Direct and comprehensive analysis of ginsenosides and diterpene alkaloids in Shenfu injection by combinatory liquid chromatography-mass spectrometric techniques.
Shenfu injection (SFI) is a widely used Chinese herbal formulation for cardiac diseases prepared from red ginseng and processed aconite root. Clinical observations and pharmacological effects on SFI have been well investigated. Chemical analysis and quality control studies of this formulation, however, are relatively limited, especially regarding toxic aconite alkaloids. In this work, 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 SFI. Highly sensitive MS allows direct analysis of injections without additional sample pretreatment required. Using diagnostic ions and fragmentation rules, we identified 23 trace diterpene alkaloids, nineteen ginseng saponins, one panaxytriol, and one 5-hydroxymethylfurfural in SFI. A LC-MS method with selected ion monitoring was then used to quantify 24 major alkaloids and ginsenosides. The method was validated in terms of linearity, accuracy and precision. Especially, the limits of quantification were low to 0.4-18ng/mL for diterpene alkaloids. The total concentrations of saponins and alkaloids were about 676-742μg/mL and 3-7μg/mL in five batches of SFI samples, respectively. Finally, cosine ratio and euclidean distance were introduced to evaluate the batch-to-batch reproducibility of SFI samples, and the results demonstrated high quality consistency. Global identification and quantification of complex constituents based on LC-MS promises wide applications in quality control and batch monitoring for herbal products.
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
Hua
H
Yang
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
Lei
L
Liu
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
Wen
W
Gao
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
Ke
K
Liu
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China. Electronic address: fleude@126.com.
Lian-Wen
LW
Qi
State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China. Electronic address: liping2004@126.com.
Ping
P
Li
D000470:Alkaloids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004224:Diterpenes / Q000737:chemistry*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D053281:Enediynes / Q000737:chemistry; D005233:Fatty Alcohols / Q000737:chemistry; D036145:Ginsenosides / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D005894:Panax / Q000737:chemistry; D015203:Reproducibility of Results; D012503:Saponins / Q000737:chemistry
D000470:Alkaloids; D004224:Diterpenes; D004365:Drugs, Chinese Herbal; D053281:Enediynes; D005233:Fatty Alcohols; D036145:Ginsenosides; D012503:Saponins; C492793:Shen-Fu; C057682:panaxytriol
Direct and comprehensive analysis; Diterpene alkaloids; Ginsenosides; Quadrupole time-of-flight mass spectrometry; Shenfu injection