Investigating the value of balance and proprioception scores to predict lower limb injuries in professional judokas.

We investigated the ability of balance and proprioception screening tests to predict lower limb injuries in professional male judokas. Fifty-three male judokas of the national teams (Mean ± SD, age 18.68 ± 3.08 years, weight 75.34 ± 11.62 kg, height 175.28 ± 7.24 cm) participated in this study. Limits of stability (LOS), single leg stability (SLS), and knee joint position sense (JPS) was recorded as the screening tests before starting the 10 month follow up. Lower limb injury was recorded all through the follow up. Fifteen lower limb sport injuries were recorded for 53 judokas during the follow-up recordings. Significant accuracy of SLS, overall bilateral ratio (AUC 0.646, 95% CI 0.452-0.839, p = 0.046), as well as JPS 60° bilateral ratio (AUC 0.657, 95% CI 0.480-0.834, p = 0.044), and LOS overall (AUC 0.696, 95% CI 0.551-0.840, p = 0.031) were revealed discriminating between injured and uninjured judokas. The optimum cut-off of SLS, overall bilateral ratio, JPS 60° bilateral ratio, and LOS overall associated with belonging to uninjured judokas group was ≤ 1.15%, ≤ 1.09%, and ≤ 1.09 respectively (sensitivity, 0.763, 0.711, 0.789 respectively; specificity, 0.600). Although the absolute unilateral balance and proprioception scores were almost the same between injured and non-injured judokas, the bilateral ratio of both these indices were different between the two groups. Lower limbs bilateral balance and proprioception asymmetries is a more important risk factor than the absolute unilateral balance and proprioception scores for sustaining lower limb injuries in professional male judokas. Medical professionals and coaches are suggested to use these findings as pre-participation screening tools identify injury-prone athletes.

Isometric and isokinetic strength of lower-limb muscles in female athletes during different phases of menstrual cycle: a causal-comparative study.

BACKGROUND: Muscle strength is affected by different stages of the menstrual cycle in women. Since the strength of the lower limb muscles plays a significant role in the occurrence of anterior cruciate ligament injury, it seems necessary to study the strength of the lower limb muscles at different stages of this cycle to take preventive measures. Therefore, this study aimed to compare the isometric and isokinetic strength of lower limb muscles in female athletes at different menstrual cycle stages. METHODS: The present study is of a causal-comparative type. Thirty-seven female athletes in anterior cruciate ligament injury high-risk disciplines participated in this study. Isometric and isokinetic knee extensors and flexors muscle strength status, including the traditional hamstrings/quadriceps (H/Q) ratios, were recorded at different stages of the menstrual cycle (follicular, ovulatory, luteal) by Biodex isokinetic dynamometry system 4. Then, the obtained results were analyzed by repeated measure analysis of variance. RESULTS: Analysis of variance with repeated measures showed isokinetic and isometric strength in the knee flexor and extensor muscles at an angular velocity of 60˚/s in the ovulatory phase are higher than the follicular and luteal ones. The strength of the muscles declined during the follicular and luteal phases (p ≤ 0.05). CONCLUSIONS: Due to the negative effect of the follicular and luteal stages of the menstrual cycle on the strength of the flexor and extensor muscles of the knee, the risk of anterior cruciate ligament injury may increase during this period. Therefore, it is recommended that all female coaches and athletes take preventive measures during this period.

The association between periodontal diseases and helicobacter pylori: an updated meta-analysis of observational studies.

INTRODUCTION: Various studies have examined the association between periodontitis and helicobacter pylori and reported conflicting results. The aimed of this systematic review and meta-analysis estimating the association between these two variables. METHODS: Electronic databases including PubMed (Medline), Scopus, Web of Sciences and Medline (Elsevier) were searched using the relevant keywords. All observational studies comparing the association between periodontitis and helicobacter pylori were considered. The Newcastle - Ottawa Quality Assessment Scale (NOS) checklist was used for assessing quality of included studies. All statistical analyses were completed using STATA (Version 16). RESULTS: Twenty-three studies with 8,638 patients (15 case-control with 2,366 patients and 8 cross-sectional with 6,272 patients) were included in this meta-analysis. After combining the selected studies, the odds of presence the Helicobacter pylori infection in patients with the periodontal disease was 2.47 (OR: 2.47; 95% CI: 2.01, 3.03; I2: 50.87%; P: 0.001). Also, the odds after combining case-control studies was 2.77 (OR: 2.77; % 95 CI: 2.11, 3.66; I2: 37.16%; P: 0.049) and after combining cross-sectional analytical ones, it was equal to 2.07 (OR: 2.07; 95% CI: 1.62, 2.65; I2: 43.25%; P: 0.050). CONCLUSION: Based on the results of this meta-analysis, the association between Helicobacter pylori infection and the periodontal disease is evident.

