Multidimensional Evaluation and Optimization of High Temperature Short Time Process of Astragalus membranaceus

doi:10.13304/j.nykjdb.2022.1049

Abstract

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

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

CLC Number:

S377

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.

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.

Figures/Tables 12

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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	—	—	—	—