Development of a validated stability-indicating HPLC- DAD method for dasabuvir and the characterization of its degradation products using LC-QToF-MS/MS

Abstract A stability-indicating HPLC-DAD method was developed and validated for the simultaneous determination of dasabuvir and its degradation products in the pharmaceutical formulation. The proposed method utilized a Symmetry® C18 (4.6 x 75 mm, 3.5 µm) column, and the mobile phase consisted of an isocratic elution of formic acid (0.1%) and acetonitrile (55:45, v/v), at a flow of 1 mL min-1; analytes were detected at 244 nm. Dasabuvir was submitted to different stress degradation conditions, such as acidic, alkaline, neutral, thermal, oxidative and photolytic, and the structural elucidation of degradation products was performed using LC-QToF-MS/MS. The HPLC-DAD stability-indicating method was validated for selectivity, linearity, limit of detection and quantification, accuracy, precision and robustness, according to ICH guidelines. Dasabuvir produced two degradation products (DP1 and DP2) from the alkaline stress conditions, which were characterized in negative ion mode. Dasabuvir was linear in the range 9.78 to 136.92 µg mL-1, and DP and DP were linear in the range 2.9 to 20.2 µg mL-1 and 1.3 to 14.9 µg mL-1, respectively. The 1 2 recovery ranged between 99.16 and 100.86%, while precision ranged from 1.02 to 2.89%. As the method can effectively separate the dasabuvir from its degradation products and quantitate them, it may be employed as a stability-indicating method for the pharmaceutical formulation.

Existence of advanced glycation / lipoxidation end-products in parenteral nutrition solutions and effects of infusion conditions on their levels

Abstract Infusion solutions must be stable from the production stage until the infusion stage. Some infusion fluids contain degradation products, known as advanced glycation end products (AGEs); however, it is unknown whether AGEs exist in parenteral nutrition solutions. We aimed to investigate this question and test the effect of infusion conditions on AGE formation in parenteral nutrition solution. Nine parenteral nutrition solutions were supplied by the pharmacy with which we collaborated. To simulate the infusion conditions, the solutions were held in a patient room with standard lighting and temperature for 24 hours. Samples were taken at the beginning (group A) and the end (24th hour, group B) of the infusion period. The degradation products were 3-deoxyglucosone, pentosidine, N-carboxymethyl lysine, and 4-hydroxynonenal, which we investigated by high-performance liquid chromatography-mass spectrometry (LC-MS) and Q-TOF LC/MS methods. Two of four degradation products, 4-hydroxynonenal and N-carboxymethyl lysine, were detected in all samples, and Group B had higher levels of both compounds compared to Group A, who showed that the quantities of these compounds increased in room conditions over time. The increase was significant for 4-hydroxynonenal (p=0.03), but not for N-carboxymethyl lysine (p=0.23). Moreover, we detected in the parenteral nutrition solutions a compound that could have been 4-hydroxy-2-butynal or furanone

The benefits of peritoneal dialysis (PD) solution with low-glucose degradation product in residual renal function and dialysis adequacy in PD patients: A meta-analysis / Beneficios de la solución de diálisis peritoneal (DP), con producto de degradación bajo en glucosa, en la función renal residual y la adecuación de la diálisis en pacientes en DP: un metanálisis

Abstract The peritoneal effects of low-glucose degradation product (GDP)-containing peritoneal dialysis (PD) solutions have been extensively described. To systematically evaluate the efficacy and safety of low GDP solution for PD patients, specifically the effect on residual renal function (RRF) and dialysis adequacy, we conducted a meta-analysis of the published randomized controlled trials (RCTs). Different databases were searched for RCTs that compared low GDP-PD solutions with conventional PD solutions in the treatment of PD patients with continuous ambulatory peritoneal dialysis (CAPD) and automated peritoneal dialysis (APD). The outcomes of RCTs should include RRF and may include small solute clearance, peritoneal transport status, nutritional status, and all-cause mortality. Seven studies (632 patients) were included. Compared with the conventional solution, low-GDP solution preserved RRF in PD patients over time (MD 0.66 mL/min, 95% CI 0.34 to 0.99; p<0.0001), particularly in one year of treatment (p<0.01), and improved weekly Kt/V (MD 0.11, 95% CI 0.05 to 0.17; p=0.0007) without an increased 4-hour D/Pcr (MD 0.00, 95% CI -0.02 to 0.02; p=1.00). Notably, the MD of RRF and urine volume between the two groups tended to decrease as time on PD progressed up to 24 months. Patients using low GDP PD solutions did not have an increased risk of all-cause mortality (MD 0.97, 95% CI 0.50 to 1.88; p=0.93). Our meta-analysis confirms that the low GDP PD solution preserves RRF, improves the dialysis adequacy without increasing the peritoneal solute transport rate and all-cause mortality. Further trials are needed to determine whether this beneficial effect can affect long-term clinical outcomes.

