ABSTRACT
OBJECTIVE: To establish a high performance liquid chromatography coupled with tandem mass spectrometry and ultraviolet detector (HPLC-UV-MS) method for determination of the mixture of Glycyrrhiza Radix extract at detoxification dosage and Semen Strychni extract at poisonous dosage, which will be applied to the characteristic ingredients profiling in rat plasma. METHODS: Sino Chrome ODS-BP column (4.6 mm×250 mm, 5 μm) was employed, which was maintained at 40℃, and the mobile phase was composed of water (0.1% formic acid) (A) and acetonitrile (B). The gradient elution program was as follows: 0.01 min 8% B, 2 min 8% B, 30 min 25% B, 32 min 30% B, 50 min 45% B, 53 min 85% B, 60 min 85% B, 61 min 8% B, and terminated at 77 min, eluted at a flow rate of 0.3 mL·min-1. A UV detector was equipped and the detection wavelength was 254 nm. ESI+ mode was selected for strychnos alkaloids with cone voltage of 30 V and capillary voltage of 2.5 V, while ESI- mode for glycyrrhiza with cone voltage of 25 V and capillary voltage of 3.5 V. Acetonitrile was used as protein precipitation agent to determine the characteristic ingredients in rat plasma. RESULTS: An HPLC-UV-MS method for simultaneous determination of liquorice and Semen Strychni extract was established and applied to measure the components absorbed into blood. The material changes of liquorice and Semen Strychni in vivo and vitro were also compared. CONCLUSION: A simple, repeatable HPLC-UV-MS method for analysis of Glycyrrhiza Radix and Semen Strychni in vivo and vitro was established. The related components are speculates, which provided the experimental basis to research the detoxification mechanism of Glycyrrhiza Radix against strychninization.
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OBJECTIVE: To establish methods for the test of chemical dyes and quantitative determination of dracorhodin perchlorate in Guanmaining tablets. METHODS: HPLC-UV-MS/MS was used for the test of chemical dyes in Guanmaining tablets, and the concentration of dracorhodin perchlorate was determined by HPLC. In the test of chemical dyes, Agilent Zorbax-SB C18 (4.6 mm × 250 mm, 5 μm) was used as stationary phase, the mixture of 0.02 mol · L-1 ammonium acetate and acetonitrile was used as mobile phase in gradient elution mode, and the UV-Vis detection wavelength was set at 520 nm. ESI+ was used as ion source, and the MS/MS spectra of the analytes were recorded to confirm the positive results. In the quantitative analysis of dracorhodin perchlorate, Phenomenex Luna C18(4.6 mm × 250 mm, 5 μm) was used as stationary phase, the mixture of acetonitrile and 0.05 mol · L-1 sodium dihydrogen phosphate solution (37:63) was used as mobile phase, and the detection wavelength was set at 440 nm. RESULTS: Among 56 batches of Guanmaining tablets, 7 batches showed positive results of sudan I, sudan IV, and/or 808 scarlet. A good linear relationship was obtained in the range of 0.408 0 - 816.6 μg · mL-1 for dracorhodin perchlorate, with the linear regression equation of y = 1.746 1 × 107ρ +2 419.9(r = 1.000 0); the average recovery was 98.37% (RSD = 1.2%, n =6). The content of dracorhodin perchlorate in Guanmaining tablets ranged from 0.01 to 0.25 mg · tablet-1. CONCLUSION: The method is simple, accurate and rapid, and can be used for the quality control of Guanmaining tablets.
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Objective: To establish the multiple fingerprints of Danhong Injection (DI) using HPLC-UV-MS method and to simultaneously determine nine kinds of the medicinal components in DI. Methods: Separation was performed on Atiantis T3 analytical column (250 mm × 4.6 mm, 5 μm) with the mobile phase of 0.05% formic acid-50% acetonitrile by gradient elution, and negative-ion SIM mode was selected for mass spectrometric detection. Results: The multiple fingerprints reflected the chemical information of Salvia miltiorrhiza and safflower in DI. The similarity of the fingerprints was higher than 0.988 in all 11 batches of DI. 5-Hydroxymethyl furfural, sodium danshensu, protocatechuic aldehyde, coumaric acid, salvianolic acid D, rosmarinic acid, salvianolic acid B, salvianolic acid A, and kaempferol-3-O-rutinoside showed the good linearity in their respective ranges of concentration, r ≥ 0.999 0.The average recovery of low-, mid-, and high-dose medicinal components (n = 9) was (99.0 ± 1.4)%, (102.0 ± 1.7)%, (99.3 ± 1.6)%, (97.6 ± 1.6)%, (100.0 ± 1.8)%, (97.9 ± 1.6)%, (100.5 ± 4.4)%, (100.6 ± 2.0)%, and (106.0 ± 4.7)%, respectively. The relative standard deviation for the method reproducibility was lower than 1.45%. The total contents of nine medicinal components in the 11 batches of DI were 2.61-3.06 mg/mL. Conclusion: This method is simple, accurate, and with good reproducibility, and the multiple fingerprints combined with the quantitative analysis could reflect the quality of DI better, which could be used to control the quality of DI.
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AIM: To develop a method of the HPLC/UV/MS/MS fingerprints of Qingkailing Injection(Radix Isatidis,Fructus Gardeniae,Flos Lonicerae Japonicae,etc.) and provide a novel conception for its quality control. METHODS: Separation was performed on Phenomenex Luna C_(18) analytical column(250 mm?4.6 mm,5 ?m).The mobile phases consisted of water containing 0.1% formic acid and acetonitrile was used as gradient elute.The flow rate was 1 mL/min.An Electrospray Ion Trap mass spectrometer was utilized for qualitative analysis and both positive and negative secondary ion scan mode were applied. RESULTS: Perfect fingerprints with excellent separation and reproducibility were obtained and 13 compounds were identified.Especially three isomers,including Neochlorogenic acid,Chlorogenic acid and Isochlorogenic acid,were differentiated successfully. CONCLUSION: The method is suitable for establishing the HPLC/MS/MS fingerprints of Qingkailing Injection and hence could be used as an effective and credible pattern for analyzing traditional Chinese medicine as well as its quality control.Moreover,based on this approch,further traditional Chinese medicine research can be performed.
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AIM To develop a comparative method for studying on the HPLC/UV/MS fingerprints of triterpene acid in the leaves and the seeds of Lagerstroemia specious L. METHODS Separation was performed on LunaC_ 18 (2.6 mm?250 mm, 5 ?)analytical column, The mobile phase was composed of acetonitrile and 0.1 % formic acid as gradient eluent. The flow rate was 1.0 ml?min -1 . RESULTS Both of corosolic acid and maslinic acid existed in the leaves and the seeds of Lagerstroemia specious L. Ursolic acid existed in the leaves and the oleanolic acid contained in the seeds of Lagerstroemia speciouswere L. CONCLUSION The method can be used for quality control of the leaves and the seeds of Lagerstroemia specious L.