目的: 建立同时测定清肝降压胶囊中10 个成分的高效液相色谱一测多评（HPLC-QAMS）方法, 并验证其准确性和可行性。方法: 采用Phenomsil C18 色谱柱（4. 6 mm×250 mm, 5 μm）, 以乙腈-0. 1% 磷酸水溶液为流动相, 梯度洗脱;流速: 1. 0 mL·min-1;柱温: 30 ℃ ;检测波长: 320 nm（检测2, 3, 5, 4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷）、250 nm（检测3′-羟基葛根素、葛根素和3'-甲氧基葛根素）、208 nm（检测泽泻醇F、泽泻醇A、24-乙酰泽泻醇A 和23-乙酰泽泻醇B）和270 nm（检测隐丹参酮和丹参酮ⅡA）。以葛根素为内参物, 建立其与其他9 种成分的相对校正因子（RCF）, 并计算含量, 实现一测多评;同时采用外标法（ESM）测定清肝降压胶囊中10 个成分的含量, 比较一测多评法计算值与外标法实测值的差异。结果: 10 个成分的线性范围分别为8. 136~162. 7、7. 488~149. 8、30. 54~610. 9、14. 11~282. 2、0. 368 0~7. 360、0. 704 0~14. 08、0. 432 0~8. 640、3. 056~61. 12、0. 512 0~10. 24、0. 848 0~16. 96 μg·mL-1, r 为0. 999 2~0. 999 9;平均回收率分别为98. 5%、98. 0%、100. 0%、100. 0%、97. 8%、98. 5%、97. 0%、99. 1%、99. 3% 和100. 1%, RSD 为1. 3%、1. 1%、0. 96%、0. 64%、1. 6%、1. 5%、1. 2%、0. 83%、1. 1% 和0. 72%;葛根素与2, 3, 5, 4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、3′-羟基葛根素、3'-甲氧基葛根素、泽泻醇F、泽泻醇A、24-乙酰泽泻醇A、23-乙酰泽泻醇B、隐丹参酮、丹参酮ⅡA 的相对校正因子分别为1. 155 1、1. 126 2、1. 276 0、0. 964 3、0. 895 9、0. 859 3、1. 366 2、0. 839 9 和0. 966 3;一测多评法计算值和外标法实测值之间无明显差异。结论: 所建立的方法简便、准确, 可用于清肝降压胶囊的质量评价研究。
Objective: To establish an HPLC-QAMS method for simultaneous determination of 10 components in Qinggan Jiangya capsules, and validate the accuracy and feasibility of the method. Methods: A Phenomsil C18（4. 6 mm×250 mm, 5 μm）column was used in the HPLC assay. Acetonitrile-0. 1% phosphoric acid aqueous solution with gradient elution was employed as the mobile phase. The flow rate was 1. 0 mL·min-1, the column temperature was maintained at 30 ℃ and the detection wavelength were set at 320 nm for 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside, 250 nm for 3'-hydroxypuerarin, puerarin and 3'-methoxypuerarin, 208 nm for alisol F, alisol A, alisol A-24-acetate and alisol B 23-acetate, and 270 nm for cryptotanshinone and tanshinone ⅡA. Puerarin was used as the internal reference substance to calculate the relative correction factors（RCFs）of others nine components, and evaluate their contents accordingly. At the same time, the external standard method（ESM）was used to quantify the ten components in Qinggan Jiangya capsules. The results of the ESM and QAMS method were compared. Results: The linear range of 10 components were 8. 136-162. 7, 7. 488-149. 8, 30. 54-610. 9, 14. 11-282. 2, 0. 368 0-7. 360, 0. 704 0-14. 08, 0. 432 0-8. 640, 3. 056-61. 12, 0. 512 0-10. 24 and 0. 848 0-16. 96 μg·mL-1（r=0. 999 2-0. 999 9）, respectively. The average recoveries were 98. 5%, 98. 0%, 99. 8%, 100. 0%, 97. 8%, 98. 5%, 97. 0%, 99. 1%, 99. 3% and 100. 1% with RSDs of 1. 3%, 1. 1%, 0. 96%, 0. 64%, 1. 6%, 1. 5%, 1. 2%, 0. 83%, 1. 1% and 0. 72%, respectively. RCFs of 2, 3, 5, 4′-tetrahydroxystilbene-2-O-β-D-glucoside, 3′-hydroxypuerarin, 3'-methoxypuerarin, alisol F, alisol A, alisol A-24-acetate, alisol B 23-acetate, cryptotanshinone and tanshinone ⅡA to puerarin were 1. 155 1, 1. 126 2, 1. 276 0, 0. 964 3, 0. 895 9, 0. 859 3, 1. 366 2, 0. 839 9 and 0. 966 3, respectively. Significant difference was not found between the calculated values of QAMS method and the measured values of ESM. Conclusion: The method is simple and accurate, and can be used for quality evaluation of Qinggan Jiangya capsules.