Resumen Los efectos peritoneales de las soluciones de diálisis peritoneal (DP) que contienen productos de degradación bajos en glucosa (PIB) se han descrito ampliamente. Para evaluar sistemáticamente la eficacia y la seguridad de la solución de PIB bajo para pacientes en DP, específicamente el efecto sobre la función renal residual (RRF) y la adecuación de la diálisis, realizamos un metanálisis de los ensayos controlados aleatorios (ECA) publicados. Se realizaron búsquedas en diferentes bases de datos de ECA que compararan la solución de DP de bajo PIB con la solución de DP convencional en el tratamiento de pacientes con EP con CAPD y APD. Los resultados de los ECA deben incluir la RRF y pueden incluir la depuración de solutos pequeños, el estado nutricional, el estado del transporte peritoneal y la mortalidad por todas las causas. Se incluyeron siete estudios (632 pacientes). En comparación con la solución convencional, la solución de bajo PIB preservó la FRR en pacientes con EP a lo largo del tiempo (DM 0,66 mL/min, IC del 95%: 0,34 a 0,99; p<0,0001), particularmente en un año de tratamiento (p<0,01), y mejoró el Kt/V semanal (DM 0,11, IC del 95%: 0,05 a 0,17; p = 0,0007), sin un aumento de D/Pcr a las 4 horas (DM 0,00, IC del 95%: -0,02 a 0,02; p = 1,00). Los pacientes que usaron una solución para DP con bajo contenido de GDP no tuvieron un mayor riesgo de mortalidad por todas las causas (DM 0,97; IC del 95%: 0,50 a 1,88; p = 0,93). Nuestro metanálisis confirma que la solución de DP de bajo PIB preserva la FRR, mejora la adecuación de la diálisis sin aumentar la tasa de transporte peritoneal de solutos y la mortalidad por todas las causas. Se necesitan más ensayos para determinar si este efecto beneficioso puede afectar los resultados clínicos a largo plazo.

Association of fibrinogen, fibrin degradation product, and D-dimer level with disease severity in hospitalized COVID-19 patients – A cross-sectional study in Eastern India

Background: Although most of the COVID-19 patients presented with mild symptoms and recovered, a considerable number of cases became serious with poor prognosis in an unpredictable manner. They mostly presented with respiratory symptoms and coagulation abnormalities with thrombosis and multi-organ failure. Hence, timely prediction of these cases with the early intervention might decrease mortality. Aims and Objectives: The objectives of this were to determine whether values of fibrinogen, fibrin degradation products (FDP), and D-dimer level correlates with disease severity in COVID-19 patients. Materials and Methods: This observational cross-sectional study was done on total 400 hospitalized COVID-19 adult patients where patients were categorized into moderate and severe cases as per guideline of Government of India. Patients with pre-existing coagulation disorder or receiving anticoagulant drugs were excluded from the study. FDP, fibrinogen, and D-dimer values of these two groups were evaluated and compared statistically to determine their significance. Results: Overall mean and standard deviation of fibrinogen, FDP, and D-dimer were 607.48 ± 177.73, 34.93 ± 29.2, and 6.23 ± 6.48 for severe category, while for moderate category disease, they were 389.77 ± 110.16, 10.79 ± 10.47, and 1.96 ± 3.3, respectively. Unpaired t-test showed that the study parameters are significantly higher in severe COVID-19 patients compared to moderate ones. Conclusion: It was concluded that elevated level of D-dimer, fibrinogen, and FDP is indicator of disease progression in COVID-19. Thus, regular estimation of these simple coagulation parameters may predict disease severity and help in adequate management.

Predictive value of serum chlorine and interleukin-6 combined with coagulation indexes on severity of severe acute pancreatitis / 中华危重病急救医学

Objective:To explore the predictive value of combined detection of serum interleukin-6 (IL-6), chloride (Cl -), D-dimer and fibrin degradation products (FDP) for severity of acute pancreatitis (AP). Methods:From December 2020 to March 2022, 132 AP patients who met the criteria for inclusion were screened for retrospective analysis from 292 AP patients admitted in emergency surgery at the First Affiliated Hospital of Zhengzhou University and they were divided into severe acute pancreatitis (SAP) group and non-SAP group, with 63 in SAP group and 69 in non-SAP group, according to classification criteria. The data including lab results, abdominal doppler ultrasound and chest and abdominal CT, etc. The bedside index for severity in acute pancreatitis (BISAP) score was calculated. Multivariate Logistic regression analysis was carried out to find the risk factors for the severity of AP patients. The receiver operator characteristic curve (ROC) was drawn to judge the clinical predictive value of each factor.Results:A total of 132 AP patients were enrolled. The serum IL-6, D-dimer, FDP levels and the BISAP score in SAP group were significantly higher than those in non-SAP group [serum IL-6 (ng/L): 62.73 (21.54, 187.47) vs. 8.22 (4.13, 14.70), D-dimer (mg/L): 5.36 (2.94, 8.25) vs. 0.94 (0.42, 2.21), FDP (mg/L): 13.54 (6.76, 22.45) vs. 3.20 (2.50, 6.10), BISAP score: 2.00 (1.00, 3.00) vs. 1.00 (0, 2.00), all P < 0.05], while the serum Cl - level was significantly lower than that of non-SAP group (mmol/L: 97.90±4.86 vs. 101.73±4.32, P < 0.05). Multivariate Logistic regression analysis showed that increased levels of IL-6 [odds ratio ( OR) = 1.02, 95% confidence interval (95% CI) was 1.01-1.04], D-dimer ( OR = 1.21, 95% CI was 1.05-1.40) and decreased Cl - level ( OR = 0.88, 95% CI was 0.79-0.98) were risk factors for SAP (all P < 0.05). The ROC curve analysis showed that the area under the ROC curve (AUC) of IL-6, Cl -, D-dimer and FDP combined to predict the severity of AP patients was larger (0.89), and the sensitivity (82.50%) and specificity (85.50%) were higher. Conclusion:Compared with single index, the combined detection of serum IL-6, Cl -, D-dimer and FDP is more precise in determining the condition of AP.

