1

Mechanical shock results in cavitation in protein solutions (the larger the volume of the liquid, the stronger the cavitation) and the formation of hydroxyl radicals.
Cavitation promotes formation of particles in the volume of the solution. The higher the protein concentration is, the greater the number of particles produced.

Randolph TW, Schiltz E, Sederstrom D, et al. Do not drop: mechanical shock in vials causes cavitation, protein aggregation, and particle formation. J Pharm Sci. 2015;104(2):602-611. doi:10.1002/jps.24259

2

Shaking and stirring of a protein (IgG1) solution resulted in the formation of different species of aggregates both qualitatively and quantitatively, and stirring was more stressful (and therefore led to increased protein aggregation) than shaking of the IgG1 solution.
The headspace in the vials had a great effect on the stability of the protein formulation during shaking.
The temperature (5 or 25 °C) was important when shaking, but not when mixing.
The addition of polysorbate prevented the aggregation of antibodies, but at certain concentrations, it resulted in increased protein aggregation.

Kiese S, Papppenberger A, Friess W, Mahler HC. Shaken, not stirred: mechanical stress testing of an IgG1 antibody. J Pharm Sci. 2008;97(10):4347-4366. doi:10.1002/jps.21328

3

Shaking pharmaceutical preparations induced the formation of structures characterized by greater disorderliness and less complexity, while at the same time increasing the gaps between the structure elements.

The differences found between the shaken and gently mixed samples may be due to the shaking-induced aggregation of large molecules or, in some cases, the inclusion of air bubbles and/or the formation of particles.

Kokornaczyk MO, Würtenberger S, Baumgartner S. Impact of succussion on pharmaceutical preparations analyzed by means of patterns from evaporated droplets. Sci Rep. 2020;10(1):570. doi:10.1038/s41598-019-57009-2

4

When IgG solution (both in water and in water-alcohol) is shaken:
- IgG macromolecule aggregation is increased;
- bubbles are generated with a different size range.
After shaking IgG solution, the process of aggregation is enhanced in a water-alcohol solvent (compared to water) due to IgG denaturation. IgG aggregates have a size of ~300 nm in water and ~900 nm in water-alcohol solutions.
IgG flotation is much more effective in water. This can be explained by the better adsorption of IgG particles (molecules and aggregates) on water bubbles compared to ethanol-water mixtures.

Bunkin NF, Shkirin AV, Ninham BW, et al. Shaking-Induced Aggregation and Flotation in Immunoglobulin Dispersions: Differences between Water and Water-Ethanol Mixtures. ACS Omega. 2020;5(24):14689-14701. doi:10.1021/acsomega.0c01444

5

1) The extent of the effect on the chemiluminescence of water and protein solutions depends on the shaking mode.
Shaking water with a frequency of 10 Hz and an amplitude of 12 mm, as well as with a frequency of 30 Hz and an amplitude of 2.3 mm, causes pronounced changes in the intensity of its chemiluminescence.
Shaking protein solutions (bovine serum albumin) at a frequency of 30 Hz for 60 seconds causes pronounced changes in the intensity of their chemiluminescence.

2) The magnitude of the effect exerted by an alternating magnetic field on the chemiluminescence of a protein solution depends on the frequency of the magnetic field and the protein concentration.
Exposure to a magnetic field with a frequency of 50 Hz and a flux density of 50 µT for 5 minutes causes pronounced changes in the chemiluminescence intensity of IgG solution with a concentration of 1 mg/ml.

Astashev ME, Serov DA, Sarimov RM, Gudkov SV. Influence of the Vibration Impact Mode on the Spontaneous Chemiluminescence of Aqueous Protein Solutions. Phys Wave Phen. 2023;31(3):189-199. doi:10.3103/S1541308X23030020

6

After mechanical treatment, the concentration of gases (molecular oxygen and carbon dioxide) in the water decreased. Neither the average size nor the number of nanoscale gas bubbles changed.
It was found that mechanical effect promotes the formation of hydrogen peroxide and hydroxyl radicals in an aqueous solution.
The generation of reactive oxygen species occurred during mixing both in a turbulent flow and in a laminar flow with Dean vortices, created in a microfluidic device (with a lower intensity).
An important and possibly decisive step in the generation of reactive oxygen species when exposed to mechanical effects is the transition of molecular oxygen from the triplet state to the singlet one.