Effects of FIFA 11 + warm-up program on kinematics and proprioception in adolescent soccer players: a parallel­group randomized control trial.

This study aimed to compare the effects of 8 weeks 11 + warm-up injury prevention program on kinematics and proprioception in adolescent male and female soccer players. Forty adolescent soccer players (20 males, 20 females) aged between 14-16 years old were randomly assigned into four groups. The experimental group performed the 11 + program for 8 weeks and the control group did their warm-up program. The kinematic variable in a cutting maneuver was measured using VICON motion analysis and ankle and knees' proprioception by joint position sense (JPS) was measured using a digital inclinometer. For kinematic variables only significant differences in knee valgus among females 11 + compared with female and male control groups were found (P < 0.05). Moreover, there were significant improvements in joint position sense variables in 11 + groups compared to control groups (P < 0.05). In conclusion, the 11 + program was proven to be a useful warm-up protocol in improving knee valgus and JPS among female and male adolescent soccer players. We suggest adding more training elements to the 11 + program that aimed to enhance the proper alignment of lower extremities which may consequently improve joint kinematics.

Composite functional movement screen score predicts injuries in youth volleyball players: a prospective cohort study.

We aimed to investigate whether composite Functional Movement Screen (FMS) test scores can predict musculoskeletal injuries (MSI) in youth volleyball players. 131 national young volleyball players (Males: n = 100, age = 16.5 years, height = 1.787 m, mass = 68.32 kg; Females: n = 31, age = 13.83 years, height = 1.684 m, mass = 65.12 kg) participated in this prospective cohort study. The FMS screen was performed before starting the season. MSI and exposure data were collected during the season via each team's certified athletic trainer. The mean FMS score and standard deviation for all volleyball players was 15.85 ± 3.31. A score of ≤ 14 was positive to predict MSI with specificity of 0.60 and sensitivity of 0.93. The odds ratio for (≤ 14/˃14) was 0.048. The relative risk for being injured was 3.46. Positive likelihood ratio was 2.34, and negative likelihood ratio was 0.11. The findings of this study demonstrated that an FMS score of ≤ 14 is an identifiable risk factor for injury in young volleyball players. The FMS can be used as a pre-season screening test to identify volleyball players who may be predisposed to sustaining MSI during the season ahead.

Proteomic Analysis of Adenovirus 5 by UHPLC-MS/MS: Development of a Robust and Reproducible Sample Preparation Workflow.

Adenoviruses (AdVs) have recently become widely used therapeutic vectors for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine. AdVs are large, nonenveloped viruses with an icosahedral capsid formed from several proteins that encloses double-stranded DNA. These proteins are the main components and key players in initial stages of infection by the virus particles, so their heterogeneity and content must be evaluated to ensure product and process consistency. Peptide mapping can provide detailed information on these proteins, e.g., their amino acid sequences and post-translational modifications (PTMs), which is crucial for the development and optimization of the manufacturing processes. However, sample preparation remains the main bottleneck for successful proteomic analysis of the viral proteins (VPs) of AdVs due to their low concentrations and vast stoichiometric ranges. To address this problem, we have developed a fast and efficient protocol for preparing samples for proteomic analysis of VPs of AdV5 that requires no cleaning step prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The approach enabled identification of 92% of amino acids in AdV5 VPs on average and quantification of 53 PTMs in a single LC-MS/MS experiment using trypsin protease. The data obtained demonstrate the method's potential utility for supporting the development of novel AdV-based gene therapy products (GTPs).

Does ipsilateral and bilateral knee strength status predict lower extremity injuries of elite judokas; a prospective cohort study.

