首页分享无患子花总皂苷提取工艺优化及抗氧化、美白活性研究

无患子花总皂苷提取工艺优化及抗氧化、美白活性研究

来源：花匠小妙招时间：2026-02-20 22:34

摘要:

以总皂苷得率为指标, 采用单因素和正交试验优化无患子花总皂苷水提取工艺, 对比无患子花水提液与无患子果皮水提液中的总皂苷、总黄酮和总酚含量。通过DPPH·、ABTS+·、O2-·清除实验以及铁离子还原能力测定评价无患子花水提液抗氧化能力；通过酪氨酸酶活性抑制实验, 探究其美白效果；通过红细胞溶血试验, 探究无患子花水提液的刺激性。结果表明：无患子花水提液的最佳提取条件为提取温度80 ℃、提取时间180 min、液料比30∶1(mL∶g), 此条件下, 总皂苷得率为12.15%。无患子花水提液的总皂苷、总黄酮和总酚含量较无患子果皮高；无患子花与果皮的水提液抗氧化能力及酪氨酸酶活性抑制能力较好, 且无患子花水提液优于果皮水提液。无患子花水提液的HD50/DI>100, 根据红细胞溶血试验刺激性分级标准, 表明无患子花水提液无刺激性。

关键词: 无患子花 / 皂苷 / 黄酮 / 抗氧化 / 酪氨酸酶

Abstract:

Single-factor and orthogonal tests were used to optimize the extraction process of total saponins from Sapindus mukorossi Gaertn. flowers, using the total saponin yield as an indicator. The total saponins, flavonoids and phenols contents in the water extracts of S. mukorossi Gaertn. flowers and pericarp were compared. Furthermore, the antioxidant activity was evaluated by DPPH·, ABTS+·, O2-· radical scavenging test, and ferric ion reducing ability assay. The whitening effect was explored by tyrosinase activity inhibition test, and the irritant properties were investigated by red blood cell haemolysis test. The results showed that the optimum extraction conditions were as follows: extraction temperature of 80 ℃, extraction time of 180 min and liquid-solid ratio of 30(mL/g). Under these conditions, the total saponins yield was 12.15%. The contents of total saponins, flavonoids and phenols in the water extract of S. mukorossi Gaertn. flower were higher than that of pericarp. The antioxidant capacity and tyrosinase activity inhibiting ability of the S. mukorossi Gaertn. flower water extract were good, which were better than that of pericarp. The HD50/DI>100 of the aqueous extract of S. mukorossi Gaertn. flowers, according to the erythrocyte haemolysis test irritation grading criteria, indicated that the aqueous extract was non-irritating.

图 1

不同提取条件对无患子花提取液中总皂苷得率的影响

Figure 1.

Effect of different extraction conditions on the yield of total saponins in soapberry flower extract

图 2

不同体积分数的无患子花和果皮水提液的抗氧化活性

a.DPPH·；b.ABTS+·；c.FRAP；d.O2-·

Figure 2.

Antioxidant activity of aqueous extract of soapberry flowers and pericarp at different volume fractions

图 3

无患子花与无患子果皮水提液的酪氨酸酶抑制率

Figure 3.

Tyrosinase inhibition rate of aqueous extract of soapberry flowers and pericarp

图 4

无患子花水提液的红细胞溶血率

Figure 4.

Erythrocyte haemolysis rate of water extract of soapberry flowers

表 1

实验体系

Table 1

Experimental system

体系待测样品/μL sample PBS/μL 酪氨酸酶/μL tyrosinase 左旋多巴溶液/μL L-dopa A 60 0 30 120 B 60 30 0 120 C 0 60 30 120 D 0 90 0 120

表 2

无患子花总皂苷的提取条件优化的正交试验结果

Table 2

Orthogonal test results of optimization of extraction conditions of total saponins from soapberry flowers

编号
No. A
液料比(mL∶g)
ratio of liquid to solid B
提取时间/min
extraction time C
提取温度/℃
extraction temperature D
空白列
blank column 总皂苷得率/%
saponins yield 1 20 100 60 1 11.07 2 20 140 80 2 10.61 3 20 180 70 3 10.72 4 25 100 80 3 12.35 5 25 140 70 1 11.82 6 25 180 60 2 12.17 7 30 100 70 2 12.47 8 30 140 60 3 12.25 9 30 180 80 1 12.79 k1 10.8002 11.9629 11.8284 11.8899 k2 12.1136 11.5600 11.6700 11.7517 k3 12.5023 11.8933 11.9176 11.7745 R 1.7021 0.4029 0.2475 0.1382

表 3

无患子花与果皮水提液中的总皂苷、总黄酮、总酚1)

Table 3

Mass fractions of total saponins, total flavonoids and total phenols in aqueous extracts of soapberry flowers and pericarp %

水提液来源总皂苷
total saponin 总黄酮
total flavonoid 总酚
total phenolic 花flowers 128.80±0.94* 28.23±0.60* 97.21±1.87* 果皮pericarp 111.76±4.10 4.42±0.09 14.19±1.03 1)*代表显著性差异, p<0.05 *represents a significant difference, p<0.05 [1]

SINGH R, SHARMA B. Biotechnological advances, phytochemical analysis and ethnomedical implications of sapindus species[M]. Springer Singapore, 2019.

