目的 建立超高效液相色谱串联质谱法(UPLC-MS/MS)同时测定昆明山海棠片中15种活性成分的含量。方法 采用Waters Cortecs T3色谱柱(2.1 mm×100 mm,1.6 μm),以体积分数0.1%甲酸乙腈溶液(A)-体积分数0.1%甲酸水溶液(B)为流动相,梯度洗脱,流速0.3 mL·min-1,柱温35 ℃,通过正负离子切换扫描,在多反应监测模式下建立昆明山海棠片中15种活性成分的含量测定方法,包括2种黄酮(儿茶素和表儿茶素)、9种二萜(雷公藤内酯三醇、雷醇内酯、雷公藤内酯二醇、雷公藤甲素、雷公藤氯内酯醇、雷公藤内酯酮、雷酚内酯、雷公藤对醌A和雷公藤对醌B)和4种三萜(去甲泽拉木醛、雷公藤红素、雷公藤内酯甲和扁塑藤素)。结果 15种待测成分分别在各自范围内线性关系良好(r2 ≥ 0.999 1),加样回收率在88.36%~115.91%范围内(RSD ≤ 4.94%),含量测定结果显示,不同企业生产的昆明山海棠片中活性成分的含量存在较大差异,可能会给制剂的临床药效和安全性带来影响。结论 建立的方法灵敏、准确,可用于昆明山海棠片中活性成分的含量测定,同时测定结果可为该制剂的质量评价提供参考和依据。
Abstract
OBJECTIVE To establish an ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of 15 active components in Tripterygium hypoglaucum tablets. METHODS Chromatographic separation was performed on a Waters Cortecs T3 column (2.1 mm × 100 mm, 1.6 μm) at a column temperature of 35 ℃. The 0.1% formic acid acetonitrile solution (A)-0.1% formic acid aqueous solution (B) was adopted as mobile phase for gradient elution. The flow rate was set at 0.3 mL·min-1. The detection was performed in multiple reaction monitoring (MRM) mode by positive and negative ion switching scan for 15 active components, including two flavonoids (catechin and epicatechin), nine diterpenoids (triptriolide, triptolidenol, tripdiolide, triptolide, tripchlorolide, triptonide, triptophenolide, triptoquinone A, and triptoquinone B), and four triterpenoids (demethylzeylasteral, celastrol, wilforlide A, and pristimerin). RESULTS The 15 investigated components showed good linearity (r2≥0.999 1) in their respective ranges, and the recoveries were in the range of 88.36%-115.91% (RSD≤4.94%). The quantitative results showed that the content of the active components in Tripterygium hypoglaucum tablets from different manufacturers varied significantly, which may have an impact on the clinical efficacy and safety of the preparations. CONCLUSION The established method is sensitive and accurate, which can be used for the determination of the active components in Tripterygium hypoglaucum tablets. The results can provide a reference and basis for the quality evaluation of this preparation.
关键词
昆明山海棠片 /
超高效液相色谱串联质谱法 /
含量测定 /
二萜 /
三萜 /
黄酮 /
质量评价
{{custom_keyword}} /
Key words
Tripterygium hypoglaucum tablet /
UPLC-MS/MS /
content determination /
diterpene /
triterpene /
flavonoid /
quality evaluation
{{custom_keyword}} /
中图分类号：
R282
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] WANG S M, WANG Y, ZHANG Z T, et al. Studies on the antifertility effect of Tripterygium hypoglaucum and its reversibility in male rats[J]. Chin Pharm J (中国药学杂志), 1989, 24(11):652-654.
[2] TANG Y. Progress in the pharmacological study of Tripterygium hypoglaucum[J]. Sichuan J Physiol Sci (四川生理科学杂志), 1999, 21(2):16-19.
[3] WAN P, WANG H B, WANG H Y, et al. The chemical combosition, pharmacologic action and clinical application of Tripterygium hypoglaucum Huteh (TH)[J]. Dermatol Venereol (皮肤病与性病), 2000, 22(4):20-21.