The association of pre-participation knee muscle strength status with lower limb injury occurrence was investigated. Knee extensors and flexors muscle strength status including the traditional hamstrings/quadriceps (H/Q), Q/Q, H/H, and the non-dominant H/Q: dominant H/Q, HQ:HQ, ratios were recorded before the 10 month judo activity. Fifteen lower limb sport injuries were recorded for 53 judokas during the follow-up questionnaires. Significant accuracy of dominant H/Q ratio 60º/s (AUC 0.702, 95% CI 0.520 to .883, p = 0.023), as well as HQ:HQ ratios 300º/s (AUC .318, 95% CI 0.138 to 0.497, p = 0.040), and 60 º/s (AUC 0.311, 95% CI 0.130 to 0.491, p = .033) were revealed discriminating between injured and uninjured judokas. The optimum cut-off of dominant H/Q ratio associated with belonging to uninjured judokas group was 43.2% (sensitivity, 0.974; specificity, 0.533). Isokinetic knee muscle dynamometry is useful for predicting the likelihood of lower limb injuries in professional judokas during competitive activity.

Pre-participation testing of ipsilateral and bilateral knee extensors and flexors strength ratios can contribute to an injury prevention program for judokas.Ipsilateral and bilateral strength ratios should be monitored at lower and higher angular isokinetic velocities.

Isokinetic muscle strength cannot be related to the odds ratio of musculoskeletal injuries in young elite wrestlers.

BACKGROUND: Wrestling is considered one of the oldest sports in the world. There is a high rate of injuries in Wrestling. To prevent injuries, it is necessary to identify the risk factors. Despite the functional importance of strength in wrestling, few studies have investigated the relationship between isokinetic strength and the rate of injuries in the sport. OBJECTIVES: The objective of the present study was to investigate the relationship between the isokinetic strength of elite wrestlers and the rate of injuries prospectively. METHOD: A total of 72 young wrestlers with at least 2 years of experience in the Tehran Wrestling Premier League participated in this study. Before the start of the competition season, the isokinetic strength of flexor and extensor muscles of the knee and shoulder were measured at different velocities by an isokinetic dynamometer. The injuries and training hours of these wrestlers were then recorded for nine months. RESULTS: The study results showed no significant relationship between isokinetic strength of flexor and extensor muscles of the knee or shoulder at different angular speeds of 60, 180, and 300°/s. There was neither significant between the ratio of the strength flexor/extensor for knee and/or shoulder in young elite wrestlers with injuries. CONCLUSION: Isokinetic strength of lower and upper limb muscles alone cannot predict wrestlers' injuries. Therefore, the use of these tests is not recommended to evaluate the risk of injury in this population.

The Effect of Shoulder Injury Prevention Program on Shoulder Isokinetic Strength in Young Male Volleyball Players.

The purpose of this study was to examine the effects of an eight-week 11 + S programme on isokinetic shoulder strength in young male volleyball players. Twenty-eight top-level young male volleyball players were recruited for this study. The players were randomly assigned into intervention (14 players) and control (14 players) groups. The intervention group performed the 11 + S as a warm-up before training at least three times a week for eight weeks. Dominant shoulder strength was measured using an isokinetic dynamometer at 60°/s and 180°/s. The functional deceleration ratio (FDR) was calculated as a functional strength profile. 20% FDR improvement after eight weeks of 11 + S at 180°/s (p = 0.003) were observed. No other statistically significant effects for shoulder strength improvement was observed (p > 0.05). Eight weeks of 11 + S warm-up programme improved the glenohumeral muscle imbalance by increasing the functional deceleration ratio (FDR) of shoulder muscles in young male volleyball players.

A novel protocol for in-depth analysis of recombinant adeno-associated virus capsid proteins using UHPLC-MS/MS.