[1] WS3-211(Z-211)-2001(Z). 国家药品监督管理局国家药品标准[S]. 2001
WS3-211(Z-211)-2001(Z). National Medical Products Administration National Drug Standards[S]. 2001
[2] 朱玉梅, 彭淑莲, 鲁卫星, 等. 清肝降压胶囊治疗高血压病临床观察[J]. 中国中医基础医学杂志, 2003, 9(4): 61
ZHU YM, PENG SL, LU WX, et al. Clinical observation on Qinggan Jiangya capsule in the treatment of hypertension[J]. Chin J Basic Med Tradit Chin Med, 2003, 9(4): 61
[3] 戴勤芳, 马丽红. 清肝降压胶囊治疗高血压临床疗效观察[J]. 中国全科医学, 2011, 14(29): 3410
DAI QF, MA LH. Effects of Qinggan Jiangya capsule in treating patients with hypertension[J]. Med China Family Phys, 2011, 14(29): 3410
[4] 吴玉兰, 何小红. 清肝降压胶囊治疗高血压的临床疗效及安全性观察[J]. 中国实用医药, 2012, 7(33): 136
WU YL, HE XH. Clinical efficacy and safety of Qinggan Jiangya capsule for hypertension[J]. China Pract Med, 2012, 7(33): 136
[5] 徐玲玲. 分析清肝降压胶囊联合硝苯地平缓释片治疗老年性高血压的临床疗效[J]. 医学信息, 2014, 27(10): 74
XU LL. Effects of Qinggan Jiangya capsule combined with extended release nifedipine tablets on senile hypertensive patients[J]. Med Inf, 2014, 27(10): 74
[6] 张晶, 张华, 刘芳勋, 等. 清肝降压胶囊联合氨氯地平治疗中老年人高血压合并睡眠障碍患者的临床观察[J]. 中国生化药物杂志, 2016, 36(1): 47
ZHANG J, ZHANG H, LIU FX, et al. Effects of Qinggan Jiangya capsule and amlodipine in treatment of middle-aged and old patients with hypertension combined with sleep disorders[J]. Chin J Biochem Pharm, 2016, 36(1): 47
[7] 冯喜群. 清肝降压胶囊联合氨氯地平治疗老年高血压合并睡眠障碍35 例临床观察[J]. 中国民族民间医药, 2016, 25(18): 92
FENG XQ. Clinical observation on Qinggan Jiangya capsule combined with amlodipine therapy treating 35 patients with old patients with hypertension combined with sleep disorders[J]. Chin J Ethnomed Ethnopharm, 2016, 25(18): 92
[8] 中华人民共和国药典2015 年版. 一部[S]. 2015: 175
ChP 2015. Vol ⅠhP 2015. Vol Ⅰ[S]. 2015: 175
[9] 尤春雪, 张振秋, 李峰, 等. HPLC 波长切换技术对葛根中8 种成分的测定及指纹图谱研究[J]. 中草药, 2013, 44(5): 616
YOU CX, ZHANG ZQ, LI F, et al. Determination and fingerprint analysis of eight constituents in Puerariae Radix by HPLC wavelength switching[J]. Chin Tradit Herb Drugs, 2013, 44(5): 616
[10] 赵路, 关琴笑, 路丽, 等. UHPLC 法同时测定不同产地葛根中5种活性成分的含量[J]. 中药材, 2015, 38(3): 473
ZHAO L, GUAN QX, LU L, et al. Simultaneous determination of five active components in Puerariae Lobatae Radix from different habitats by UHPLC[J]. J Chin Med Mater, 2015, 38(3): 473
[11] 赵万里, 黄小强, 许文, 等. RP-HPLC-DAD 同时测定泽泻中11个三萜类成分[J]. 中草药, 2016, 47(16): 2933
ZHAO WL, HUANG XQ, XU W, et al. Simultaneous determination of 11 triterpenoids in Alismatis Rhizoma by RP-HPLC-DAD[J]. Chin Tradit Herb Drugs, 2016, 47(16): 2933
[12] 曹柳, 李青苗, 方清茂, 等. 3 种炮制方法对泽泻中4 种三萜类成分的影响[J]. 中成药, 2016, 38(9): 1994
CAO L, LI QM, FANG QM, et al. Effects of three processing methods on four triterpenoids in Alismatis Rhizoma[J]. Chin Tradit Pat Med, 2016, 38(9): 1994
[13] 翟宏宇, 单柏宇, 王海洋, 等. 一测多评法测定保心宁片中6 个丹参类指标性成分的含量[J]. 药物分析杂志, 2018, 38(6): 973
ZHAI HY, SHAN BY, WANG HY, et al. Simultaneous determination of six index components in Baoxinning tablets by a single marker[J]. Chin J Pharm Anal, 2018, 38(6): 973
[14] 朱培芳, 赵紫伟, 赵荣华. 高效液相色谱法测定12 个市售地何首乌中有效成分的含量[J]. 中华中医药杂志, 2014, 29(11): 3605
ZHU PF, ZHAO ZW, ZHAO RH. Determination the effective component of 12 commercial polygonum multiflorum by HPLC[J]. China J Tadit Chin Med Pharm, 2014, 29(11): 3605
[15] 童凯, 李昭玲, 闫燊, 等. 川牛膝酒炙和盐炙前后HPLC 化学指纹图谱及其主要药效成分量变化研究[J]. 中草药, 2016, 47(4): 580
TONG K, LI ZL, YAN S, et al. HPLC fingerprint of crude Cyathula officinalis and its processed products stir-fried with wine and salt and variation of their index components[J]. Chin Tradit Herb Drugs, 2016, 47(4): 580
[16] 王珏, 朱育凤, 姚毅, 等. HPLC 同时测定何首乌中5 种成分及指纹图谱的建立[J]. 中国中医药信息杂志, 2019, 26(6): 75
WANG J, ZHU YF, YAO Y, et al. Simultaneous determination of five components in Polygoni Multiflori Radix by HPLC and establishment of fingerprints[J]. Chin J Inf Tradit Chin Med, 2019, 26(6): 75
[17] 程沛, 韩东岐, 胡伟慧, 等. 高效液相色谱法同时测定丹参中10个水溶性和4 种脂溶性成分的含量[J]. 药物分析杂志, 2015, 35(6): 991
CHENG P, HAN DQ, HU WH, et al. Simultaneous determination of ten hydrophilic compounds and four lipophilic compounds in Radix Salviae Miltiorrhizae by HPLC[J]. Chin J Pharm Anal, 2015, 35(6): 991
