Nursing workload assessment in an intensive care unit: A retrospective observational study using the Nursing Activities Score.

BACKGROUND: Nursing Activities Score (NAS) is a promising tool for calculating the nursing workload in intensive care units (ICU). However, data on intensive care nursing activities in Portugal are practically non-existent. AIM: To assess the nursing workload in a Portuguese ICU using the NAS. STUDY DESIGN: Retrospective cohort study developed throughout the analysis of the electronic health record database from 56 adult patients admitted to a six-bed Portuguese ICU between 1 June-31 August 2020. The nursing workload was assessed by the Portuguese version of the NAS. The study was approved by the Hospital Council Board and Ethics Committee. The study report followed the STROBE guidelines. RESULTS: The average occupancy rate was 73.55% (±16.60%). The average nursing workload per participant was 67.52 (±10.91) points. There was a correlation between the occupancy rate and the nursing workload. In 35.78% of the days, the nursing workload was higher than the available human resources, overloading nurse staffing/team. CONCLUSIONS: The nursing workload reported follows the trend of the international studies and the results reinforce the importance of adjusting the nursing staffing to the complexity of nursing care in this ICU. This study highlighted periods of nursing workload that could compromise patient safety. RELEVANCE TO CLINICAL PRACTICE: This was one of the first studies carried out with the NAS after its cross-cultural adaptation and validation for the Portuguese population. The nursing workload at the patient level was higher in the first 24 h of ICU stays. Because of the 'administrative and management activities' related to the 'patient discharge procedures', the last 24 h of ICU stays also presented high levels of nursing workload. The implementation of a nurse-to-patient ratio of 1:1 may contribute to safer nurse staffing and to improve patient safety in this Tertiary (level 3) ICU.

Usability of Functional Electrical Stimulation in Upper Limb Rehabilitation in Post-Stroke Patients: A Narrative Review.

Stroke leads to significant impairment in upper limb (UL) function. The goal of rehabilitation is the reestablishment of pre-stroke motor stroke skills by stimulating neuroplasticity. Among several rehabilitation approaches, functional electrical stimulation (FES) is highlighted in stroke rehabilitation guidelines as a supplementary therapy alongside the standard care modalities. The aim of this study is to present a comprehensive review regarding the usability of FES in post-stroke UL rehabilitation. Specifically, the factors related to UL rehabilitation that should be considered in FES usability, as well a critical review of the outcomes used to assess FES usability, are presented. This review reinforces the FES as a promising tool to induce neuroplastic modifications in post-stroke rehabilitation by enabling the possibility of delivering intensive periods of treatment with comparatively less demand on human resources. However, the lack of studies evaluating FES usability through motor control outcomes, specifically movement quality indicators, combined with user satisfaction limits the definition of FES optimal therapeutical window for different UL functional tasks. FES systems capable of integrating postural control muscles involving other anatomic regions, such as the trunk, during reaching tasks are required to improve UL function in post-stroke patients.

Optimal multi-field functional electrical stimulation parameters for the "drinking task - reaching phase" and related upper limb kinematics repeatability in post stroke subjects.

BACKGROUND: No specific guidelines for the management of functional electrical stimulation (FES) parameters in post stroke patients have been defined yet, despite its frequent use. The purpose of this study is to characterize the optimal FES parameters that assist the reaching phase of drinking task ("drinking task - reaching phase") on post stroke subjects and to analyze the related upper limb (UL) movement quality indicators repeatability. METHODS: An observational study with a test and re-test design involving ten post stroke subjects with UL dysfunction was performed. End-point and joint kinematics of contralesional UL were assessed during the "drinking task - reaching phase" with FES through a test and retest design. FES parameters were adjusted to improve UL function according to a consensus between physiotherapists and patients' perspective. FINDINGS: It was possible to establish reliable FES parameters that assisted the "drinking task - reaching phase". All FES parameters presented high to very high repeatability and led to moderate to very high repeatability in almost UL movement quality indicators during the "drinking task - reaching phase". INTERPRETATION: These findings show that the main characteristics of FES parameters that improves patient perception of change are quite stable, which facilitate its implementation in clinical practice by allowing consistence between intervention sessions.

Optimal Multifield Functional Electrical Stimulation Parameters for the "Turn on the Light" Task and Related Upper Limb Kinematics Repeatability in Poststroke Subjects.

