COMPARISON OF DIFFERENT METHODS OF CENTRIFUGATION FOR OPTIMUM PREPARATION OF PLATELET RICH PLASMA (PRP)& FRAMING A CUSTOMIZED PRP PROTOCOL-AN INNOVATIVE APPROACH

Introduction:The rationale behind the therapeutic potential of the high concentration of platelets in PRP (Platelet Rich Plasma) for intervention procedures of soft tissue injuries is often challenged by the ability of the centrifugation methods followed in extracting the high quality PRP with prompt contented quantity too.This study aims to unravel the basic science of PRP preparation involved in centrifugation protocols, spin methods, speed and time duration of centrifugation to obtain consistent therapeutic platelets yield. Materials and Methods:30 participants were subjected to intervention procedures using PRP. For therapeutic purpose 3 centrifugation protocol types are followed. Ø Protocol 1-Conventional Old protocol Ø Protocol 2-Customised New protocol Ø Protocol 3-Spin Reversal of the routine -hard spin followed by soft spin The Platelets and WBC concentration and composition in the final autologous PRP sample harvested as a result of double centrifugation method are compared for quantity and efficiency. Results:The maximum PCF-3.21 times above baseline and PRR -64.21% are through new customized spin proving its efficiency. As determined by one-way ANOVA, there is statistically significant difference between the PROTOCOL groups (1,2,3) for the variables PRP platelet count, PRP Leucocytes, PCF, PRR. A Tukey post hoc test revealed that there is statistically significant value for Protocol 2 and Protocol 3 compared to the Protocol 1 for PRP platelet count, PCF and PRR whereas for PRP Leucocytes, there was a statistically significant difference between the Protocol 2 and Protocol 3 which again implicates the necessity of spin reversal in yielding better quantity of leucocytes. Conclusion:To produce PRP samples with consistent and reproducible compositionsof platelets and leucocytes with better quality control standard through a detailed, precise and stepwise description of the manufacturing protocol has been explained in our study. Our perspective is in standardizing a safe, simple protocol that can be followed to obtain an optimal consistent platelet yield without the use of commercial kits, which has been proved statistically too.

Development of Simultaneous Analysis Method for Multi-Compounds Content of New Shilajit Using HPLC-UV and the Cognitive Enhancing Effect: Mongolian Shilajit

Shilajit has a longstanding use as an anti-aging and memory enhancing drug. It is known to have excellent anti-bacterial effects and is believed to be effective for cognitive enhancement, but is difficult to standardize because of the lack of quality control standards. This study, for the first time, proposes a quality control standard using a simultaneous analytical method for the drug’s multi-compound content using high-performance liquid chromatography-ultraviolet detection (HPLC-UV) as an aid for the internationalization of Mongolian Shilajit. Phenolic compounds 1-6 were isolated from Mongolian Shilajit extract using bioassay-guided isolation, and the isolated compounds were evaluated for cognitive-related anti-Alzheimer’s disease (AD) activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), β-site amyloid precursor protein-cleaving enzyme 1 (BACE1), and advanced glycation end-product (AGE) formation assays. The isolated compounds showed good effects for each activity. In addition, the isolated compounds were successfully quantified using a validated quantitative HPLC analysis method. As a result, the isolated compounds were suggested as standard marker compounds for Mongolian Shilajit. Also, we proved that the original material of Mongolian Shilajit is a lichen named Xanthoparmelia somloensis (Gyel.) Hale using HPLC-UV, ultra-high-performance liquid chromatography-electrospray ionization/hybrid linear trap-quadruple-orbitrap-high-resolution mass spectrometry (UHPLC-ESI/LTQ-HRMS).

A study on the preparation and quality standard of Saifula granules

Objective: To establish the decocting and formulation technology of Saifula granules, and establish a reliable TLC identification quality control standard. Methods: Orthogonal test was used to optimize the water extraction process and TLC was used for qualitative identification. Results: To optimize the process parameters of SFL granules, first, the content of total flavonoids and the total extraction rate were taken as the indexes. Through single factor and orthogonal test, the optimal extraction process was determined as follows: extraction time 1 h, solvent multiple 1:10, extract three times. After the process parameters were determined, in order to further explain the chemical composition of SFL extract, 15 compounds in SFL extract were identified by UHPLC-Q-Orbitrap-MS, 13 batches of SFL extracts were prepared, and their fingerprints were analyzed. The results showed that there were 28 common characteristic peaks in the 13 batches of SFL extracts. The similarity evaluation results of fingerprints showed that the similarities of 13 batches of SFL extracts were greater than 0.9. An HPLC-DAD method for the simultaneous determination of gallic acid, corilagin, and ellagic acid was established. The method has good repeatability, stability, and accurate results. Conclusion: The standard can comprehensively reflect the material basis of SFL granules, and the above methods are accurate, easy to operate, stable and feasible, and provide scientific basis and basis for the development, promotion, and clinical application of SFL granules.

