1756 Key wordssanqishangyao tabletsuplccharacteristic chromatogramprincipal components analysishydroxysafflor yellow Apaeoniflorinneoeriocitrinnaringi

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1 * UPLC UPLC ACQUITY UPLC BEH C mm 2. 1 mm 1. 7 μm A- 0. 1% B 0 ~ 10 min 4% A 12% A10 ~ 22 min 12% A 16% A22 ~ 35 min 16% A 25% A 0. 2 ml min nm 40 / Chempattern TM PCA Score Loading UPLC 8 A 4 PCA UPLC UPLC A R917 A UPLC fingerprint analysis of Sanqishangyao tablets HU Xiao - ru 1 YANG Mei - li 1 2 SUN Lei 1 WANG Ming - juan 1 LI Ai - hua 3 DAI Zhong 1* MA Shuang - cheng 1 1. National Institutes for Food and Drug Control Beijing China2. Baoding Institute for Food and Drug Control Baoding China 3. Shenyang Green Pharmaceutical Co. Ltd. Shenyang China Abstract ObjectiveTo establish a UPLC chromatographic fingerprint analysis method of Sanqishangyao tablets. MethodsThe characteristic fingerprint was performed by HPLC and characteristic peaks were identified by LC - MS. The samples were separated on the ACQUITY UPLC BEH - C 18 column50 mm 2. 1 mm 1. 7 μm and e- luted with a gradient elution0-10 min 4% A 12% A10-22 min 12% A 16% A22-35 min 16% A 25% Aof acetonitrile as the mobile phase A and the solution containing 0. 1% formic acid as the mobile phase B at a flow rate of 0. 2 ml min - 1. The detection wavelength was 230 nm and the column temperature was 40. According to the chempattern TM advanced chemometrics solution a common pattern was built and statistically analyzed by principal components analysispca. The Score plot and the Loading plot were obtained with the retention time and the peak area for each main chromatographic peak as the variable. Four characteristic peaks in the fingerprints were identified with applying electrospray ionizationesisource in the positive /negative ion mode. ResultsThe characteristic fingerprint of Sanqishangyao tablets was established with naringin as the reference peak. Four characteristic peaks were identified as hydroxysafflor yellow A neoeriocitrin naringin paeoniflorin. Eight co - possessing peaks were designated as characteristic common peaks. The PCA showed the chemical compositions in the 27 samples were obviously different. The main difference of the chemical compositions was the content of neoeriocitrin naringin and paeoniflorin. ConclusionThe fingerprint method can reflect the actual quality with good stability and reproducibility and be applied in the quality control of Sanqishangyao tablets. * Tel E - maildaizhong@ nicpbp. org. cn Tel E - mailhuxru2008@ 126. com

2 1756 Key wordssanqishangyao tabletsuplccharacteristic chromatogramprincipal components analysishydroxysafflor yellow Apaeoniflorinneoeriocitrinnaringin UPLC 1 Agilent 6320 Ion Trap LC - MS Agilent 1200 DAD ACQUITY UPLC 1-2 Waters Empower2 Mettler XS105DU Millipore 2010 A % % % 2000ZFB Fisher Millipore Tab 1 1 Sources of Sanqishangyao tablets sample No. source batch sample No. source batch sample No. source batch 1 Anhui Inner Mongolia Shanghai Changchun Shandong Sichuan Gansu Shandong Sichuan Heilongjiang Shanxi Sichuan Heilongjiang Shanxi Shaanxi Jilin Shaanxi Shaanxi Jilin Shaanxi Shaanxi Jilin Shaanxi Yunnan Heilongjiang Shanghai Yunnan μm ACQUITY UPLC BEH C mm 1 ml 0. 4 mg 2. 1 mm 1. 7 μm A- 0. 1% B 0 ~ 10 min 4% A 12% A10 ~ 2 g 22 min 12% A 16% A22 ~35 min 16% A 25% A 20 ml ml min nm 40 W 40 khz 30 min 1 μl UPLC 1

3 Fig 1 HPLC chromatograms of methanol extract of Sanqishangyao tablets and medical materials A. Sanqishangyao tablets B. naringin ml min V ESI + ESI nm m /z 50 ~ MPa 2 2 ESI + A ESI - B ESI + CHPLC - UV230 nm D Fig2 Total ion chromatorgramtic ESI + A TICESI - B /MS ion chromatogramesi + Cand HPLC - UV230 nmcharacteristic fingerprintdof Sanqishangyao tablets RSD 1. 1% ~ 2. 2%

4 h RSD 1. 2% ~ % RSD 1. 7% ~ 3. 5% h UPLC Fig 3 UPLC chromatograms of 27 batches of Sanqishangyao tablets A. 1 ~ 9 samples 1-9 B. 10 ~ 18 samples C. 19 ~ 27 samples 19-27

