灵莲花颗粒微生物限度检查方法适用性研究

廖辉; 贺婵娟; 杨红霞; 张璐

doi:10.2095-3593.2026.050016

中国药物评价 >

2026 , Vol. 43 >Issue 3: 189 - 189-195

DOI: https://doi.org/10.2095-3593.2026.050016

评价技术与方法

灵莲花颗粒微生物限度检查方法适用性研究

廖辉 ,
贺婵娟 ,
杨红霞 ,
张璐

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1.湖州市食品药品检验研究院，浙江湖州 313000；
2.华东理工大学材料科学与工程学院，上海 201424

廖辉，男，硕士，工程师，研究方向：药物分析及质量标准研究、药品微生物方法开发及应用研究

收稿日期: 2025-11-17

修回日期: 2026-04-07

录用日期: 2026-05-19

网络出版日期: 2026-08-12

基金资助

浙江省药品监督管理局科技计划项目（2024005）

收起

Study on the Suitability of the Microbial Limit Test Method for Linglianhua Granules

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1.Huzhou Institute for Food and Drug Control， Zhejiang Huzhou 313000， China；
2.School of Materials Science and Engineering， East China University of Science and Technology， Shanghai 201424， China

Received date: 2025-11-17

Revised date: 2026-04-07

Accepted date: 2026-05-19

Online published: 2026-08-12

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摘要

目的：建立灵莲花颗粒的微生物限度检查方法并验证其适用性，为准确评价该制剂的微生物污染提供依据。方法：依据《中华人民共和国药典》（2025年版）通则，进行微生物计数法及控制菌检查法的适用性试验。计数法采用平皿法（稀释度1∶10~1∶200）及含中和剂（1%卵磷脂+3%聚山梨酯80+0.5% PVP）的平皿法（1∶10、1∶50），测定对金黄色葡萄球菌、铜绿假单胞菌、枯草芽孢杆菌、白色念珠菌及黑曲霉的回收率；控制菌检查分别采用常规法、培养基稀释法及中和剂法，检测耐胆盐革兰阴性菌、大肠埃希菌及沙门菌。结果：平皿法在1∶100稀释时需氧菌总数回收率比值为0.82~1.05，在1∶50稀释时霉菌和酵母菌总数回收率比值为0.82~1.05；使用中和剂后（1∶50），需氧菌总数和霉菌酵母菌总数的回收率比值均提升至0.84~1.07。控制菌检查中，耐胆盐革兰阴性菌和大肠埃希菌在常规法与中和剂法中均能检出；沙门菌仅在培养基稀释法（10 g+190 mL TSB）或中和剂法（10 g+含中和剂90 mL TSB）中检出，常规法无法消除其抑制。结论：所建方法经验证适用于灵莲花颗粒微生物限度检查。中和剂系统可有效消除抑菌作用且无毒性，推荐需氧菌总数采用稀释平皿法（1∶100）或中和剂法（1∶50），霉菌和酵母菌总数采用中和剂法（1∶50）；对于控制菌检查，耐胆盐革兰阴性菌及大肠埃希菌可采用常规法；沙门菌则需采用培养基稀释法或中和剂法。该方法准确可靠，可用于产品质量控制。

关键词： ; font-size:medium; ">灵莲花颗粒；微生物限度；方法适用性；抑菌活性；回收率比值

本文引用格式

廖辉 , 贺婵娟 , 杨红霞 , 张璐 . 灵莲花颗粒微生物限度检查方法适用性研究[J]. 中国药物评价, 2026 , 43(3) : 189 -189-195 . DOI: 10.2095-3593.2026.050016