OBJECTIVE: To characterize the optimal functional electrical stimulation (FES) parameters that assist the turn on the light task (TOTL) on poststroke participants and to analyze the related upper limb (UL) kinematics repeatability. DESIGN: Cross-sectional study. SETTING: Human movement research center. PARTICIPANTS: Poststroke individuals (N=11) with history of a single unilateral stroke that resulted in a motor control dysfunction of the contralesional UL. INTERVENTIONS: FES based on surface multifield technology applied to the contralesional wrist and finger extensors during the TOTL. MAIN OUTCOME MEASURES: FES outcome metrics (virtual electrodes, stimulation duration, intensity) and kinematic metrics (end-point kinematics [absolute and relative duration, mean and peak velocities, relative instant of peak velocity, index of curvature, number of movement units] and joint kinematics [shoulder, elbow, wrist end position and range of movement]). Outcome measures were assessed 2 times with a 72-hour maximum time interval. CONCLUSION: It was possible to establish reliable FES parameters that assisted the TOTL on poststroke participants. These stimulation parameters led to high to very high repeatability in terms of UL kinematics for most of the cases.

Solid-State Form Characterization of Riparin I.

Riparin I is an alkamide with potential anxiolytic activity in preclinical studies. The characterization and understanding of solid-state properties play an importance role in drug development. For this work, the solid state of five riparin I batches (RIP-1, RIP-2, RIP-3, RIP-4, and RIP-5), obtained by the same synthesis process, were characterized by Scanning Electron Microscopy (SEM), Differential Scanning Calorimetry (DSC), DSC-photovisual, Thermogravimetry (TG), Fourier Transform Infrared (FTIR), Pyrolysis (Pyr-GC/MS), X-ray Powder Diffraction (PXRD), and Solid-State Nuclear Magnetic Resonance (ssNMR) techniques. Batches of riparin I with different crystal habits resulting in crystallization impurities were observed, which can be attributed to the presence of triethylamine. The main differences were observed by DSC, PXRD, and ssNMR analysis. DSC curves of RIP-2 and RIP-3 presented endothermic peaks at different temperatures of fusion, which can be attributed to the mixture of different crystalline forms. PXRD and ssNMR results confirmed crystallinity differences. The results offer evidence of the importance of controlling the reproducibility of the synthesis in order to obtain the adequate morphology for therapeutic efficacy and avoiding future problems in quality control of riparin I products.

Effect of corn steep liquor (CSL) and cassava wastewater (CW) on chitin and chitosan production by Cunninghamella elegans and their physicochemical characteristics and cytotoxicity.

Microbiological processes were used for chitin and chitosan production with Cunninghamella elegans UCP/WFCC 0542 grown in different concentrations of two agro-industrial wastes, corn steep liquor (CSL) and cassava wastewater (CW) established using a 2² full factorial design. The polysaccharides were extracted by alkali-acid treatment and characterized by infrared spectroscopy, viscosity, thermal analysis, elemental analysis, scanning electron microscopy and X-ray diffraction. The cytotoxicity of chitosan was evaluated for signs of vascular change on the chorioallantoic membrane of chicken eggs. The highest biomass (9.93 g/L) was obtained in trial 3 (5% CW, 8% CSL), the greatest chitin and chitosan yields were 89.39 mg/g and 57.82 mg/g, respectively, and both were obtained in trial 2 (10% CW, 4% CSL). Chitin and chitosan showed a degree of deacetylation of 40.98% and 88.24%, and a crystalline index of 35.80% and 23.82%, respectively, and chitosan showed low molecular weight (LMW 5.2 × 10³ Da). Chitin and chitosan can be considered non-irritating, due to the fact they do not promote vascular change. It was demonstrated that CSL and CW are effective renewable agroindustrial alternative substrates for the production of chitin and chitosan.

The influence of shoulder position during multi-joint exercises in the relative scapular muscles activity in symptomatic and asymptomatic conditions.

BACKGROUND: Scapular muscles changes, as increased upper trapezius activity and decreased middle and lower trapezius and serratus anterior muscle activity, have been demonstrated in shoulder pain specific or non-specific conditions. Shoulder external rotation exercises have been recommended to improve scapular activity in shoulder pain. OBJECTIVE: To evaluate the relative scapular muscles activity during multi-joint exercises combining shoulder external rotation, trunk rotation and scapular squeeze. METHODS: Forty-one participants with and without shoulder pain were assessed in a cross-sectional study. They performed isometric multi-joint exercises at 0∘ and 90∘ of shoulder abduction with and without support. The relative activity of upper, middle, and lower trapezius and serratus anterior (upper/middle and lower portions) was measured through electromyography. The scapular muscular balance was assessed by the ratio between relative activity of the upper trapezius and the other muscles. RESULTS: Both groups presented similar results. The exercise at 90∘ abduction led to increased relative muscle activity against maximal voluntary contraction in both groups for upper trapezius (with support: 4% MVIC, p= 0.001 or 15% MVIC, p< 0.0001; and without support: 11% MVIC, p< 0.0001 or 13%, p< 0.0001, for asymptomatic and symptomatic group, respectively) and lower trapezius (with support: 66% MVIC, p< 0.0001 or 62% MVIC, p< 0.0001, for asymptomatic and symptomatic group.