5 A LC - MS Tab 2 2 Retention time and identified compounds of the characteristic peaks in HPLC fingerprints M r peak No. retention time/min identified compound precursor ion/m /z fragment ionsm /z λ max /nm Ahydro * xysafflor yellow A M - H 2 O + H M - 2H 2 O M + H + H M - glu + H M - glu - H M + H - 2 O + H * paeoniflorin M - Ar - COO M - glu + H M + H M - 2glu + H M - H 2 O - 2glu M - H - H M - CO + H M - 2H 2 O * neoeriocitrin M + H + H M - CO - CO + H M M - H - rha + H M - Orha + H M rha - C 7 H 6 O 2 + H M - rha - glu + H * naringin M - H 2 O + H M - 2H 2 O M + H + H M - 3H 2 O + H M - C 7 H M + Na + O M - H M - rha + H M - rha - H 2 O + H M - rha - glu + H * compound structures were identified by comparison with authentic standards UPLC 3 4 Chempattern TM PCA 27 Score PCA Loading 4 Score ~ min ~ 10 min UPLC Loading UPLC

6 Fig4 PCA analysis of the fingerprints of Sanqishangyao tablets A. common fingerprint pattern B. 1 2 Score 1 ~ 27 Score plot of 1-2 principal components based on the raw datasamples 1-27 C. 1 2 Loading 1-12 Loading plot of 1-2 principal components based on the raw datapeaks Ahydroxysafflor yellow A 9. paeoniflorin 11. neoeriocitrin 12. naringin UPLC HPLC UPLC μl A UPLC 2

7 Fig 5 Comparison between chromatograms of outlier sample 8 24 and 10 and common patterns for Sanqishangyao tablets A. 8 sample 8upperand fingerprint common patternb. 24 sample 24upperand fingerprint common patternc. 10 sample 10upperand fingerprint common pattern

8 1762 1FENG Jia - tao JIN Yu WANG Jin - cheng et al. Development of full - quantified fingerprint for J. Chin Hosp Pharm J quality control of traditional Chinese medicines J. Chin J Chromatogr Xie PS Chen SB Liang YZ et al. Chromatographic fingerprint a- nalysis a rational approach for quality assessment of traditional Chinese herbal medicinej. J Chromatogr A LUO Zheng - liang FENG Xiu - zhen. 12SHI Jun - min GUAN Jia ZHANG Qing - HPLC method for the determination of the Sanqi Shangyao tablets wen et al. Application of ultra performance liquid of paeoniflorin and naringinhplc chromatography in pharmaceutical analysis J. Guid Chin Med J. Chin J Pharm Anal YAO Yu - duan FU Yu JIANG Ping et al. Determination of notoginsenoside R 1 ginsenoside Rg 1 and Rb 1 in Sanqishangyao tablets by HPLC 3 J. Chin Pharm QIU Shao - hua ZHOU Li - zhang QIU Jie - fen. Determination of paeoniflorin in Sanqishangyao tablets by HPLC 9SUN Yan - tao ZHAO Lei WEI Hong - li et al. Simultaneous determination for the content of 4 kinds of constituents in Sanqishangyao capsules by HPLC 4 J. Herb. Med LI Xiao - hong XIONG Zhi - li YU Ming - 3ChP Vol Ⅰ S yang. High performance liquid chromatographic finger- 4WANG Jian - fei. Determination of aconitine mesaconitine prints of ethanol and cyclohexane extracts of Rhizoma Drynariae and hypaconitine in Sanqi Shangyao tablets from different factories by and quantitative analysis of index components based on principal HPLCHPLC component analysis J. Chin Tradit Pat Med YANG Mei - li DAI Zhong HU Xiao - ru J. Chin J Chromatogr et al. UPLC simultaneous determination of the content for 11Xu SJ Yang L Tian RT et al. Species differentiation and quality 4 constituents in Sanqi Shangyao tabletsuplc assessment of Radix Paeoniae Rubra Chi - shaoby means of 4 J. Chin J Pharm Anal high - performance liquid chromatographic fingerprint J. J Chromatogr A

中草药 Chinese Traditional and Herbal Drugs 第 5 卷第 1 期 201 年 1 月 51 其他试剂均为分析纯对照品栀子苷 ( 批号 1109-201115) 甘草苷 ( 批号 1110-201106) 连翘酯苷

中草药 Chinese Traditional and Herbal Drugs 第 5 卷第 1 期 201 年 1 月 51 其他试剂均为分析纯对照品栀子苷 ( 批号 1109-201115) 甘草苷 ( 批号 1110-201106) 连翘酯苷 50 中草药 Chinese Traditional and Herbal Drugs 第 5 卷第 1 期 201 年 1 月凉膈散汤剂 HPLC-DAD 指纹图谱研究 * 胡建兰, 邵萌, 余林中南方医科大学中医药学院, 广东广州 510515 摘要 : 目的研究凉膈散汤剂 HPLC-DAD 指纹图谱, 建