Abstract

Objective： To establish and validate a microbial limit test method for Linglianhua granules， providing a basis for accurately assessing microbial contamination in this preparation. Methods：In accordance with General Chapters of the Chinese Pharmacopoeia (2025 edition)， suitability tests for microbial enumeration and tests for specified microorganisms were conducted. For microbial enumeration， the plate count method (dilution 1∶10 to 1∶200) and the plate method containing neutralizers (1% lecithin， 3% polysorbate 80， 0.5% PVP) at dilutions of 1∶10 and 1∶50 were used to determine the recovery rates of Staphylococcus aureus， Pseudomonas aeruginosa， Bacillus subtilis， Candida albicans， and Aspergillus niger. For the tests for specified microorganisms， the conventional method， the medium dilution method， and the neutralizer method were employed to detect bile-tolerant Gram-negative bacteria， Escherichia coli， and Salmonella. Results： Using the plate method， the recovery ratio for the total aerobic bacterial count at a 1∶100 dilution ranged from 0.82 to 1.05， while that for total molds and yeasts at a 1∶50 dilution ranged from 0.82 to 1.05. After applying the neutralizer (at 1∶50 dilution)， the recovery ratios increased to a range of 0.84 to 1.07. In tests for specified microorganisms， bile-tolerant Gram-negative bacteria and Escherichia coli， were detected by both the conventional and neutralizer methods. Salmonella was detectable only with the medium dilution method (10 g+190 mL TSB) or the neutralizer method (10 g+90 mL TSB containing neutralizers)， as inhibition could not be eliminated by the conventional method. Conclusion： The established method was validated and is suitable for the microbiological limit testing of Linglianhua granules. The neutralizer system effectively eliminated antimicrobial effects without introducing toxicity. It is recommended to use the dilution plate method (1∶100) or the neutralizer method (1∶50) for the total aerobic microbial count， and the neutralizer method (1∶50) for the total combined yeasts and molds count. The conventional method was found to be suitable for testing bile-tolerant Gram-negative bacteria and Escherichia coli， whereas Salmonella requires the medium dilution method or the neutralizer method. This method is accurate， reliable， and applicable for product quality control.

Key words： ; font-size:medium; ">Linglianhua granules； Microbial limit； Method suitability； Antimicrobial activity； Recovery ratio

