目的 优化大孔树脂纯化菟丝子总黄酮工艺。方法 采用单因素实验,以总黄酮、金丝桃苷的吸附率和解析率为指标,筛选最佳大孔树脂型号。以总黄酮、金丝桃苷吸附量为指标,考察吸附时间、漏点浓度、上样液pH对纯化工艺的影响。以总黄酮、金丝桃苷洗脱率为指标,考察洗脱液流速、洗脱液体积、洗脱液浓度对纯化工艺的影响。在此基础上,以总黄酮与金丝桃苷含量的归一值为指标,利用Box-Behnken响应面法优化并确立最佳工艺。结果 菟丝子总黄酮纯化的最佳树脂型号为AB-8型,最佳工艺条件为:上样液pH 3.9、洗脱液体积19.6BV,洗脱液浓度74%,验证实验的总OD值为0.982 5,与模型预测值接近。纯化前后,总黄酮含量由18.95 mg·g-1提高至85.68 mg·g-1,金丝桃苷含量由4.12 mg·g-1提高至16.85 mg·g-1。结论 优选的纯化工艺方法稳定、可行,可用于菟丝子总黄酮的纯化。
Abstract
OBJECTIVE To optimize the purification process of total flavonoids from Cuscuta chinensis by macroporous resin. METHODS Single factor experiment was used to select the best type of macroporous resin based on the adsorption rate and resolution rate of total flavonoids and hyperoside. The effects of adsorption time, concentration of leakage point and pH on the purification process were investigated with the adsorption amount of total flavonoids and hyperoside as indexes. The effects of eluent flow rate, eluent volume and eluent concentration on the purification process were investigated with the elution rate of total flavonoids and hyperoside as indexes. On this basis, the Box-Behnken response surface methodology was used to optimize the optimum process using the normalized value of total flavonoids and hyperoside content as the index. RESULTS The optimal resin type for the purification of total flavonoids from Cuscuta chinensis was AB-8, and the optimal process conditions were as follows:pH 3.9, eluent volume 19.6BV, eluent concentration 74%. The total OD value of validation test was 0.982 5, which was close to the predicted value. Before and after purification, the content of total flavonoids was increased from 18.95 mg·g-1 to 85.68 mg·g-1, and the content of hyperoside increased from 4.12 mg·g-1 to 16.85 mg·g-1. CONCLUSIONS The optimized purification technology is stable and feasible, and could be used for the purification of total flavonoids from Cuscuta chinensis.
关键词
大孔树脂 /
Box-Behnken设计-响应面法 /
纯化工艺 /
菟丝子总黄酮 /
总评归一法
{{custom_keyword}} /
Key words
macroporous resin /
Box-Behnken design-response surface methodology /
purification process /
dodder-seed total flavonoid /
general evaluation normalization method
{{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] Ch.P(2020)Vol Ⅰ(中国药典2020年版.一部)[S]. 2020:309-310.
[2] SUN Y L, LI Z Q, FENG G Y, et al. Optimization of extraction technology of total flavonoids from Cuscuta seed by ultrasonic assisted compound enzyme[J]. Mod Chin Med(中国现代中药), 2017, 19(9):1296-1300.
[3] LIU Q, TIAN Y C, BIN B, et al. Research progress on protective effect of dodder flavonoids on reproductive system[J]. Chin Med Guide(中医药导报), 2020, 26(12):148-151,156.
[4] ZHU B J, ZHANG T, CHEN Y. Progress in clinical application and pharmacology of cuscuta lucid eye effect[J]. J Pharm Pract(药学实践杂志), 2020, 38(5):409-412.
[5] WANG X M, XI X T, LIU H Y, et al. Effects of dodder flavonoids on apoptosis and apoptosis-related proteins of vascular smooth muscle cells[J]. Chin J Gerontol(中国老年学杂志), 2015, 35(20):5742-5744.
[6] YE C L, CHEN Y, LI H X, et al. Study on antioxidant activity and inhibition of tumor cell proliferation of Polysaccharides from Cuscuta chinensis[J]. J Henan Univ Technol (Nat Sci Ed)(河南工业大学学报自然科学版), 2020, 41(5):73-78.