Changes and clinical significance of the levels of peripheral blood fibrinogen and its degradation products in patients with multiple myeloma / 中国医师进修杂志

Objective:To explore the level changes and clinical significance of peripheral blood fibrinogen (FIB) and its degradation products (FDP) in patients with multiple myeloma (MM).Methods:One hundred and two MM patients who treated in Tongling People's Hospital from January 2015 to April 2018 were selected and divided into good prognosis group and poor prognosis group according to the prognosis. The correlation between the levels of FIB and FDP in peripheral blood and prognosis and the predictive value of poor prognosis were analyzed, and the survival was analyzed.Results:The ratio of tumor cells in bone marrow, international staging system (ISS) stage, and the levels of serum hemoglobin, albumin, creatinine, and β2-microglobulin in good prognosis group and poor prognosis group had significant differences ( P<0.05). The levels of peripheral blood FIB and FDP in the poor prognosis group and after 3 cycles of chemotherapy were higher than those in the good prognosis group: before chemotherapy: (4.71 ± 0.68) g/L vs. (4.02 ± 0.65) g/L, (50.56 ± 9.14) mg/L vs. (37.52 ± 8.25) mg/L; after 3 cycles of chemotherapy: (4.15 ± 0.62) g/L vs. (3.42 ± 0.53) g/L, (42.28 ± 9.51) mg/L vs. (6.59 ± 1.60) mg/L, there were statistical differences ( P<0.05). Controlled other factors after chemotherapy, the peripheral blood FIB and FDP of 1 cycle, 3 cycles of chemotherapy were still significantly related to the prognosis ( P<0.05). The area under the curve value of the combination of peripheral blood FIB and FDP before chemotherapy was 0.852, which was greater than independent detection of any index. The 2-year survival rate of patients with high levels of FIB and FDP were lower than those of patients with low levels ( P<0.05). Conclusions:Peripheral blood FIB and FDP of MM patients are independent risk factors that affect the prognosis. An increase in their levels indicates a poor prognosis. Clinical treatment can be actively improved according to the changes in the levels of the two to ensure the maximum benefit for patients.

Coagulation and fibrinolysis parameters and mortality in patients with ischemic stroke: a 10-year follow-up study / 国际脑血管病杂志

Objective:To investigate the long-term death of patients with ischemic stroke and its influencing factors.Methods:Based on the data of patients with ischemic stroke in the multi-center oral fibrinogen-lowering drug secondary prevention database, the follow-up patient information and the cause of death were registered through the epidemiological investigation method, and then compared with the baseline data of patients in the original database.Results:A total of 278 patients completed the follow-up, and 166 were in lumbrokinase group and 112 were in control group. There were 124 deaths (44.6%) within 10 years, of which 92 (74.2%) were vascular deaths. In the lumbrokinase group, 74 patients (44.6%) died of all causes and 55 (33.1%) died of vascular diseases; in the control group, 50 (44.6%) died of all causes and 37 (33.0%) died of vascular diseases. Cox proportional risk model analysis showed that lumbrokinase treatment had no significant effect on the 10-year survival rate of patients with ischemic stroke. The analysis of death influencing factors showed that the baseline international normalized ratio (INR) was significantly associated with the 10-year non-vascular death risk of patients (hazard ratio [ HR] 1.98, 95% confidence interval [ CI] 1.21-3.25; P=0.006). The greater the decrease of tissue plasminogen activator (tPA) within half a year, the lower the 10-year all-cause mortality risk ( HR 0.94, 95% CI 0.90-0.99; P=0.011); the greater the decrease in INR within one year , the lower the 10-year vascular death risk ( HR 0.41, 95% CI 0.17-0.96; P=0.040); the greater the decrease of D-dimer within one year , the higher the risk of the 10-year vascular death ( HR 1.37, 95% CI 1.02-1.83; P=0.034). The greater the decrease of INR in patients with ischemic stroke within one year, the higher the 10-year non-vascular death risk ( HR 2.15, 95% CI 1.29-3.59; P=0.004). Conclusions:The 10-year mortality rate of patients with ischemic stroke is higher, and about 3/4 are vascular deaths. The fibrinogen-lowering treatment in the acute stage has no significant effect on the 10-year all-cause mortality of patients with ischemic stroke. The greater the decrease of tPA in half a year, the lower the all-cause mortality; the greater the decrease of D-dimer level at baseline and within 1 year, the higher the 10-year vascular death; the greater the decrease of INR at baseline and within 1 year, the higher the 10-year non-vascular death risk.