[4] YAO Y, YANG Q L. Survey of research on Tripterygium hypoglaucum[J]. Lishizhen Med Mater Med Res (时珍国医国药), 2001, 12(12):1129-1131.
[5] WANG Y, LONG C F. The advances in Chinese medical herbs (Triptergium hypoglaucum Hutch)[J]. Med Recapit (医学综述), 2006, 12(11):691-692.
[6] LI S Y, ZHANG C, HE L, et al. Clinical application of Tripterygium hypoglaucum[J]. Chin J Ethnomed Ethnopharm (中国民族民间医药杂志), 2006(83):325-330.
[7] ZHANG Y, FANG Y F, WANG Y. Research progress on pharmacological action and clinical application of Tripterygium hypoglaucum[J]. People′s Mil Surg (人民军医), 2008, 51(7):463.
[8] HAN J T. Research progress on the pharmacological effects and clinical application of Tripterygium hypoglaucum[J]. J Mod Med Health (现代医药卫生), 2009, 25(16):2459-2460.
[9] PAN Z J, LI Z W, ZHANG Z H, et al. Research progress on chemical composition and activity of THH[J]. Appl Chem Ind (应用化工), 2016, 45(2):8-10.
[10] Ch.P (2020) VolⅠ(中国药典2020版.一部)[S]. 2020:1133-1134.
[11] LU X Y. Determination of wilforlide A in Tripterygium hypoglaucum hutch tablets by HPLC[J]. China Pharm (中国药业), 2012, 21(12):52-53.
[12] WANG A W, WANG N N, LIU S, et al. Determination of triptolide, wilforlide and total triptolides in various Tripterygium wilfordii preparations[J]. Cent South Pharm (中南药学), 2015, 13(10):1081-1085.
[13] FENG X L, DING Y X, WANG Z, et al. Determination of five avtive components in various Tripterygium wilfordii preparations by HPLC-MS/MS[J]. Chin J Hosp Pharm (中国医院药学杂志), 2022, 42(5):478-484.
[14] ZHANG F, LI Y C. Progress in structure modification of triptolide[J]. Acta Pharm Sin (药学学报), 2004, 39(10):857-864.
[15] WANG C, LI C, MA J, et al. Bioactive 18(4→3)-abeo-abietanoid derivatives from the leaves of Tripterygium wilfordii[J]. RSC Adv, 2015, 5(38):30046-30052.
[16] LI Z, LI Y C. Structural diversity and structure-activity relationship —new drug research and development of Tripterygium wilfordii Hook.f.[J]. Prog Chem (化学进展), 2009, 21(12):2483-2491.
[17] HONGQUAN D, YOSHIHISA T, HIROSHI M, et al. Immunosuppressive terpenoids from extracts of Tripterygium wilfordii[J]. Chem Inform, 2010, 33(4):8413-8424.
[18] NI L, MA J, LI C, et al. Novel rearranged and highly oxygenated abietane diterpenoids from the leaves of Tripterygium wilfordii[J]. Tetrahedron Lett, 2015, 56(10):1239-1243.
[19] MA P C, LV X Y, HE C H, et al. Structural study of triptriolide isolated from Tripterygium wilfordii[J]. Acta Batanica Sin (植物学报), 1991, 33(5):370-377.
[20] WANG Q, WANG Y D, ZHENG X W, et al. Quality analysis of Leigongteng Duogan Tablets based on national drug sampling and inspection work[J]. Mod Chin Med (中国现代中药), 2020, 22(9):1534-1538.
[21] WANG Z J, STEVEN Y, CHEN S, et al. Bioavailability of wilforlide A in mice and its concentration determination using an HPLC-APCI-MS/MS method[J]. J Chromatogr B, 2018, 1090:65-72.
[22] MIAO K L, ZHANG X K, DONG Y. Studies on triterpenoids constituents of Tripterygium wilfordii Hook. f.[J]. Nat Prod Res Dev (天然产物研究与开发), 2000, 12(4):1-7.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
PDF(1181 KB)
