多维度评价及优化黄芪瞬时高温灭菌工艺

doi:10.13304/j.nykjdb.2022.1049

摘要/Abstract

摘要：

为优化黄芪瞬时高温灭菌（high temperature short time，HTST）工艺参数并多维度考察其对黄芪质量的影响，基于指标相关性的权重系数（criteria importance though intercrieria correlation，CRITIC）法采用正交设计优化黄芪灭菌工艺参数，以灭菌温度、灭菌时间、药材粉碎粒度为考察因素，以灭菌率、5种化合物含量及1，1-二苯基-2-苦基肼自由基（1，1-diphenyl-2-picrylhydrazyl，DPPH·）清除率为考察指标，通过直观及方差分析评价灭菌对3个考察指标的影响；并运用液质联用技术（liquid chromatography-mass spectrometry，LC-MS）确认指纹图谱共有峰的结构，以偏最小二乘回归分析法分析共有峰与抗氧化活性的谱效关系。结果显示，灭菌温度是具有显著影响的因素（P<0.05），最佳灭菌工艺参数为灭菌温度（170±2）℃，灭菌时间5 s，粉碎粒度50目；按优化工艺灭菌后的3批样品微生物水平均符合药典规定，5种化合物含量及DPPH·清除率与灭菌前比较无明显变化；灭菌前后指纹图谱相似度均大于0.900；谱效学分析相关性结果与化合物单体抗氧化的试验结果及其结构特点基本一致。综上所述，瞬时高温灭菌对黄芪中微生物具有明显杀灭作用且对其质量无明显影响，表明该方法适用于黄芪药材灭菌。

关键词: 黄芪, 瞬时高温灭菌, 指标相关性权重系数, 正交设计, 谱效关系

Abstract:

In order to optimize the high temperature short time （HTST） process parameters of Astragalusmembranaceus and investigate the effects on the quality of Astragalusmembranaceus from many angles， based on the criteria importance though intercrieria correlation （CRITIC） method， orthogonal design was used to optimize the sterilization process parameters of Astragalusmembranaceus. The sterilization temperature， sterilization time and fineness of pulverization of the medicinal materials were inspected， and the sterilization rate， the content of 5 compounds and the free radical clearance rate of 1，1-diphenyl-2-picrylhydrazyl （DPPH·） were determinated.The effects of sterilization on the 3 indexes were evaluated by intuitive and variance analysis. The structure of common peaks was confirmed by liquid chromatography-mass spectrometry （LC-MS）. Partial least square regression analysis was used to analyze the relationship between flavonoids and antioxidant activity of common peaks. The results showed that the sterilization temperature was a significant factor （P<0.05）. The optimal sterilization process parameters were determined as： sterilization temperature （170±2） ℃， sterilization time 5 s， fineness of pulverization 50 mesh. The microbial standard test of the 3 batches of samples after sterilization according to the optimized process all met the requirements of Pharmacopoeia of the People’s Republic of China. The contents of 5 compounds and DPPH? clearance rate had no significant change compared with that before sterilization. The similarity of fingerprints before and after sterilization was greater than 0.900， and the correlation results of spectral dynamic analysis were basically consistent with the experimental results and structural characteristics of the compounds. In conclusion， high temperature short time had obvious killing effect on microorganism in Astragalus membranaceus， but had no obvious effect on their quality， indicating that this method was suitable for the sterilization of Astragalus membranaceus.

Key words: Astragalusmembranaceus, high temperature short time, criteria importance though intercrieria correlation （CRITIC）, orthogonal design, spectral effect relationship

中图分类号:

S377

时晓宇, 焦连庆, 于敏, 田义新, 焦安妮, 栾依琳. 多维度评价及优化黄芪瞬时高温灭菌工艺[J]. 中国农业科技导报, 2024, 26(7): 223-233.

Xiaoyu SHI, Lianqing JIAO, Min YU, Yixin TIAN, Anni JIAO, Yilin LUAN. Multidimensional Evaluation and Optimization of High Temperature Short Time Process of Astragalus membranaceus[J]. Journal of Agricultural Science and Technology, 2024, 26(7): 223-233.

