目的 明确苦参总黄酮抗菌活性的化学成分,建立高效液相色谱法同时测定7种黄酮成分的方法。方法 采用硅胶柱、ODS柱色谱等方法进行分离纯化,综合理化性质和波谱学数据进行结构鉴定。HPLC同时测定7种黄酮成分,采用Dikma C18色谱柱,甲醇-水梯度洗脱,检测波长280 nm。结果 从苦参总黄酮抗菌提取物中分离鉴定7个化合物,分别为苦参啶(1)、槐属二氢黄酮G (2)、苦参酮(3)、异脱水淫羊藿素(4)、异黄腐醇(5)、芒柄花素(6)和三叶豆紫檀苷(7)。上述7种成分依次在质量浓度8.70~87.00、44.25~442.50、128.10~1 281.00、9.40~94.00、48.40~484.00、14.20~142.00、25.70~257.00 μg·mL-1内线性关系良好,r值均大于0.999 0,加样回收率均符合要求。其中,苦参酮、异黄腐醇、槐属二氢黄酮G含量相对高,分别占提取物的(16.24±0.02)%、(7.60±0.02)%、(6.59±0.03)%。结论 苦参总黄酮抗菌作用的主要化学成分可能是苦参酮、异黄腐醇、槐属二氢黄酮G等。所建立的HPLC测定法简便、准确,可用于同时测定苦参总黄酮中7种成分。
Abstract
OBJECTIVE To study the chemical constituents of the total flavonoids from Sophora flavescens and establish a method for simultaneous determination of seven compounds. METHODS The compounds were isolated by chromatography on silica gel and ODS column and their structures were elucidated by spectroscopic analysis. The samples were analyzed on a Dikma C18 column (4.6 mm×250 mm,5 μm); gradient elution was performed using mobile phase composed of methanol (A)and water (B); the detection was carried out using a photodiode array detector at 280 nm. RESULTS Seven compounds were isolated and their structures were identified as kuratidine (1), sophoraflavanone G (2), kurarinone (3), isoanhydroicaritin (4), isoxanthohumol (5), formononetin (6), and trifolirhizin (7). The calibration curve was linear within 8.70-87.00, 44.25-442.50, 128.10-1 281.00, 9.40-94.00, 48.40-484.00, 14.20-142.00, and 25.70-257.00 μg·mL-1 for kuraridine, sophoraflavanone G, kurarinone, isoanhydroicaritin, isoxanthohumol, formononetin, and trifolirhizin, respectively (r>0.999 0), and the extraction recoveries varied from 95% to 105%. CONCLUSION The main chemical components contributing to antibacterial activity of total flavonoids may be sophoraflavanone G, kurarinone, and isoxanthohumol. The method is simple, rapid, accurate, and can be used simultaneously to determine the contents of the seven active ingredients.
关键词
苦参总黄酮 /
苦参酮 /
槐属二氢黄酮G /
抗菌活性 /
高效液相色谱法 /
分离纯化 /
结构鉴定
{{custom_keyword}} /
Key words
total flavonoid /
kurarinone /
sophoraflavanone G /
antibacterial activity /
HPLC /
isolation purification /
structure identification
{{custom_keyword}} /
中图分类号：
R284
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] FAN H, ZHUANG X, DUAN X Y, et al. Therapeutic effects of colonic coated compound lightyellow sophora root capsules in patients with ulcerative colitis and mechanisms underlying such effects[J]. World Chin J Dig (世界华人消化杂志), 2009, 17(23):2429-2432.
[2] FAN H, ZHANG L J, ZHONG M, et al. Intervention effect of compound Sophorae Flavescentis decoction on DOR-β-arrestin1-Bcl-2 signal transduction pathway in rats with ulcerative colitis[J]. World Chin J Dig (世界华人消化杂志), 2013, 21(8):647-654.
[3] LI C L, CHEN Z S. Clinical effects and influence of Jianpiqingchang recipe on interleukin-4 and interleukin-8 expression in serum of ulcerative colitis patients[J]. Chin J Integr Trad West Med Dig(中国中西医结合消化杂志), 2007, 15(5):285-288.
[4] XIE X L, SHENG Y M, ZENG M, et al. ffect of different administrations of Yuchangning on ulcerative colitisin rats induced by TNBS[J].Chin Pharm J(中国药学杂志), 2010, 45(12):906-908.
[5] FAN H, LIAO Y, CHEN X Y, et al. Oxymatrine attenuates ulcerative colitis by modulating the β2AR-β-arrestin2-NF-κB signaling pathway in rats[J]. World Chin J Dig (世界华人消化杂志), 2010, 18(22):2308-2316.