Postural control during turn on the light task assisted by functional electrical stimulation in post stroke subjects.

Postural control mechanisms have a determinant role in reaching tasks and are typically impaired in post-stroke patients. Functional electrical stimulation (FES) has been demonstrated to be a promising therapy for improving upper limb (UL) function. However, according to our knowledge, no study has evaluated FES influence on postural control. This study aims to evaluate the influence of FES UL assistance, during turning on the light task, in the related postural control mechanisms. An observational study involving ten post-stroke subjects with UL dysfunction was performed. Early and anticipatory postural adjustments (EPAs and APAs, respectively), the weight shift, the center of pressure and the center of mass (CoM) displacement were analyzed during the turning on the light task with and without the FES assistance. FES parameters were adjusted to improve UL function according to a consensus between physiotherapists' and patients' perspectives. The ANOVA repeated measures, Paired sample t and McNemar tests were used to compare postural control between the assisted and non-assisted conditions. When the task was assisted by FES, the number of participants that presented APAs increased (p = 0.031). UL FES assistance during turning on the light task can improve postural control in neurological patients with UL impairments.

A preformulation study of a new medicine for Chagas disease treatment: physicochemical characterization, thermal stability, and compatibility of benznidazole.

This work aimed the studies of physicochemical characterization, thermal stability, and compatibility of benznidazole (BNZ) drug by spectroscopy (NMR, IR), thermoanalytical (differential thermal analysis, differential scanning calorimetry, and thermogravimetry), and chromatographic (HPLC) techniques, beyond the analytical tools of Van't Hoff equation and Ozawa model. The compatibility study was conducted by binary mixtures (1:1, w/w) of the drug with microcrystalline cellulose 102 and 250, anhydrous lactose, and sodium starch glycolate. The physicochemical characterization confirmed data reported in scientific literature, guaranteeing authenticity of the analyzed raw material. The drug melts at 191.68°C (∆H, 119.71 J g(-1)), characteristic of a non-polymorphic raw material, and a main stage decomposition at 233.76-319.35°C (∆m, 43.32%) occurred, ending the study with almost all mass volatilized. The quantification of drug purity demonstrated a correlation of 99.63% between the data obtained by chromatographic (99.20%) and thermoanalytical technique (99.56%). The Arrhenius equation and Ozawa model showed a zero-order kinetic behavior for the drug decomposition, and a calculated provisional validity time was 2.37 years at 25°C. The compatibility study evidenced two possible chemical incompatibilities between BNZ and the tested excipients, both associated by the authors to the reaction of the BNZ's amine and a polymer carbohydrate's carbonile, being maillard reactions. The BNZ reaction with anhydrous lactose is more pronounced than with the sodium starch glycolate because the lactose has more free hydroxyl groups to undergo reduction by the drug. In this sense, this work guides the development of a new solid pharmaceutical product for Chagas disease treatment, with defined quality control parameters and physicochemical stability.

Does the cleat model interfere with ankle sprain risk factors in artificial grass?

BACKGROUND: The cleats-surface interaction has been described as a possible risk factor for lateral ankle sprain. However, their interaction is still unknown in individuals with chronic ankle instability. The purpose of this study was to determine the influence of different soccer cleats on kinematic, kinetic and neuromuscular ankle variables on artificial grass in soccer players with and without chronic ankle instability. METHODS: Eighty-two amateur athletes divided in two groups: 40 with chronic ankle instability and 42 without chronic ankle instability. All subjects performed 2 series of 6 consecutive crossover jumps with dominant foot, each one with one of the four models of cleats (Turf, Artificial grass, Hard and Firm ground). Cleat and group main effect and interactions of kinematic, kinetic and neuromuscular variables were analyzed according to factorial repeated measures ANOVA. FINDINGS: No statistically significant cleat and group main effect and interactions were identified in kinematic, kinetic and electromyographic magnitude of the peroneal muscles. A main effect of the group was observed for peroneus longus activation time for TF model (p = 0.010). INTERPRETATION: In soccer players, the contributor variables for ankle sprain were not influenced by the kind of soccer cleat used in a functional jump test on artificial grass. However, players with chronic ankle instability present delayed postural adjustments in peroneus longus with the TF model compared to players without chronic ankle instability.