Gudkov SV, Penkov NV, Baimler IV, et al. Effect of Mechanical Shaking on the Physicochemical Properties of Aqueous Solutions. Int J Mol Sci. 2020;21(21):8033. doi:10.3390/ijms21218033

7

Atomizing of water enriched the resulting microdroplets with hydrogen peroxide.

Lee JK, Walker KL, Han HS, et al. Spontaneous generation of hydrogen peroxide from aqueous microdroplets. Proc Natl Acad Sci U S A. 2019;116(39):19294-19298. doi:10.1073/pnas.1911883116

8

Potentiated solutions (solutions resulting from successive dilutions with vibration treatment) of Mercury (II) increase the effective concentration (activity) of mercury ions when performing inversion voltammetry.

Petrov SI, Epstein OI. Effect of potentiated solutions on mercury(II) signal in inversion voltammetry. Bull Exp Biol Med. 2003;135 Suppl 7:99-101. doi:10.1023/a:1024707519510

9

Use of high dilutions of superconductor components (prepared with exposure of the solution to the vibration effect) in the manufacture of superconducting ceramics improved the properties of the final product.

Kamentsev KE, Bush AA. The superconducting properties of YBa2Cu3Oy ceramics fabricated using ultrahigh dilution technology. Ceram Int. 2022;48(21):32196-32204. doi:10.1016/j.ceramint.2022.07.161

10

The use of high-dilutions of piezoelectric powder (prepared with exposure of the solution to the vibration effect) in the manufacture of piezoelectric ceramics changed the properties of the final product.

Spitsin A, Bush AA, Kamentsev KE. Piezoelectric and dielectric properties of Bi3TiNbO9 prepared by hot pressing from powders activated using the serial dilution method. Sci Rep. 2020;10:22198. doi:10.1038/s41598-020-78826-w

11

Long-term (several hours and/or days) changes in the spectral parameters were detected after shaking, ultraviolet irradiation, and other effects on the aqueous medium.

Belovolova L, GlushkovM. Vinogradov E, Babintsev V, Golovanov V.Ultraviolet fluorescence of water and highly diluted aqueous media. Phys Wave Phen. 2009;17:21-31. doi:10.3103/S1541308X0901004X.

12

Subsonic frequency fluctuations changed the redox potential of water.

Styrkas AD, Nikishina NG. Mechanochemical processes in water. High Energy Chem. 2007;41:396-402. doi:10.1134/S0018143907060021

13

Shaking lead to changes in the spectra of UHD of europium acetate.
Processing of water with a microfluidic system significantly affected its properties: intermolecular binding is changed

Slatinskaya OV, Pyrkov YN, Filatova SA, Guryev DA, Penkov NV. Study of the Effect of Europium Acetate on the Intermolecular Properties of Water. Front Phys. 2021;9:641110. doi:10.3389/fphy.2021.641110

14

Mechanical treatment of water increased its electrical conductivity and heat capacity, and decreased its viscosity.

VPT – Vortex Process Technology. Watreco. https://www.watreco.com/technology. Accessed December 6, 2023.

15

Along with the traditional rhythmic shaking, other physical effects influenced the physical-chemical properties of water and aqueous solutions.

Gudkov SV, Lyakhov GA, Pustovoy VI, Shcherbakov IA. Influence of Mechanical Effects on the Hydrogen Peroxide Concentration in Aqueous Solutions. Phys Wave Phen. 2019;27(2):141-144. doi:10.3103/S1541308X19020092

Gudkov SV, Lyakhov GA, Pustovoy VI, Shcherbakov IA. Vibration–Vortex Mechanism of Radical-Reaction Activation in an Aqueous Solution: Physical Analogies. Phys Wave Phen. 2021;19(2):108-113. doi: 10.3103/S1541308X21020060

Shcherbakov IA. Influence of External Impacts on the Properties of Aqueous Solutions. Phys Wave Phen. 2021;29(2):89-93. doi:10.3103/S1541308X21020114

Shcherbakov IA. Specific features of the concentration dependences of impurities in condensed media. Phys Wave Phen. 2020;28(2):83-87. doi:10.3103/S1541308X20020156

16

Shaking during the preparation of serial dilutions of hydrated fullerene affected its properties (in particular, the buffer capacity and the degree of heterogeneity in droplets). The properties of solutions during dilutions changed according to complex and nonlinear laws, so the final dilutions could not be identical to the initial substance and water in their characteristics.