RATIONALE: In-depth characterization of the three capsid viral proteins (VPs 1, 2, and 3) of adeno-associated viruses (AAVs) is immediately needed to ensure the consistency in gene therapy products and processes. These proteins are typically present at very low concentrations in matrices containing high concentrations of excipients and salts. Thus, there is a need for convenient methods for sample preparation before proteomic analysis. The aim of this study was to meet this need by developing a fast, reliable approach for isolating VPs in a manner enabling their efficient digestion and in-depth characterization using liquid chromatography-mass spectrometry (LC-MS). METHODS: VPs from Anc80 were precipitated with different organic solvents, and the resulting precipitates were dissolved in either sodium deoxycholate (SDC) and N-dodecyl-beta-D-maltoside (DDM) or guanidine hydrochloride (Gu-HCl). The peptides obtained by the following enzymatic digestion by either trypsin or Asp-N were analyzed using LC-MS/MS. RESULTS: We found that precipitation with chloroform/methanol/water results in fast, efficient preparation of VP samples, allowing 100% and 99.2% amino acid sequence coverage of VP1 for trypsin and Asp-N digestion, respectively. This also allowed complete sequence confirmation of VP1, VP2, and VP3 of Anc80, as well as characterization of the amino acid sequences of the N- and C-terminal regions of each VP, together with their post-translational modifications (PTMs). CONCLUSIONS: The presented method enables fast, reliable, and relatively cheap sample preparation for identifying AAV serotypes and characterizing the heterogeneity of capsid viral proteins, including their PTMs.

The effect of a shoulder injury prevention programme on proprioception and dynamic stability of young volleyball players; a randomized controlled trial.

BACKGROUND: The "FIFA 11 + Shoulder" programme has been reported to reduce the incidence of upper extremity injuries among soccer goalkeepers. It has also been recommended for overhead sports. The purpose of this study was therefore to investigate the effect of an 8-week "FIFA 11 + shoulder" (11 + S) programme on shoulder joint position sense (JPS), threshold to detect passive motion (TTDPM) and upper quarter Y Balance Test in young male volleyball players. METHODS: Thirty-two healthy young elite male volleyball players (17.49 ± 1.47 years) participated in this quasi-experimental study. Participants, recruited from two clubs participating in Iranian premier league, were randomly assigned into two groups; (1) the intervention group who performed the "FIFA 11 + shoulder" programme as their warm up protocol, three times per week, and (2) the control group who kept their routine warm up protocol meanwhile. Proprioception tests including JPS and TTDPM of internal and external rotator muscles of the dominant shoulder were recorded via the isokinetic system pro 4. The upper quarter Y Balance Test determined the shoulder dynamic stability. RESULTS: No statistically significant differences were observed for JPS and TTDPM of shoulder internal and external rotator muscles; shoulder stability however significantly increased only in the intervention group (p = 0.03, ηp2=0.02). CONCLUSION: Upper quarter dynamic stability improvement due to the 11+S programme leads to volleyball players' performance and may therefore contribute to a reduction in risk of sustaining injury if applied long-term. TRIAL REGISTRATION: The trial was retrospectively registered atIranian Registry of Clinical Trials with the number of IRCT20201030049193N1 at 04/12/2020.

The Effect of the "11+ Kids" Program on the Isokinetic Strength of Young Football Players.

PURPOSE: The "11+ Kids" injury-prevention program has been shown to reduce injuries and related costs in youth football players less than 14 y of age. A major argument to convince coaches to use this exercise-based injury-prevention program is a potential performance enhancement of the players. Therefore, this study investigated the effects of the "11+ Kids" program on isokinetic strength. METHODS: Two teams were randomly assigned to the intervention and control groups. The intervention group replaced their warm-up by the "11+ Kids" and the control group warmed up as usual. Two days before and after the 10-wk intervention, isokinetic strength of the hip adductors and abductors, knee flexors and extensors, and ankle invertors and evertors was tested. RESULTS: Thirty-one players (mean age 11.5 [0.8] y) completed the study. The intervention group showed large improvements in all isokinetic strength measures (P < .001 for all measures; Cohen d = 0.8-1.4), whereas the control group only showed negligible to medium positive effects (P values ranging from .006 to .718; Cohen d = -0.1 to 0.7). The intervention was beneficial compared with the control group regarding isokinetic strength of the hip adductors (P < .001), knee flexors (P = .002), and ankle evertors (P < .001) and invertors (P = .005). CONCLUSIONS: Given the relatively short intervention period of 10 wk, the observed improvements relate to a practically meaningful effect of the intervention. The gain in strength may improve players' performance and may contribute to a reduction of injury risk in the long-term application.

Proprioception is not associated with lower extremity injuries in U21 high-level football players.