图/表 12

参考文献 27

1	国家药典委员会. 中华人民共和国药典: 2020年版[M]. 北京: 中国医药科技出版社, 2020: 1088.
2	张蔷, 高文远, 满淑丽. 黄芪中有效成分药理活性的研究进展[J]. 中国中药杂志,2012, 37(21): 3203-3207.
	ZHANG Q, GAO W Y, MAN S L. Chemical composition and pharmacological activities of Astragali Radix [J]. China J. Chin. Materia Med., 2012, 37(21): 3203-3207.
3	ZHENG Y J, REN W Y, ZHANG L N, et al.. A review of the pharmacological action of astragalus polysaccharide [J/OL]. Front. Pharmacol., 2020,11:349 [2022-11-01]. .
4	LIU P, ZHAO H, LUO Y. Anti-aging implications of Astragalus membranaceus (Huangqi): a well-known chinese tonic [J]. Aging Dis., 2017, 8(6): 868-886.
5	滕宝霞, 牟建平, 贺晓文. 5种不同灭菌方式对黄芪饮片中毛蕊异黄酮葡萄糖苷含量的影响[J]. 甘肃医药, 2019, 38(6):544-546.
6	孔祥山,邵晓慧, 林海岩. 中药制剂灭菌技术的应用初探[J].山东医药工业, 2002, 21(1): 39-40.
7	胡彦君, 王雅琪, 伍振峰, 等. 臭氧灭菌技术在中药及其制剂应用中的研究进展[J]. 中国中药杂志, 2015, 40(16): 3137-3141.
	HU Y J, WANG Y Q, WU Z F, et al.. Preliminary study on suitability of ozone sterilization in traditional Chinese medicine and its preparation [J]. China J. Chin. Materia Med., 2015, 40(16): 3137-3141.
8	张立雯, 林玲, 梁伟洪,等. 干热、湿热、辐照对龙胆和秦艽化学成分和灭菌效果的比较研究[J]. 中药新药与临床药理,2016, 27(5): 692-697.
	ZHANG L W, LIN L, LIANG W H, et al.. Effects of dry heat, moist heat and cobalt-60 sterilization on chemical composition and microbial killing of radix gentianae and radix gentianae macrophyllae [J]. Trad. Chin. Drug Res. Clinical Pharmacol., 2016, 27(5): 692-697.
9	林彤, 毕福钧, 吕渭升, 等. 中药的辐照灭菌现状与监管[J].中国药学杂志, 2019, 54(17): 1442-1447.
	LIN T, BI F J, LYU W S, et al.. Status and control of radicidation of tranditional Chinese medicine [J]. Chin. Pharm. J., 2019, 54(17): 1442-1447.
10	尚海宾, 陶海涛, 乔晓芳. 中药高温瞬时灭菌设备的智能化设计与性能确认[J]. 流程工业, 2022(7): 58-61.
11	黄定轩. 基于客观信息熵的多因素权重分配方法[J]. 系统工程理论方法应用, 2003, 12(4): 321-324.
	HUANG D X. Means of weights allocation with multi-factors based on impersonal message entropy [J]. Syst. Eng. Theory Methodol. Appl.,2003,12(4):321-324.
12	中华人民共和国药典委员会. 中华人民共和国药典(四部) [M]. 北京: 中国医药科技出版社, 2020: 160-179.
13	康超超, 王学成, 伍振峰, 等. 当归原生粉乙醇灭菌工艺优化及其品质比较研究 [J]. 中草药, 2019, 50(6): 1341-1347.
	KANG C C, WANG X C, WU Z F, et al.. Optimization of ethanol sterilization technology for Angelicae Sinensis Radix and comparison of its quality [J]. China Tradit. Herb. Drugs, 2019, 50(6): 1341-1347.
14	陈伯丛,王汝上,罗德祥.不同产地黄芪总黄酮HPLC指纹图谱研究[J].北方药学,2016,13(7):8-10.
	CHEN B C, WANG R S, LUO D X. HPLC fingerprinting of flavonoids in radix astragali [J]. J. North Pharmacy, 2016,13(7):8-10.
15	SHARMA O P, BHAT T K. DPPH antioxidant assay revisited [J]. Food Chem., 2009,113(4): 1202-1205.