Yablonskaya O, Buravleva E, Novikov K, Voeikov V. Peculiarities of the physicochemical properties of hydrated C60 fullerene solutions in a wide range of dilutions. Front Pharmacol. 2021;9:627265. doi:10.3389/fphy.2021.627265

17

In a set of dilutions (C2-C33) prepared using serial dilution with shaking, the impedance was determined using dielectric spectroscopy in the range of 20 Hz – 10 MHz. It was shown that with an increase in the dilution level, the average resistance decreased, and the entire series of dilutions had a certain pattern that persisted up to 30 days. This means that the result does not depend on the concentration of the solute, but on the sample preparation procedure. This is probably due to the emergence of reactive oxygen and nitrogen species during mechanical shaking of the solution, and the products of subsequent complex chemical reactions.

Lobyshev VI. Non-Monotonous Impedance Patterns of Diclofenac Centesimal Potentiated Solutions and Their Evolution. Water. 2022. doi:10.14294/WATER.2022.S4

18

The publication combines the results of ultrahigh dilution studies carried out by five independent research groups using nuclear magnetic resonance (NMR). They have shown an increase in the T1 relaxation time and the T1/T2 ratio with a decrease in the T2 relaxation time in UHD. Changes in the relaxation time gradually increased with increasing the dilution level, and they completely disappeared after the heating-cooling cycle. These results can be explained by the formation of growing nanostructures with the participation of nanobubbles and other elements coming from the environment.

Demangeat JL. Water proton NMR relaxation revisited: Ultrahighly diluted aqueous solutions beyond Avogadro’s limit prepared by iterative centesimal dilution under shaking cannot be considered as pure solvent. J Mol Liq. 2022;360:119500. doi:10.1016/j.molliq.2022.119500

19

The photon emission (PE) intensity in the Na/K-bicarbonate solution increased 2-3-fold when hydrated fullerenes were added in doses equivalent to 10^-13, 10^-15, and 10^-19-10^-20 М.
The addition of intermediate doses affected the PE of Na/K-bicarbonate significantly less.

Voeikov VL, Do Ming HA, Mukhitova OG, et al. Activated bicarbonate solutions as models of confined ontic open system and prototypes of living respiring systems. Int J Des Nat Ecodyn. 2010;5(1):30-38. doi:10.2495/DNE-V5-N1-30-38

20

The procedure of serial dilution of the initial solution with shaking at each step (potentization) changes the collective properties of the aqueous solution, and these properties are stored by the system for a long time.
The properties of potentized solutions are explained more by the mechanochemical modification of water and aqueous solutions of substances as a result of shaking, rather than by the dilution process.
Multiple serial dilution of solutions accompanied by intensive shaking leads to the formation of chemically active compounds and self-organization in the solution, which manifests itself in the formation of mesoparticles and non-monotonic changes in physical-chemical properties.

Lobyshev VI. Biological activity of low and ultra-low concentrations. Biophysics. 2022;67:523-533. doi:10.1134/S0006350922040145

21

Repeated shaking of antiserum dilutions prepared using vigorous shaking leads to an increase in the value of the T1/T2 ratio in dilutions.

Demangeat JL. Gas nanobubbles and aqueous nanostructures: the crucial role of dynamization. Homeopathy. 2015;104(2):101-115. doi:10.1016/j.homp.2015.02.001

22

In 1990s-2020s, several research groups used NMR to show an increase in the T1/T2 ratio in dilution samples prepared with vigorous shaking.

Demangeat JL. Water proton NMR relaxation revisited: Ultrahighly diluted aqueous solutions beyond Avogadro’s limit prepared by iterative centesimal dilution under shaking cannot be considered as pure solvent. J Mol Liq. 2022;360:119500. doi:10.1016/j.molliq.2022.119500