[6] YANG J H, LIU H X, HUANG C S, et al. Reasearch advances of prenly flavones[J]. Technol Dev Chem Ind (化工技术与开发), 2015,44(3):14-30.
[7] YANG J L, ZHANG Y. Influence of prenylation at C-8 of ring A of flavonoid on bioactivity of parent compound[J]. Chin Tradit Herb Drugs(中草药), 2014,45(16):2418-2421.
[8] ZHAO P, ZHANG Y J, HIROBUMI Y, et al. Recent advance on the chemistry, bioactivity and biosynthesis of prenylated flavonoids from sophora flavecens[J]. Nat Prod Res Dev(天然产物研究与开发), 2004, 26(2):172-178.
[9] YANG Z X, LI X, SHAN B S, et al. Recent advance of total flavonoids from sophora flavecens[J]. Chin Tradit Pat Med(中成药), 2016, 38(5):1120-1124.
[10] YANG Z X, SHAN B S, LI X, et al. Pharmacological activity and structure-activity relationship of sophoraflavanone G[J]. Chin Med Mat(中药材), 2016, 39(2):457-461.
[11] DU S M. Study of Jieyanqing tablets used to locate in the colon by oral administration[D]. Haerbin:Heilongjiang University of Chinese Medicine, 2010.
[12] DU S M, MA L Q, SUN J J, et al. Study on in vitro antibacterial activity of extracts from Sophora flavescens Ait. [J]. Acta Chin Med Pharm (中医药学报),2010,38(3):74-75.
[13] ZHANG R J. Study on flavonoids extraction, analysis and correlative characters from Radix Sophorae Flavescentis[D]. Jinan:Shandong University,2008.
[14] JUNG M J, KANG S S, JUNG H A, et al. Isolation of flavonoids and a cerebroside from the stem bark of albizzia julibrissin[J]. Arch Pharm Res, 2004, 27(6):593-599.
[15] KANG T H, SEONG S J, KO W G, et al. Cyotoxic lavandulyl flavanones from Sophora flavescens[J]. J Nat Prod, 2000, 63(5):680-681.
[16] HATAYAMA K, KOMATSU M. Studies on the constituents of Sophora species Ⅴ. constituents of the root of Sophora angustifolia SIEB. et ZUCC .[J] .Chem Pharm Bull,1971, 19(10):2126-2131.
[17] KOMATSU M, TOMIMORI T, HATAYAMA K, et al. Studies on the constituents of Sophora species. Ⅳ. constituents of the root of Sophora angustifolia siebet zucc. (1)[J].Yakugaku Zasshi, 1970, 90(4):463-468.
[18] WILHELM H, WESSJOHANN L A. An efficient synthesis of the phytoestrogen 8-prenylnaringen in from xanthohum ol by a novel demethylation process[J]. Tetrahedron, 2006, 62(29):6961- 6966.
[19] MA Q, LEI H M, ZHOU Y X, et al. Studys on chemical constituents of Trifolium pratense [J]. Chin Pharm J(中国药学杂志),2005,40(14):1057-1059.
[20] WEN M, MA Y B, MAO X J, et al. Studies on chemical constiuents from flowers of Sophora viciifolia Hance[J]. Chin Pharm J(中国药学杂志),2010,45(19):1451-1454.
[21] TSUCHIYA H, SATO M, FUJIWARA S, et al. Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant staphylococcus aureus [J]. J Ethnopharmacol, 1996, 50(1):27-34.
[22] CHA J D, MOON S E, KIM J Y, et al. Antibacterial activity of sophoraflavanone G isolated from the roots of Sophora flavescens against methicillin-resistant Staphylococcus aureus[J]. Phytother Res, 2009, 23(9):1326-1331.
[23] KIM C S, PARK S N, AHH S J, et al. Antimicrobial effect of sophoraflavanone G isolated from Sophora flavescens against mutans streptococci[J]. Anaerobe, 2013, 19(2):17-21.
[24] LIU Y, MENG J, CHEN L. Study on HPLC fingerprint of flavonoids in Radix Sophorae[J].Tradit Chin Drug Res Clin Pharm (中药新药与临床药理), 2013, 24(3):282-285.
[25] MA H Y, ZHOU W S, CHU F J, et al. HPLC fingerprint of flavonoids in Sophora flavescens and determination of five components[J]. China J Chin Meter Med(中国中药杂志), 2013, 38(16):2690-2695.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
国家自然科学基金面上资助项目(81274091);黑龙江省自然科学基金资助项目(H2016057);国家教育部春晖计划资助项目(Z2008-1-15016);黑龙江省自然科学基金资助项目(D200744);黑龙江中医药大学“优秀创新人才支持计划”资助项目(2012)
{{custom_fund}}
PDF(713 KB)
