ± 5 ppm (M ? 56) and m/z 305.2224 ± 5 ppm (M + 16) revealed the presence of desbutyl‐bupivacaine and three distinct hydroxylated bupivacaine metabolites. Post‐acquisition analysis allowed us to produce semi‐quantitative evaluations of the concentration–time profiles for bupicavaine metabolites. Copyright © 2015 John Wiley & Sons, Ltd.     

Simultaneous determination of cyadox and its metabolites in plasma by high‐performance liquid chromatography tandem mass spectrometry

Cyadox is a novel antimicrobial growth‐promoter of the quinoxalines. For food safety and pharmacokinetic studies, a convenient, sensitive and reproducible LC‐ESI‐MS/MS method was developed for the simultaneous determination of cyadox and its major metabolites, quinoxaline‐2‐carboxylic acid, 1,4‐bisdesoxycyadox, cyadox‐1‐monoxide and cyadox‐4‐monoxide in chicken plasma. Plasma sample was subjected to a simple deproteinisation with acetonitrile. Analysis was performed on a C₁₈ column by detection with mass spectrometry in multiple reaction monitoring mode. A gradient elution program with 0.2% formic acid, methanol and acetonitrile was performed at a flow rate of 0.2 mL/min. The decision limits (CCαs) of five analytes in plasma ranged from 1.0 to 4.0 μg/L, and the detection capabilities (CCβs) were <10 μg/L. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The extraction recoveries of five analytes were between 87.4 and 93.9% in plasma at the spiked levels of 5 (10)–200 μg/L with the relative standard deviations <10% for each analyte. The developed method demonstrated a satisfactory applicability in real plasma samples.     

Analysis of saikosaponins in rat plasma by anionic adducts-based liquid chromatography tandem mass spectrometry method

Saikosaponins (SSs) are a class of triterpene saponins with a wide spectrum of bioactivities. A sensitive liquid chromatography tandem mass spectrometry (LC‐MS/MS) method was developed for simultaneous determination of saikosaponin a, saikosaponin c, saikosaponin d and saikosaponin b₂ in rat plasma. Plasma samples were prepared by liquid–liquid extraction. The analytes and the internal standard (IS) digoxin were well separated on an octadecyl column using gradient elution and analyzed by monitoring the fragmentation transition pair of anionic adducts to deprotonated molecules in negative‐mode electrospray. By neutral loss of HCOOH, the transition pairs of m/z 825 → 779 for SSa, SSd, SSb₂ and the IS, and m/z 971 → 925 for SSc were sensitive for MS/MS detection with the lower limits of quantification in the range of 0.20–0.40 ng/mL. Method validation experiments were performed, including selectivity, precision, accuracy, linearity, matrix effect, recovery and stability. The validated method was further applied to determine the pharmacokinetics parameters of SSa, c and d in rats following a single oral administration of the extract of chaihu (the dried roots of Bupleurum chinense DC). Copyright © 2011 John Wiley & Sons, Ltd.     

Simultaneous high‐performance liquid chromatography with tandem mass spectrometry quantification of six bioactive components in rat plasma after oral administration of Yougui pill

Yougui pills are a classic Chinese medicine that shows significant effects on nerve regeneration and neuroprotection in modern pharmacological studies. With a complex formula, Yougui pills have faced significant challenges in the fields of bioanalysis and pharmacokinetics in animals and human studies. In the present study, a specific and accurate high‐performance liquid chromatography with tandem mass spectrometry method was developed and validated for the quantitative determination of the six bioactive components in rat plasma after oral administration of Yougui pills. Chromatographic separation was performed on a C18 column with a gradient elution system. Samples were analysed using positive ion mode with multiple reaction monitoring mode. The assay showed good linearity for all six bioactive components in the dynamic range of 0.50 to 50 ng/mL with acceptable intra‐ and inter‐batch accuracy and precision. The lower limits of quantification were 0.50 ng/mL for all six bioactive components. The method was successfully applied to rat pharmacokinetics after oral administration of Yougui pills. All six bioactive components were detected in rat plasma, including songorine, benzoylhypaconitine, benzoylmesaconitine, neoline, karacoline, and sweroside, while some other target compounds were not detected, such as rhmannioside A, loganin, and cornuside I. After oral administration of Yougui pills at a dose of 2500 mg/kg, all six bioactive components were rapidly absorbed, resulting in t_max values less than 1 h and relative lower C_max values. The t_1/2 values for songorine, benzoylhypaconitine, benzoylmesaconitine, neoline, karacoline, and sweroside were calculated to be 2.62 ± 0.67, 2.11 ± 0.45, 1.94 ± 0.35, 1.88 ± 0.31, 2.07 ± 0.44, and 1.59 ± 0.30 h, which indicated that Yougui pills should be taken in multiple oral doses over a relatively short period.     