Different Cleat Models Do Not Influence Side Hop Test Performance of Soccer Players with and Without Chronic Ankle Instability.

The lateral ankle sprain is one of the most common sport injury, representing 10-30% of all musculoskeletal disorders. The lateral ankle sprain is induced by sport gestures involving changes of direction and landing manoeuvres and constitutes a risk factor for the occurrence of chronic ankle instability. Although cleat models and performance have been already explored, no study has evaluated this relationship in athletes with chronic ankle instability. Therefore, the purpose of the study was to analyse the influence of different soccer cleat models on Side Hop Test performance of athletes with and without chronic ankle instability. Thirty-nine athletes were divided into two groups, a chronic ankle instability group (n = 20) and a healthy group (n = 19). Each participant performed the Side Hop Test, executing 10 consecutive jumps on dry artificial grass with 4 cleat models. The Qualisys System and two force platforms were used to analyse the test runtime, the distance travelled and the mean velocity. No statistically significant interaction was observed between the group and the cleat model for all variables evaluated. In addition, no differences were observed between models or groups. In this specific test, performance does not seem to be influenced by different cleat models on dry artificial grass in athletes with and without chronic ankle instability.

Influence of Cleats-Surface Interaction on the Performance and Risk of Injury in Soccer: A Systematic Review.

OBJECTIVE: To review the influence of cleats-surface interaction on the performance and risk of injury in soccer athletes. DESIGN: Systematic review. DATA SOURCES: Scopus, Web of science, PubMed, and B-on. ELIGIBILITY CRITERIA: Full experimental and original papers, written in English that studied the influence of soccer cleats on sports performance and injury risk in artificial or natural grass. RESULTS: Twenty-three articles were included in this review: nine related to performance and fourteen to injury risk. On artificial grass, the soft ground model on dry and wet conditions and the turf model in wet conditions are related to worse performance. Compared to rounded studs, bladed ones improve performance during changes of directions in both natural and synthetic grass. Cleat models presenting better traction on the stance leg improve ball velocity while those presenting a homogeneous pressure across the foot promote better kicking accuracy. Bladed studs can be considered less secure by increasing plantar pressure on lateral border. The turf model decrease peak plantar pressure compared to other studded models. CONCLUSION: The soft ground model provides lower performance especially on artificial grass, while the turf model provides a high protective effect in both fields.

Influence of prolonged wearing of unstable shoes on upright standing postural control.

OBJECTIVE: To study the influence of prolonged wearing of unstable shoes on standing postural control in prolonged standing workers. METHODS: The participants were divided into two groups: one wore unstable shoes while the other wore conventional shoes for 8weeks. Stabilometry parameters related to centre of pressure (CoP), rambling (RM) and trembling (TR) as well as the total agonist/antagonist muscle activity, antagonist co-activation and reciprocal activation were evaluated during upright standing, before and after the 8weeks period. In both moments, the subjects were evaluated wearing the unstable shoes and in barefoot. RESULTS: The unstable shoe condition presented increased CoP displacement related variables and decreased co-activation command compared to barefoot before and after the intervention. The prolonged wearing of unstable shoes led to: (1) reduction of medial-lateral CoP root mean square and area; (2) decreased anteroposterior RM displacement; (3) increased anteroposterior RM mean velocity and mediolateral RM displacement; (4) decreased anteroposterior TR RMS; and (5) increased thigh antagonist co-activation in the unstable shoe condition. CONCLUSION: The unstable shoe condition is associated to a higher destabilising effect that leads to a selection of more efficient and accurate postural commands compared to barefoot. Prolonged wearing of unstable shoes provides increased effectiveness and performance of the postural control system, while wearing of unstable shoes in upright standing, that are reflected by changes in CoP related variables and by a reorganisation of postural control commands.

Model Analytical Development for Physical, Chemical, and Biological Characterization of Momordica charantia Vegetable Drug.

