The present study was designed to develop and validate a rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with tandem mass spectrometry(UHPLC-MS/MS) method for the simultaneous deter...The present study was designed to develop and validate a rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with tandem mass spectrometry(UHPLC-MS/MS) method for the simultaneous determination of five major active constituents in the traditional Chinese medicinal preparation Xingxiong injection(XXI) in rat plasma, including quercetin 3-O-rutinoside(QCR), kaempferol 3-O-rutinoside(KFR), isorhamnetin 3-O-rutinoside(ISR), bilobalide(BB), and ligustrazine(LGT). The plasma samples were pretreated by protein precipitation with acetonitrile. The chromatographic separation was achieved on a Waters Symmetry C18 analytical column(2.1 mm × 100 mm, 3.5 mm) with a mobile phase of 0.1% aqueous formic acid(A)-acetonitrile(B). Quantitation of the five bioactive constituents was achieved. Naringin was used as the internal standard(IS). All the calibration curves showed good linearity(r > 0.996) over the concentration range, with the lowest limit of quantification(LLOQ) between 2-18 ng·m L^(-1). The intraand inter-day accuracy and precision of the analytes were both within acceptable limits. Moreover, satisfactory extraction recoveries(90.92%-104.03%) were obtained by protein precipitation. The validated method was successfully applied to a pharmacokinetic study of XXI in rats after intravenous administration at three doses. The pharmacokinetic parameters of the five compounds varied in a dose-dependent manner within the tested dosage range. The present study was the first report of pharmacokinetic study for XXI.展开更多
Xingxiong injection(XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacologic...Xingxiong injection(XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacological studies, chemical analysis and quality control studies on this formula are relatively limited. In the present study, a high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in XXI. According to the fragmentation rules and previous reports, thirty ginkgo flavonoids, four ginkgo terpene lactones, and one alkaloid were identified. A high performance liquid chromatography coupled with triple quadrupole mass spectrometry(HPLC-QQQ MS) method was then applied to quantify ten major constituents in XXI. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. The total contents of ginkgo flavonoids were about 22.05-25.51 μg·mL-1 and the ginkgo terpene lactones amounts were about 4.41-8.70 μg·m L-1 in six batches of XXI samples, respectively. Furthermore, cosine ratio algorithm and distance measurements were employed to evaluate the similarity of XXI samples, and the results demonstrated a high-quality consistency. This work could provide comprehensive information on the quality control of Xingxiong injection, which be helpful in the establishment of a rational quality control standard.展开更多
基金financially supported from National Natural Science Foundation of China(Nos.81673569,and 81473343)Program for New Century Excellent Talents in University(No.NECT-13-1034)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘The present study was designed to develop and validate a rapid, sensitive, and reliable ultra-high performance liquid chromatography coupled with tandem mass spectrometry(UHPLC-MS/MS) method for the simultaneous determination of five major active constituents in the traditional Chinese medicinal preparation Xingxiong injection(XXI) in rat plasma, including quercetin 3-O-rutinoside(QCR), kaempferol 3-O-rutinoside(KFR), isorhamnetin 3-O-rutinoside(ISR), bilobalide(BB), and ligustrazine(LGT). The plasma samples were pretreated by protein precipitation with acetonitrile. The chromatographic separation was achieved on a Waters Symmetry C18 analytical column(2.1 mm × 100 mm, 3.5 mm) with a mobile phase of 0.1% aqueous formic acid(A)-acetonitrile(B). Quantitation of the five bioactive constituents was achieved. Naringin was used as the internal standard(IS). All the calibration curves showed good linearity(r > 0.996) over the concentration range, with the lowest limit of quantification(LLOQ) between 2-18 ng·m L^(-1). The intraand inter-day accuracy and precision of the analytes were both within acceptable limits. Moreover, satisfactory extraction recoveries(90.92%-104.03%) were obtained by protein precipitation. The validated method was successfully applied to a pharmacokinetic study of XXI in rats after intravenous administration at three doses. The pharmacokinetic parameters of the five compounds varied in a dose-dependent manner within the tested dosage range. The present study was the first report of pharmacokinetic study for XXI.
基金supported by the National Natural Science Foundation of China(Nos.81473343 and 81202898)Project in the National Science&Technology Pillar Program during the Twelfth Five-year Plan Period(No.2012BAI29B07)+2 种基金Program for New Century Excellent Talents in University(No.NECT-13-1034)Natural Science Foundation of Jiangsu Province(No.BK20130652)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Xingxiong injection(XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacological studies, chemical analysis and quality control studies on this formula are relatively limited. In the present study, a high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in XXI. According to the fragmentation rules and previous reports, thirty ginkgo flavonoids, four ginkgo terpene lactones, and one alkaloid were identified. A high performance liquid chromatography coupled with triple quadrupole mass spectrometry(HPLC-QQQ MS) method was then applied to quantify ten major constituents in XXI. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. The total contents of ginkgo flavonoids were about 22.05-25.51 μg·mL-1 and the ginkgo terpene lactones amounts were about 4.41-8.70 μg·m L-1 in six batches of XXI samples, respectively. Furthermore, cosine ratio algorithm and distance measurements were employed to evaluate the similarity of XXI samples, and the results demonstrated a high-quality consistency. This work could provide comprehensive information on the quality control of Xingxiong injection, which be helpful in the establishment of a rational quality control standard.