参考文献

[1] 常章富. 简谈如何用好中成药(二)[J]. 中国执业药师， 2009， 6(4)：47-50.
     [2] 路遥，姚瑶，白文佩，等. 灵莲花颗粒对不同睡眠质量更年期综合征患者的疗效研究[J]. 中国药物滥用防治杂志， 2021， 27(6)：907-910.
     [3] Vu N， Lou JR， Kupiec TC. Quality control analytical methods： microbial limit tests for nonsterile pharmaceuticals， Part 1 [J]. Int J Pharm Compd， 2014， 18(3)：213-221.
     [4] Vu N， Lou JR， Kupiec TC. Quality control： microbial limit tests for nonsterile pharmaceuticals， part 2 [J]Int J Pharm Compd， 2014， 18(4)：305-310.
     [5] 范一灵，朱冉，杨燕，等. 2025年版《中国药典》微生物标准的变化与发展趋势[J]. 中国药品标准，2025，26(1)：93-98.
     [6] 庞云娟，刘康连，梁晓玲，等. 药品微生物限度检查方法适用性的研究进展[J]. 药物分析杂志，2024，44(8)：1285-1292.
     [7] Lombard B， Cornu M， Lahellec C， et al. Experimental evaluation of different precision criteria applicable to microbiological counting methods[J]. J AOAC Int， 2006， 89(1)：11A-13A.
     [8] 许华玉，杜芸，钱文静，等. 药品微生物限度检查法中细菌和真菌计数方法的验证试验[J]. 中国中药杂志，2005，30(24)：1918-1920.
     [9] Wang Lirong， Chen Shujing， Liu Suyi， et al. A comprehensive review of ethnopharmacology， chemical constituents， pharmacological effects， pharmacokinetics， toxicology， and quality control of gardeniae fructus[J]. J Ethnopharmacol. 2023， 320：117397.
     [10] 李海波，马金凤，庞倩倩，等. 栀子的化学成分研究[J]. 中草药， 2020， 51(22)：5687-5697.
     [11] 刘亭亭，王萌. 女贞子化学成分与药理作用研究进展[J]. 中国实验方剂学杂志， 2014， 20(14)：228-234.
     [12] Xi Yanli， Hu Lianxin， Chen Xiang， et al. Antibacterial and anti-inflammatory polysaccharide from fructus ligustri lucidi incorporated in pva/pectin hydrogels accelerate wound healing[J]. Molecules， 2024， 29(7)：1423.
     [13] 程敏，胡正海. 墨旱莲的生物学和化学成分研究进展[J]. 中草药， 2010， 41(12)：2116-2118.
     [14] Md S， Md SI， Sadiya BR， et al. Bioactivity assessment of analgesic and anti-inflammatory properties of eclipta prostrata extract on rat models[J]. Asian J Adv Res Rep， 2025， 19(5)：226-236.
     [15] Yu Shujuan， Yu Jinhai， He Fei， et al. New antibacterial thiophenes from Eclipta prostrata[J]. Fitoterapia， 2020， 142：104471.
     [16] agar T， Frlan R， Koevar Glava N. Using subcritical water to obtain polyphenol-rich extracts with antimicrobial properties[J]. Antibiotics (Basel)， 2024， 13(4)：334.
     [17] Cueva C， Moreno-Arribas MV， Martín-álvarez PJ， et al. Antimicrobial activity of phenolic acids against commensal， probiotic and pathogenic bacteria[J]. Res Microbiol， 2010， 161(5)：372-382.
     [18] Justino AB， Peres PABM， Saito N， et al. Quercetin-conjugated gold-decorated simonkolleite nanohybrids： insights into oxidative stress and antibacterial activity[J]. J Trace Elem Med Biol， 2025， 91：127709.
     [19] Zhao Yuchao， Xia Chunfeng， Jiang Chengkun， et al. Comparison of the content of active ingredients and antioxidant， antibacterial and anti-inflammatory activities in different parts of Fructus Choerospondiatis[J]. Sci Rep， 2024， 14(1)：28670.
     [20] Cruz RC， Werneck SMC， Oliveira CS， et al. Influence of different media， incubation times， and temperatures for determining the MICs of seven antifungal agents against paracoccidioides brasiliensis by microdilution[J]. J Clin Microbiol， 2013， 51(2)：436-443.
     [21] 赵新霞，杨淑先，王海波，等. 聚山梨酯80对药品微生物限度检查的影响[J]. 中国药品标准， 2019， 20(3)：224-230.
     [22] Katerji A， Saleh T， Yaser B， et al. Evaluation of new formulations for neutralizing antimicrobial preservatives in pharmaceutical preparations[J]. Heliyon， 2023， 9(3)：e14555.
     [23] 廖祥茹，陈晨，李旸，等. 聚山梨酯80在微生物限度检查中的应用[J]. 中国药师， 2014， 17(10)：1776-1777.
     [24] Shi Xuechao， Wang Yimei， Hou Shuxian， et al. Synthesis of bimetallic compounds derived from Polyvinylpyrrolidone modified-FeMo2S3 and ZnMo2S3 Mo-clusters and their antibacterial application against water-borne microorganisms[J]. Tungsten， 2025， 7(3)：1-18.
     [25] Gradiar Centa U， Miheli M， Sternia M， et al. Tackling microbial adhesion to surfaces by adding mesoporous SiO2 nanoparticles to nanocomposite based on PVDF-HFP and PVP polymers[J]. Surf Interfaces， 2025， 56：105713.
     [26] Garcia AM， Bizeto MA， Ferrari VB， et al. Direct evaluation of microbial growth dynamics and colloidal stability of silver nanoparticles stabilized by poly(vinyl pyrrolidone) and poly(vinyl alcohol)[J]. J Nanopart Res， 2020， 22(80)：3974-3983.

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