Quality Control and Safety Evaluation of Culture Medium for Human Assisted Reproduction / 中国医疗器械杂志

OBJECTIVE@#In this paper, the key points of quality control and safety evaluation of human assisted reproductive medium were summarized to provide reference for the establishment of relevant standards and quality control in the future.@*METHODS@#Through literature research, the key factors of quality control and risk control of human assisted reproductive medium were summarized, and the problems in clinical transformation were discussed.@*RESULTS@#It is very important for the development of human assisted reproduction technology to study the active ingredients and their harmful degradation products and drugs in the culture medium of assisted reproduction.@*CONCLUSIONS@#At present, the biggest challenge is to effectively control the quality of the culture medium for human assisted reproduction, establish corresponding inspection methods and quality standards for the key components, ensure the safety and effectiveness during the product shelf life, and thus improve the success rate of human assisted reproduction technology.

Evaluation of coagulation parameters: Coronavirus disease 2019 (COVID-19) between survivors and nonsurvivors

SUMMARY OBJECTIVE: This study aims to investigate and compare the coagulation parameters of coronavirus disease 2019 (COVID-19) patients with mortal and nonmortal conditions. METHODS: In this study, 511 patients diagnosed with COVID-19 were included. Information about 31 deceased and 480 recovered COVID-19 patients was obtained from the hospital information management system and analyzed retrospectively. Whether there was a correlation between coagulation parameters between the mortal and nonmortal patients was analyzed. Descriptive analyses on general characteristics of the study population were performed. Visual (probability plots and histograms) and analytical methods (Kolmogorov-Smirnov and Shapiro-Wilk test) were used to test the normal distribution. Analyses were performed using the SPSS statistical software package. RESULTS: Out of 511 patients, 219 (42.9%) were females and 292 (57.1%) were males. There was no statistically significant difference between males and females in terms of mortality (p=0.521). In total, the median age was 67 (22). The median age was 74 (13) in the nonsurvivor group and 67 (22) in the survivor group, and the difference was statistically significant (p=0.007). The D-dimer, prothrombin time, international normalized ratio, neutrophil, and lymphocyte median age values with p-values, in the recovered and deceased patient groups were: 1070 (2129), 1990 (7513) μg FEU/L, p=0.005; 12.6 (2.10), 13.3 (2.1), p=0.014; 1.17 (0.21), 1.22 (0.19), p=0.028; 5.51 (6.15), 8.54 (7.05), p=0.001; and 0.99 (0.96), 0.64 (0.84), p=0.037, respectively, with statistically significant differences. CONCLUSIONS: As a result of this study, D-dimer, prothrombin time, and international normalized ratio increase were found to be associated with mortality. These parameters need to be closely monitored during the patient follow-up.

Roles of certain biochemical and hematological parameters in predicting mortality and ICU admission in COVID-19 patients

SUMMARY OBJECTIVE: In this study, we aimed to retrospectively analyze the roles of certain hematological and biochemical parameters in predicting mortality and intensive care unit admission in patients diagnosed with coronavirus disease 2019 (COVID-19). METHODS: We analyzed the complete blood count and biochemical parameters of 186 COVID-19 patients by using the polymerase chain reaction test. Whether these parameters can be used to predict intensive care unit admission and mortality in the COVID-19 patients was investigated. RESULTS: The complete blood count and biochemical parameters of COVID-19 patients and in those admitted to intensive care unit were compared. The red cell distribution width, ferritin, lactate dehydrogenase, D-dimer, C-reactive protein, prothrombin time, and creatinine levels were found to be the most significant parameters. We found that these parameters are significant for predicting not only intensive care unit admission, but also the mortality of the patients admitted to the intensive care unit. CONCLUSIONS: We determined that the most effective parameters to predict intensive care unit admission and mortality in COVID-19 patients are ferritin, lactate dehydrogenase, D-dimer, C-reactive protein, red cell distribution width, creatinine, and intensive care unit. Close monitoring of these parameters and early intervention in alterations are of vital importance.

Comparisons of serum procalcitonin and fibrinogen degradation product levels between type 2 diabetes mellitus patients with Escherichia coli bloodstream and urinary tract infections / 中国基层医药