Vaccinium macrocarpon (Cranberry)-Based Dietary Supplements: Variation in Mass Uniformity, Proanthocyanidin Dosage and Anthocyanin Profile Demonstrates Quality Control Standard Needed

Vaccinium macrocarpon (syn. American Cranberry) is employed in dietary supplements (DS) with the aim to improve urinary tract well-being. This property is linked to the antiadhesion-activity of proanthocyanidins (PACs) against uropathogenic-bacteria. However, the current European legislation has been criticized for being weak and ineffective. Indeed, recent scientific works report mislabeled, contaminated, and adulterated supplements containing dangerous or unknown compounds, or sold at toxic doses. In this work, we analysed 24 DS that claim to contain cranberry, and to have a specific dosage of PACs. Our tests included the control of the good manufacturing practice according to the European Pharmacopoeia, and the verification of the claimed dosage of PACs. Moreover, in order to confirm the real presence of cranberry in DS, chemical fingerprinting via HPLC-UV/Vis-MS/MS was employed. Our results showed that 17 DS did not comply with the uniformity test of dosage forms, and only five contained cranberry. Finally, 16 DS claimed an incorrect amount of PACs. These data suggest that several cranberry-based DS are present in the European market with insufficient quality controls. Considering that often DS are self-prescribed by consumer relying on their claim, the data obtained in this work should encourage more controls and stricter rules.

Quality evaluation of Lycium barbarum (wolfberry) from different regions in China based on polysaccharide structure, yield and bioactivities

Background Lycium barbarum (wolfberry) has been widely cultivated in China, particularly in northwest regions. However, the fruit size and taste of L. barbarum from different habitats are quite different. Traditionally, only the fruit of L. barbarum produced in Ningxia province is recorded as an authentic herb, although the detailed mechanism responsible for this remains obscure. Polysaccharides are considered major active ingredients in L. barbarum which is crucial for its quality evaluation. Methods In this study, we assessed the yield, monosaccharide composition, molecular weight, and conformation of L. barbarum polysaccharides (LBPs) collected from different regions of China. The antioxidant and immune activities of LBPs were also determined as its quality indicator. Results Our results showed that the similarity values of monosaccharide composition were larger than 0.926, and the Mw of the two fractions (peaks 1–2) in LBPs were ranging from 1.36 × 106 to 2.01 × 106 (peak 1), and 6.85 × 104 to 10.30 × 104 (peak 2) which indicated that the structure of LBPs were similar. In addition, results showed that there was no significant difference in antioxidant and immune activities of nine LBPs from different regions. However, the yield of LBPs from Qinghai Province (low atmospheric temperature, high altitude) was significantly lower (p < 0.05) than those collected from Xinjiang and Ningxia province. Conclusions These data suggested that the L. barbarum produced in Ningxia and Xinjiang maybe more suitable as materials for medicines and functional foods. This study also provides a reference for improving the quality control standard of LBPs.

The antimicrobial activity of alcoholic extracts of leaves and bark from two varieties of ”Sangre de Drago” compared with the antimicrobial activity present in the latex of the same species

Background: This research was carried out in order to study the antimicrobial effectiveness of crude latex of two varieties of "Sangre de Drago": Croton lechleri Muller Arg. and Croton urucurana Baill and compare that effectiveness to the antimicrobial activity of the alcoholic extracts of its leaves and bark. Methods: The activity of the alcoholic extracts and latex were evaluated against bacterial strains of Staphylococcus epidermidis, Bacillus subtilis and Escherichia coli. The extraction of the alcoholic extracts (20% Tincture) of the leaves, bark and latex from the two Croton species was carried out by maceration using 70% alcohol as a menstruum, at room temperature, for 2 to 7 days, with shaking at least twice a day. A 20% tincture was obtained, from which the physical and chemical parameters were determined as indicated by the Ecuadorian Quality Control Standard for natural medicinal products. Results: It was found that both the alcoholic extracts of the plant material and the crude latex indicate antimicrobial activity for S. epidermidis, moderate antimicrobial activity for B. subtilis and no antimicrobial activity for E. coli. The moderate antimicrobial activity against B. subtilis, at doses of 125 p.p.m., is in line with the findings of previous studies by other authors. Conclusions: he antimicrobial activity of the latex of the two species against S. epidermidis is not registered in literature and, the negative antimicrobial activity for E. coli does not agree with what has been reported by previous studies.

Evaluation of two Sigma Transwab® systems for Maintenance of Viability of pathogenic Candida spp. Using the Clinical and Laboratory Standards Institute M40-A2 Standard

ABSTRACTA key aspect to routine microbiology processes include the retrieval, transport and maintenance of specimens. Swab transport systems (STS) can be utilised for their low cost, ease of use and their ability to recover and maintain specimens over long durations. An increase in healthcare complications due to fungal infections raises the requirement for STS to efficiently recover and preserve pathogens of yeast origin. The Clinical and Laboratory Standards Institute (CLSI) M40-A2 protocol is used to assess the compliance of STS to a quality control standard but at present does not include the recovery of yeast. The aim of this study was to compare the results of two commercial STS and their ability to recover and maintain viability of five clinical and reference strains of Candida spp., including C. auris, when stored at room temperature and 4°C, over 48 h, using the qualitative roll plate method. Findings from this study indicate that the STS used in this study are suitable for the collection and maintenance of the Candida spp. tested, and is very suitable for the recovery of clinical C. auris.

Quality Control Audit in the Republic of Tajikistan: Present Status and Perspective Standardization

The article describes the development and implementation of quality control standard auditor’s work in the Republic of Tajikistan based on the International Standards on Auditing.