[7] QIN J J, QIAN H Q, WEI J, et al. Extraction technology and antioxidant activity of total flavonoids from Cuscuta[J]. Sci Technol Food Ind(食品工业科技), 2019, 40(23):151-157.
[8] SHAO J Z, GUO X L, ZHANG J, et al. Clinical application and dosage of Dodder[J]. Jilin Tradit Chin Med(吉林中医药), 2019, 39(1):36-39.
[9] SONG X Y, JIANG Y T. Study on pharmacological action and treatment of dysmenorrhea of total flavonoids of Dodder[J]. Med Inf(医学信息), 2020, 33(8):29-31.
[10] ZHANG X Y, SUN X M, SONG H, et al. Study on extraction methods and estrogen activity of total flavonoids from Cuscuta chinensis[J]. Chin Pharm J(中国药学杂志), 2021, 56(1):23-28.
[11] DING J X, LI W L, BAI J, et al. Study on spectral correlation of estrogenoid activity of Cuscuta chinensis[J]. Chin Pharm J(中国药学杂志), 2013, 48(5):337-340.
[12] JI R F. Quality evaluation of flavonoids in cuscuta Granule[D]. Jinan:Shandong University, 2019.
[13] AHMAD A, TANDON S, XUAN T D, et al. A Review on phytoconstituents and biological activities of Cuscuta species[J]. Biomed Pharm, 2017, 92(8):772-795.
[14] WANG X X, FAN G M, WEI F M, et al. Hyperoside protects rat ovarian granulosa cells against hydrogen peroxide-induced injury by sonic hedgehog signaling pathway[J]. Chem Biolog Interact, 2019, 9(1):310.
[15] KIM Y H, KANG M K, LEE E J, et al. Astragalin inhibits cigarette smoke-induced pulmonary thrombosis and alveolar inflammation and disrupts PAR activation and oxidative stress-responsive MAPK-signaling. [J]. Inter J Mol Sci, 2021, 22(7):3692.
[16] WANG R, LUO Q, FENG Y. The antioxidant capacity of kaempferol was determined by DPPH, ABTS and FRAP[J]. Guangzhou Chem Ind(广州化工), 2021, 49(3):58-59,63.
[17] FENG Y L, LU L P, QU G Y. Advances in antitumor activity of quercetin derivatives[J]. China J Chin Mater Med(中国中药杂志), 2020, 45(15):3565-3574.
[18] ZHU M, ZHAO L M, WANG P, et al. Isorhamnetin inhibits ovalbumin-induced lung inflammation in asthmatic mice[J]. Chin J Pathophysiol(中国病理生理杂志), 2021, 37(1):106-111.
[19] DI J L, SHAN W T, ZHAO J, et al. Effects of apigenin combined with adriamycin on proliferation, apoptosis and cell cycle of Hepatocellular Carcinoma HepG2 cells[J]. J Tradit Chin Med(中医药学报), 2021, 49(5):31-34.
[20] WU W, CUI G H, LU L. Optimization of multiple indicators in empirical design:star design and the application of general evaluation "normalized value"[J]. Chin Pharm J(中国药学杂志), 2000, 35(8):26-29.
[21] MIAO M S, PENG M F, YAN X L, et al. Effect of total flavonoids from Cuscuta on letrozole-induced polycystic ovary syndrome in rats[J]. Chin J Exp Form(中国实验方剂学杂志), 2019, 25(16):17-23.
[22] LIU G M, BAI X, TANG Y B, et al. Research progress of preventing male infertility by Cuscuta seed[J]. J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报), 2021, 23(4):162-166.
[23] MIAO M S, PENG M F, ZHU Z W, et al. Effects of dodder total flavone on polycystic ovary syndrome rat models induced by DHEA combined HCG[J]. Saudi J Biol Sci, 2019, 26(4):821-827.
[24] ZHENG Y B, YANG X X, BAO Y R, et al. Study on anti-liver cancer mechanism of total flavonoids from Dodder based on PI3K/PTEN/VEGF[J]. Chin J Hosp Pharm(中国医院药学杂志), 2018, 38(14):1462-1465.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
黑龙江省重点研发计划指导类项目资助(GZ20210092)
{{custom_fund}}
PDF(1799 KB)