Objective:To compare serum procalcitonin and fibrinogen degradation product levels between type 2 diabetes mellitus patients with Escherichia coli bloodstream and urinary tract infections. Methods:The clinical data of 82 type 2 diabetes mellitus patients with Escherichia coli infections who received treatment between December 2014 and December 2019 in the First Affiliated Hospital of Datong University (The Fifth People's Hospital of Datong) were retrospectively analyzed. These patients were assigned to bloodstream infection ( n = 40) and urinary tract infection ( n = 42) according to the way of Escherichia coli infection. Serum procalcitonin and fibrinogen degradation product levels, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, C-reactive protein, white blood cell count, D-Dimer level, antithrombin III activity, and electrolytes were determined and compared between the two groups. Correlation between procalcitonin and other variables was analyzed. Multiple linear regression analysis was performed with procalcitonin level as a dependent variable and other relevant indexes as independent variables. Results:Body temperature, white blood cell count, neutrophil count, monocyte count, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, procalcitonin level, C-reactive protein level, fibrinogen degradation product level, and D-Dimer level in the bloodstream injection group were (39.49 ± 0.64) ℃, (14.92 ± 11.78) × 10 9/L, (13.39 ± 11.60) × 10 9/L, (0.72 ± 0.36) ×10 9/L, (14.86 ± 10.52), (199.15 ± 160.69), (22.81 ± 17.86) μg/L, (133.44 ± 63.63) mg/L, (49.71 ± 41.44) mg/L, (16.56 ± 12.20) mg/L, respectively, which were significantly higher than those in the urinary tract infection group [(37.12 ± 1.20) ℃, (9.04 ± 3.95) × 10 9/L, (6.25 ± 4.02) × 10 9/L, (0.42 ± 0.29) × 10 9/L, (3.67 ± 3.34), (120.01 ± 44.08), (4.46 ± 8.69) μg/L, (39.22 ± 22.16) mg/L, (3.81 ± 3.41) mg/L, (0.84 ± 0.75) mg/L), t = 7.356, 2.578, 3.162, 2.958, 5.538, 2.591, 2.810, 4.825, 2.902, 2.375, all P < 0.05]. Platelet count, lymphocyte count, blood sodium level and antithrombin Ⅲ activity in the bloodstream infection group were (167.50 ± 104.93) × 10 9/L, (1.06 ± 0.58) × 10 9/L, (130.89 ± 6.50) mmol/L, (57.88 ± 16.28)% , which were significantly lower than those in the urinary tract infection group [(239.40 ± 82.52)× 10 9/L, (2.14 ± 0.71) × 10 9/L, (138.46 ± 5.96) mmol/L, (90.11 ± 8.90)%, t = -2.853, -6.313, -4.046, -7.350, all P < 0.05]. Correlation analysis revealed that serum procalcitonin level was positively correlated with body temperature ( r = 0.387), white blood cell count ( r = 0.355), neutrophil count ( r = 0.368), C-reactive protein ( r = 0.605), fibrinogen degradation product level ( r = 0.616), D-Dimer level ( r = 0.486) (all P < 0.05), and it was negatively correlated with sodium level ( r = -0.319) and antithrombin Ⅲ activity ( r = -0.465) (both P < 0.05). Multiple linear regression analysis results revealed that fibrinogen degradation product level and body temperature were greatly correlated with procalcitonin level. Conclusion:Inflammatory indicators procalcitonin level, body temperature, white blood cell count, neutrophil count, C-reactive protein, fibrinogen degradation product level and D-Dimer level were remarkably higher in type 2 diabetes mellitus patients with Escherichia coli bloodstream infection than those in type 2 diabetes mellitus patients with Escherichia coli urinary tract infection. Procalcitonin level was greatly correlated with body temperature and fibrinogen degradation product level.

Forced degradation studies and stability-indicating liquid chromatography method for determination of tirofiban hydrochloride and synthetic impurities / Estudo da degradação forçada e método indicativo da estabilidade por cromatografía líquida para determinação de cloridrato de tirofibana e impurezas sintéticas

SUMMARY This study aimed to develop and validate a stability-indicating liquid chromatography method for the determination of tirofiban hydrochloride and two synthetic impurities (impurity A and impurity C). The method utilizes a RP-18 column (250 mm x 4.6 mm; 5 µm) with the PDA detector for quantitation. A mixture of triethylamine 0.1% (acidified to pH 5.5 with phosphoric acid) and acetonitrile was used as the mobile phase at a flow rate of 1 mL min-1 with gradient elution. The method presented satisfactory linearity, precision, accuracy and robustness, as well as low limits of detection and quantification, which demonstrate sensitivity in the determination of tirofiban and impurities A and C. It was selective for the determination of the drug and impurities analysed, without interference of the degradation products generated under forced conditions, demonstrating the stability-indicating capacity of the proposed method. Tirofiban showed to be practically stable to oxidative (30% H2O2 for 24 h) and thermal (75 °C for 24 h) conditions, but presented degradation to UVA light and acid hydrolysis, obeying the first order kinetics for both. In this way, it can be used as a stability-indicating method in the quality control of the raw material of tirofiban hydrochloride, as well as of the finished product. The obtained results demonstrate the importance of deepening the studies in this area, to guarantee the quality of commercialized pharmaceutical products.

RESUMO Este estudo teve como objetivo desenvolver e validar método indicativo da estabilidade por cromatografía líquida para determinação de cloridrato de tirofibana e duas impurezas de síntese (impureza A e impureza C). O método utilizou coluna de fase reversa RP-18 (250 mm x 4,6 mm; 5 µm) e detector PDA para quantificação. A fase móvel foi composta por uma mistura de trietilamina 0,1% (acidificada com ácido fosfórico para pH 5,5) e acetonitrila, à vazão de 1 mL/min, no modo gradiente. O método apresentou linearidade, precisão, exatidão, robustez, bem como baixos limites de detecção e quantificação, demonstrando sensibilidade na determinação da tirofibana e impurezas A e C. O método apresentou seletividade na determinação do fármaco e das impurezas, sem interferência dos produtos de degradação gerados na degradação forçada da tirofibana, demonstrando sua capacidade indicativa de estabilidade. O fármaco apresentou-se estável a oxidação (H2O2 30% por 24 h) e a degradação térmica (75 °C por 24 h), mas degradou frente à luz UVA e hidrolise ácida, obedecendo cinética de primeira ordem para ambas. Dessa forma, pode ser utilizado como um método indicativo de estabilidade no controle de qualidade da matéria -prima do cloridrato de tirofibana, bem como no produto acabado. Os resultados obtidos demonstram a importância de aprofundar os estudos na área, com intuito de garantir a qualidade dos produtos farmacêuticos comercializados.

