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Effects of drying method and excipient on the structure and physical stability of protein solids: Freeze drying vs. spray freeze drying.

ABSTRACT: This study aims to determine the impacts of drying method and excipient on changes in protein structure and physical stability of model protein solids. Protein solids containing one of two model proteins (lysozyme or myoglobin) were produced with or without excipients (sucrose or mannitol) using freeze drying or spray freeze drying (SFD). The protein powders were then characterized using solid-state Fourier transform infrared spectroscopy (ssFTIR), differential scanning calorimetry (DSC), circular dichroism spectrometry (CD), size exclusion chromatography (SEC), BET surface area measurements and solid-state hydrogen deuterium exchange with mass spectrometry (ssHDX-MS). ssFTIR and CD could identify little to no difference in structure of the proteins in the formulation. ssHDX-MS was able to identify the population heterogeneity, which was undetectable by conventional characterization techniques of ssFTIR and CD. ssHDX-MS metrics such as D_max and peak area showed a good correlation with the protein physical instability (loss of the monomeric peak area by size exclusion chromatography) in 90-day stability studies conducted at 40 °C for lysozyme. Higher specific surface area was associated with greater loss in monomer content for myoglobin-mannitol formulations as compared to myoglobin-only formulations. Spray freeze drying seems a viable manufacturing technique for protein solids with appropriate optimization of formulations. The differences observed within the formulations and between the processes using ssHDX-MS, BET surface area measurements and SEC in this study provide an insight into the influence of drying methods and excipients on protein physical stability.

SUBMITTER: Mutukuri TT

PROVIDER: S-EPMC7856218 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Microcapsules of ciriguela peel extracts obtained by ultrasound-assisted extraction were prepared by spray drying, whose results were compared with those of freeze-drying as a control. The effects of spray-drying air temperature, feed flow rate and ratio of encapsulating agents (maltodextrin and arabic gum) were studied. Encapsulation efficiency, moisture content, total phenolic compounds (TPC), water activity, hygroscopicity, solubility, colorimetric parameters, phenolic profile by HPLC/DAD, simulated gastrointestinal digestion and morphology of spray-dried and freeze-dried microcapsules were evaluated, as well as their stability of TPC during 90 days storage at 7 and 25 °C. Spray-dried extract showed higher encapsulation efficiency (98.83%) and TPC (476.82 mg GAE g-1) than freeze-dried extract. The most abundant compounds in the liquid extract of ciriguela peel flour were rutin, epicatechin gallate, chlorogenic acid and quercetin. Rutin and myricetin were the major flavonoids in the spray-dried extract, while quercetin and kaempferol were in the freeze-dried one. The simulated gastrointestinal digestion test of microencapsulated extracts revealed the highest TPC contents after the gastric phase and the lowest one after the intestinal one. Rutin was the most abundant compound after the digestion of both spray-dried (68.74 µg g-1) and freeze-dried (93.98 µg g-1) extracts. Spray-dried microcapsules were of spherical shape, freeze-dried products of irregular structures. Spray-dried microcapsules had higher phenolic compounds contents after 90 days of storage at 7 °C compared to those stored at 25 °C, while the lyophilized ones showed no significant difference between the two storage temperatures. The ciriguela agro-industrial residue can be considered an interesting alternative source of phenolic compounds that could be used, in the form of bioactive compounds-rich powders, as an ingredient in pharmaceutical, cosmetic and food industries.

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Effects of drying method and excipient on the structure and physical stability of protein solids: Freeze drying vs. spray freeze drying.

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