Football is a contact sport with a significant risk of injury. Although proprioception is well studied in rehabilitation, little is known about the association between proprioception and the occurrence of sport injuries. The purpose of this study was to look into the association between ankle and knee proprioception and lower extremity injuries in young football players. Seventy-three football players from the highest U-21 league in Iran volunteered to participate in this study. Before the start of the 2017-2018 competitive season, joint position sense was measured at 30°, 60° and 90° knee flexion and at 10° and 15° ankle dorsiflexion, and inversion using the Biodex Isokinetic pro 4 system. The teams' medical staff recorded football-related lower extremity injuries. We used mixed effects Cox regression models to calculate hazard ratios (HRs) with 95% CIs, acknowledging the clustered data structure. Twenty-two players (30.1%) suffered one or more lower extremity injuries during the season. None of the proprioception measures examined was significantly associated with the risk of lower extremity injuries. Based on these results of our sample, joint position sense does not seem to be associated with lower extremity injuries in young male football players.

The 11+ Kids warm-up programme to prevent injuries in young Iranian male high-level football (soccer) players: A cluster-randomised controlled trial.

OBJECTIVE: To assess the effectiveness of the 11+ Kids warm-up programme regarding injury reduction in male high-level children's football players. DESIGN: Cluster-randomised controlled trial. METHOD: Male youth football teams of Iran's high-level football schools were invited to participate. Inclusion criteria were: teams are competing in the highest league of their province; players are between 7 and 14 years old; regular training takes place at least twice per week. Teams were excluded if they used an injury prevention measure. Participating clubs were randomised to an intervention (INT, N = 20 teams) and a control group (CON, N = 22 teams), stratified by the number of teams and the age group. The groups were blinded against each other. The follow-up period was one season (9 months). INT replaced their warm-up by 11+ Kids. CON performed a standard warm-up programme. The primary outcome was the injury incidence density (injuries per 1000 h of football exposure), compared between groups by incidence rate ratios (RR). RESULTS: In total, 64,047 h of football exposure of 962 players (INT = 443 players, 31,934 h of football, CON = 519 players, 32,113 h of football) were recorded. During the study, 90 (INT = 30; CON = 60) injuries occurred. The overall injury incidence density in INT was reduced by 50% compared to CON (RR 0.50; 95%-CI 0.32, 0.78). No injuries occurred during the execution of the intervention exercises. CONCLUSIONS: The 11+ Kids reduces injuries in high-level children's football players, thus supporting player health and potentially performance and player development.

The association between the isokinetic muscle strength and lower extremity injuries in young male football players.

BACKGROUND: Validating any screening test to predict and prevent football injuries is in need of identifying related risk factors through prospective designs. In spite of the extensive use of strength testing in football players, there are limited studies investigating the relationship between isokinetic muscle strength and injury risk in young football players. The present study aimed to evaluate the relationship between isokinetic strength and the risk of lower extremity injury among Iranian young football players. METHOD: ology: seventy three U-21 football players participated in this study. Isokinetic strength of hip, knee and ankle muscles were measured using the Isokinetic system pro 4. Injuries and exposure in training and matches were registered prospectively by club medical staff for one season. RESULTS: Significant relationships were revealed between the isokinetic strength of hip abductor and adductor muscles, and isokinetic strength ratio of hip abductor/adductor muscles at an angular speed of 30°/sec, the isokinetic strength of hip abductor muscles at 90°/sec, and isokinetic strength of knee flexor and extensor muscles at 60°/sec and knee flexor/extensor strength ratio at angular velocities of 60°/sec with the injury occurrence among football players. CONCLUSION: lower extremity isokinetic strength indices are associated with injuries in young male football players.

A Multi-layered Quantitative In Vivo Expression Atlas of the Podocyte Unravels Kidney Disease Candidate Genes.

Damage to and loss of glomerular podocytes has been identified as the culprit lesion in progressive kidney diseases. Here, we combine mass spectrometry-based proteomics with mRNA sequencing, bioinformatics, and hypothesis-driven studies to provide a comprehensive and quantitative map of mammalian podocytes that identifies unanticipated signaling pathways. Comparison of the in vivo datasets with proteomics data from podocyte cell cultures showed a limited value of available cell culture models. Moreover, in vivo stable isotope labeling by amino acids uncovered surprisingly rapid synthesis of mitochondrial proteins under steady-state conditions that was perturbed under autophagy-deficient, disease-susceptible conditions. Integration of acquired omics dimensions suggested FARP1 as a candidate essential for podocyte function, which could be substantiated by genetic analysis in humans and knockdown experiments in zebrafish. This work exemplifies how the integration of multi-omics datasets can identify a framework of cell-type-specific features relevant for organ health and disease.