16	YANG B, ZHAO M M, SHI J, et al.. Effect of ultrasonic treatment on the recovery and DPPH radical scavenging activity of polysaccharides from longan fruit pericarp [J]. Food Chem., 2008,106(2):685-690.
17	王存琴,陈颖,汪雷,等. HPLC同时测定大叶冬青叶中10种黄酮成分的含量及抗氧化活性研究[J].天然产物研究与开发,2019,31(4):557-565.
	WANG C Q, CHEN Y, WANG L,et al.. Determination of 10 polyphenol components by HPLC and antioxidant activity in the leaves of Ilex latifolia [J]. Nat. Prod. Res. Dev., 2019,31 (4):557-565.
18	王强,张湛,杨彬,等.基于AHP-CRITIC混合加权法的正交试验优选神气汤提取工艺[J].中国药业,2021,30(19):51-55.
	WANG Q, ZHANG Z, YANG B, et al..Optimization of extraction process of Shenqi decoction by the orthogonal test based on AHP-CRITIC mixed weighting method [J]. China Pharm., 2021, 30(19):51-55.
19	冯伟红,李春,信伟梅,等.生物测定法用于中药质量评价的探索研究——以夏枯草抗氧化活性与总酚酸含量相关性的研究为例[J].中国中药杂志,2016,41(14):2660-2668.
	FENG W H, LI C, XIN W M, et al.. Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example [J]. China J. Chin. Materia Med., 2016,41(14):2660-2668.
20	张靖. 黄芪药对及复方的化学成分质谱学鉴定及多成分定量研究[D]. 广州: 广州中医药大学, 2015.
	ZHANG J. The study on mass spectrometry identification and multi-component quantitative studies of Huangqi herb pair and its compound preparation [D]. Guangzhou: Guangzhou University of Chinese Medicine, 2015.
21	谭雪霞, 朱宏明, 唐铖, 等. HPLC-MS鉴定黄芪水煎液中黄酮类成分[J]. 现代药物与临床, 2021, 36(2): 231-235.
	TAN X X, ZHU H M, TANG C, et al.. HPLC-MS identification on flavonoids in Astragali radix decoction [J]. Drugs Clinic, 2021, 36(2): 231-235.
22	周颖, 黄锐. 超高温瞬时灭菌工艺优化[J]. 中国乳业, 2020(2): 81-84.
23	袁武会. 参术止带糖浆的灭菌工艺研究[J]. 中国药业,2013,22(2): 25-27.
24	李振豪, 李顺仓, 王杰, 等. 不同灭菌法对婴儿健脾散成分及微生物的影响[J]. 流程工业, 2020(6): 56-59.
25	张婷, 焦连庆, 刘融融, 等. 基于多指标分析优化苦丁茶冬青瞬时高温灭菌工艺[J]. 食品工业科技, 2023, 44(8):205-211.
	ZHANG T, JIAO L Q, LIU R R, et al.. Optimize the instantaneous high temperature sterilization process of Ilex kudingcha based on multiple index analysis [J]. Sci. Technol. Food Ind, 2023, 44(8): 205-211 .
26	姚静, 杨晓宁, 朱平, 等. HPLC-CAD指纹图谱结合化学计量学评价不同产地黄芪质量[J]. 中成药, 2022, 44(10): 3214-3219.
	YAO J, YANG X N, ZHU P, et al.. Quality evaluation of Astragali Radix from different producing areas by HPLC⁃ CAD fingerprints combined with chemometrics [J]. Chin. Tradit. Patent Med., 2022, 44(10): 3214-3219.
27	陈星宇, 谭魏, 汪虹, 等. 黄芪黄酮抗氧化活性的构效关系分析[J]. 广州化工, 2021, 49(24): 26-30.
	CHEN X Y, TAN W, WANG H, et al.. Structure-activity relationship analysis on antioxidant activity of astragalus flavonoidss [J]. Guangzhou Chem. Ind., 2021,49(24):26-30.