Fibrinogen and D-dimer variances and anticoagulation recommendations in Covid-19: current literature review

SUMMARY INTRODUCTION Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a newly described virus responsible for the outbreak of the coronavirus disease 2019 (Covid-19), named by the World Health Organization (WHO) in February/2020. Patients with Covid-19 have an incidence of acute respiratory distress syndrome (ARDS) of 15.9-29% and sepsis is observed in all deceased patients. Moreover, disseminated intravascular coagulation (DIC) is one of the major underlying causes of death among these patients. In patients with DIC, there is a decrease in fibrinogen and an increase in D-dimer levels. Some studies have shown that fibrinogen and one of its end products, D-dimer, might have a predictive value for mortality in patients with non-Covid sepsis secondary to complications of DIC. Therefore, anticoagulation, considering its mortality benefits in cases of non-Covid sepsis, may also have an important role in the treatment of Covid-19. METHODS We reviewed the literature of all studies published by April 2020 on patients infected with Covid-19. Our review was limited to D-dimer and fibrinogen changes and anticoagulation recommendations. RESULTS Anticoagulation therapy can be started following the DIC diagnosis in Covid-19 patients despite the bleeding risks. In addition, the current evidence suggests a routine use of anticoagulation, particularly in patients with higher D-dimer levels (> 3.0 μg/mL). CONCLUSION Covid-19 is a systemic, hypercoagulable disease requiring more studies concerning treatment. Aanticoagulation is still an issue to be studied, but D-dimer rise and disease severity are the indicative factors to start treatment as soon as possible.

RESUMO INTRODUÇÃO O coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2) é o vírus responsável pelo surto recentemente batizado de doença pelo coronavirus 2019 (Covid-19) pela Organização Mundial de Saúde (OMS) em fevereiro/2020. Os doentes com Covid-19 têm uma incidência de síndrome de dificuldade respiratória aguda (SDRA) de 15,9-29% e sepse é observada em todos os pacientes que vêm a óbito. Além disso, a coagulação intravascular disseminada (DIC) é uma das principais causas subjacentes de morte entre esses pacientes. Em pacientes com DIC, ocorre com uma diminuição do fibrinogênio e um aumento dos níveis de dímero D. Alguns estudos mostraram que o fibrinogênio e um dos seus produtos finais, o dímero D, podem ter um valor preditivo para a mortalidade em pacientes com sepse não relacionada à Covid-19 decorrente de complicações da DIC. Portanto, a anticoagulação, considerando seus benefícios quanto à mortalidade na sepse não relacionada à Covid-19, pode também ter um papel importante no tratamento da Covid-19. MÉTODOS Realizamos uma revisão de todos os estudos publicados até abril de 2020 sobre pacientes infectados com Covid-19. A nossa revisão limitou-se a alterações no dímero D, nos fibrinogênios e recomendações de anticoagulantes. RESULTADOS A terapêutica anticoagulante pode ser iniciada após o diagnóstico de DIC em pacientes com Covid-19 apesar dos riscos de hemorragia. Além disso, a evidência atual sugere o uso rotineiro da anticoagulação, principalmente em pacientes com níveis mais elevados de dímero D (> 3, 0 µg/mL). CONCLUSÃO A Covid-19 é uma doença sistêmica e hipercoagulável que requer mais estudos em relação ao tratamento. A anticoagulação ainda é uma questão a ser estudada, mas o aumento de dímeros D e a gravidade da doença são os fatores indicativos para o início do tratamento o mais rápido possível.

High D-dimer levels are associated with prostate cancer

RESUMO OBJETIVO O câncer de próstata é uma das neoplasias mais comuns em homens. Os principais fatores de risco para a ativação da coagulação e trombose são malignidade e idade mais avançada. O risco de trombose pode estar associado ao aumento do nível dos marcadores de coagulação, tais como o fibrinogênio e D-dímero. O objetivo deste estudo é avaliar a relação entre os marcadores de coagulação e o câncer de próstata. METODOLOGIA Este estudo prospectivo incluiu os pacientes que foram submetidos à biópsia de próstata transretal guiada por ultrassonografia e que passaram por cirurgia da próstata entre janeiro de 2015 e janeiro de 2016. Os níveis no plasma de antígeno prostático específico (PSA), PSA livre (fPSA), porcentagem de fPSA, D-dímero e fibrinogênio foram medidos antes dos procedimentos. Os pacientes foram divididos em dois grupos de acordo com os resultados de patologia. Os pacientes com hiperplasia benigna da próstata foram colocados no grupo 1 e os pacientes com câncer de próstata no grupo 2. RESULTADOS No total, 76 pacientes foram incluídos neste estudo. Houve um total de 53 pacientes no grupo 1 e 23 pacientes no grupo 2. A idade média dos pacientes e os níveis de PSA, fPSA, fibrinogênio e D-dímero foram, respectivamente, 65.33 ± 7.47 anos, 8.21 ± 4.59, 1.41 ± 0.74 ng/ml, 309.75 ± 80.46 mg/dl e 0.42 ± 0.39 µg/ml no grupo 1. No grupo 2, a idade média dos pacientes e os níveis de PSA, fPSA, fibrinogênio e D-dímero foram, respectivamente, 66.08 ± 6.7 anos, 145.69 ± 509.35, 7.32 ± 15 ng/ml, 312.16 ± 69.48 mg/dl, 1.09 ± 2.11 µg/ml. Biópsia da próstata e cirurgia transuretal foram realizadas em 64 (%84,21) e 12 (%15,79) pacientes, respectivamente. CONCLUSÃO O presente estudo demonstrou que os níveis de D-dímero no plasma foram maiores em pacientes com câncer de próstata. Novos estudos com um maior número de pacientes são necessários para definir a relação entre câncer de próstata e distúrbios de coagulação.