[2]

曾庆钱, 郑良豹, 黄意成, 等. 无患子的本草考证及研究进展[J]. 林业与环境科学, 2018, 34(4): 168-175.

DOI: 10.3969/j.issn.1006-4427.2018.04.028

ZENG Q Q, ZHENG L B, HUANG Y C, et al. Herbal textual and research progress of Sapindus mukorossi[J]. Forestry and Environmental Science, 2018, 34(4): 168-175.

DOI: 10.3969/j.issn.1006-4427.2018.04.028 [3]

GOYAL S. Medicinal plants of the genus Sapindus(Sapindaceae)-A review of their botany, phytochemistry, biological activity and traditional uses[J]. Journal of Drug Delivery and Therapeutics, 2014, 4(5): 7-20.

[4]

SHARMA A, SATI S C, SATI O P, et al. A new triterpenoid saponin and antimicrobial activity of ethanolic extract from Sapindus mukorossi Gaertn. [J/OL]. Journal of Chemistry, 2013: 218510[2023-06-12]. https://doi.org/10.1155/2013/218510.

[5]

林清霞, 郑德勇. 泡沫分离法分离纯化无患子皂苷的研究[J]. 生物质化学工程, 2013, 47(3): 34-38.

DOI: 10.3969/j.issn.1673-5854.2013.03.007

LIN Q X, ZHENG D Y. Foam separation and purification of saponin from Sapindus mukorossi[J]. Biomass Chemical Engineering, 2013, 47(3): 34-38.

DOI: 10.3969/j.issn.1673-5854.2013.03.007 [6]

SHAH M, PARVEEN Z, KHAN M R. Evaluation of antioxidant, anti-inflammatory, analgesic and antipyretic activities of the stem bark of Sapindus mukorossi[J/OL]. BMC Complementary and Alternative Medicine, 2017, 17: 526[2023-06-12]. https://doi.org/10.1186/s12906-017-2042-3.

[7]

JIN K, SUN Y, ZHOU S. Sapindus saponins inhibits tyrosinase & glutathione-induced melanin deposition by obstructing DOPA and cysteyl derivatives transformation pathways[C]//2021 International Symposium on Biomedical Engineering and Computational Biology, 2021: 1-6.

[8]

雷发玲, 肖蕾, 么春艳, 等. 无患子皂苷抗炎抗菌功效研究[J]. 中国野生植物资源, 2020, 39(7): 9-13.

DOI: 10.3969/j.issn.1006-9690.2020.07.002

LEI F L, XIAO L, YAO C Y, et al. Anti-inflammatory and antimicrobial effects of saponin from Sapindus mukorossi[J]. Chinese Wild Plant Resources, 2020, 39(7): 9-13.

DOI: 10.3969/j.issn.1006-9690.2020.07.002 [9]

HENG W, LING Z, NA W, et al. Analysis of the bioactive components of Sapindus saponins[J]. Industrial Crops and Products, 2014, 61: 422-429.

DOI: 10.1016/j.indcrop.2014.07.026 [10]

LIU M, CHEN Y L, KOU Y H, et al. Aqueous extract of Sapindus mukorossi induced cell death of A549 cells and exhibited antitumor property in vivo[J/OL]. Scientific Reports, 2018: 4831[2023-06-12]. https://doi.org/10.1038/s41598-018-23096-w.

[11]

CHEN I C, CHEN M T, CHUNG T W. Analysis of antioxidant property of the extract of saponin by experiment design methodology[C/OL]//IOP Conference Series: Earth and Environmental Science. IOP Publishing, 2020, 594(1): 012002[2023-06-12]. DOI10.1088/1755-1315/594/1/012002.

[12]

赵志敏, 南艳平, 唐青涛, 等. 无患子总皂苷抗炎及抑制酪氨酸酶活性研究[J]. 时珍国医国药, 2014, 25(7): 1592-1595.

https://www.cnki.com.cn/Article/CJFDTOTAL-SZGY201407022.htm

ZHAO Z M, NAN Y P, TANG Y Q T, et al. Anti-inflammatory activity and inhibitory activity on tyrosinase of soapnut saponin from Sapindus mukorossi Gaertn[J]. Lishizhen Medicine and Materia Medica Research, 2014, 25(7): 1592-1595.

https://www.cnki.com.cn/Article/CJFDTOTAL-SZGY201407022.htm [13]

陈雷, 赵春琳, 周生学. 无患子总皂苷体外抑制酪氨酸酶活性的研究[J]. 安徽农业科学, 2015, 43(21): 56-58.