Simultaneous quantitation of two diastereoisomers of lobaplatin in rat plasma by supercritical fluid chromatography with tandem mass spectrometry and its application to a pharmacokinetic study

Lobaplatin, consisting of two diastereoisomers, is a third‐generation platinum antineoplastic agent that has shown encouraging anticancer activity in a variety of tumor types. To investigate any stereospecificity in the pharmacokinetics of lobaplatin, a novel, simple, rapid and sensitive supercritical fluid chromatography with tandem mass spectrometry method was developed for the simultaneous quantitation of lobaplatin diastereoisomers in rat plasma. After a simple protein precipitation with methanol, the analytes and dexpantoprazole (internal standard) were chromatographed on an Acquity UPC² system with a Chiralcel OZ‐RH column using a mobile phase consisting of carbon dioxide and methanol (65:35, v/v) at 40°C over 6 min. The assay was linear over a concentration range of 25–15 000 ng/mL for both diastereoisomers using 100 μL of rat plasma for sample preparation. The lower limit of quantification was 25 ng/mL for both compounds, which was sufficient to detect the diastereoisomers in the incurred samples within this study. Intra‐ and inter‐day precisions were below 11.8% and the accuracies were below 4.5%. The validated method was successfully applied to a pharmacokinetic study after an intravenous administration of 7.6 mg/kg lobaplatin to rats. There was no apparent stereospecificity in the pharmacokinetics between the two diastereoisomers of lobaplatin.     

A highly sensitive method (supercritical fluid chromatography coupled with ion mobility mass spectrometry) for determination of multiple compounds in radix curcumae

A highly sensitive method was developed for simultaneously separating and identifying multiple compounds in radix curcumae. The determination of these compounds was achieved by combining supercritical fluid chromatography with drift tube ion mobility quadrupole time-of-flight MS. Related parameters were optimized: the RX-SIL column was used as the stationary phase, methanol was selected as the organic modifier, back pressure was 120 bar, back temperature was 60°C, the mobile phase flow rate was 1.75 mL/min, the makeup solvent was 0.2% formic acid/methanol with a flow rate of 0.7 mL/min. Under optimal conditions, multipolar compounds were separated. Furthermore, these compounds were identified by the values of collision sectional areas. The established method was verified by related parameters and exhibited good linearity, sensitivity, precision and accuracy. It could be extended to analyze other curcuminoids and sesquiterpenoids in natural products.     

Direct online extraction and determination by supercritical fluid extraction with chromatography and mass spectrometry of targeted carotenoids from red Habanero peppers (Capsicum chinense Jacq.)

Recently, supercritical fluid chromatography coupled to mass spectrometry has gained attention as a fast and useful technology applied to the carotenoids analysis. However, no reports are available in the literature on the direct online extraction and determination by supercritical fluid extraction with chromatography and mass spectrometry. The aim of this research was the development of an online method coupling supercritical fluid extraction and supercritical fluid chromatography for a detailed targeted native carotenoids characterization in red habanero peppers. The online nature of the system, compared to offline approaches, improves run‐to‐run precision, enables the setting of batch‐type applications, and reduces the risks of sample contamination. The extraction has been optimized using different temperatures, starting from 40°C up to 80°C. Multiple extractions, until depletion, were performed on the same sample to evaluate the extraction yield. The range of the first extraction yield, carried out at 80°C, which was the best extraction temperature, was 37.4–65.4%, with a %CV range of 2–12. Twenty‐one targeted analytes were extracted and identified by the developed methodology in less than 17 min, including free, monoesters, and diesters carotenoids, in a very fast and efficient way. Quantification of the β‐carotene was carried out by using the optimized conditions.     

Highly sensitive and accurate profiling of carotenoids by supercritical fluid chromatography coupled with mass spectrometry

We attempted to establish a high‐speed and high‐resolution profiling method for a carotenoid mixture as a highly selective and highly sensitive detection method; the analysis was carried out by supercritical fluid chromatography (SFC) coupled with mass spectrometry (MS). When an octadecyl‐bonded silica (ODS) particle‐packed column was used for separation, seven carotenoids including structural isomers were successfully separated within 15 min. This result indicated not only improved separation but also improved throughput compared to the separation and throughput in RP‐HPLC. The use of a monolithic ODS column resulted in additional improvement in both the resolution and the throughput; the analysis time was reduced to 4 min by increasing the flow rate. Furthermore, carotenoids in biological samples containing the complex matrices were separated effectively by using several monolithic columns whose back pressure was very low. The mass spectrometer allowed us to perform a more sensitive analysis than UV detection; the detection limit of each carotenoid was 50 pg or below. This is the first report of carotenoid analysis carried out by SFC‐MS. The profiling method developed in this study will be a powerful tool for carrying out accurate profiling of biological samples.     