Clinical significance of the level of fibrin degradation products in drowning patients without cardiac arrest / Journal of Rural Medicine

Objective: This study aimed to retrospectively investigate the clinical significance of the level of fibrin degradation products in drowning patients without cardiac arrest.Patients and Methods: All drowning patients who were transported to our department from January 2011 to December 2019 were retrospectively investigated through a medical chart review and included as subjects in the present study. The exclusion criteria were the occurrence of cardiac arrest before patient arrival to our department and lack of measurement of the fibrin degradation product level on arrival. The subjects were divided into two groups: early discharge group, which included patients who were discharged within 3 days, and late discharge group, which included patients who were discharged after 3 days.Results: The early discharge group included 10 subjects and the late discharge group included 39 subjects. No significant differences were observed in age, sex, proportion of freshwater drowning cases, proportion of alcohol drinkers, vital signs, blood gas analysis findings, proportion of lung lesions, or survival rate between the two groups. The levels of glucose and fibrin degradation products on arrival were significantly greater in the early discharge group than in the late discharge group. A multivariate analysis showed that the only significant predictor of early discharge was the fibrin degradation product level among variables identified in a univariate analysis.Conclusion: This is the first study to show that the level of fibrin degradation products on arrival can predict early or late discharge in drowning patients without cardiac arrest before arriving to the hospital.

Identification of amino alcohols related substances of atenolol by LC-Q-TOF/MS / 中国药科大学学报

@#To identify the amino alcohols related substances in atenolol. The related substances in atenolol and its stressed samples were pre-column derivatized with 9-fluorenylmethyl chloroformate. The separation was carried out on an Inertsil ODS-SP column (250 mm×4.6 mm, 5 μm) with linear gradient elution by methanol-ammonium acetate solution as the mobile phases. Electrospray positive ionization high-resolution Q-TOF/MS was used for the determination of the accurate masses and elemental compositions of the parent and fragment ions of these related substance derivatives. The structures of all the detected substances were identified by spectral analysis and synthetic analysis. Under the established conditions, atenolol and its amino alcohols related substances were well separated, and a total of 14 impurity peaks were detected and identified, of which 12 were related substances and 2 were derivatization reaction by-products. The established LC-MS method provides a reference for the examination and quality control of atenolol related substances.

Diagnostic Significance of Fibrin Degradation Products and D-Dimer in Patients With Breast Cancer-Related Lymphedema

OBJECTIVE: To find out whether levels of fibrin degradation products (FDP) and D-dimer are increased in breast cancer-related lymphedema (BCRL) as in many vascular diseases. FDP and D-dimer have been used in blood tests to help differentiate deep vein thrombosis in the diagnosis of lymphedema. Levels of FDP and D-dimer are often elevated in patients with BCRL. METHODS: Patients with BCRL (group I), non-lymphedema after breast cancer treatment (group II), and deep venous thrombosis (group III) from January 2012 to December 2016 were enrolled. Levels of FDP and D-dimer were measured in all groups and compared among groups. RESULTS: Mean values of FDP and D-dimer of group I were 5.614±12.387 and 1.179±2.408 μg/μL, respectively. These were significantly higher than their upper normal limits set in our institution. Levels of FDP or D-dimer were not significantly different between group I and group II. However, values of FDP and D-dimer in group III were significantly higher than those in group I. CONCLUSION: Values of FDP and D-dimer were much higher in patients with thrombotic disease than those in patients with lymphedema. Thus, FDP and D-dimer can be used to differentiate between DVT and lymphedema. However, elevated levels of FDP or D-dimer cannot indicate the occurrence of lymphedema.

Identification of the related substances of cyclosporin A by LC-MS techniques / 中国药科大学学报

@#To identify the related substances of cyclosporin A by LC-MS techniques, the separation of cyclosporin A and its related substances was carried out on a Hypersil BDS C18(100 mm×4. 6 mm, 2. 4 μm)column with isocratic elution by a mixture of acetonitrile-water-MTBE and formic acid(430 ∶520 ∶50 ∶1)as the mobile phase. Cyclosporin A and its 29 related substances(9 process related and 20 degradants)were well separated under the established conditions. Among them, 13 were listed in EP and the rest 16 were unknown products having not been reported before. Electrospray positive ionization high resolution TOF/MS was used for the determination of the accurate mass and elemental composition of parent ions of all the components, and triple quadrupoles tandem mass was employed for the product mass spectra determination. Thence, the structures of all the 29 detected substances were successfully characterized through spectra elucidation and the fragmentation pathways analysis. The established LC-MS method was successfully employed for the separation and identification of the related substances of cyclosporin A and it is useful for its fermentation processes and quality control.