Long-term effects of the 11+ warm-up injury prevention programme on physical performance in adolescent male football players: a cluster-randomised controlled trial.

This study examined the long-term effects of the 11+ on physical performance in adolescent male football (soccer) players. Eighty-two 14- to 16-year-old male football players (11+ = 42 players, control = 40 players) participated. Teams were randomised to control (CON) and intervention (INT) groups. INT applied the 11+ injury prevention programme for 30 weeks at least twice a week as a warm-up. CON performed their standard warm-up. Motor performance tests were conducted 1 week prior and 1 week after the competition season. We used magnitude-based inferences and linear mixed-effects models to analyse performance test results. INT showed superior results compared to CON in the vertical jump height 7.5% (95%-CI 4.4%, 10.7%), the Bosco 15-s-jump test 7.2% (95%-CI 2.2%, 12.4%), and the Illinois agility test -2.6% (95%-CI -4.1%, -1.1%). Possibly beneficial effects in favour of INT were found in the 9.1 m sprint test -3.1% (95%-CI -6.1%, 0.1%). Possibly harmful effects (i.e. in favour of CON) were observed in the dribbling test 2.8% (95%-CI -0.8%, 6.4%). The 11+ warm-up programme can improve different performance measures in football players. Coaches might implement additional dribbling drills next to the 11+ to achieve improvements observed in dribbling ability when using a regular warm-up programme.

Hydrophobic Interaction Chromatography for Bottom-Up Proteomics Analysis of Single Proteins and Protein Complexes.

Hydrophobic interaction chromatography (HIC) is a robust standard analytical method to purify proteins while preserving their biological activity. It is widely used to study post-translational modifications of proteins and drug-protein interactions. In the current manuscript we employed HIC to separate proteins, followed by bottom-up LC-MS/MS experiments. We used this approach to fractionate antibody species followed by comprehensive peptide mapping as well as to study protein complexes in human cells. HIC-reversed-phase chromatography (RPC)-mass spectrometry (MS) is a powerful alternative to fractionate proteins for bottom-up proteomics experiments making use of their distinct hydrophobic properties.

Degradation of protein translation machinery by amino acid starvation-induced macroautophagy.

Macroautophagy is regarded as a nonspecific bulk degradation process of cytoplasmic material within the lysosome. However, the process has mainly been studied by nonspecific bulk degradation assays using radiolabeling. In the present study we monitor protein turnover and degradation by global, unbiased approaches relying on quantitative mass spectrometry-based proteomics. Macroautophagy is induced by rapamycin treatment, and by amino acid and glucose starvation in differentially, metabolically labeled cells. Protein dynamics are linked to image-based models of autophagosome turnover. Depending on the inducing stimulus, protein as well as organelle turnover differ. Amino acid starvation-induced macroautophagy leads to selective degradation of proteins important for protein translation. Thus, protein dynamics reflect cellular conditions in the respective treatment indicating stimulus-specific pathways in stress-induced macroautophagy.

Fast and easy phosphopeptide fractionation by combinatorial ERLIC-SCX solid-phase extraction for in-depth phosphoproteome analysis.

Mass spectrometry-based phosphoproteomic analysis is a powerful method for gaining a global, unbiased understanding of cellular signaling. Its accuracy and comprehensiveness stands or falls with the quality and choice of the applied phosphopeptide prefractionation strategy. This protocol covers a powerful but simple and rapid strategy for phosphopeptide prefractionation. The combinatorial use of two distinct chromatographic techniques that address the inverse physicochemical properties of peptides allows for superior fractionation efficiency of multiple phosphorylated peptides. In the first step, multiphosphorylated peptides are separated according to the number of negatively charged phosphosites by electrostatic repulsion-hydrophilic interaction chromatography (ERLIC). A subsequent strong cation exchange (SCX) step separates mostly singly phosphorylated peptides in the ERLIC flow-through according to their positive charge. The presented strategy is inexpensive and adaptable to large and small amounts of starting material, and it allows highly multiplexed sample preparation. Because of its implementation as solid-phase extraction, the entire workflow takes only 2 h to complete.