开发一种测定粉末状物质中休眠生物存活率的技术

I. A. Gaidashev, Yuliana Gennadievna Nikolaeva, A. V. Syroeshkin
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摘要

为了实现制药生物技术和含有冻干细菌制剂的益生菌质量控制的目的,以及确保生物安全的目的,需要有快速测定休眠形式活力的方法。本文介绍了一种测定干粉物质活力的方法。本文提出了冻干的 Kalanchoe daigermontiana 粉末、干燥的盐蒿休眠卵和干燥的 Lycopodium sp.建议将 SARS-CoV-2 病毒样颗粒(VLPs)悬浮液作为无细胞参考材料。研究中使用了以下元素分析方法:能量色散装置上的 X 射线荧光光谱法(XRF)、电热原子化原子吸收光谱法和泽曼背景校正法(GZ-AAS)。自由水含量是通过核磁共振自旋回波测定的。还使用了一种新方法来记录厘米和毫米波长范围内宽带无线电发射的动力学变化,测量持续时间长达 20 分钟。制备了 K. daigermontiana 和 A. salina 的活体和灭活粉末制剂,并通过元素组成、含水量和自旋-自旋弛豫时间(T2)对其进行了表征。对于这些活体和失活制剂,已确定其内在热辐射的通量密度会随着活力的丧失而降低一个数量级。未来,所获得的结果将有助于快速检测冻干疗法(从药用植物到稳定肠道微生物菌群的细菌混合物)的活力。关键词:毫米波发射、X 射线荧光分析、核磁共振自旋回波、纳米颗粒、粉末状物质、药用植物原料
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Development of a technique for determining the viability of dormant organisms in powdered substances
The purposes of pharmaceutical biotechnology and quality control of probiotics containing lyophilized bacterial preparations, as well as the purpose of ensuring biological safety, require methods for rapid determination of the viability of dormant forms. This paper describes an approach to determining the viability of dry powdered substances. Lyophilized powder of Kalanchoe daigermontiana, dry dormant eggs of Artemia salina, and dry powder of Lycopodium sp. spores were proposed as model objects. Suspensions of virus-like particles (VLPs) to SARS-CoV-2 were proposed as a cell-free reference material. The following methods of elemental analysis were used in the study: X-ray fluorescence spectroscopy (XRF) on an energy-dispersive device, atomic absorption spectroscopy with electrothermal atomization, and Zeeman background correction (GZ-AAS). The free water content was determined by NMR spin echo. A new method was also used for recording the kinetics of changes in broadband radio emission in the centimeter and millimeter wavelength ranges, with a measurement duration of up to 20 minutes. Live and inactivated powdered preparations of K. daigermontiana and A. salina were prepared and characterized by elemental composition, water content, and spin-spin relaxation time (T2). For these live and inactivated preparations, it was determined that the flux density of intrinsic thermal radio emission decreases by an order of magnitude with the loss of viability. In the future, the results obtained will allow for a rapid test of the viability of lyophilized therapeutics (from medicinal plants to bacterial mixtures that stabilize the intestinal microflora). Keywords: millimeter emission, X-ray fluorescence analysis, NMR spin echo, nanoparticles, powdered substances, medicinal plant raw materials
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