时间Time/min	乙腈 Acetonitrile	0.2%甲酸水 0.2% formic acid water
0—40	15%~60%	85%~40%
40—55	60%~75%	40%~25%
55—56	75%~15%	25%~85%
56—60	15%	85%

时间Time/min	乙腈 Acetonitrile	0.2%甲酸水 0.2% formic acid water
0—40	15%~60%	85%~40%
40—55	60%~75%	40%~25%
55—56	75%~15%	25%~85%
56—60	15%	85%

样品编号Sample number	毛蕊异黄酮苷 Calycosin-7-glucoside	毛蕊异黄酮 Calycosin	山奈酚 Kaempferol	芒柄花苷 Ononin	刺芒柄花素Formononetin	总含量 Total content
1	0.040	0.003	0.032	0.021	0.011	0.107
2	0.057	0.004	0.041	0.035	0.012	0.149
3	0.058	0.004	0.044	0.036	0.013	0.155
4	0.037	0.003	0.029	0.034	0.010	0.113
5	0.036	0.004	0.034	0.030	0.012	0.116
6	0.058	0.004	0.036	0.034	0.013	0.145
7	0.038	0.003	0.034	0.033	0.010	0.118
8	0.037	0.003	0.034	0.024	0.011	0.109
9	0.044	0.004	0.037	0.020	0.013	0.118

样品编号Sample number	毛蕊异黄酮苷 Calycosin-7-glucoside	毛蕊异黄酮 Calycosin	山奈酚 Kaempferol	芒柄花苷 Ononin	刺芒柄花素Formononetin	总含量 Total content
1	0.040	0.003	0.032	0.021	0.011	0.107
2	0.057	0.004	0.041	0.035	0.012	0.149
3	0.058	0.004	0.044	0.036	0.013	0.155
4	0.037	0.003	0.029	0.034	0.010	0.113
5	0.036	0.004	0.034	0.030	0.012	0.116
6	0.058	0.004	0.036	0.034	0.013	0.145
7	0.038	0.003	0.034	0.033	0.010	0.118
8	0.037	0.003	0.034	0.024	0.011	0.109
9	0.044	0.004	0.037	0.020	0.013	0.118

样品编号 Sample number		灭菌温度 Sterilization temperature/℃	灭菌时间 Sterilization time/s	粉碎粒度/目 Fineness of pulverization/mesh	空白 Blank	灭菌率 Sterilization rate/%	5种化合物总含量 Total content of 5 compounds/%	DPPH• 清除率 DPPH•clearance rate/%	综合评分Synthesize score
1		170±2	10	80		96.95	0.107	52.29	0.866
2		170±2	7	50		95.50	0.149	62.89	0.964
3		170±2	5	24		96.75	0.155	67.79	0.999
4		160±2	10	24		87.20	0.113	53.96	0.832
5		160±2	7	80		87.15	0.116	57.95	0.852
6		160±2	5	50		90.80	0.145	60.20	0.923
7		150±2	10	50		81.85	0.118	58.88	0.831
8		150±2	7	24		83.50	0.109	55.53	0.813
9		150±2	5	80		80.93	0.118	59.01	0.827
灭菌率 Sterilization rate/%	K1	96.333	88.600	88.277	90.350	—	—	—	—
	K2	88.383	88.717	89.383	87.877	—	—	—	—
	K3	82.093	89.493	89.150	88.583	—	—	—	—
	极差R	14.240	0.893	1.106	2.473	—	—	—	—
5种化合物总含量 Total content of 5 compounds/%	K1	0.137	0.113	0.114	0.120	—	—	—	—
	K2	0.125	0.125	0.137	0.127	—	—	—	—
	K3	0.115	0.139	0.126	0.130	—	—	—	—
	极差R	0.022	0.026	0.023	0.010	—	—	—	—
DPPH•清除率 DPPH•clearance rate/%	K1	60.990	55.043	56.417	56.007	—	—	—	—
	K2	57.370	58.790	60.657	58.620	—	—	—	—
	K3	57.807	62.333	59.093	61.540	—	—	—	—
	极差R	3.620	7.290	4.240	5.533	—	—	—	—
综合评分 Synthesize score	K1	0.943	0.876	0.848	0.867	—	—	—	—
	K2	0.869	0.876	0.939	0.874	—	—	—	—
	K3	0.857	0.916	0.881	0.927	—	—	—	—
	极差R	0.086	0.040	0.091	0.060	—	—	—	—