Liquid chromatography tandem mass spectrometry with triple stage fragmentation for highly selective analysis and pharmacokinetics of alarelin in rat plasma

Alarelin, a gonadotropin‐releasing hormone analogue, is widely used in China for the treatment of endometriosis and uterine leiomyoma. In order to investigate its pharmacokinetic behavior and support the preclinical application of new formulations, we have developed a novel and highly selective bioanalytical method to determine alarelin in rat plasma based on liquid chromatography tandem mass spectrometry with triple stage fragmentation. After sample preparation by protein precipitation followed by reversed phase solid phase extraction, alarelin and triptorelin (internal standard) were chromatographed on an Ascentis^® Express C18 column (50 mm × 4.6 mm, 2.7 µm) using gradient elution with 0.1% formic acid in water and acetonitrile at a flow rate of 1 mL/min. Detection was by positive mode electrospray ionization followed by triple stage fragmentation using the transitions at m/z 584.6→249.1→221.0 for alarelin and 656.5→249.1→176.0 for triptorelin, The assay was linear in the concentration range 0.3‐10 ng/mL with excellent precision and accuracy. It was successfully applied to a pharmacokinetic study in rats administered a dose of 13.5 µg/kg alarelin by intramuscular injection. The results show that the triple stage fragmentation strategy allows highly selective analysis of alarelin and has the potential to be widely applied to the bioassay of other peptidic drugs.     

Pharmacokinetic and metabolism studies of bavachinin through ultra‐high‐performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry

Bavachinin (BVC), one of the main bioactive prenylated flavonoids derived from Psoralea corylifolia Linn, has a wide variety of pharmacological effects, such as antiangiogenic, antitumor, antiallergic, anti‐inflammatory and antibacterial activities, especially as a pan‐peroxisome proliferator‐activated receptors agonist. A rapid and sensitive method for quantifying BVC in rat plasma was developed and validated through ultra‐high‐performance liquid chromatography coupled with electrospray‐ionization tandem mass spectrometry. Furthermore, a complete metabolic investigation of BVC was performed through ultra‐high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry. In the pharmacokinetic analysis, BVC exhibited rapid oral absorption (T_max = 0.68 ± 0.21 h), good elimination (T_1/2 = 2.27 ± 1.63 h) following oral administration and poor absolute bioavailability (5.27%). Moreover, 11 metabolites of BVC in plasma, urine, bile and feces were characterized. The main metabolic pathways of BVC involved isomeriszation, glucuronidation, sulfonation, hydroxylation, methoxylation and reduction. In conclusion, the present study provides a sensitive quantitative method with a lower limit of quantification of 1 ng/mL and an improved comprehension of the physiological disposition of BVC.     

Identification of lipophilic components in Citri Reticulatae Pericarpium cultivars by supercritical CO2 fluid extraction with ultra‐high‐performance liquid chromatography–Q Exactive Orbitrap tandem mass spectrometry

To systematically identify the lipophilic constituents of Citri Reticulatae Pericarpium from different cultivars, supercritical CO₂ fluid extraction and ultra‐high‐performance liquid chromatography–Q Exactive Orbitrap tandem mass spectrometry were integrated for the component analysis of 18 batches of Citri Reticulatae Pericarpium from 12 cultivars for the first time. A total of 57 components from the supercritical CO₂ fluid extracts were demonstrably or tentatively identified by the obtained parent peaks, fragment peaks, and retention times. In total, two flavonoids, six organic acids, nine coumarins, three aldehydes, seven esters, three terpenes, one limonoid, and five other compounds were detected for the first time; notably, coumarin components have not yet been reported in Citri Reticulatae Pericarpium. Furthermore, the extract constituents differed between cultivars. In particular, organic acids were more abundant in Citrus reticulata “Chachi” than in other cultivars, and pterostilbene was exclusively found in Citrus reticulata “Yichangju”. The results showed that a greater variety of compounds in Citri Reticulatae Pericarpium could be extracted by supercritical CO₂ fluid extraction and detected by ultra‐high‐performance liquid chromatography–Q Exactive Orbitrap tandem mass spectrometry. This study provides a more scientific basis for further analysis of the pharmacological activity and quality of Citri Reticulatae Pericarpium components from different cultivars.     

