ABSTRACT
OBJECTIVE：To identify t he chemical constituents of Fengliaoxing fengshi dieda wine. METHODS ：An ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry （UPLC-Q-Exactive- MS）technique was used for identifying chemical constituents of Fengliaoxing fengshi dieda wine. The determination was performed on Thermo Accucore aQ RP 18 column with mobile phase consisted of 0.1％ formic acid-methanol （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was 30 ℃，and the sample size was 2 μL. HESI source was adopted，ion mode was positive and negative ion mode ，nitrogen as sheath gas and auxiliary gas. The positive ion mode had spray voltage of 3.5 kV， capillary heating temperature of 350 ℃，sheath gas pressure of 35 psi，auxiliary gas pressure of 15 arb，and ion source heating temperature of 320 ℃. The negative ion mode had spray voltage of 3.2 kV，capillary heating temperature of 350 ℃， sheath gas pressure of 35 psi，auxiliary gas pressure of 15 arb，and ion source heating temperature of 300 ℃. The mass axis was calibrated by external standard method （mass error less than 5 ppm）. The scanning range of the first mass spectrometry was m/z 80.0-1 200.0 （the resolution was 70 000），the scanning range of secondary mass spectrometry was m/z 80.0-1 200.0（the resolution was 17 500），and the collision voltage was 20，40，60 eV. Retrieved from CNKI ，VIP，PubMed and other database ，the chemical constituents information of each Chinese traditional medicine in Fengliaoxing fengshi dieda wine were collected to establish chemical constituents database. The structure of the compounds was identified on the basis of above constituents database ，the relevant literature ，retention time of reference substance and MS fragmentation regularity. RESULTS & CONCLUSIONS ： Fifty-nine compounds were identified in Fengliaoxing fengshi dieda wine ，including 12 flavones（e.g. neoeriocitrin ，hesperidin.），8 alkaloids （e.g. baogongteng C or erycibellin ， ephedrine，pseudoephedrine）， 9 organic acids （e.g. chlorogenic acid ， cryptochlorogenic acid ，neochlorogenic acid ），7 coumarins（e.g. xanthotoxol）， 4 esters， 4 amino acids ， and 15 other categories（including volatile oils ，terpenes，amides）. UPLC- 5103YX11SF37（17）] Q-Exactive-MS technology can quickly and accurately identify the chemical constituents in Fengliaoxing fengshi dieda wine.
ABSTRACT
OBJECTIVE：To establish an HPLC fingerprint for Fengliaoxing fengshi dieda wine ，and to determine the contents of 10 effective constituents. METHODS ：HPLC method was adopted. The determination was performed on Inertsil ODS- 3 C18 column with mobile phase consisted of acetonitrile - 0.1％ phosphoric acid water （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 25 ℃ ，and detection wavelength was set at 210 nm（10-15 min，hydrochloride ephedrine ， hydrochloride pseudoephedrine ），300 nm（70-120 min，cinnamaldehyde），345 nm（15-70 min，neochlorogenic acid ，scopolin， chlorogenic acid ，cryptochlorogenic acid ，scopoletin，isochlorogenic acid A ，isochlorogenic acid C ）. The sample size was 10 μL. Using scopoletin peak as reference ，HPLC fingerprints of 10 batches of samples were drawn. The similarity evaluation was performed by using Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）to confirm common peak. RESULTS：There were 18 common peaks in HPLC chromatograms of 10 batches of samples ，and the similarity was above 0.980. Totally 12 components including hydrochloride ephedrine ， hydrochloride pseudoephedrine ， neochlorogenic acid ， scopolin， chlorogenic acid ，cryptochlorogenic acid ，scopoletin，isochlorogenic acid B ，narigin，isochlorogenic acid A ，isochlorogenic acid C and cinnamaldehyde widentified. The linear range of hydrochloride ephedrine ，hydrochloride pseudoephedrine ，neochlorogenic acid， scopolin， chlorogenic acid ， cryptochlorogenic acid ， scopoletin， isochlorogenic acid A ， isochlorogenic acid C and cinnamaldehyde were 2.475-247.5，2.600-260.0，3.820-382.0，3.900-390.0，3.060-306.0，4.760-476.0，4.540-454.0，4.900-490.0， 4.540-454.0，7.590-759.0 μg/mL（r＞0.999 0）. The limits of quantitation were 0.320 0，0.350 0，0.021 4，0.024 3，0.036 8，0.025 7， 0.152 1，0.042 9，0.025 1，0.350 3 μg/mL，respectively；the limits of detection were 0.160 0，0.180 0，0.007 9，0.004 0，0.001 2， 0.007 3，0.076 0，0.001 4，0.008 1，0.201 4 μg/mL，respectively. RSDs of precision ，stability and reproducibility tests were all lower than 2％. The recoveries were 95.03％-106.85％（RSD were 0.67％-2.68％，n＝6）．The contents were 0.013 3- 0.214 1 mg/mL. CONCLUSIONS：Established fingerprint is stable ，accurate and specific ，and can be used for quality control of Fengliaoxing fengshi dieda wine ；the content determination method is rapid ，accurate and reliable ，and can be used for simultaneous determination of 10 components.