DOI: 10.3969/j.issn.0517-6611.2015.21.020

CHEN L, ZHAO C L, ZHOU S X. Research on the inhibition of tyrosinase activity in vitro by total saponins from Sapindus[J]. Anhui Journal of Agricultural Sciences, 2015, 43(21): 56-58.

DOI: 10.3969/j.issn.0517-6611.2015.21.020 [14]

徐圆圆, 周思维, 陈仲, 等. 无患子不同器官中的总皂苷和总黄酮含量[J]. 南京林业大学学报(自然科学版), 2021, 45(4): 83-89.

https://www.cnki.com.cn/Article/CJFDTOTAL-NJLY202104010.htm

XU Y Y, ZHOU S W, CHEN Z, et al. Contents of the total saponins and total flavonoids in different organs of Sapindus mukorossi[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2021, 45(4): 83-89.

https://www.cnki.com.cn/Article/CJFDTOTAL-NJLY202104010.htm [15]

钱天元, 孙亭, 李成平, 等. 无患子花色素理化性质研究[J]. 食品工业科技, 2017, 38(17): 212-215, 234.

https://www.cnki.com.cn/Article/CJFDTOTAL-SPKJ201717040.htm

QIAN T Y, SUN T, LI C P, , et al. Study on the physicochemical properties of pigment from flower of Sapindus mukurossi[J]. Science and Technology of Food Industry, 2017, 38(17): 212-215, 234.

https://www.cnki.com.cn/Article/CJFDTOTAL-SPKJ201717040.htm [16]

阳春苗, 王秋玲, 梁芳, 等. 响应面法优化无患子皂苷超声提取工艺研究[J]. 生物化工, 2021, 7(1): 1-4, 10.

DOI: 10.3969/j.issn.2096-0387.2021.01.001

YANG C M, WANG Q L, LIANG F, et al. Optimization of ultrasonic extraction technology of saponins from Sapindus mukorossi Gaertn. by response surface analysis[J]. Biological Chemical Engineering, 2021, 7(1): 1-4, 10.

DOI: 10.3969/j.issn.2096-0387.2021.01.001 [17]

WANG B, QU J, LUO S, et al. Optimization of ultrasound-assisted extraction of flavonoids from olive(Olea europaea) leaves, and evaluation of their antioxidant and anticancer activities[J/OL]. Molecules, 2018, 23(10): 2513[2023-06-12]. https://doi.org/10.3390/molecules23102513.

[18]

XU S J, FANG D, TIAN X Q, et al. Subcritical water extraction of bioactive compounds from waste cotton(Gossypium hirsutum L.) flowers[J/OL]. Industrial Crops and Products, 2021, 164: 113369[2023-06-12]. https://doi.org/10.1016/j.indcrop.2021.113369.

[19]

WANG Y J, ZHAO L, ZHANG R Y, et al. Optimization of ultrasound-assisted extraction by response surface methodology, antioxidant capacity, and tyrosinase inhibitory activity of anthocyanins from red rice bran[J]. Food Science & Nutrition, 2020, 8(2): 921-932.

[20]

谢红. 酸枣仁皂苷的分离纯化及生物活性研究[D]. 广州: 华南理工大学, 2021.

XIE H. Study on separation, purification and biological activity of Zizyphi Spinosae Semen saponins[D]. Guangzhou: South China University of Technology, 2021.

[21]

上海日用化学品行业协会. T/SHRH 015-2018 化妆品-酪氨酸酶活性抑制实验方法[S]. 上海: 上海日用化学品行业协会, 2018.

Shanghai Daily Chemistry Trade Association. T/SHRH 015-2018 Cosmetics-tyrosinase activity inhibition test method[S]. Shanghai: Shanghai Daily Chemistry Trade Association, 2018.

[22]

俞婕, 祝愿, 鲍红洁. 红细胞溶血法用于体外评价日化产品和原料刺激性的研究[J]. 中国洗涤用品工业, 2020(12): 36-41.

DOI: 10.3969/j.issn.1672-2701.2020.12.003

YU J, ZHU Y, BAO H J. Study on the in-vitro RBC hemolysis assay for the irritation evaluation of daily chemical products and raw materials[J]. China Cleaning Industry, 2020(12): 36-41.