On-line capillary supercritical fluid chromatography-inductively coupled plasma mass spectrometry for the analysis of organometallic compounds

Supercritical fluid chromatography with inductively coupled plasma-mass spectrometric detection is a new and useful technique for the analysis of organometallic compounds. An interface has been developed for coupling these techniques. The interface is compact, easy to assembel, and causes minimal disruption to either instrument. A series of organotin, organoarsenic, and organorion compounds has been used to evaluate the interface and ICPMS as a detector for SFC. The effect of analyte concentration and restrictor temperature on peak intensity has been investigated and the effect of the mobile phase on the plasma has also been studied to assess background interference from changes in the plasma.     

Quantification of homoegonol in rat plasma using liquid chromatography–tandem mass spectrometry and its pharmacokinetics application

Homoegonol is a biologically active neolignan isolated from Styrax species with cytotoxic, antimicrobial, anti‐inflammatory and anti‐asthma activities. For the quantification of homoegonol in rat plasma, a selective and sensitive liquid chromatography–tandem mass spectrometric method was developed and validated for the first time using protein precipitation with methanol as a sample clean‐up procedure. The analytes were separated in an Atlantis dC₁₈ column using a gradient elution of methanol and 0.1% formic acid, and mass‐to‐charge ratios were determined in selective reaction monitoring mode using tandem mass spectrometry with m/z 343.12 > 296.97 for homoegonol and m/z 517.30 > 282.90 for udenafil (internal standard). The standard curve was linear over the concentration ranges of 1 ? 500 ng/mL using a 30 μL rat plasma sample. The coefficient of variation and relative error for intra‐ and inter‐assay at four quality control levels were 3.9–10.0 and ‐3.3–2.7%, respectively. The overall recovery of homoegonol from rat plasma using protein precipitation was 99.7 ± 7.7%. The pharmacokinetics parameters of homoegonol were dose‐independent after both intravenous (1, 2.5 and 5 mg/kg doses) and oral (5, 10 and 20 mg/kg doses) administration in male Sprague–Dawley rats. Copyright © 2014 John Wiley & Sons, Ltd.     

Determination of multi‐class antibiotic residues in compost by microwave‐enhanced accelerated solvent extraction and ultra performance convergence chromatography with tandem mass spectrometry

This work reports the development and application of a multi‐class compound analysis method for the determination of 20 antibiotic residues in compost. Samples were processed by microwave‐enhanced accelerated solvent extraction at 120°C for 7.5 min. Salting‐out homogeneous liquid‐liquid extraction was used to remove water and water‐soluble impurities from the extract before ultra performance convergence chromatography with tandem mass spectrometry analysis. By using the supercritical fluid (carbon dioxide) and organic solvent (methanol) as the mobile phase, the 20 antibiotics and the internal standard were well separated in 8.2 min without obvious matrix effect. Method validation was performed and good trueness (relative error in the range of ±5.0%) and precision (inter‐ and intraday relative standard deviations < 10.8%) were obtained. Method detection and quantitation limits were 0.8–1.9 and 2.7–7.1 ng/g, respectively. Recoveries were assessed at three concentration levels (10, 60, and 400 ng/g) and acceptable mean values (70.4–111.9%) were found. This method has also been used to analyze real samples, and the average concentrations of antibiotics (excepting the concentrations < method quantitation limits) were determined up to 123.6 ng/g. The results showed the method could be helpful for the analysis of multi‐class antibiotics in environmental samples.     

Method development and validation for the determination of biogenic amines in soy sauce using supercritical fluid chromatography coupled with single quadrupole mass spectrometry

Biogenic amines have been reported in many foods such as fish, meat, and soy sauce. The consumption of foods containing high concentrations of biogenic amines has been associated with health hazards. In this study, a green and efficient method using supercritical fluid chromatography coupled with single quadrupole mass spectrometry was developed for determination of biogenic amines in soy sauce. The chromatographic and mass spectrometry conditions were systematically optimized in terms of selectivity and peak shape. Nine biogenic amines were well separated within 25 min on a Cosmosil 5HP column using 5% (v/v) water and 0.2% (v/v) ammonia solution in methanol as mobile phase additives at a backpressure of 120 bar and temperature of 40°C. The established method was fully validated regarding the linearity, sensitivity, precision, and accuracy. The limits of detection and limits of quantification ranged from 0.03 to 10.50 μg/mL and 0.10 to 23.1 μg/mL, respectively. The relative standard deviations for intra‐ and interday precisions were all lower than 9.36% and the recoveries ranged from 75.82 to 99.63% and 80.10 to 99.89% for two levels of standards spiked in soy sauce, respectively. Finally, the established method was successfully applied to the quantitative analysis of biogenic amines in soy sauce.