Changes of coagulation and fibrinolysis system in children with Henoch-Schonlein purpura nephritis and its clinical significance / 中国基层医药

Objective@#To investigate the changes of coagulation and fibrinolysis system in children with Henoch-Schonlein purpura nephritis and its clinical significance.@*Methods@#From January 2013 to December 2016, 73 children with Henoch-Schonlein purpura inthe First People's Hospital of Huzhouwere selected as the Henoch-Schonlein purpura group, 73 children with Henoch-Schonlein purpura nephritis were selected as the Henoch-Schonlein purpura nephritis group, and 73 healthy children were selected as the control group.Antithrombin III activity (AT III), fibrinogen degradation products (FDP), plasma D-dimer (D-D), prothrombin activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA), fibrinogen (FIB) levels and 24-hour urinary protein excretion were measured in three groups.@*Results@#The levels of FIB, AT III%, FDP, D-D, PAI-1 and t-PA in the Henoch-Schonlein purpura group and the Henoch-Schonlein purpura nephritis group[(2.89±0.76)g/L, (3.51±0.81)g/L; (145.72±8.46)%, (163.24±9.05)%; (1.31±0.67)mg/L, (1.54±0.72)mg/L; (0.87±0.52)mg/L, (1.18±0.67)mg/L; (66.47±2.58)ng/L, (91.02±3.24)ng/L; (16.34±0.98)μg/L, (12.35±1.06)μg/L]were higher than those in the control group[(1.88±0.54)g/L, (119.48±8.92)%, (0.92±0.33)mg/L, (0.32±0.18)mg/L, (31.25±3.02)ng/L, (6.82±0.75)μg/L](t=9.256, 18.236, 4.462, 8.540, 75.760, 65.912; 14.306, 29.423, 6.688, 10.591, 115.297, 36.387, all P<0.05). The levels of FIB, AT III%, FDP, D-D and PAI-1 in theHenoch-Schonlein purpura nephritis group were higher than those in the Henoch-Schonlein purpura group (t=4.769, 12.083, 1.998, 3.123, 50.644, all P<0.05), and t-PA in the Henoch-Schonlein purpura nephritis group was lower than that in Henoch-Schonlein purpura group (t=23.165, P<0.05). The 24-hour urinary protein excretion in theHenoch-Schonlein purpura nephritis group[(1.48±0.89)g/24h]was higher than that in the Henoch-Schonlein purpura group[(0.11±0.02)g/24h] and control group[(0.10±0.05)g/24h](t=13.149, 13.227, all P<0.05). There was no statistically significant difference between Henoch-Schonlein purpura group and control group (t=1.587, P>0.05). The 24-hour urinary protein excretion was positively correlated with AT III%, FDP, D-D and IL-33 in patients with Henoch-Schonlein purpura nephritis (r=0.502, 0.546, 0.483, all P<0.05), but not correlated with FIB, PAI-1 and t-PA (r=0.189, 0.213, -0.175, all P>0.05).@*Conclusion@#Patients with purpuric nephritis are in a state of hypercoagulability and hyperfibrinolysis, and coagulation and fibrinolysis disorders are closely related to renal damage in patients.

Applications of computed tomography and ischemic modified albumin and D-D Levels in disease assessment of patients with acute pulmonary embolism / 中国医师杂志

Objective@#To investigate the clinical application values of computed tomography (CT), ischemic modified albumin (IMA) and D-dimer (D-D) levels in the disease assessment of patients with acute pulmonary embolism (APE).@*Methods@#From June 2015 to June 2018, 100 suspected APE patients in our hospital were selected as the study subjects, after the CT " gold standard" inspection, the 80 patients diagnosed with APE were as APE group, including 38 cases in high-risk group and 42 cases in low-risk group: 20 non APE cases and 60 healthy volunteers at the same time were selected as control group. The serum IMA level was detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA), and the plasma D-D level was detected by immunoturbidimetry. Receiver operating characteristc (ROC) curve was used to analyze the diagnostic values of IMA and D-D for APE disease.@*Results@#The levels of IMA and D-D in APE group were significantly higher than those in non APE group and control group (P<0.05); the levels of IMA and D-D in non APE group were significantly higher than those in control group (P<0.05), while those in high-risk group were significantly higher than those in low-risk group (P<0.05); CT examination after admission was as the gold standard for diagnosing APE disease; ROC curve showed that, the area under curve (AUC) of D-D and IMA in diagnosing APE was 0.875 and 0.763, respectively, with corresponding sensitivity 90.01% and 93.87%, specificity 52.21% and 95.65% respectively; the AUC of IMA combined D-D level in diagnosing APE was 0.834, the sensitivity was 95.87%, and the specificity was 78.69%; the AUC of D-D and IMA in diagnosing high-risk APE was 0.950 and 0.914, respectively, with the corresponding sensitivity 97.21% and 93.98%, specificity 31.58% and 76.98%, respectively. The AUC of IMA combined D-D level in diagnosing high-risk APE was 0.958, with sensitivity 96.39%, specificity 76.87%. Compared with CT results, in the 80 patients diagnosed with APE, there were 4 patients with negative IMA and 6 patients with negative D-D, and the control group samples test showed negative IMA and D-D levels were all negative. The consistency of CT combine with D-D ( Kappa=0.734, P=0.000), and CT combine with IMA (Kappa=0.819, P=0.000) were good.@*Conclusions@#According to the results of CT examination, IMA combined with D-D level has a good sensitivity and specificity in the diagnosis of APE, which can effectively improve the diagnostic rate of APE diseases, and provide an important basis for clinical rapid and reliable detection and treatment of APE diseases.