Momordica charantia is a species cultivated throughout the world and widely used in folk medicine, and its medicinal benefits are well documented, especially its pharmacological properties, including antimicrobial activities. Analytical methods have been used to aid in the characterization of compounds derived from plant drug extracts and their products. This paper developed a methodological model to evaluate the integrity of the vegetable drug M. charantia in different particle sizes, using different analytical methods. M. charantia was collected in the semiarid region of Paraíba, Brazil. The herbal medicine raw material derived from the leaves and fruits in different particle sizes was analyzed using thermoanalytical techniques as thermogravimetry (TG) and differential thermal analysis (DTA), pyrolysis coupled to gas chromatography/mass spectrometry (PYR-GC/MS), and nuclear magnetic resonance ((1)H NMR), in addition to the determination of antimicrobial activity. The different particle surface area among the samples was differentiated by the techniques. DTA and TG were used for assessing thermal and kinetic parameters and PYR-GC/MS was used for degradation products chromatographic identification through the pyrograms. The infusions obtained from the fruit and leaves of Momordica charantia presented antimicrobial activity.

Simultaneous determination of ranitidine and metronidazole in human plasma using high performance liquid chromatography with diode array detection.

The development and validation of a simple method for the simultaneous determination of ranitidine and metronidazole in human plasma is described. Plasma samples (250 microL) were deproteinized by precipitation with 60% perchloric acid, centrifuged and the supernatant directly injected into the HPLC. Separation was achieved in isocratic mode with a Shimpak C(18) column and a mobile phase consisting of 10mM potassium dihydrogen phosphate pH 3.5:acetonitrile (90:10, v/v) with UV detection at 315 nm. The method showed good selectivity and sensitivity. Good and consistent recovery for metronidazole and ranitidine was obtained: 96.22+/-3.52 and 95.00+/-4.50% for ranitidine (25-1000 ng/mL) and metronidazole (60-10,000 ng/mL), respectively (n=3). With this one-step sample preparation method, both ranitidine and metronidazole could be quantified simultaneously in human plasma with good precision (R.S.D.<15%) and accuracy (bias values below 15%). The limit of quantification for ranitidine and metronidazole were 20 and 40 ng/mL plasma, respectively.

Action of silver nanoparticles towards biological systems: cytotoxicity evaluation using hen's egg test and inhibition of Streptococcus mutans biofilm formation.

This study aimed to evaluate the cytotoxicity and bactericidal properties of four silver nanoparticle (AgNP) colloids and their ability to inhibit Streptococcus mutans biofilm formation on dental enamel. The cytotoxicity of AgNPs was evaluated based on signs of vascular change on the chorioallantoic membrane using the hen's egg test (HET-CAM). Bactericidal properties and inhibition of S. mutans biofilm formation were determined using a parallel-flow cell system and a dichromatic fluorescent stain. The percentage of viable cells was calculated from regression data generated from a viability standard. AgNP colloids proved to be non-irritating, as they were unable to promote vasoconstriction, haemorrhage or coagulation. AgNP colloids inhibited S. mutans biofilm formation on dental enamel, and cell viability measured by fluorescence was 0% for samples S1, S2, S3 and S4 and 36.5% for the positive control (diluted 30% silver diamine fluoride). AgNPs are new products with a low production cost because they have a lower concentration of silver, with low toxicity and an effective bactericidal effect against a cariogenic oral bacterium. Moreover, they do not promote colour change in dental enamel, which is an aesthetic advantage compared with traditional silver products.

Influence of long-term wearing of unstable shoes on compensatory control of posture: an electromyography-based analysis.

PURPOSE: This study investigated the influence of long-term wearing of unstable shoes (WUS) on compensatory postural adjustments (CPA) to an external perturbation. METHODS: Participants were divided into two groups: one wore unstable shoes while the other wore conventional shoes for 8 weeks. The ground reaction force signal was used to calculate the anterior-posterior (AP) displacement of the centre of pressure (CoP) and the electromyographic signal of gastrocnemius medialis (GM), tibialis anterior (TA), rectus femoris (RF) and biceps femoris (BF) muscles was used to assess individual muscle activity, antagonist co-activation and reciprocal activation at the joint (TA/GM and RF/(BF+GM) pairs) and muscle group levels (ventral (TA+RF)/dorsal (GM+BF) pair) within time intervals typical for CPA. The electromyographic signal was also used to assess muscle latency. The variables described were evaluated before and after the 8-week period while wearing the unstable shoes and barefoot. RESULTS: Long-term WUS led to: an increase of BF activity in both conditions (barefoot and wearing the unstable shoes); a decrease of GM activity; an increase of antagonist co-activation and a decrease of reciprocal activation level at the TA/GM and ventral/dorsal pairs in the unstable shoe condition. Additionally, WUS led to a decrease in CoP displacement. However, no differences were observed in muscle onset and offset. CONCLUSION: Results suggest that the prolonged use of unstable shoes leads to increased ankle and muscle groups' antagonist co-activation levels and higher performance by the postural control system.