DOI: 10.3969/j.issn.1672-2701.2020.12.003 [23]

韦蒙, 许新恒, 李俊龙, 等. 滇重楼茎叶总皂苷提取工艺优化及其体外抗氧化活性分析[J]. 天然产物研究与开发, 2015, 27(10): 1794-1800.

https://www.cnki.com.cn/Article/CJFDTOTAL-TRCW201510021.htm

WEI M, XU X H, LI J L, et al. Extraction and antioxidant activity of total saponins from stems and leaves of Paris polyphylla var. yunnanensis (Franch.) Hand. -Mazz[J]. Natural Product Research and Developmen, 2015, 27(10): 1794-1800.

https://www.cnki.com.cn/Article/CJFDTOTAL-TRCW201510021.htm [24]

张翠. 无患子总皂苷制备工艺研究[D]. 无锡: 江南大学, 2012.

ZHANG C. The production of total sapindus-saponins from Sapindus mukurossi Gaertn[D]. Wuxi: Jiangnan University, 2018.

[25]

刘俊宏, 邱智东. 无患子总皂苷提取工艺优化及其对酪氨酸酶的抑制活性[J]. 江苏农业科学, 2020, 48(9): 228-232.

https://www.cnki.com.cn/Article/CJFDTOTAL-JSNY202009044.htm

LIU J H, QIU Z D. Optimisation of the extraction process of total saponins from Sapindus mukurossi Gaertn and its inhibitory activity on tyrosinase[J]. Jiangsu Agricultural Sciences, 2020, 48(9): 228-232.

https://www.cnki.com.cn/Article/CJFDTOTAL-JSNY202009044.htm [26]

黄素梅, 王敬文, 杜孟浩, 等. 无患子总皂苷的提取工艺研究[J]. 安徽农业科学, 2010, 38(1): 354-356.

DOI: 10.3969/j.issn.0517-6611.2010.01.144

HUANG S M, WANG J W, DU M H, et al. Research on the extraction technology of total saponins from Sapindus mukorossi Gaerth[J]. Anhui Journal of Agricultural Sciences, 2010, 38(1): 354-356.

DOI: 10.3969/j.issn.0517-6611.2010.01.144 [27]

HU X, TANG J, ZHANG G, et al. Optimization of extraction process and antioxidant activities of saponins from Camellia fascicularis leaves[J]. Journal of Food Measurement and Characterization, 2021, 15: 1889-1898.

DOI: 10.1007/s11694-020-00754-0 [28]

LI J, ZU Y G, FU Y J, et al. Optimization of microwave-assisted extraction of triterpene saponins from defatted residue of yellow horn(Xanthoceras sorbifolia Bunge. ) kernel and evaluation of its antioxidant activity[J]. Innovative Food Science & Emerging Technologies, 2010, 11(4): 637-643.

[29]

LIM J G, PARK H M, YOON K S. Analysis of saponin composition and comparison of the antioxidant activity of various parts of the quinoa plant(Chenopodium quinoa Willd. )[J]. Food Science & Nutrition, 2020, 8(1): 694-702.

[30]

EL IDRISSI Y, ELOUAFY Y, EL MOUDDEN H, et al. Evaluation of antioxidant and antimicrobial activity of saponin extracts from different parts of Argania spinosa L. Skeels[J/OL]. Progress In Microbes & Molecular Biology, 2023, 6(1)[2023-07-12]. DOI: 10.36877/pmmb.0000338.

[31]

SILVA R M G, MARTINS G R, NUCCI L M B, et al. Antiglycation, antioxidant, antiacne, and photoprotective activities of crude extracts and triterpene saponin fraction of Sapindus saponaria L. fruits: An in vitro study[J]. Asian Pacific Journal of Tropical Biomedicine, 2022, 12(9): 391-399.

DOI: 10.4103/2221-1691.354430 [32]

CHANG T S. An updated review of tyrosinase inhibitors[J]. International Journal of Molecular Sciences, 2009, 10(6): 2440-2475.

[33]

WEI M P, QIU J D, LI L, et al. Saponin fraction from Sapindus mukorossi Gaertn as a novel cosmetic additive: Extraction, biological evaluation, analysis of anti-acne mechanism and toxicity prediction[J/OL]. Journal of Ethnopharmacology, 2021, 268: 113552[2023-06-12]. https://doi.org/10.1016/j.jep.2020.113552.

[34]

RATZ-ŁYKO A, ARCT J, MAJEWSKI S, et al. Influence of polyphenols on the physiological processes in the skin[J]. Phytotherapy Research, 2015, 29(4): 509-517.

[35]

ZHAO W, YANG A, WANG J, et al. Potential application of natural bioactive compounds as skin-whitening agents: A review[J]. Journal of Cosmetic Dermatology, 2022, 21(12): 6669-6687.

[36]

ALVES E N, DE FARIAS PRESGRAVE R, et al. A reassessment of the in vitro RBC haemolysis assay with defibrinated sheep blood for the determination of the ocular irritation potential of cosmetic products: Comparison with the in vivo Draize rabbit test[J]. Alternatives to Laboratory Animals, 2008, 36(3): 275-284.