Correlation of thermal analysis and pyrolysis coupled to GC-MS in the characterization of tacrolimus.

In recent years, thermal analysis has assumed major role in the pharmaceutical industry because it can be used to evaluate the stability both in the control of raw materials and the finished product, having employment potential in the development and characterization of new products and assessment processes. Tacrolimus (TCR) is a macrolide lactone with potent immunosuppressive activity. The purpose of this study was to characterize tacrolimus raw material using Thermal analysis and Pyrolysis coupled to Gas chromatography-Mass spectrometry (Pyr-GC-MS). It was analyzed four samples of tacrolimus named TCR A, B, C and D. Thermal analysis experiments was performed in Shimadzu equipment, under nitrogen and synthetic air atmosphere in different heating rate. Pyrolysis analysis was conducted in isothermal conditions of 300°C and 400°C coupled to GC-MS, in which the mass spectrometer was operated in scan mode to detect ions in the range of mass of m/z 25-900. The thermal studies by DSC, DTA and DSC-Photovisual showed desolvation process for all tacrolimus raw materials and TG-dynamical demonstrated two pseudo-polymorphic forms (monohydrate and sesquihydrate) of tacrolimus. It was observed good correlation between the stoichiometric mass losses of the TG-dynamical and identification of product ion in Pyr-GC/MS technique. It was possible to correlate the five pyrolytic product ions with the Ozawa kinetic analysis from the thermal decomposition of TG-dynamical. The thermal studies (DSC, DSC-Photovisual, DTA and TG-dynamical) were applied in the thermal characterization of the raw materials of tacrolimus which showed pseudo-polymorphic forms, which must be monitored by pharmaceutical industry, avoiding future problems in pharmaceutical process, chemical stability and bioavailability of the tacrolimus product.

Influence of wearing an unstable shoe construction on compensatory control of posture.

This study investigated the influence of wearing unstable shoe construction (WUS) on compensatory postural adjustments (CPA) associated with external perturbations. Thirty-two subjects stood on a force platform resisting an anterior-posterior horizontal force applied to a pelvic belt via a cable, which was suddenly released. They stood under two conditions: barefoot and WUS. The electromyographic (EMG) activity of gastrocnemius medialis, tibialis anterior, rectus femoris, biceps femoris, rectus abdominis, and erector spinae muscles and the center of pressure (CoP) displacement were acquired to study CPA. The EMG signal was used to assess individual muscle activity and latency, antagonist co-activation and reciprocal activation at joint and muscle group levels. Compared to barefoot, WUS led to: (1) increased gastrocnemius medialis activity, (2) increased total agonist activity, (3) decreased antagonist co-activation at the ankle joint and muscle group levels, (4) increased reciprocal activation at the ankle joint and muscle group levels, and (5) decrease in all muscle latencies. No differences were observed in CoP displacement between conditions. These findings demonstrate that WUS led to a reorganization of the postural control system associated to improved performance of some components of postural control responses.

Short-term stability studies of ampicillin and cephalexin in aqueous solution and human plasma: Application of least squares method in Arrhenius equation.

A limited number of studies with application of the Arrhenius equation have been reported to drugs and biopharmaceuticals in biological fluids at frozen temperatures. This paper describes stability studies of ampicillin and cephalexin in aqueous solution and human plasma applying the Arrhenius law for determination of adequate temperature and time of storage of these drugs using appropriate statistical analysis. Stability studies of the beta-lactams in human plasma were conducted at temperatures of 20°C, 2°C, -20°C and also during four cycles of freeze-thawing. Chromatographic separation was achieved using a Shimpak C(18) column, acetonitrile as organic modifier and detection at 215nm. LC-UV-MS/MS was used to demonstrate the conversion of ampicillin into two diastereomeric forms of ampicilloic acid. Stability studies demonstrated degradation greater than 10% for ampicillin in human plasma at 20°C, 2°C and -20°C after 15h, 2.7days, 11days and for cephalexin at the same temperatures after 14h, 3.4days and 19days, respectively, and after the fourth cycle of freezing-thawing. The Arrhenius plot showed good prediction for the ideal temperature and time of storage for ampicillin (52days) and cephalexin (151days) at a temperature of -40°C, but statistical analysis (least squares method) must be applied to avoid incorrect extrapolations and estimated values out uncertainty limits.